Vol. 5 No. 1 February 2021
68
Vol. 5 No. 1 February 2021
Transcript of Vol. 5 No. 1 February 2021
Vol. 5 No. 1 February 2021
Journal of Applied Agricultural Science and Technology (JAAST)
Journal of Applied Agricultural Science and Technology (JAAST) is an international journal, focuses on applied agricultural science and applied agricultural technology in particular: agricultural mechanization, food sciences, food technology, agricultural information technology, agricultural economics, agricultural statistics, bioinformatics, farm structure, farm power, agricultural machinery, irrigation and drainage, land and water resources engineering, renewable energy, environment, crop production, and crop protection. JAAST has been ACCREDITED by the Ministry of Research, Technology and Higher Education (RistekDikti) of The Republic of Indonesia as an achievement for the peer-reviewed journal. The recognition published in Director Decree No.10/E/KPT/2019 April 4, 2019, effective from Vol 1 No 1 2017. This journal provides immediate open access to its content on the principle that making research freely available to the public supports a greater global exchange of knowledge. The journal can be accessed at www.kinfopolitani.com Benefits of open access for the author, include:
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EDITORIAL TEAM Editor in Chief Dr. Edi Syafri, ST, M.Si. Scopus ID: 57196348984 Politeknik Pertanian Negeri Payakumbuh, Indonesia Editorial Board Members: 1. Assoc. Prof. Samsuzana Abd Aziz. Scopus ID: 57192693512
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Jember University, Indonesia 12. Jiban Shresta.
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Politeknik Pertanian Negeri Payakumbuh, Indonesia Editorial Assistant: Yulia Chyntia Hariati, A.Md.T Editorial Office Department of Agricultural Enginering, Politeknik Pertanian Negeri Payakumbuh Jl. Raya Negara Km.7 Tanjung Pati 26271 Kec. Harau Kab Limapuluh Kota Sumatera Barat https://www.jaast.org email: [email protected]
ACKNOWLEDGMENT On the behalf of editorial board, we would like to thank the reviewer for the contributions and efforts, without which it would be imposible to maintain the standard of peer-reviewed journal in this issue.
Prof. Yayan Sanjaya, Universitas Pendidikan Indonesia, Indonesia
Prof. Reni Desmiarti, Bung Hatta University, Indonesia
Dr. Haryuni, Universitas Tunas Pembangunan Surakarta, Indonesia
Ir. Darwin H Pangaribuan, M.Sc, Ph.D, University of Lampung, Indonesia
Dr. Turhadi, MSi, Pusat Penelitian Bioteknologi dan Bioindustri Indonesia, Indonesia
Dr. Mochamad Asrofi, ST., Jember University, Indonesia
Jhon Hardy Purba, Universitas Panji Sakti, Indonesia
Parwito, SP, MP, Universitas Ratu Samban, Indonesia
Ir. Irwan A, M.Si, Politeknik Pertanian Negeri Payakumbuh, Indonesia
Jamaluddin, M.Si, Politeknik Pertanian Negeri Payakumbuh, Indonesia
TABLE OF CONTENT
Volume 5, Number 1, Page 1-63 February 2021 Articles Growth and Yield Performance of Aromatic Fine Rice as Influenced by Varieties and Fertilizer Managements Shams Islam, Md. Al Mamun Or Roshid, Md. Shafiqul Islam Sikdar, Md. Sohrab Hossain ............................................................................................... 1 Identification of Local Rice Genotypes from Deli Serdang, North Sumatera, Indonesia to Drought Stress Condition, Irawati Chaniago, Noverina Chaniago, Irfan Suliansyah, Nalwida Rozen ........... 13 Antifungal Activity of Essential Oils of Leaves, Rhizomes Oils and Fraction Wild Ginger Elettariopsis Slahmong Ck Lim Inhibit The Colony Growth of Sclerotium Rolfsii Nurmansyah, Herwita Idris, Nasril Nasir .............................................................. 28 Stability and Toxicity Test of Angkak Pigment Powder from Sago Hampas- Rice Flour Substrate as Natural Dyes Dian Pramana Putra, Novelina, Alfi Asben ........................................................... 38 The Design and Building of Medium Capacity Drying House for Bokar Sri Aulia Novita, Hendra, Perdana Putera, Fithra Herdian, Muhammad Makky, Khandra Fahmi ...................................................................... 50 Editor's Corner The Prospect of Knowledge Growing System (KGS) for Plant Disease Early Detection System Ika Noer Syamsiana ................................................................................................ 62
Journal of Applied Agricultural Science and Technology E-ISSN: 2621-25285(1): 1-12 (2021) ISSN: 2621-4709
Received January 27, 2021; Accepted March 2, 2021; Published March 10, 2021 https://doi.org/10.32530/jaast.v5i1.6 This is an open access article under the CC BY-SA 4.0 license https://creativecommons.org/licenses/by-sa/4.0
1
GROWTH AND YIELD PERFORMANCE OF AROMATIC FINE RICE AS INFLUENCED BY VARIETIES AND FERTILIZER MANAGEMENTS
Shams Shaila Islam*,1, Md. Al Mamun Or Roshid1, Md. Shafiqul Islam Sikdar1, Ahmed Khairul Hasan2, Md. Sohrab Hossain1
1Department of Agronomy, Faculty of Agriculture, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh
2Department of Agronomy, Faculty of Agriculture, Bangladesh Agricultural University, Mymensing, Bangladesh
*Corresponding authorEmail: [email protected]
Abstract. This Research was conducted to investigate the effect of fertilizer management on growth and yield performance of aromatic fine rice varieties. The experiment consisted of two factors were aromatic fine rice and fertilizer management. There were four varieties namely Kalizira, Kataribhog, Tulshimala and BRRI Dhan34 with four fertilizer treatments recommended dose of fertilizers (T1), cowdung @ 10 tha-1 (T2), 50% of recommended dose of fertilizers + 50% cowdung(T3), 75% of recommended dose of fertilizers + 50% cowdung (T4). The result showed that BRRI Dhan 34 significantly superior for effective tillers number/ hill (18.46), panicles length (26.67cm), grains number/ panicle (146.30), harvest index (40.73 %), grain protein content (6.23%), grain yield (2.79 tha-1), straw yield (4.06 tha-1), and biological yield (6.85 tha-1). Among fertilizer management, the highest effective tillers number/hill (16.20), grains number/ panicle (142.45, panicles length (26.66 cm),1000 grain weight (13.75), grain protein content (10.2%), biological yield (6.30), were obtained from T3 treatment. Based on interaction effect showed that the highest effective tillers number/hill (18.36), grains number/ panicle (155.33), panicles length (26.73 cm), grain protein content (10.80%), biological yield (7.85) was found with BRRI Dhan 34 combined with T3 treatment. However, grain yield increased with the increase in nitrogen levels. Together tillers number/hill, grains number/panicle, grain protein content, harvest index, grain yield was the main responsible yield contributing characters to improve the yield quality of aromatic fine rice. Keywords: aromatic fine rice; cowdung; fertilizer management; yield performance
1. Introduction
Rice (Oryza sativa L.) is an essential cereal crop nourishing more than half of the world’s
populations supplying 50 to 80% off regular caloric consumption (Amirjani, 2011). About 75.01%
of the total cropped area of Bangladesh is used for rice production where annual production of
34.71 million tons from 11.28 million hectares of land (BBS, 2015). Different rice genotypes have
different characters. Some have special appeal for their aroma and scent. The major identified
aromatic varieties in Bangladesh are Kalizira, Chinigura, Kataribhog, BR5, Bashful, BRRI Dhan
34, BRRI Dhan 37, BRRI Dhan 38 (Bashmoti type), Khaskani, Badshabhog, Dudshagar,
Tulshimala, Khirshabhog, Horibhog, Parbatjira, Khasha, Modhumadab, Tilkapur, Chinikanai,
Khirkon, and Shakhorkora. Though productivity of aromatic fine rice is very low but its demand
for internal consumption and for export is increasing day by day (Haque et al., 2012). Basmati
Islam et al. JAAST 5(1): 1–12 (2021)
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(aromatic) rice has extremely high demand and it occupies a unique place in the world rice market
(Singh et al., 2018). Dinajpur region is a native area of some indigenous aromatic rice varieties.
About 30% of rice land in Dinajpur is covered by aromatic rice varieties during the ‘Aman’ season
(Alam et al., 2002). Aromatic fine rice has high market value, due to price and taste like polau,
khir, firny, payesh and exporting can bring a considerable amount of foreign exchange for the
country (Paul et al., 2016).
The use of organic inputs such as crop residues, manures and compost have great potential
to improve soil productivity and crop yield through the improvement of the physical, chemical,
and microbiological properties of the soil as well as nutrient supply (Stone & Elioff, 1998).
According to Tanimu (2013), cowdung manure contains the three major plant nutrients, nitrogen,
phosphorus, and potassium (NPK), as well as many essential nutrients such as Ca, Mg, S, Zn, B,
Cu, Mn, etc. However higher yields depend on the rational and effective application of chemical
fertilizers (Plucknett & Smith, 1986). But many researchers have reported a significant response
of rice production depends on (organic + inorganic fertilizers) in different soils in Bangladesh
(Uddin et al., 2018). So, the use of a judicious combination of organic and inorganic fertilizers is
very important for rice production in a tropical country (Esfahani et al., 2019).
The indigenous aromatic rice varieties, however, poor yielders having a poor response to
fertilizer application (Mohioddin et al., 2014). As the use of organic manures plays an important
role to enhance the fertilizer use efficiency, reduce the cost of nutrient supply, increase production.
Researchers observed aromatic rice gradually losing their aroma and qualities due to lack of soil
organic matter and the use of imbalance chemical fertilizers. Therefore, the objectives of the
research article were to compare the growth and yield performance of aromatic fine rice varieties
with different fertilizer managements and to identify the most responsible yield contributing traits
for higher production of aromatic fine rice yield.
2. Methods
The experiment was conducted at the Agronomy Research Field of Hajee Mohammad
Danesh Science and Technology University, Dinajpur, Bangladesh during (July to December
2017). Experimental site was characterized under the sub-tropical weather and climate by three
distinct seasons with medium high land containing soil pH 5.6, organic carbon 0.45%,
temperatures differed from July to December (33°C to 25°C), humidity (72% to 82%), and rainfall
(296.2 mm-10 mm). This location contained latitude, longitude, and elevation from sea level
(25.13o N, 88.23oE, 37.5 m). Two factors were included in the experiment namely, Factor-A (Four
varieties namely), Kalizira, Kataribhog, Tulshimala, and BRRI Dhan34. Factor-B (4 fertilizer
doses): Recommended fertilizer doses (T1), Cowdung @ 10 t/ha (T2), 50% recommended
fertilizers doses + 50% cowdung (T3) and 75% of recommended fertilizers doses + 50% cowdung
Islam et al. JAAST 5(1): 1–12 (2021)
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(T4). The experiment was laid out in a Randomized Complete Block Design with three replications
where the size of each unit plot was 2.5m x 2.0 m. Before sowing in the nursery, seeds were soaked
in water for 24 hrs. After that kept into gunny bags in dark condition. When seeds sprouted, sown
in wet seedbed on 4th July 2017. At 36 days old, 3 seedlings/ hill were transplanted on 9th August
2017 according to the experimental design. Fertilizers like TSP(P2O5) and MoP (K2O) were
applied at final land preparation. The nitrogen (urea) was applied as an experimental specification.
One-third urea was applied during final land preparation, and rest urea in two equal installments
at 33 and 66 DAT, respectively. Agronomic actions e.g., weed and insect control were done
manually. Insect pests were controlled by the application of 20 ml per 1 L Cypermethrin 10% w/v
EC and 50 ml per 1 L Benfuracarb 20% w/v EC with water.
2.1. Data collection
Data on individual plant parameters was recorded from selected hills/plots. Grain yield,
straw yield, biological yield, and harvest index were recorded from the whole plot at harvesting
time and qualitative traits were recorded from selected grain. Collected parameters were plant
height (cm), tillers number/ hill, effective tillers/ hill, panicles length(cm), grains number/panicle,
1000-grain weight, grain protein content (%), grain yield (t/ ha), straw yield (t/ ha), biological yield
(t/ ha) and harvest index (%).
2.2. Statistical Analysis
Data were analysed statistically as per the design used following the analysis of variance
(ANOVA) technique and the mean differences were adjusted with DMRT at a 5% level of
significance using the statistical computer package program, (MSTAT-C) following Russel
(1986).
3. Results and Discussions
3.1. Plant height on varieties and fertilizer treatments
Tables 1 and 2 showed that varieties and fertilizer treatments were significantly influenced
by plant height. It was observed that Tulshimala produced the tallest plant 161.44cm and the
smallest 144.55cm by BRRI Dhan34. It was evident that plant height differed significantly from
varieties due to genetic variation, nutrient uptake, photosynthesis rate, etc. This result agreed with
(Jiang, et al., 2003). The tallest plant 155.96 cm was observed at T1 while the shortest 149.32 cm
was observed at T2 fertilizer management treatments.
3.2. Effective tillers number/ hill on genotypes and fertilizer treatments
Tables 1 and 2 showed that effective tillers number/ hill was significantly influenced by
genotypes and fertilizer managements. Highest (18.46) was found from BRRI Dhan34 which was
followed by Tulshimala (16.36) and Kalizira (16.16). Whereas, lowest (14.06) from Kataribhog
Islam et al. JAAST 5(1): 1–12 (2021)
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varieties. Chandel et al. (2010) and Sarkar et al. (2014) reported that tillers number/ hill differed
significantly for different varieties due to genetic and varietal variation that might be influenced
by heredity. The highest tillers number/ hill was obtained 16.20 at T3 treatment. Adequacy of
nitrogen application probably favored the cellular activities during yield and development which
lead to increase tillers number/hill. In the present experiment with nitrogen at higher level
produced identically higher effective tillers.
Table 1. Varietal effect on the yield and yield contributing characters of aromatic rice
Variety Plant height (cm)
Tiller numbers/hill
(no)
Panicles length (cm)
Grains/ panicle
(no)
1000- grain
weight (g)
Grain protein content
(%)
Straw yield
Biological yield
(tha-1)
V1 145.12c 16.16b 24.56c 133.67c 12.44c 6.12b 3.07c 5.10c V2 155.24b 14.06c 25.65b 133.56c 14.05a 6.08b 3.50b 5.73b V3 161.44a 16.36b 24.45c 136.45b 12.60c 6.23b 3.06c 4.98c V4 144.55c 18.46a 26.67a 146.30a 13.37b 7.17a 4.06a 6.85a Level of Significance ** ** ** ** ** ** ** **
LSD 3.167 0.633 0.519 1.15 0.445 1.12 0.334 0.379 CV (%) 2.53 5.06 2.63 0.95 4.10 0.93 0.98 8.43
Here V1 means variety, figure bearing same, or no letter (s) do not differ significantly at 5% level of significance by Duncan's Multiple Range Test, * = Significant at 5% level of significance, ** = Significant at 1% level of significance
3.3. Panicle length (cm) on varieties and fertilizer doses
Panicle length was significantly influenced due to varieties and fertilizer doses. The results
indicated that the longest 26.67cm by BRRI Dhan34 and the shortest 24.45 cm produced by
Tulshimala (Table 1). Panicles length with varieties differed significantly among each other due
to their differences in genetic variation reported by (Chandel et al. 2010; Islam et al., 2013).
Panicle length was significantly influenced by fertilizer management. Results showed that the
longest 26.66 cm was produced at T3 while the shortest 23.66 cm was obtained from T2 fertilizer
management treatment (Table 2).
3.4. Grains number/panicle on varieties and fertilizer doses
Results showed that grains number/panicle was significantly influenced by varieties. BRRI
Dhan34 produced statistically the highest 146.30 no. and lowest 133.56no. was found in
Kataribhog (Table 1). Reza et al. (2016) reported that grains number/panicle influenced
significantly due to varieties. Grains number/ panicle varied significantly due to differences in
nitrogen levels. Grains number/ panicle increased using T4 treatment i.e., increasing fertilizer
management. Table 2 showed the statistically highest grains number/panicle 142.45no was
recorded from T3 while lowest from T2 treatment. An adequate supply of nitrogen contributed to
grain formation that probably increased grains number/ panicle.
Islam et al. JAAST 5(1): 1–12 (2021)
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3.5. 1000 grain weight (g) on varieties and fertilizer doses
Table 1 showed that the highest 1000 grain weight 14.05 gm was obtained from Kataribhog
and the lowest 12.60 gm was from Tulshimala. It was evident that variation in 1000 grain weight
might be due to differences in the size of the grains that are partly controlled by the genetic make-
up of the studied varieties. Islam et al. (2016) reported that 1000 grain weight is stable, important
yield determining the character and influenced by the environment that differed significantly
among the cultivars due to genetic make-up which supported the present experimental result. A
similar result was reported by (Djaman et al., 2016). Table 2 showed that the influence of different
treatments of organic, inorganic, and their combinations on 1000 grain weight (g) was significant.
The highest 1000 grain weight 13.75 gm was obtained from T3 and lowest 12.94 gm from T4
treatments.
Table 2. Effect of fertilizer management on the yield and yield contributing traits of aromatic fine rice.
Here F means fertilizer, figure bearing same or no letter (s) do not differ significantly at 5% level of significance by Duncan's Multiple Range Test, * = Significant at 5% level of significance, ** = Significant at 1% level of significance
3.6. Grain Protein Content (%) on varieties and fertilizer managements
Grain protein content was significantly varied due to varieties (Table 1). The highest 7.17%
was obtained from BRRI Dhan34 and the lowest 6.08% was found from Kataribhog. This result
was liked by Aziz et al. (2017) who recorded variable protein percentage among varieties. Table
2 showed that the influence of different treatments of organic, inorganic, and their combinations
on grain protein content was significant. The highest grain protein content obtained 10.2 % T3 and
lowest 9.67% from T2 fertilizer management treatments.
Fertilizer Plant height (cm)
Effective tiller
number/ hill (no)
Panicles length (cm)
Grains/ panicle
(no)
1000- grain
weight (g)
Grain protein content
(%)
Straw yield (tha-1)
Biological yield (tha-1)
F1/T1 155.96a 14.59c 25.55b 142.05a 13.42a 9.92b 3.67a 6.27a
F2/T2 149.32b 14.57c 23.66c 135.88c 13.23a 9.67b 3.18b 4.75b
F3/T3 149.56b 16.20a 26.66a 142.45a 13.75a 10.2a 3.66a 6.30a
F4/T4 151.43b 15.45b 25.42b 140.60b 12.94b 10.0a 3.16b 4.90b
Level of Significance ** ** ** ** **
**
** **
LSD 3.167 0.644 0.528 1.2 0.445 0.566 0.612 0.388
CV (%) 2.45 5.35 2.67 0.97 4.25 2.77 3.67 8.32
Islam et al. JAAST 5(1): 1–12 (2021)
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Table 3. Interaction effect of varieties and fertilizer managements on the growth and yield contributing traits of aromatic fine rice.
Interaction (Variety × Fertilizer)
Plant height (cm)
Effective tillers/ hill
(no)
Panicles/ length (cm)
Grains/ panicle
(no)
1000-grain
weight (g)
Grain protein content
(%)
Straw yield (tha-1)
Biological yield (tha-1)
V1F1 155.42d 16.21c 22.55c 131.34g 10.86d 8.20c 3.35f 5.74b
V1F2 141.89f 15.45d 21.41c 135.77f 11.35c 8.45c 2.62k 4.35c
V1F3 142.10f 15.76d 24.02b 137.77e 11.53c 8.33c 3.33f 5.76b
V1F4 148.93e 17.06b 19.38d 133.77g 10.43d 8.67c 2.85j 4.45c
V2F1 158.35c 14.94f 24.09b 132.77g 14.35a 8.23c 3.64e 6.24b
V2F2 153.87d 15.84d 22.18c 137.11e 14.05a 8.50c 3.26f 5.12b
V2F3 155.62d 16.42c 24.55b 136.26e 14.35a 8.68c 3.75d 6.58b
V2F4 155.03d 17.25b 20.35d 135.43f 13.74b 8.88c 2.92i 4.65c
V3F1 165.71a 16.42c 22.55c 136.10e 11.16c 9.23b 3.10h 5.20b V3F2 159.75c 15.54d 21.65c 141.00d 10.55d 9.02b 2.95i 4.61c
V3F3 161.42b 16.78b 24.55a 137.66e 11.25c 8.80 3.20g 5.40b
V3F4 161.44b 17.07b 21.10c 137.66e 10.45f 9.34b 2.95i 4.46c
V4F1 150.14e 16.84c 25.56a 139.00d 13.35b 8.80c 4.55a 7.85a
V4F2 149.21e 16.60c 23.23b 153.00b 12.82b 10.00a 3.88c 5.48b
V4F3 146.61e 18.36a 26.73a 155.33a 12.37b 10.80a 4.45b 7.85a
V4F4 138.16g 18.04a 23.10b 149.77c 13.75ab 10.20a 3.85c 6.15b
Level of Significance * ** ns ** ** ** ** **
LSD 6.333 1.268 1.058 2.20 0.789 2.12 2.10 0.765 CV (%) 2.53 5.08 2.62 0.98 4.14 0.99 0.92 8.35
Here, V= Variety and F= Fertilizer; figure bearing, same or no letter (s) do not differ significantly at 5% level of significance by Duncan's Multiple Range Test, * = Significant at 5% Level of significance, ** = Significant at 1% level of significance, ns= Non- significant.
3.7. Grain yield (t/ ha) on varieties and fertilizer doses
Results showed that grain yield had significant variation on varieties (Figure 1). The
highest 2.79 tha-1 was achieved from BRRI Dhan34 and the lowest 1.92 tha-1 from Tulshimala. The
highest yield in BRRI Dhan34 may be due to higher effective tillers number/hill, panicles length,
grains/ panicle, and grain protein content. These findings were very much like (Kabir et al., 2004).
Scented rice responded significantly to different fertilizer management practices. Dry matter
production during the crop growth period and its translocation to panicles are the major
determinants of grain yield of rice. Further, grain yield of a genotype depends largely on the total
dry matter accumulation and its distribution after anthesis, as the major portion of the dry matter
produced during the post anthesis period is translocated to the panicles. Grain yield increased with
the application of T3 fertilizer management treatment.
Islam et al. JAAST 5(1): 1–12 (2021)
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Results in Figure 2 showed that the highest grain yield (2.64 tha-1) from T3 hence lowest
(1.57 tha-1) from T2 and T4 treatments. A significant improvement in grain yields due to the
combined application of organic and inorganic fertilizer management (Manzoor et al., 2006). The
efficiency of inorganic fertilizers might be increased when it was applied along with organic
manures and brought a beneficial effect on rice grain yield due to an increase in effective tillers/
ha as reported by Srinivas et al. (2010).
Figure 1. Varietal effect on the grain yield of aromatic fine rice
Figure 2. Effect of fertilizer on the grain yield of aromatic fine rice
Figure 3. Interaction effect of varieties and fertilizer managements on the grain yield of aromatic
fine rice
00.51
1.52
2.53
V1 V2 V3 V4
2.03 2.231.92
2.79
Grai
n yi
eld(
t/ha
)
Varieties
0
1
2
3
F1 F2 F3 F4
2.6
1.57
2.64
1.74
Gra
in y
ield
(t/h
a)
Fertilizer
0
0.5
1
1.5
2
2.5
3
3.5
V1F1
V1F2
V1F3
V1F4
V2F1
V2F2
V2F3
V2F4
V3F1
V3F2
V3F3
V3F4
V4F1
V4F2
V4F3
V4F4
2.39
1.73
2.43
1.6
2.6
1.86
2.83
1.73
2.1
1.66
2.2
1.51
3.3
1.6
3.4
2.3
Grai
n yi
eld(
t/ha
)
Interaction of varieties and Fertilizer doses
Islam et al. JAAST 5(1): 1–12 (2021)
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3.8. Straw yield (t/ ha) on varieties and fertilizer doses
Table 1 showed that the highest straw yield 4.06 tha-1 was produced by BRRI Dhan34 and
lowest 3.06 tha-1 by Tulshimala. The effect of the nitrogen level was found to be highly significant
in respect of straw yield (Table 2). The highest 3.67 tha-1 was produced at T1 treatment. Nitrogen
influenced vegetative growth in terms of plant height and tillers number/ hill which resulted in
increased straw yield. Among, the treatments higher straw yield at T1 and the lowest recorded at
T4 treatment. Combination result (Table 3) showed that v4F1 gave highest result. This agreed with
the findings of (Das et al., 2009; Bahadur et al., 2013; Mannan et al., 2013; Meena et al., 2016).
3.9. Biological yield (tha-1) on varieties and fertilizer doses
The varietal effect on biological yield was significant. Table 1 showed the highest 6.85 tha-
1 from BRRI Dhan34 and the lowest 4.98 tha-1 was recorded from Tulshimala. This result was
supported by (Islam et al., 2014; Singh et al., 2017). The effect of bio fertilizer and nitrogen was
found significant in terms of biological yield (Table 2). The highest 6.30 tha-1 was found from T3
and the lowest 4.75 tha-1 was recorded from T2 treatment. Nitrogen level positively influenced
grain yield and straw yield which increased biological yield.
3.10. Harvest index (%) on varieties and fertilizer doses
The harvest index was significantly influenced by varieties. From Table 1 and Figure 4, it
was evident that the highest 40.73% from BRRI Dhan34 and the lowest 38.55 % was recorded
from Tulshimala. The highest 42.9% was found from F3 and the lowest 33.05% was recorded from
F2 treatment (Figure 5). Gill and Aulakh (2018) reported that genotype had a great influence on
the harvest index.
Figure 4. Varietal effect on the harvest index of aromatic fine rice
37
37.5
38
38.5
39
39.5
40
40.5
41
V1 V2 V3 V4
39.8
38.9138.55
40.73
Harv
est i
ndex
(%)
varieties
Islam et al. JAAST 5(1): 1–12 (2021)
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Figure 5. Effect of fertilizer on the harvest index of aromatic rice
Figure 6. Interaction effect of varieties and fertilizer on the harvest index of aromatic rice
3.11. Interaction effect of varieties and fertilizer managements
Table 3 showed that interaction between varieties and nutrient management was significant
on yield and yield components of aromatic fine rice. Highest plant height 165.71cm from V3F1,
highest tiller numbers/plant 18.36 from V4F3, longest panicles length 26.73 cm from V4F3,
longest grains number/panicle 155.33 from V4F3, maximum 1000 grain weight 14.35gm from
V2F3; highest grain protein content 10.80% from V4F3; highest biological yield 7.85 tha-1
fromV4F3; highest grain yield 3.4 tha-1 observed from V4F3 (Figure 3); highest straw yield 4.55
tha-1 from V4F1; the highest harvest index 43.31% from V4F3(Figure 6). Maximum yield and yield
contributing traits found from BRRI Dhan34+interaction with 50% of recommended dose of
fertilizers + 50% cowdung, and BRRI Dhan 34 +75% of recommended dose of fertilizers + 50%
cowdung.
0
10
20
30
40
50
F1 F2 F3 F4
41.47
33.05
42.9
35.51
Har
vest
inde
x(%
)
Fertilizer
0
5
10
15
20
25
30
35
40
45
V1F1
V1F2
V1F3
V1F4
V2F1
V2F2
V2F3
V2F4
V3F1
V3F2
V3F3
V3F4
V4F1
V4F2
V4F3
V4F4
41.6439.7742.19
35.96
41.67
36.33
43
37.240.38
3640.74
33.86
42.04
29.2
43.31
37.4
Harv
est i
ndex
(%)
Interaction of varieties and Fertilizer doses
Islam et al. JAAST 5(1): 1–12 (2021)
10
Besides, lowest plant height 141.89 cm from V1F1, lowest tiller numbers/plant 14.94 from
V2F1; longest panicles length 19.38 cm from V1F4; lowest grains number/panicle 131.34 no. from
V1F1, minimum1000 grain weight 10.43 gm from V1F4; lowest grain protein content 8.20 % from
V1F1; lowest grain yield 1.51 tha-1 observed from V3F4; lowest straw yield 2.62 tha-1 from V1F2;
lowest harvest index 29.20 % from V4F2. Whereas most of the minimum yield contributing traits
found from Kalizira + Recommendation fertilizer doses; and Kalizira + Cowdung @ 10 tha-1. All
the findings are shown (Table 3).
4. Conclusion
From the present study, it can be concluded that yield contributing characters like effective
tillers number/ hill (18.46), longest panicles length (26.67cm), highest grains number/ panicle
(146.30 no), grain protein content (6.23%), grain yield (2.79 tha-1), highest straw yield (4.06 tha-
1), highest biological yield (6.85 tha-1), highest harvest index (40.73 %) was achieved from
aromatic fine rice BRRI Dhan34. Highest effective tiller (16.20 no), highest panicles length
(26.66cm), grains number/panicle (142.45 no), 1000 grain weight (13.75), grain protein content
(10.2%) and biological yield (6.30 tha-1) obtained from 50% of recommended dose of fertilizers +
50% cowdung treatment combination. At the same time, the highest grain yield (3.4 tha-1) was
found between the interaction of BRRI dhan34 with (50% recommended fertilizers doses + 50%
cowdung) treatment. Besides, tillers number/hill, grains number/panicle, grain protein content,
harvest index, grain yield main yield contributing characters to improve the grain yield of aromatic
fine.
Acknowledgments
This research work was supported by the IRT (Institute of Research and Training) Center,
project code (5921) of Hajee Mohammad Daesh Science and Technology University Dinajpur,
Bangladesh for funding research and providing all necessary supports. The authors are grateful to
the Agronomy Department, Faculty of Agriculture, Hajee Mohammad Daesh Science and
Technology University Dinajpur, Bangladesh also for kindly providing the aromatic fine rice
seeds.
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Journal of Applied Agricultural Science and Technology E-ISSN: 2621-2528 5(1): 13-27 (2021) ISSN: 2621-4709
Received November 10, 2020; Accepted March 8, 2021; Published March 10, 2021 https://doi.org/10.32530/jaast.v5i1.4
This is an open access article under the CC BY-SA 4.0 license https://creativecommons.org/licenses/by-sa/4.0
13
IDENTIFICATION OF LOCAL RICE GENOTYPES FROM DELI SERDANG, NORTH SUMATERA, INDONESIA TO DROUGHT STRESS CONDITION
Noverina Chaniago1,2, Irfan Suliansyah3, Irawati Chaniago*,3, Nalwida Rozen3
1Department of Agricultural Science, Faculty of Agriculture, Universitas Andalas, Indonesia
2Department of Agrotechnology, Faculty of Agriculture, Universitas Islam Sumatera Utara, Indonesia 3Department of Agrotechnology, Faculty of Agriculture, Universitas Andalas, Indonesia
*Corresponding author
Email: [email protected] Abstract. Deli Serdang regency in North Sumatera, Indonesia has a high diversity of local rice that has adapted to various climatic and edaphic conditions including drought. Studies on tolerance level of various local rice genotypes to drought are important to be carried out. This will help plant breeders with germplasms to support rice breeding program. Polyethylene glycol (PEG) is a water-soluble compound with high osmotic pressure and unlikely to have specific interaction with biological, chemicals and often to be used in the studies of drought stress in plants. This experiment was aimed to determine tolerance level of local rice genotypes from Deli Serdang, North Sumatera, Indonesia to drought stress. A completely randomized design (CRD) with 3 replicates were used in this study. The first factor was 23 local rice genotypes and 4 tolerance genotypes to drought stress and the second factor was PEG 6000 concentration i.e 0 and 20% (w/v). The percentage of germination, plant height, length and number of roots protruding from paraffin-wax layer, seedling fresh and dry weight, index of tolerance, probability of resistance, and leaf proline content were measured in this study. Based on morphological and physiological characters that measured in this study six local rice genotypes, namely Gemuruh, Ramos Merah, Arias, Sialus, Silayur, and Sirabut were categorized as tolerance to drought stress. These resistant local rice genotypes were potential to be used for further drought stress studies in paddy field. Keywords: local rice; Deli Serdang; drought stress; tolerant
1. Introduction
A landrace is a very valuable genetic asset and need to be managed properly. Landrace has
been cultivated for generations and has adapted well to various climatic conditions. Deli Serdang
Regency in North Sumatera, Indoneasi has a diversity of rice local that found in several villages.
However, management and utilization of the rice’s germplasm are still very low which, in the long
run, may result in a reduction and extinction of the germplasm. This low management and
utilization of local rice is resulted from low availability if the local rice as well as limited number
of farmers growing the rice. Therefore, attempts should be made to preserve this germplasm
collection such as characterization of the existing local rice and improving the characters to
become new superior varieties (Neeraja et al., 2006).
The distribution of plant species across places and various environmental conditions is often
determined by exposure to environmentally-driven stress such as drought. Plant response to
environmental stress may vary and occurs at different organization levels, from morphological,
anatomical, cellular, biochemical, and molecular levels (Manavalan et al., 2009; Muñoz & Quiles,
14 Chaniago et al. JAAST 5(1): 13–27 (2021)
2013). Drought is one of the most serious limiting factor for rice growth and has resulted in a
significant decline in rice productivity (Sabar & Arif, 2014). Drought may affect various stages of
plant growth and development such as seedling vigor (Singh et al., 1999), root depth and density
(Ludlow & Muchow, 1990; Manavalan et al., 2009), and flowering (Bolaños & Edmeades, 1996).
Plant morphological changes such as lengthen roots occurred during drought stress. Roots grew
deeper and reached soil depth for over 20 cm below the surface (Allah et al., 2010) and this to help
plants to reach water for their growth during the shortage of water (Sujinah & Jamil, 2016).
Proline content has been widely used as an indicator of plant response to drought (Barunawati
et al., 2016). Proline is an amino acid that plays important role in preserving nitrogen, acting as an
osmo-regulator, and protecting certain enzymes. Proline levels were found to increase in response
to drought stress in the various crops, such as maize (Yang et al., 2014), wheat (Barunawati et al.,
2016) and rice (Purbajanti et al., 2017; Zain et al., 2014). Proline keeps cell turgor pressure and
supporting root growth during drought (Zivcak et al., 2016).
Polyethylene Glycol (PEG) has been widely used in the study of water stress in plants including
rice (Choi et al., 2000; Swapna & Shylaraj, 2017). PEG is an inert chemical compound, non-toxic,
and has a high molecular weight (Jiang & Lafitte, 2007), increases the osmotic potential of growing
medium that may reduce the amount of water intake of seedlings (Jatoi et al., 2014). Studies in
screening of local rice genotypes from Deli Serdang, North Sumatera, Indonesia to drought stress
has not yet reported. The experiment reported here used PEG to mimic water stress during the
growth and development of rice that collected from Deli Serdang, North Sumatera, Indonesia. The
experiment aimed to determine tolerance level of local rice genotypes from Deli Serdang, North
Sumatera, Indonesia to drought stress. Some morphological and physiological characters were
observed in this study.
2. Methods
2.1. Rice Genotype Materials
Experiments were carried out at Laboratory of Plant Physiology and glasshouse of Faculty of
Agriculture, Universitas Islam Sumatera Utara, Medan, from February to April 2020. Twenty-
three (23) local rice genotypes from Deli Serdang, North Sumetera, Indonesia were subjected to
the experiment. Four (4) rice varieties categorized as tolerant to drought stress were included as
control. A two-way factorial design with 27 rice genotypes and two levels of PEG concentrations
were assigned. Local rice genotypes were Kuku Balam (G1), Siudang (G2), Pandan Wangi (G3),
Sigambiri Merah (G4), Sigantang (G5), Sibelacan (G6), Gemuruh (G7), Sipingkol (G8), Beras
Hitam (G9), Sipirok (G10), Merah Wangi (G11), Serang (G12), Ramos Putih (G13), Ramos Merah
(G14), Arias (G15), Maraisi Merah (G16), Sigambiri Putih (G17), Sijambi (G18), Tambur Kersik
15 Chaniago et al. JAAST 5(1): 13–27 (2021)
(G19), Sialus (G20), Silayur (G21), Sirabut (G22), Sigimbal (G23). In this study, four rice varieties i.e
Inpago 8 (G24), Inpago 10 (G25), Inpago 11(G26) dan Inpari 39 (G27) as categorized as tolerant to
drought stress were used as control.
2.2. Germination Test
Surface sterilization of rice seeds. Rice seeds were washed with running tap water. The seeds
were then washed with distilled water for three times followed by washing in 70% ethanol for 30
seconds. The seeds were then washed with 2% NaOCl for 15 minutes. The sterilised seeds were
immediately washed 4 times with distilled water to get rid of all traces of sterilant and were left to
dry at room temperature.
Seed germination. The rice sterilized seeds were soaked in 20% PEG solution (PEG 6000,
Sigma-Aldrich, Steinheim, Germany) for 60 minutes. Ten seeds were then placed in a 60-mm
diameter Petri dish with 2 mL of 20% PEG and were kept for germination at room temperature for
7 days. The control treatment group used sterile water to wet the germinating medium (0% PEG).
Germination percentage was counted at 7 days after planting.
2.3. Early Growth of Rice with PEG
Early growth of rice seedling was observed in a double pot-growing media. Pre-germinated
rice was transferred onto pots containing paraffin and vaseline mixture (6:4, w/w). The mixture
was preheated at 70°C and was then poured into a perforated-base plastic pot and let to dry. The
solid paraffin and vaseline mixture of 3 mm thick was used as a basal bedding of potting mixture
onto which 200 g of soil and sand mixture (1:1; w/w) was added. Two pre-germinated rice
seedlings with a 1-cm-long radicle were planted at each pot. Another plastic pot containing 45 mL
of Hoagland’s nutrient solution (Harper & Nicholas, 1976) was prepared. The pot containing rice
seedlings was carefully stacked into a nutrient solution pot so that the base of the seedling pot did
not reach the nutrient solution. The pots were carefully placed at a wooden rack in a glass house
and were kept for 4 weeks. Each pot was watered every day with 7 mL of distilled water.
2.4. Measurement and Data Collection
Data on plant responses to drought stress were recorded at 28 days after transplanting. Data
collection includes plant height (PH), number of roots protruding from the paraffin layer (RN),
length of roots protruding from the paraffin layer (RL), and plant fresh and dry weight (following
hot-air dried at 70°C for 48 hours to get a constant weight of plant tissue), stress tolerant index
(Iriany et al., 2005), resistance probability, and leaf proline content. The probability of resistance
(%) is calculated using a curve of normal standard graph. Total values of area below the curve
were used for the calculation following (Sauro, 2007).
16 Chaniago et al. JAAST 5(1): 13–27 (2021)
The measurement of proline content was conducted as follows: 0.5 g of leaves were extracted
with 3 mL of 3% 5-sulphosalicylic acid in a mortar. The leaf extract was then centrifuged at
21,000× g for 15 min. The clear supernatant was carefully poured into a glass vial prior to adding
2 mL of 5-sulphosalicylic acid to the residue. The mixture was centrifuged for a second time. The
second supernatant was poured onto the first supernatant and was thoroughly mixed (so-called a
final supernatant). Two mL of the final supernatant was mixed with 2 mL of 3% ninhydrin reagent
and 2 mL of acetic acid glacial. The mixture was then heated at 100°C for 1 hour in a water-bath
and was cooled in an ice bath. Four mL of toluene was added to the mixture and was mixed for 15
seconds. Absorbance value of the mixture was measured at 520 nm in a spectrophotometer. The
proline content of the leaf was expressed in μM/g FW (fresh weight of leaves) (Larkunthod et al.,
2018).
The tolerance index (TI) was obtained using the following equation (1):
!" = !"!# %
!"$%" (1)
where Yd and Yn respectively represent observed variables under drought and normal conditions.
Hyd is the highest observed variable under drought conditions. TI ˃ 0.5 = tolerant and TI ˂ 0.5 =
susceptible (Fernandez, 1993, as cited in Iriany et al., 2005).
2.5. Data Analysis
Analysis of variance was applied to the data and mean separation was calculated according to
Duncan’s New Multiple Range Test at 5% level. If only the effect of PEG was found to be
significant, then mean separation was calculated as for t-Dunnet at 5%.
Table 1. Analysis of variance of 23 landrace rice of Deli Serdang district and 4 rice varieties tolerant to drought (germination percentage (GP), plant height (PH), length of roots protruding from the paraffin layer (RL), and number of roots protruding from the paraffin layer (RN), fresh weight (FW) and dry weight (DW) of plants
Source of Variance
GP PH RL RN FW DW
Genotype (G) 16.05* 4.75* 20.83* 2.94 * 21.03* 16.18* PEG (P) 855.95* 3.83* 19.90* 1.75 ns 539.87* 83.97* G x P 36.45* 0.17ns 14.18* 6.17* 27.45* 17.67*
Remarks: * (significant); ns (not significant)
3. Results and Discussion
The summary of analysis of variance on various observations demonstrated different responses
to water stress induced by 20% PEG. Responses of germination percentage (GP), plant height
(PH), length of roots protruding from the paraffin layer (RL), and the number of roots protruding
from the paraffin layer (RN), fresh weight (FW), and dry weight (DW) of plants are presented in
Table 1. Rice genotypes (G) significantly affected all variables observed. Water stress (P) affected
all variables but the number of roots protruding from the paraffin layer (RN). The interaction
17 Chaniago et al. JAAST 5(1): 13–27 (2021)
between rice genotypes and water stress only affected germination percentage (GP), length of roots
protruding from the paraffin layer (RL), and the number of roots protruding from the paraffin layer
(RN), fresh weight (FW), and dry weight (DW) of rice plants.
3.1. Germination Percentage
Various germination percentages resulted in different tolerance and probability of resistance
towards water stress under the experimental condition (Table 2).
All rice genotypes showed 100% of germination in 0% PEG except for G1, G2, and G4. These
three rice genotypes showed very low germination which might be resulted from low seed viability
as the seeds had been stored for 6 months after collecting from the field. In contrast, treatment of
20% PEG reduced germination and the response varied within genotypes. interestingly our study
showed that there were 4 local rice genotypes (G14, G16, G21, and G22), germinated similar or higher
than that of tolerant rice varieties. These 4 local rice genotypes demonstrated their potential to be
tolerant to drought with a tolerant index of >0.5.
Table 2. Germination percentage, tolerant index, and the resistant probability of 23 genotypes of local rice in Deli Serdang district and 4 varieties of drought-tolerant in response to PEG
Genotype Germination percentage (%)
Genotype Mean
Tolerance Index
Criteria Resistant Probability
PEG 0 % PEG 20 % (%) Kuku Balam (G1) 20.00 m 13.33 n 16.67 m 0.10 Susceptible 20.80 Siudang (G2) 33.33 k 13.33 n 23.33 l 0.06 Susceptible 6.20 Pandan Wangi (G3) 100.00 a 53.33 i 76.67 e 0.31 Susceptible 60.95 Sigambiri merah (G4) 53.33 i 20.00 m 36.67 k 0.08 Susceptible 13.50 Sigantang (G5) 100.00 a 33.33 k 66.67 g 0.12 Susceptible 24.45 Sibelacan (G6) 100.00 a 60..00 h 80.00 d 0.38 Susceptible 64.60 Gemuruh (G7) 100.00 a 53.00 i 76.67 e 0.30 Susceptible 42.70 Sipingkol (G8) 93.33 b 40.00 j 66.67 g 0.18 Susceptible 39.05 Padi Hitam (G9) 100.00 a 26.66 l 63.33 h 0.07 Susceptible 9.85 Sipirok (G10) 80.00 e 13.33 n 46.67 j 0.02 Susceptible 2.55 Merah Wangi (G11) 100.00 a 53.33 i 76.67 e 0.30 Susceptible 46.35 Serang (G12) 100.00 a 60.00 h 80.00 d 0..38 Susceptible 68.25 Ramos Putih (G13) 100.00 a 53.33 i 76.67 e 0.30 Susceptible 50.00 Ramos Merah (G14) 80.00 d 93..33 b 88.33 b 0.55 Tolerant 75.55 Arias (G15) 100.00 a 33.33 k 66.67 g 0.12 Susceptible 28.10 Maraisi (G16) 100.00 a 73.33 f 86.67 b 0.57 Tolerant 79.20 Sigambiri Putih (G17) 100.00 a 40.00 j 70.00 f 0.17 Susceptible 31.75 Sijambi (G18) 100.00 a 53.33 i 76.67 e 0.30 Susceptible 53.65 Tambur Kersik (G19) 100.00 a 40.00 j 70.00 f 0.17 Susceptible 35.40 Sialus (G20) 93.33 b 26.66 l 60.00 i 0.08 Susceptible 17.15 Silayur (G21) 100.00 a 73.33 f 86.67 b 0.57 Tolerant 82.25 Sirabut (G22) 100.00 a 73.33 f 86.67 b 0.57 Tolerant 86.50 Sigimbal (G23) 100.00 a 53.33 i 76.67 e 0.30 Susceptible 57.30 Inpago 8 (G24)* 100.00 a 86.66 c 93.33 a 0.80 Tolerant 97.45 Inpago 10 (G25)* 100.00 a 73.33 f 86.67 b 0.57 Tolerant 90.15 Inpago 11 (G26)* 100.00 a 66.66 g 83.33 c 0.48 Susceptible 71.90 Inpari 39 (G27)* 100.00 a 73.33 f 86.67 b 0.57 Tolerant 93.80 Mean PEG 90.99 a 50.12 b
Remarks: * (Control varieties: drought-tolerant), CV = 12.59% Mean values within the same column followed by similar small letter are not significantly different at 5% DMRT. Criteria of tolerant index: Ti > 0.5 = tolerant and Ti< 0.5 = susceptible
18 Chaniago et al. JAAST 5(1): 13–27 (2021)
3.2. Plant Height
Water stress created by the application of 20% PEG did not affect the plant height of all rice
genotypes tested (Table 3). All genotypes were tolerant to water stress except for G1 and G22.
The tolerant genotypes had a resistant probability of 4.05-93.80%. However, there were 8 local
rice genotypes (G10, G12, G4, G8, G21, G3, G13, and G5) with a resistant probability of >60% and
higher than that of 3 tolerant rice varieties (G24, G26, and G27).
Table 3. Plant height, tolerant index, and the resistant probability of 23 genotypes of local rice of Deli Serdang district and 4 varieties of drought-tolerant in response to PEG, 4 weeks after planting
Genotype Plant height (cm) Genotype Tolerant Index
Criteria Resistant Probability
(%) PEG 0 % PEG 20 % Mean
Kuku Balam (G1) 30.00 12.66 21.33 k 0.13 Susceptible 2.55 Siudang (G2) 35.83 34.33 35.08 ab 0.80 Tolerant 3.05 Pandan Wangi (G3) 35.00 37.73 36.37 b 0.99 Tolerant 75.55 Sigambiri merah (G4) 31.00 39.16 35.08 b 1.20 Tolerant 90.15 Sigantang (G5) 33.66 36.16 34.92 b 0.94 Tolerant 64.60 Sibelacan (G6) 34.33 34.83 34.58 b 0.86 Tolerant 53.65 Gemuruh (G7) 29.00 30.66 29.83 ef 0.79 Tolerant 31.75 Sipingkol (G8) 28.33 36.33 32.33 cd 1.13 Tolerant 86.50 Padi Hitam (G9) 30.33 32.66 31.50 de 0.85 Tolerant 50.00 Sipirok (G10) 28.00 40.66 34.33 bc 1.44 Tolerant 97.45 Merah Wangi (G11) 25.66 26.50 26.08 j 0.66 Tolerant 24.45 Serang (G12) 34.00 41..00 37.50 a 1.20 Tolerant 93.80 Ramos Putih (G13) 30.75 34.66 27.58 ghij 0.95 Tolerant 68.25 Ramos Merah (G14) 29.00 25.66 27.33 hij 0.55 Tolerant 13.50 Arias (G15) 30.00 34.50 27.25 ij 0.96 Tolerant 71.90 Maraisi (G16) 31.33 30.83 31.08 def 0.73 Tolerant 28.10 Sigambiri Putih (G17) 29.33 31.66 30.50 def 0.83 Tolerant 42.70 Sijambi (G18) 24.33 29.00 26.67 j 0.84 Tolerant 46.35 Tambur Kersik (G19) 32.33 27.00 29.67 efg 0.54 Tolerant 9.80 Sialus (G20) 29.66 32.66 31.17 def 0.87 Tolerant 57.30 Silayur (G21) 26.00 33.00 29.50 efgh 1.02 Tolerant 79.20 Sirabut (G22) 30.00 13.50 19.50 k 0.15 Susceptible 6.20 Sigimbal (G23) 33.66 30.00 31.83 de 0.65 Tolerant 20.80 Inpago 8 (G24)* 26.33 31.66 29.00 fghi 0.92 Tolerant 60.95 Inpago 10 (G25)* 27.00 33.83 30.42 def 1.03 Tolerant 82.85 Inpago 11 (G26)* 26.00 24..83 25.42 j 0.57 Tolerant 17.15 Inpari 39 (G27)* 24.66 28.33 26.50 j 0.79 Tolerant 35.40 Mean PEG 29.09 a 31.09 b
Remarks: * (Control varieties: drought-tolerant), CV = 21.64% Mean values within the same column followed by similar small letter are not significantly different at 5% DMRT. Criteria of tolerant index: Ti > 0.5 = tolerant and Ti< 0.5 = susceptible.
3.3. Length and Number of Roots Protruding from Paraffin Layer
Data of length and number of roots protruding from paraffin layer 4 weeks after planting are
presented in Table 4. Roots of local rice from the 20% PEG treatment group were shorter than that
of 0% PEG group. Within the treatment group of 20% PEG, local rice genotype Arias (G15)
demonstrated the longest root, in contrast to genotype Inpago 8 (G24) with the shortest root, 11.0
and 3.0 cm respectively. Water stress through the application of 20% PEG reduced root length and
the number of roots of rice variety Inpago 8 (G24), 40 and 75% respectively. Rice var. Inpago 8 is
19 Chaniago et al. JAAST 5(1): 13–27 (2021)
one of drought-tolerant rice varieties used for this experiment. Among all 27 rice genotypes tested,
there were only 6 genotypes showing water stress tolerance. They were genotypes G2, G4, G7, G8,
G9, and G15. It was interesting to note that 3 rice varieties tolerant to water stress (G25, G26, and
G27) had a lower probability to be drought-tolerant with a probability of <60%.
The number of roots protruding from paraffin layer varied between the rice genotypes. The
highest number of roots was observed from local rice genotypes G10 and G17 with 4 roots observed
at each pot. All 4 drought-tolerant rice varieties had a tolerant index lower than 0.5 according to
the number of roots protruding from paraffin layer. Therefore, these four rice varieties fell into a
category of susceptible to drought according to their root growth.
3.4. Plant fresh and dry weight
In general, PEG-induced water stress reduced plant fresh and dry weight of local rice
genotypes and 4 drought-tolerant rice varieties (Table 5). According to plant fresh weight, all 23
local rice genotypes were susceptible to drought with tolerant index was lower than 0.5. However,
seven genotypes showed a >60% probability of drought-resistant according to plant fresh weight,
i.eG10 (93.80%), G15 (90.15%), G14 (86.50%), G8 (82.85%), G5 (79.20%), G1 (75.55%) and G12
(71.90%). A different response was observed in plant dry weight. Six rice genotypes, G3, G6, G11,
G15, G20 dan G22 were tolerant to water stress. Eight local rice genotypes had a probability to be
resistant to drought (G6, G22, G15, G20, G11, G3, G2, and G21). Interestingly, G24 (Inpago 8), a
drought-tolerant rice variety was considered susceptible according to plant fresh and dry weight.
3.5. Proline Content
PEG-induced water stress increased proline content in the leaf of local rice genotypes tested
(Table 6). The highest proline content was observed form rice genotype G21 (6,404 µM/g),
followed by G19 (5,052 µM/g), G10 (4,517 µM/g), G14 (4,188 µM/g), G1 (4,169 µM/g) and G13
(4,047 µM/g). The proline content of these six local genotypes was higher than that of four
drought-tolerant rice varieties.
All local rice genotypes studied were tolerant to water stress under the experimental
condition except for genotypes G2, G4, G5, and G16 according to the value of tolerant index.
However, a different response was observed for the percentage of drought resistance probability.
There were nine local rice genotypes with >60% drought resistance probability, i.e. G21 (97.45%),
G19 (93.80%), G1 (90.15%), G14 (82.85%), G10 (79.20%), G13 (75.55%), G7 (71.90%), G20 (64.60%)
and G3 (60.95%). Interestingly, these nine local genotypes showed higher probability of drought
tolerance than that of two drought-tolerant varieties, Inpago 11 (G26) and Inpari 39 (G27). All
genotypes tested except for Sigambiri Merah and Sigantang increased their proline content in
response to drought stress. In addition, genotypes Sigambiri Merah and Sigantang showed proline
20 Chaniago et al. JAAST 5(1): 13–27 (2021)
content lower than that of other rice genotype tested, 0.23 and 0.18 respectively. Therefore, one of
the criteria used to select resistant genotypes to drought at this research is high level of proline.
Seed germination mostly depends on the metabolic activities and the stimulation of growth
hormones in seeds. Water availability is necessary for seed germination and insufficient water will
inhibit the germination process. However, to some extent, drought-resistant seeds may germinate
and grow under water stress conditions. The research reported here found significant differences
in the germination rate of local rice tested. Four local rice genotypes were found to be tolerant to
water stress with a germination rate higher than that of four drought-tolerant varieties, i.e. Ramos
Merah (93.33%) followed by Silayur, Sirabut, and Marasi with a 73.33% germination rate for each
genotype. Rice seed treated with PEG absorbed less water as PEG increased the osmotic potential
of the growing media which resulted a reduction in germination percentage (Jatoi et al., 2014).
The higher the concentration of PEG applied the lower the germination rate of two varieties of
black rice tested (Nurmalasari, 2018).
The plant height of local rice was not affected by PEG under this experimental condition.
Therefore, plant height would not be a good character to evaluate the drought resistance of rice
genotype tested. However, it was interesting to note that plant height was higher in PEG-treated
rice than the control group. This may result from a fast response to water once the seedlings were
removed to the glass house for further growth. Slow germination process under water stress would
reserve some of the resources from the endosperm and be used for later growth. In the other hands,
rice seeds from the control treatment group undergone a high rate of metabolic process during
germination in the presence of enough water. When these seedlings were removed from the
glasshouse, they may have needed more energy and resources to adapt to the new environment
with certain stagnancy in early rapid growth. Research has shown that drought-tolerant rice grew
quite well with a relatively good plant height (Kumar et al., 2014; Larkunthod et al., 2018; Mejri
et al., 2016).
Commonly, plant increases root cell division and elongation to reach water table in the soil
under water stress. This has been known as one mechanism to adapt to the water deficiency (Allah
et al., 2010). A significant difference is in the root length and the number of roots protruding from
paraffin layer at 4 WAP was recorded between the PEG and the control treatment group. Rice
genotype with a potential to be tolerant to drought had longer roots protruding from the paraffin
though less amount of roots was recorded. Therefore, the amount of root protruding from the
paraffin was not good enough to be used as an indicator for drought tolerance.
21 Chaniago et al. JAAST 5(1): 13–27 (2021)
Table 4. Length of roots and number of roots protruding from the paraffin layer, tolerant index, and resistant probability of 23 genotypes of local rice of Deli Serdang district and 4 varieties of drought-tolerant in response to PEG, 4 weeks after planting
Genotype Root length (cm) PEG (0%) PEG (20%)
Genotype Mean
Tolerant Index
Crite-ria
Resistant Prob (%)
Root number PEG 0% PEG(20%)
Genotype Mean
Tolerant Index
Crite-ria
Resistant Prob (%)
Kuku Balam (G1) 0.3 opq 0.0 q 0.15 j 0.00 S 2.55 2.00 d 0.00 f 1.00 e 0.00 S 2.55
Siudang (G2) 0.5 no 6.0 g 3.25 e 6.55 T 97.45 2.00 d 3.00 c 2.50 cd 1.13 T 82.85
Pandan Wangi (G3) 0.5 nop 1.0 lm 0.75 hij 0.18 S 68.25 5.00 a 3.00 c 4.00 a 0.45 S 53.65
Sigambiri merah (G4) 0.2 pq 3.0 j 1.60 fg 4.09 T 93.80 1.00 e 2.00 d 1.50 de 1.00 T 71.90
Sigantang (G5) 0.0 q 0.3 opq 0.15 j 0.00 S 6.20 0.00 f 1.00 e 0.50 g 0.00 S 6.20
Sibelacan (G6) 8.0 e 1.0 lm 4.50 d 0.01 S 24.45 2.00 d 3.00 c 2.50 cd 1.13 T 86.50
Gemuruh (G7) 0.7 mno 5.5 h 3.10 e 3.92 T 90.15 1.00 e 2.00 d 1.50 de 1.00 T 75.55
Sipingkol (G8) 6.0 g 8.23 e 7.12 b 1.03 T 82.85 4.00 b 2.00 d 3.00 b 0.25 S 39.05
Padi Hitam (G9) 11.0 c 10.0 d 10.50 a 0.82 T 79.20 2.00 d 1.00 e 1.50 de 0.13 S 31.75
Sipirok (G10) 1.2 l 1.0 lm 1.10 gh 0.07 S 60.95 2.00 d 4.00 b 3.00 b 2.00 T 90.15
Merah Wangi (G11) 0.5 nop 0.5 nop 0.50 hij 0.04 S 46.35 1.00 e 1.00 e 1.00 e 0.25 S 42.70
Serang (G12) 2.0 k 2.0 k 2.00 f 0.01 S 28.10 3.00 c 1.00 e 2.00 d 0.08 S 28.10
Ramos Putih (G13) 0.5 nop 0.4 nopq 0.45 ij 0.02 S 39.05 1.00 e 1.00 e 1.00 e 0.25 S 46.35
Ramos Merah (G14) 0.5 nop 1.0 lm 0.75 hij 0.18 S 71.90 1.00 e 3.00 c 2.00 d 2.25 T 93.80
Arias (G15) 10.0 d 11.0 c 10.50 a 1.10 T 86.50 3.00 c 3.00 c 3.00 b 0.75 T 64.60 Maraisi (G16) 1.0 lm 0.8 lmn 0.90 hi 0.05 S 53.65 1.00 e 2.00 d 1.50 de 1.00 T 79.20
Sigambiri Putih (G17) 0.5 nop 1.0 lm 0.75 hij 0.18 S 75.55 1.00 e 4.00 b 2.50 cd 4.00 T 97.45
Sijambi (G18) 1.0 lm 0.8 lmn 0.90 hi 0.06 S 57.30 3.00 c 3.00 c 3.00 b 0.75 T 68.25
Tambur Kersik (G19) 20.0 a 0.0 q 10.00 a 0.00 S 9.85 1.00 e 0.00 f 0.50 f 0.00 S 9.85
Sialus (G20) 7.0 f 1.0 lm 4.00 d 0.01 S 31.75 5.00 a 2.00 d 3.50 ab 0.20 S 35.40
Silayur (G21) 0.5 nop 0.5 nop 0.0 hij 0.04 S 50.00 2.00 d 2.00 d 2.00 d 0.50 T 57.30
Sirabut (G22) 12.0 b 0.0 q 6.00 c 0.00 S 13.50 2.00 d 0.00 f 1.00 e 0.00 S 13.50
Sigimbal (G23) 0.5 nop 0.3 opq 0.40 ij 0.01 S 35.40 2.00 d 2.00 d 2.00 d 0.50 T 60.95
Inpago 8 (G24)* 5.0 i 3.0 j 4.00 d 0.16 S 64.60 4.00 b 1.00 e 2.50 cd 0.06 S 24.45
Inpago 10 (G25)* 1.0 lm 0.5 nop 0.75 hij 0.02 S 42.70 1.00 e 1.00 e 1.00 e 0.25 S 50.00
Inpago 11 (G26)* 0.5 nop 0.0 q 0.25 j 0.00 S 17.15 1.00 e 0.00 f 0.50 f 0.00 S 17.15
Inpari 39 (G27)* 0.5 nop 0 q 0.25 j 0.00 S 20.80 1.00 e 0.00 f 0.50 f 0.00 S 20.80
Mean PEG 3.39 a 2.18 b 2.00 1.74 Remarks: * (Control varieties: drought-tolerant). Mean values within the same column followed by similar small letter are not significantly different at 5% DMRT. Criteria of tolerant index: Ti > 0.5 = tolerant (T) and Ti < 0.5 = susceptible (S). CV for root length = 21.85%; CV for root number = 26.68%
22 Chaniago et al. JAAST 5(1): 13–27 (2021)
Table 5. Plant fresh and dry weight, tolerant index, and resistant probability of 23 genotypes of local rice of Deli Serdang district and 4 varieties of drought-tolerant in response to PEG, 4 weeks after planting
Genotype Fresh weight (g) PEG (0%) PEG (20%)
Genotype Mean
Tolerant Index
Crite-ria
Resistant Prob (% )
Dry weight (g) PEG 0% PEG 20%
Genotype Mean
Tolerant Index
Crite-ria
Resistant Prob (%)
Kuku Balam (G1) 0.46 c 0.16 q 0.31 b 0.29 S 75.55 0.16 b 0.10 g 0.13 a 0.42 S 53.65
Siudang (G2) 0.25 l 0.13 u 0.19 ij 0.25 S 60.95 0.12 e 0.09 h 0.11 c 0.45 S 64.60
Pandan Wangi (G3) 0.16 q 0.11 w 0.14 m 0.15 S 17.15 0.08 i 0.08 i 0.08 k 0.53 T 68.25
Sigambiri merah (G4) 0.32 g 0.10 w 0.21 gh 0.21 S 46.35 0.12 e 0.07 j 0.10 cd 0.27 S 35.40
Sigantang (G5) 0.48 b 0.10 w 0.29 cd 0.29 S 79.20 0.17 a 0.08 i 0.13 a 0.25 S 24.45
Sibelacan (G6) 0.21 o 0.13 u 0.17 kl 0.22 S 50.00 0.08 i 0.11 f 0.10 cd 1.01 T 93.80
Gemuruh (G7) 0.17 q 0.10 w 0.14 m 0.14 S 13.50 0.07 j 0.05 l 0.06 fg 0.24 S 20.80
Sipingkol (G8) 0.30 i 0.14 t 0.22 fg 0.31 S 82.85 0.13 d 0.09 h 0.11 c 0.42 S 57.30
Padi Hitam (G9) 0.26 jk 0.12 v 0.19 ij 0.23 S 53.65 0.13 d 0.07 j 0.10 ij 0.25 S 28.10
Sipirok (G10) 0.55 a 0.10 w 0.33 a 0.33 S 93.80 0.17 a 0.06 k 0.12 ab 0.14 S 9.85
Merah Wangi (G11) 0.17 q 0.11 w 0.14 m 0.15 S 20.80 0.06 k 0.07 j 0.07 f 0.54 T 75.55
Serang (G12) 0.43 e 0.10 w 0.26 e 0.27 S 71.90 0.16 b 0.08 i 0.12 ab 0.27 S 39.05
Ramos Putih (G13) 0.35 f 0.11 w 0.23 f 0.25 S 64.60 0.09 h 0.06 k 0.08 ef 0.27 S 42.70
Ramos Merah (G14) 0.45 d 0.11 w 0.28 d 0.31 S 86.50 0.17 a 0.09 h 0.13 a 0.32 S 46.35
Arias (G15) 0.23 m 0.16 q 0.20 hj 0.31 S 90.15 0.12 e 0.12 e 0.12 ab 0.80 T 86.50 Maraisi (G16) 0.26 k 0.11 w 0.19 ijk 0.20 S 39.05 0.13 d 0.07 j 0.10 cd 0.25 S 31.75
Sigambiri Putih (G17) 0.31 h 0.12 v 0.22 g 0.26 S 68.25 0.10 g 0.05 l 0.08 ef 0.17 S 13.50
Sijambi (G18) 0.22 n 0.10 w 0.16 l 0.16 S 24.45 0.13 d 0.05 l 0.09 e 0.13 S 6.20
Tambur Kersik (G19) 0.27 j 0.10 w 0.19 ijk 0.19 S 31.75 0.07 j 0.06 k 0.07 f 0.34 S 50.00
Sialus (G20) 0.10 w 0.10 w 0.10 no 0.10 S 2.55 0.04 m 0.06 k 0.05 g 0.60 T 79.20
Silayur (G21) 0.10 w 0.10 w 0.10 no 0.10 S 6.20 0.04 m 0.05 l 0.05 g 0.42 S 60.95
Sirabut (G22) 0.17 q 0.15 s 0.16 l 0.24 S 57.30 0.10 g 0.11 f 0.11 c 0.81 T 90.15
Sigimbal (G23) 0.22 n 0.10 w 0.16 l 0.16 S 28.10 0.10 g 0.03 n 0.07 f 0.06 S 2.55
Inpago 8 (G24)* 0.22 n 0.12 v 0.17 kl 0.20 S 42.70 0.08 i 0.05 l 0.07 f 0.21 S 17.15
Inpago 10 (G25)* 0.23 m 0.11 w 0.17 kl 0.19 S 35.40 0.08 i 0.08 i 0.08 ef 0.53 T 71.90
Inpago 11 (G26)* 0.13 u 0.11 w 0.12 n 0.13 S 9.85 0.06 k 0.08 i 0.07 f 0.71 T 82.85
Inpari 39 (G27)* 0.16 q 0.20 p 0.18 jkl 0.36 S 97.45 0.06 k 0.15 c 0.11 c 2.50 T 97.45
Mean PEG 0.27 a 0.12 b 0.11 a 0.08 b Remarks: * (Control varieties: drought-tolerant). Mean values within the same column followed by similar small letter are not significantly different at 5% DMRT.
Criteria of tolerant index: Ti > 0.5 = tolerant (T) and Ti < 0.5 = susceptible (S), CV for plant fresh weight = 21.02%; CV for plant dry weight = 21.70
23 Chaniago et al. JAAST 5(1): 13–27 (2021)
Table 6. Proline content, tolerant index, and resistant probability of 23 genotypes of local rice of Deli Serdang district and 4 varieties of drought-tolerant in response to PEG, 4 weeks after planting
Genotype Proline content (µM/g) Genotype Tolerant Index
Criteria Resistant Probability
(%) PEG 0 % PEG 20 % Mean
Kuku Balam (G1) 1.424 4.169 2.79 1.91 Tolerant 90.15 Siudang (G2) 1.671 2.094 1.88 0.40 Susceptible 13.50 Pandan Wangi (G3) 2.028 3.672 2.85 1.04 Tolerant 60.95 Sigambiri merah (G4) 1.869 1.671 1.77 0.23 Susceptible 6.20 Sigantang (G5) 2.047 1.549 1.79 0.18 Susceptible 2.55 Sibelacan (G6) 1.963 2.911 2.43 0.67 Tolerant 35.40 Gemuruh (G7) 1.474 3.324 2.39 1.17 Tolerant 71.90 Sipingkol (G8) 1.587 2.601 2.09 0.67 Tolerant 39.05 Padi Hitam (G9) 1.268 2.113 1.69 0.55 Tolerant 28.10 Sipirok (G10) 2.113 4.517 3.31 1.51 Tolerant 79.20 Merah Wangi (G11) 1.399 2.263 1.83 0.57 Tolerant 31.75 Serang (G12) 2.686 2.019 2.35 0.24 Susceptible 9.85 Ramos Putih (G13) 1.887 4.047 2.96 1.36 Tolerant 75.55 Ramos Merah (G14) 1.794 4.188 2.99 1.53 Tolerant 82.85 Arias (G15) 1.352 2.911 2.13 0.98 Tolerant 53.65 Maraisi (G16) 1.812 2.160 1.98 0.40 Susceptible 17.15 Sigambiri Putih (G17) 1.681 2.986 2.33 0.83 Tolerant 46.35 Sijambi (G18) 1.427 2.225 1.82 0.54 Tolerant 24.45 Tambur Kersik (G19) 1.747 5.052 3.39 2.28 Tolerant 93.80 Sialus (G20) 1.418 3.155 2.28 1.09 Tolerant 64.60 Silayur (G21) 1.578 6.404 3.99 4.06 Tolerant 97.45 Sirabut (G22) 2.620 3.700 3.16 0.82 Tolerant 42.70 Sigimbal (G23) 1.578 3.005 2.37 0.89 Tolerant 50.00 Inpago 8 (G24)* 1.427 3.164 2.29 1.09 Tolerant 68.25 Inpago 10 (G25)* 1.455 3.803 2.62 1.55 Tolerant 86.50 Inpago 11 (G26)* 1.596 2.272 1.93 0.50 Tolerant 20.80 Inpari 39 (G27)* 1.887 3.512 2.69 1.02 Tolerant 57.30 Mean PEG 1.733 3.166
Remarks: * (Control varieties: drought-tolerant), CV = 13.09%
Many rice seedlings with high amount of roots protruding from paraffin did not
reach the Hoagland’s solution and died. We found 6 local rice genotypes tolerant to water
stress with root longer than that of drought-tolerant varieties control group. The longest
root (11 cm) was recorded in genotype Arias with 3 roots protruded from paraffin layer.
Our finding was in accordance with previous study that rice var. Batang Piaman, Cisokan,
and Ceredek had longer roots in response to water stress (Rahmadianti et al., 2017). A
similar phenomenon was reported in sorghum. A higher amount of roots protruding form
paraffin layer was observed from a 20% PEG 6000 treatment group compared to control
treatment group. Water deficiency affected root length of sorghum. Five out of 10 tested
sorghum genotypes were drought-tolerant based on their ability to protrude from paraffin
layer (Chaniago et al., 2017).
A significant reduction in plant fresh and dry wight was recorded from the 20%
PEG treatment group. Six local rice were tolerant to drought according to plant dry
24 Chaniago et al. JAAST 5(1): 13–27 (2021)
weight. It was Pandan Wangi, Sibelacan, Merah Wangi, Arias, Sialus dan Sirabut. Water
stress resulted a reduction in plant growth as reflected by reduction in plant weight
(Larkunthod et al., 2018; Sulistyo et al., 2016). Water deficiency affects plant
physiological process leading to morphological changes. Plants respond through a
reduction in leaf transpiration rate and stomatal enclosure due to loss of cell turgidity
(Taiz & Zeiger, 2006). Stomatal enclosure inhibits gas exchange, such as CO2 and O2,
between plants and atmosphere through stomata (Liu et al., 2004) which will in turn
reduce the photosynthesis and other physiological processes in the plants and will reduce
biomass in the plant tissue (Sujinah & Jamil, 2016). Reduction in the growth and
development of rice plants is impaired by physiological and environmental tensions.
Climate changes and shortage of water has become a major global issue in food
production. Drought may limit plant growth through alteration in physiological and
biochemical process at various levels from cellular to a whole plant (Rahim et al., 2020).
Water balance during cropping seasons determines plant growth and yield. Insufficient
amount of water reduced yield and farmers’ economic return (Dwiratna et al., 2018).
However, a certain microorganism such as arbuscular mycorrhizae fungi could facilitate
plant roots to develop better and acquire soil water table. Some species of arbuscular
mycorrhizae fungi has demonstrated to be directly involved in the infection and intensify
root growth of citronella plants in dry area of West Sumatera (Armansyah et al., 2018).
The proline content increased in response to water stress. Some genotypes showed
a significant increase in proline content. Local rice genotype Silayur increased its proline
as much as 75.35% over non-stressed control treatment. Increase in proline content in
other genotypes varied and 9 genotypes showed an increase for over 50%. Water stress
as induced by PEG caused negative effect to plants’ growth rate and cell enlargement
through reduction in the rates of plant hormones and turgor pressure (Inostroza et al.,
2015). Furthermore, drought stress may result in damage of cell membrane through
stimulation of free radical formation in cells. Cell membrane impairment has also been
used as a major parameter for cellular response to water stress (Sharifi et al., 2012; Rahim
et al., 2020). However, a simple method of determining proline content is considered
enough to determine plant response to water stress. Proline is an amino acid produced by
plants exposed to drought stress and function as osmo-protectant to adjust cell osmolality
(Nurmalasari, 2018). Increased level of proline enables drought-stressed plants to keep
low water potentials, and play major role in maintaining cell turgor pressure and root
growth (Zivcak et al., 2016).
25 Chaniago et al. JAAST 5(1): 13–27 (2021)
Various criteria might be used to determine whether rice genotypes are susceptible
or tolerant to PEG-induced drought. Our data demonstrate various response to different
criterion. For instance, local rice genotype Sirabut was classified as susceptible according
to its plant height and root length. In contrast, genotype Sirabut showed to be tolerant in
term of its germination percentage, plant dry weight, and proline content of leaf. The
highest proline content was observed from genotype Sirabut. Therefore, we combined
some criteria such as germination percentage, root length, plant dry weight, and proline
content to determine to local rice genotype tolerant to drought.
4. Conclusions
Local rice of Deli Serdang subjected to PEG-induced water stress under the
experimental condition revealed that 6 (six) genotypes were found to be tolerant to
drought according to the tolerant index and drought-resistant probability, and high proline
content. The genotypes were Gemuruh, Ramos Merah, Arias, Sialus, Silayur, and Sirabut.
These six local rice genotypes will be used for further experiment in search for drought
resistant rice genotypes.
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Armansyah, Anwar, A., Syarif, A., Yusniwati, & Febriamansyah, R. (2018). Exploration and identification of the indigenous Arbuscular Mycorrhizae Fungi (AMF) in the rhizosphere of citronella (Andropogon nardus L.) in the dry land regions in West Sumatra Province, Indonesia. International Journal on Advanced Science, Engineering and Information Technology, 8(1), 85–92. https://doi.org/10.18517/ijaseit.8.1.2363
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Journal of Applied Agricultural Science and Technology E-ISSN: 2621-2528 5(1): 28-37 (2021) ISSN: 2621-4709
Received January 30, 2021; Accepted March 5, 2021; Published March 10, 2021 https://doi.org/10.32530/jaast.v5i1.8 This is an open access article under the CC BY-SA 4.0 license https://creativecommons.org/licenses/by-sa/4.0
28
ANTIFUNGAL ACTIVITY OF ESSENTIAL OILS OF LEAVES, RHIZOMES OILS AND FRACTION WILD GINGER Elettariopsis slahmong CK Lim INHIBIT THE
COLONY GROWTH OF Sclerotium rolfsii
Nurmansyah*,1, Herwita Idris1, Nasril Nasir2
1Research Instalation of Bogor Research Institute for Spice and Medicinal Plants, Laing Solok, Indonesia
2Biology Departement, Faculty of Mathematics and Natural Sciences, Andalas University, Padang, Indonesia
*Corresponding author
Email: [email protected]
Abstract. This study aims to see the effectiveness of essential oils of leaves, rhizomes and fraction of wild ginger Ellettariopsis slahmong CK Lim against the pathogenic fungus Sclerotium rolfsii which causes rot disease of the stem base of peanut plants an in vitro.The study consisted of two sub activities: (a) inhibited of colony diameter using Patato Dextrosa Agar (PDA) medium and (b) inhibited of colony biomass using Potato dextrose Broth (PDB) medium, the treatments tested were leaf essential oil and rhizome of wild ginger and fractions A1, B2, C3 and D4, with concentration levels (0, 100, 250 and 500 ppm). Experiments (a) and (b) were arranged in the form of a Completely Randomized Design (CRD) in factorial each with 4 replications. The results showed that the leaf essential oil and rhizome of wild ginger and its fractions had the potential to be used as a vegetable fungicide. The A1 fraction has the best antifungal effectiveness compared to the B2 fraction, leaf oil, rhizomes and other fractions, with the highest inhibition of diameter and biomass of S rolfsii colony the 49.47% and 51.46%. Essential oils of leaves and rhizome oil are not statistically significantly different, but in numerically leaf oil are better than rhizome oil. The C3 fraction showed the lowest colony diameter inhibition and biomass of 34.70% and 36.95%. The best concentration level in inhibition the growth of S rolfsii mushroom is 500 ppm, with inhibition of the diameter and biomass of the colony by 81.74% and 84.25%. Keywords: essential oil; rhizomes; fraction; wild ginger; sclerotum rolfsii
1. Introduction
Wild Ginger Elettariopsis slahmong C.K. Lim, is a wild plant of the family zinggiberaceae
which was first discovered in Thailand by CK Lim this plant, it has a very strong pungent odor
called the smelly leavis or stink bug plant (Picheansoonth & Yupparach, 2007). There are several
other species of the genus Elettariopsis that have been studied are E elan C.K. Lim, E exserta
(Scort) Bak, E smithiae Y. K. Kam, E triloba (Gagneb) Loes, E manophylla and E wandokthong
which are new species from Thailand (Picheansoonthon & Yupparach, 2010). Wild Ginger
contains essential oils from leaves, rhizomes and roots obtained by hydro destilation (Wong et al,
2006). This wild ginger plant is widely used as a spice and salad in Malaysia, Thailand and
Indonesia (Magdaulih & Nasir, 2014).
These wild ginger plants can also be found in West Sumatra as wild plants in the forest of
Bonjol Pasaman Regency, Kinali Regency of West Pasaman, Anai Valley Padang Pariaman
Regency, Lubuak Basuang Agam Regency and Aia Angek Sijunjung Regency. Wild ginger plants
Nurmansyah et al. JAAST 5(1): 28 –37 (2021) 29
contain essential oils from leaves and rhizomes and their roots. Essential oils and volatile
compounds from this plant are insecticidal against the Drosophila melanogaster insect which is a
vector of blood diseases in banana plants (Nasir et al., 2014).
Research on the effect of wild ginger essential oil on plant pathogenic fungi has not been
widely tested, essential oils from leaves can control the pathogen Colletotrichum gloesporioides
that causes Antrachnose disease in the red dragon fruit plant Hylocereus polyrhizuz. At the
concentration level of 1000 ppm leaf oil can suppress the growth of pathogenic C gleosporioides
100% (Nasir & Nurmansyah, 2016), it is also effective against Phytophthora palmivora fungi
(Nasir, 2017).
The main content of wild ginger leaf oil is 2-decanoic acid 48.04%, nonanoic acid 9.18%,
2-octenal 8.97%, nonanal 2.96%, octanal 1.2% and 75 other components below 1% (Nasir &
Nurmansyah, 2016), The main components of rhizomes and roots are 2-Tridecenal 39.81%, 2-
decanoic acid 26.3%, 2 octanal 7.56%, hexadecanoic acid 2.86%, nonanoic acid 3.85% and 70
other components below 1% (Nasir, 2016).
Sclerotium rolfsii is a pathogenic fungus that attacks various plants including peanuts,
tomatoes, chili, papaya, orchids and soursop. This pathogenic fungus is also known to attack
patchouli plants (Sukamto & Wahyuno, 2013), S rolfsii is a soil borne pathogen such as Botrytis
cinerea, Pythium sp, Rhizoctonia solani, Sclerotinia minor dan Verticillium dahlia (Sumartini,
2012; Thiessen & Woodward, 2012)
The yield decrease caused by S. rolfsii fungus attack on peanut plants can reach 44.51 percent
(Buhaira & Asniwita, 2009). Each component has not been tested, therefore further research is
recommended. Antifungal essential oil chemical components that are able to penetrate the fungal
cell walls, thereby causing interference with the metabolic processes in the cell so that it interferes
with cell growth, at certain concentrations will result in the death of the fungal cells.
Control of this pathogen can be done with leaf oil fungicide and cinnamon stick
(Cinnamomum burmanii) with a concentration level of 500 ppm able to suppress the growth of the
colony 100% (Nurmansyah, 2014) and Piper aduncum oil at a concentration level of 500 ppm
able to control the pathogen S rolfsii reached 92.77% (Nurmansyah, 2016). Extract of garlic can
control the patogen S rolfsii 92.66% at concentration 5% (Supriyono, 2011). The cironella and
bamboo piper oil botanical fungicides have a good enough ability to suppress the stem rot disease
of peanut with an emphasis percentage of 86.38 and 93.21% at interval application one time a
week ( Idris et al, 2020).
Based on the description above, then the use of wild ginger pesticides to control plant
diseases has enough potential, considering that these plants are wild plants and will be more
beneficial if they can be developed as plants that have economic value for controlling pests and
Nurmansyah et al. JAAST 5(1): 28 –37 (2021) 30
plant diseases. This study aims to look at the effectiveness of leaf oil fungicides, wild ginger
rhizomes (Ellettariopsis slahmong CK Lim) and their fraction of Sclerotium rolfsii fungi that cause
root rot disease in the peanut an in vitro.
2. Methods
The study was conducted from April to December 2018 in the pest and disease laboratory of the
Laing Solok Experiment Garden in West Sumatra.
2.1 Distillation and fractionation
Wild ginger oil is obtained by distillation of leaves and rhizomes, which are withered for
4 days for the leaves first, while the rhizomes are dried for about 10 days, the ingredients are
chopped before being distilled first. Distillation is carried out using a kettle protip Balittro system
of steam, the results of the distillation of leaf oil and rhizomes are mixed and fractionated at the
Faculty of Agriculture of Kogoshima University Japan (by means of multilevel distillation), seven
fractions are obtained and there are quite a large volume number of four fractions namely the A1,
B2, C3 and D4 fractions. Leaves, rhizomes and fractions obtained in the identification of chemical
components contained using GC-MS in Andalas University Laboratory Padang.
2.2 Pathogen isolation
Sclerotium rolfsii isolates were obtained by isolating from peanut plants that were attacked
by stem rot rot in the Payo area of Tanjung Harapan Solok sub-district. Isolates were identified
and propagated Potato Dextrosa Agar (PDA) media as the source of the inoculum to be tested,
isolates were used 5 days old in PDA media.
2.3 Antifungal power testing
2.3.1 Pression of Sclerotium rolfsii colony diameter
Tests are carried out by mixing until homogeneous the treatment material into a sterile
PDA medium, according to the treatment and concentration tested before freezing (45ºC), then
poured into petridish and allow to harden, after hardening the inoculation of fungi, fungal mat
from the Sclerotium rolfsii fungus is cut with a sterile corkbore diameter of 6 mm, placed in the
middle of the treated medium, then incubated in an incubator of 28ºC for 4 days. The experiment
was arranged in the form of a Completely Randomized Design (CRD) in factorial each of 4
replications, the treatments were: Leaf oil and roots of wild Ginger and fractions A1, B2, C3 and
D4 as factors I, concentration levels (0, 100, 250 and 500 ppm) as factor II.
2.3.2 Emphasis of colony biomass
Tests using Patato Dextrose Broth (PDB) liquid medium, as much as 25 ml of the medium
are input into each test tube, then sterilized in an autoclave, after sterile is cooled and then put the
treatment material to be tested according to concentration, then do inoculation of fungi test, the
fungal mat of the Sclerotium rolfsii mushroom was cut with a sterile corkbore of 6 mm in diameter,
Nurmansyah et al. JAAST 5(1): 28 –37 (2021) 31
and inserted into the treated medium, then incubated in an incubator of 28ºC for 4 days. The
experiments were arranged in the form of a Completely Randomized Design (CRD) in factorial
each of 4 replications. The treatments were: Leaf oil and wild ginger root fractions A1, B2, C3 and
D4 as factors I, concentration levels (0, 100, 250 and 500 ppm) as factor II. Furthermore, the
growth of fungal colonies was taken and dried in an oven at 80ºC for 48 hours, then the biomass
was weighed. Inhibition or suppression of colony diameter and biomass growth, calculated by the
formula (Pandey et al, 1982 in (Noveriza & Miftakhurohmah, 2010)
𝑥 = !"#!× 100% (1)
X = Percentage inhibition growth of diameter/biomass colony a = Diameter/biomass growth of treatment b = Diameter/biomass growth of control (untreated)
3. Results and Discussion
The results showed that leaf oil, rhizome and fraction of wild ginger (Elettariopsis slahmong) are
antifungal and can inhibit the growth of the fungus Sclerotium rolfsii which causes rot disease of
the stem base of the peanut plant. A1 and B2 fractions showed better antifungal effectiveness
compared to essential oils of leaves and rhizomes and fractions of C2 and D4 (Table 1)
Table 1. Effect of leaf, rhyzom and fraction of essential oils of E. slahmong and concentration level against Sclerotium rolfsii colony diameter growth (4 DAI)
Treatments Colony diametre (mm) Inhibition (%) Botanical pesticide Leaf oils Rhizome oils Fraction A1 Fraction B2 Fraction C3 Fraction D4
44.18 44.69 42.37 43.19 54.75 45.00
47.31 c
46.71 cd 49.47 a 48.50 b 34.70 e 46.34 d
Concentration level 0 ppm (Control +) 100 ppm 250 ppm 500 ppm Control (without treatments)
81.75 53.71 32.08 15.25 83.75
2.69 d 36.92 c 61.69 b 81.74 a 0.00 -
CV (%) - 1.98 Note. The numbers followed by the same letter are not significantly different according to DMRT. Test at 5% level. DAI (days after inoculations). Control+ (Solvents and emulsifiers)
From Table 1 it can be seen that the leaf essential oil with rhizome is not statistically
significantly different, but the leaf oil rate is more fungicidal than rhizome oil, so the fraction of
D4 with rhizome oil is also not statistically significantly different.
The higher of concentration level the smaller the diameter of the test mushroom colony.
At a concentration of 500 ppm the suppression of S. rolfsii fungi colony diameter growth reached
81.74%.
Nurmansyah et al. JAAST 5(1): 28 –37 (2021) 32
The interaction of essential oils of leaves, rhizomes and fraction of wild ginger with a
concentration level showed that the higher the concentration level the higher the inhibitory effect
on the diameter growth of the test fungi colonies (Table 2).
Table 2. Interactions of leaf oil, rhizome and E. slahmong fraction with levels concentration on growth of Sclerotium rolfsii colony diameter (4 days after inoculation)
Treatments Colony diameter (mm) Inhibition growth (%) Leaf oil s(Md)
0 ppm 100 ppm 250 ppm 500 ppm
81.75 52.50 29.50 13.00
2.39 m 37.31 j
64.77 fg 84.78 b
Rhizome oils(Mr) 0 ppm
100 ppm 250 ppm 500 ppm
81.75 53.50 30.00 13.50
2.39 m 36.11 jk 64.18 g 83.88 bc
Fraction A1 0 ppm
100 ppm 250 ppm 500 ppm
81.75 48.75 27.50 11.50
2.39 m 41.78 i 67.16 de 86.26 a
Fraction B2 0 ppm
100 ppm 250 ppm 500 ppm
81.75 50.00 28.50 12.50
2.39 m 40.59 i 65.97 ef 84.78 b
Fraction C3 0 ppm
100 ppm 250 ppm 500 ppm
81.75 63.50 47.25 26.50
2.39 m 24.18 l 43.58 h 68.35 d
Fraction D4 0 ppm
100 ppm 250 ppm 500 ppm
81.75 54.00 29.75 14.50
2.39 m 35.52 k 64.48 fg 82.68 c
Note. The numbers followed by the same letter are not significantly different according to DMRT. Test at 5% level.
From Table 2, it can be seen that the concentration level of 100 ppm of wild ginger essential
oil and the fraction has shown an emphasis on the growth of S. rolfsii mushroom colony diameter
with growth suppression ranging from 24.18 - 41.78% and at a concentration level of 500 ppm A1
fraction has been able suppress the growth of test fungi colonies by 86.26%, while in leaf essential
oil the growth of new colonies increased to 84.78% at the same concentration level.
Test results on Sclerotium rolfsii fungi biomass on Patato Dextrosa Broth media showed
that fungal biomass growth suppression was quite effective, colony biomass in the treatment of
A1 fraction showed the highest suppression of colony biomass growth compared to B2, leaf oil,
Nurmansyah et al. JAAST 5(1): 28 –37 (2021) 33
rhizome, and D4 fractions, C3 fraction showed effective suppression of biomass colonies in A1
treatment. The Suppression of the lowest colony biomass growth (Table 3).
Table 3. Effects of essential oils of leaves, rhizomes and E. slahmong fraction as well as levels concentration of Sclerotium rolfsii colony biomass growth (4 DAI)
Treatments Colony biomass (mm) Inhibition growth (%) Botanical Pesticide Leaf oils Rhizome oils Fraction A1 Fraction B2 Fraction C3 Fraction D4
79.62 80.75 76.94 78.19 99.87 80.93
49.76 c 49.17 cd 51.46 a 50.67 b 36.95 e 48.93 d
Concentration level 0 ppm (Control +) 100 ppm 250 ppm 500 ppm Control (without treatments)
153.25 94.70 58.04 24.87 158.50
3.31 d 40.29 c 63.43 b 84.25 a
0.00 CV (%) - 1.79
Note. The numbers followed by the same letter are not significantly different according to DMRT. Test at 5% level. CV (coefficient variation). Control+ (Solvents and emulsifiers) The interaction of essential oils of leaves, rhizomes and wild ginger fraction with the
concentration level, showed that the higher the concentration level, the higher the inhibitory power
of the biomass colony of the test fungus. At the concentration level of 500 ppm A1 and B2
fractions, no significant difference was seen in suppressing the growth of S.rolfsii fungi colony
biomass, with growth suppression reaching 88.10% and 87.22%, but significantly different from
leaf oil, rhizomes and other fractions . Leaf oil, rhizome and D4 fraction also did not show a
statistical difference, but significantly different from the C3 fraction with the lowest growth
suppression of 72.55% at the same concentration level (Table 4).
From Table 1-4 shows that the fraction A1, A2, leaf oil and wild ginger rhizome have a
high antifungal power against the S rolfsii fungi significantly different from the C3 fraction and
the D4 fraction. The main components contained in wild ginger leaf oil are 2-Decanoic acid
(48.04%), nonanoic acid (9.18%), 2-octenal (8.97%), hexanoic acid (2.46%), 6-tetradecene
(2.35%), nonanal (2.06), ascobic acid (1.66%), heptanoic acid (1.61%) and octanal (1.20%). In the
main content of rhizome oil are 2-Tridecenal (39.81%), 2-decanoic acid (26.39%), 2-octenal
(7.56%), hexadecanoic acid (2.86%), nonanoic acid (3, 85), 2-deconyl acetate (2.31%), Eucalyptol
(2.13%). In the A1 component, the main components are 2-Decanoic acid (27.24%), 2-octenal
(17.01%), decenal (12.40%) and nonanoic acid (1.37%). The main content of B2 fraction was
Decanal (38.31%), octanal (8.42%), 6-tetradecene (3.24%), 2-octenoic acid 2.03%), and octenal
(1.56%). C3 fraction containing 2-Tridecenal (26.57%), decanal (25.65%), 2-octenal (7.75%), 2-
propenoic acid (5.11%), 2-decenyl acetate (5) , 44%), benzaldehyde (3.87%), and dedecenal
Nurmansyah et al. JAAST 5(1): 28 –37 (2021) 34
(2.35%) and the D4 fraction of the uatam content was 2-Tridecenal (19.41%), 2-dimethyl
(3cloropropyl) sililoxymethyltetra (16.99%) , 2-octenal (16.08%), 1-ethyl-1- (undec-10-enyl) oxy-
1-silacyclopenta (15.99%), decanal (2.87%), octanal (2.84% ), 6-tridecene 2.79% (Nasir &
Nurmansyah, 2016; Nasir, 2016).. Each component has not been tested, therefore further research
is recommended. Antifungal essential oil chemical components that are able to penetrate the fungal
cell walls, thereby causing interference with the metabolic processes in the cell so that it interferes
with cell growth, at certain concentrations will result in the death of the fungal cells.
Table 4. Interactions of leaf oil, rhizome and E. slahmong fraction with concentration levels on growth of Sclerotium rolfsii colony biomass (4 days after inoculation)
Treatments Colony biomass (mm) Inhibition growth (%) Leaf oils (Md)
0 ppm 100 ppm 250 ppm 500 ppm
153.25 90.00 53.50 21.75
3.31 l 43.21 i 66.24 f
86.28 bc Rhizome oils (Mt)
0 ppm 100 ppm 250 ppm 500 ppm
153.25 93.00 54.25 22.50
0.31 l 41.63 j 65.93 f 85.81 c
Fraction A1 0 ppm
100 ppm 250 ppm 500 ppm
153.25 86.50 49.00 19.00
3.31 l 45.42 g 69.08 e 88.01 a
Fraction B2 0 ppm
100 ppm 250 ppm 500 ppm
153.25 88.75 50.50 20.25
3,31 l 44.00 hi 68.13 e 87.22 ab
Fraction C3 0 ppm
100 ppm 250 ppm 500 ppm
153.25 116.25 87.00 43.00
3.31 l 26.65 k 45.27 gh 72.55 d
Fraction D4 0 ppm
100 ppm 250 ppm 500 ppm
153.25 93.75 54.00 22.75
3.31 l 40.85 j 65.93 f 85.65 c
Note. The numbers followed by the same letter are not significantly different according to DMRT. Test at 5% level.
From the above results it is clear that decanoic acid is more fungicidal than decanal,
(Kumar et al, 2011), reporting that decanoic acid (capric acid) and its esters have been used in
medical science and are the best antifungal. The decanal and nonanal are antifungal components
that can inhibit the growth of sclerotia of Sclerotinia sclerotiorum that causes stem rot disease
Nurmansyah et al. JAAST 5(1): 28 –37 (2021) 35
from the canola and stem rot from the sunflower. Decanoic acid and nonanoic acid at a
concentration level of 100 ppm have been able to kill spores from the fungus basidiomycetes that
cause brown root disease, pentanoic acid and hexanoic acid are effective against test fungi at a
concentration level of 1000 ppm (Schmidt, 1984 in Clausen et al, 2010). Essential oils from wild
Ginger leaves can control the pathogen Colletotrichum gloesporioides that causes Antrachnose
disease in the red dragon fruit plant Hylocereus polyrhizuz. At the concentration level of 1000 ppm
leaf oil can suppress the growth of pathogenic C. gleosporioides fungal colonies 100% (Nasir &
Nurmansyah, 2016).
According to Lim in (Picheansoonth & Yupparach, 2007), stated that the results of the
Elletaropsis spp oil analysis of the components inside are almost the same as slight differences
such as E. elan, the main components are monoterpenes, geraniol 71.6%, comphane and
phelandrena. E. smithiae, is the main component of monoterpenes, geraniol 38.10%, neral 29.10%,
comphane and fancylacetate. E. triloba main components are 16,16% citral, limonane,
phellandrena and terpene acetate. The active compounds contained in E. slahmong oil according
to CK Lim are terpenoid components, (E)-2-octenal (46,3%) and (E)-2- decenal 36,8% rhizomes
and roots with the main content (E) -2-decenal 79.4% (Wong et al, 2006).
Botanical fungicides that have been tested and are effective against S. rolfsii fungi are
essential oils from Casia vera (Cinnamomum burmanii) leaves and twigs with a concentration level
of 500 ppm which can suppress the growth of S. rolfsii colony 100% (Nurmansyah, 2014) and
essential oils from several wild sirih such as sirih hantu, sirih cambai, sirih kaduak and sirih are
quite effective in suppressing the growth of this fungus (Nurmansyah, 2012). Bamboo piper
essential oil (Piper aduncum) at a concentration level of 500 ppm is able to control the pathogen
S rolfsii reaching 92.77% (Nurmansyah, 2016). This wild ginger fungicide has the bioprospek to
be developed because the results are not significantly different from the bamboo piper and
cinnamon fungicides.
4. Conclusions
The results of the study concluded that leaf oil and rhizome of wild ginger Elettariopsis
slahmomong and their fractions have the potential to be used as botanical fungicides. The A1
fraction has the best antifungal effectiveness compared to leaf oil, rhizome and other fractions,
leaf oil and rhizome oil are not statistically significantly different, but in terms of leaf oil is better
than rhizome oil. The best concentration level in this study was 500 ppm with the highest emphasis
on diameter and biomass growth of 81.74% and 84.25%.
References
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Protection against Mold and Sapstain. Forest Products Journal, 60(3), 301–304. Idris, Nurmansyah, Helfi Gustia, A. I. Ramadhan. (2020). The Potential and Effectiveness of
Three Botanical Fungicides to Control Stem Rot Disease in Peanuts. Technogy Report of Kannsai Univercity, 62(4), 1745–1752.
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Magdaulih, E., & Nasir, N. (2014). Antifungal Activity of Essential Oil of Cymbopogon nardus L. and Elettariopsis slahmong Lim. against Colletotrichum sp. on Red Dragon Fruit (Hylocereus polyrhizus). J. Bio. UA, 3(2), 97–102.
Nasir, Dharma. A, Efdi. M, & Yuhendra, E. F. (2014). Natural product of wild Zingiberaceae Elettariopsis slahmong_ biopesticide to control the vector of banana blood disease bacterium in West Sumatera. Research of Pharmaceutical Biological and Chemical Science, 5(5), 1250–1256.
Nasir. (2017). Essential oils from Elettariopsis slahmong C. K. Lim and Cinnamomum burmanii [Nees & T. Nees] Bl. inhibit the colony growth of Phythopthora palmivora of cocoa. Scholars Research Library Der Pharmacia Lettre, 9(4), 95–107.
Nasir & Nurmansyah. (2016). Leaf Essential Oil of Wild Zingiberaceae Elettariopsis slahmong CK Lim to Control Antrachnose Disease in Red Dragon Fruit Hylocereus polyrhizus. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(5), 2463–2471.
Nasir, N. (2016). Rhizome essential oil and fractions of Elettariopsis slahmong CK. Lim against Colletotricum gloesporioides in red dragon fruit Hylocereus polyrhizus. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(2164), 2164–2171.
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Picheansoonthon, C., & Yupparach, P. (2010). Further Study on the Elettariopsis Baker (Zingiberaceae) in Thailand－a New Species and a New Record. Taiwania, 55(4), 335–341.
Sukamto & Wahyuno, D. (2013). Identifikasi dan karakterisasi Sclerotium rolfsii Sacc . penyebab penyakit busuk batang nilam (Pogostemon cablin Benth). Bul. Littro, 24(1), 35–41.
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Journal of Applied Agricultural Science and Technology E-ISSN: 2621-25285(1): 38-49 (2021) ISSN: 2621-4709
Received February 2, 2021; Accepted March 10, 2021; Published March 10, 2021 https://doi.org/10.32530/jaast.v5i1.9 This is an open access article under the CC BY-SA 4.0 license https://creativecommons.org/licenses/by-sa/4.0
38
STABILITY AND TOXICITY TEST OF ANGKAK PIGMENT POWDER FROM SAGO HAMPAS- RICE FLOUR SUBSTRATE AS NATURAL DYES
Dian Pramana Putra1, Novelina2, Alfi Asben*,2
1Faculty of Agricultural, Ekasakti University, Padang, Indonesia 2Faculty of Agricultural Technology, Andalas University, Padang, Indonesia
*Coresponden authorEmail: [email protected]
Abstract. Sago hampas is waste from processing sago starch. Sago hampas is generally thrown away without any further treatment. Sago hampas contain nutrients that can be used as a substrate for fermentation of angkak. The study aims to determine the stability and toxicity of angkak pigment powder from sago hampas-rice flour substrate. The study used exploratory research design through experiments in the laboratory. This study used the UV-vis spectrophotometer method in observing the stability of the Angkak powder pigment and the brine shrimps method of angkak pigment powder toxicity test. The results showed that the level of solubility of angkak pigment powder will increase at higher temperatures. Stability of angkak pigment powder tends to decrease with longer heating, the higher of heating temperature and the longer of irradiation. Angkak pigment powder are more stable at neutral and alkaline pH compared with acidic pH. And it is not toxic to experimental animals with LC50 value of angkak pigment powder of 2,897.05 ppm. The conclusion of this study is that temperature affects the intensity of the angkak pigment. Angkak is unstable along with heating time, heating temperature and longer of irradiation. Keywords: monascus purpureus; angkak pigment powder; pigment stability; pigment toxicity
1. Introduction
Sago hampas is one of the agricultural industrial wastes whose utilization is still not well
managed. Sago hampas can be used as a fermentation substrate in making Angkak. According to
Asben (2012) the percentage of the main ingredients of sago hampas are hemicellulose 14%,
cellulose 21%, fat 2%, crude protein 1%, lignin 6%, starch 51%, and others 5%. Therefore sago
hampas can be used as angkak pigment fermentation substrate because it contains carbon, nitrogen
and minerals needed by microbes as nutrients during the fermentation process. Angkak is a
fermented product by the Monascus purpureus. Monascus is known to be an important fungus in
producing red pigment. The pigment produced by M. purpureus is very stable and safe to use as a
food additive. Monascus purpureus has been widely used to produce pigments through
fermentation processes both on solid substrates and on liquid substrates (Sheu et al. 2000 in
Yuliani, 2014).
Angkak is generally made using rice as a fermentation substrate. For this reason, it is
necessary to use other ingredients as angkak pigment substrate, one of which is sago hampas.
Research on the use of sago hampas as a fermentation substrate has been carried out by Asben and
Kasim (2015) and Asben et al. (2017), but no extraction of the sago residue pigment was carried
out. Generally Angkak which is sold in the market is still in solid form which is the result of
Putra et al. JAAST 5(1): 38 –49 (2021)
39
fermentation and is still mixed with the fungi of Monascus itself. So that its use will affect the
consistency of the product to be dyed. It is necessary to separate the Angkak pigment from the
fermentation substrate such as the research of Putra et al. (2018) which extracts the Angkak
pigment from the sago hampas substrate. Angkak has a consistent color but is less stable to
physical and chemical influences such as heat, UV-rays and sunlight, can mix with other natural
dyes with food ingredients, non-toxic and not a carcinogen (Hesseltine, 1965 and Stainkraus, 1983
in Jenie et al., 1994). According to Triana and Nurhidayat (2009), angkak pigments are also not
carcinogenic, besides the presence of lovastatin in angak can reduce cholesterol and triglycerides.
It is necessary to study the stability and toxicity of angkak pigment powder before applying it to
food.
Research related to the pigment stability of Angkak has been carried out by several
researchers, such as Priatni (2015) who conducted research on encapsulation studies and pigment
stability from fermented rice extracts. Jenie et al. (1997) conducted a study on the production of
Angkak Concentrate and Pigment Powder from Monascus purpureus and its stability during
storage. Furthermore Kaur et al. (2009) conducted a study of production and evaluation of the
physicochemical properties of red pigment from Monascus purpureus MTCC 410. In a study conducted by Putra et al. (2018), in fermentation of Angkak sago hampas
produced Angkak pigment powder. In this study, Angkak pigments were extracted using an
ultrasonic bath with a long extraction time as a treatment. in his research there was no test on the
stability of the resulting Angkak pigments. This study aims to determine the stability of angkak
pigment powder from physical and chemical influences and determine the toxicity of angkak
pigment powder produced.
2. Methods
2.1. Materials
The raw material used in the form of angkak pigment powder obtained from the extraction
of angkak powder from sago hampas and rice flour. Chemicals used include distilled water,
methanol, DMSO, buffered citrate pH 3, 4 and 5, buffered phosphate pH 6, 7 and 8 and carbonate
Na pH 9 and others. Experimental animals namely Artemian salina Leach shrimp larvae.
The tools used are pH meter (Delta OHM HD 2105.2, Italy), spectrophotometer (Shimadzu
UV-1800), cabinet dryer (Corsiar Manufacturing), ovens (Philip Harris Ltd and Memmert),
analytical scales (Kern ABJ 220-4M), lamps incandescent 20 watts, aquariums, water pumps,
glassware such as goblets, test tubes and others.
2.2. Experimental Design
The design in this study is based on research by Putra et al. (2018) with differences in the
extraction time of Angkak pigments. Furthermore, the best results from his research were
Putra et al. JAAST 5(1): 38 –49 (2021)
40
continued with testing the stability of the Angkak pigment. The research design used was
explorative research methods through experiments in the laboratory. Data was collected by direct
observation after the research object was given treatment, then conducted a series of tests.
2.3. Experiments
Preparation and refreshment of M. purpureus and preparation of sago hampas (Asben &
Kasim, 2015)
Sago hampas is dried up to 10% moisture content and size is reduced by using a blender then
filtered with a sieve to obtain 40-60 mesh size. sago hampas is ready to be used as a fermentation
substrate. The pure culture of M. purpureus is refreshed on the agar sloping PDA media. Incubated
at room temperature for 21 days. Ascospores / conidia are released on the sloping surface by giving
5 mL of sterile distilled water. Continue to be crushed with the ose so that the askospora is released
and is suspended in aquades. The number of spores is calculated with a heamocytometer. Culture
is ready to use.
Angkak pigmen production from sago hampas-rice flour substrat (Asben & Kasim, 2015)
Fermentation uses a mixture of sago hampas with a particle size of 40-60 mesh and rice flour
with a ratio of sago hampas : rice flour (1: 1) in a sterile erlenmeyer. Next added 2.5% glucose
solution to ± 50% substrate water content. Sterilization with an autoclave of 121 oC for 20 minutes.
Substrate is allowed to cool in laminar air flow, then put inoculum as much as 10% of the total
substrate, the substrate is stirred until homogeneous. Then incubated at room temperature 28-30 oC for 14 days. Then the fermentation results are dried for 72 hours at 40-45 oC until the water
content of the powder becomes ± 7%. Angkak crushed and filtered using a 100 mesh size sieve,
then extraction and observation were carried out.
Angkak pigmen extraction and encapsulation (Putra et al. 2018)
Angkak pigment extraction process is carried out by following the procedure as follows.
Angkak that has been prepared previously and added 60% ethanol with a ratio of 1:10.
Furthermore, it was extracted using an ultrasonic bath according to the extraction time 40 minutes.
The extraction results were filtered with Whatman paper No 1. Then the resulting filtrate was
evaporated using a rotary vacuum evaporator solvent until the pigment angkak extract was
obtained. The extract is then encapsulated. The encapsulation process is carried out using
maltodextrin as a filler with a concentration of 10%. Angkak pigment extract was added to the
maltodextrin solution, then homogenized and dried using a freeze dryer until the water content
reached ± 5%. Furthermore, an analysis of angkak pigment powder was carried out.
2.4. Analysis
Observations made on the encapsulated red yeast rice extracted include: Test the solubility
of red yeast rice pigment powder, Test the effect of temperature, pH, heating time and exposure
Putra et al. JAAST 5(1): 38 –49 (2021)
41
time on the stability of the pigment powder (Jenie et al., 1997); and the toxicity test of Angkak
Pigment Powder according to the Brine Shrimps method (Hernindya et al., 2014).
3. Results and Discussion
3.1. Solubility of Angkak pigment powder
This test was conducted on the best product from the research of the extraction time of
angkak pigments and the types of solvents used in the extraction of angkak pigments. The results
of the analysis on the ease of solubility of angkak pigment powder can be seen in Figure 1.
The solubility test of Angkak pigment powder is done by dissolving Angkak pigment powder
in water with several different temperatures. Angkak pigment powder solubility test needs to be
done to determine the solubility level of red angkak pigment in water. Increased color intensity of
Angkak pigments due to differences in water temperature occurs in all types of pigments. The
absorbance value of red pigment increased asorbansi from 0.728 to 0.865.
Based on the graph above it is known that an increase in temperature will increase the
intensity of the yellow, orange and red pigments in Angkak pigment powder. Thus, it can be seen
that the pigment is more soluble at high temperatures. This happens because of the kinetic energy
from heat. According to Jenie et al., (1994), the kinetic energy that occurs due to heating at high
temperatures can cause the pigment to decompose faster and dissolve in water.
Figure 1. Graph of solubility of Angkak pigment powder in water
The ease of Angkak pigment powder in cold water is caused by the use of maltodextrin as
filler material in the encapsulation of Angkak pigment, one of the properties of maltodextrin is
soluble in cold water. This is in accordance with the statements of Blanchard and Katz (1995),
explaining further the properties possessed by maltodextrin such as experiencing rapid dispersion,
having high solubility properties and forming films, forming low hygroscopic properties, low
browning properties, able to inhibit crystallization and has a strong bond.
0.728
0.767
0.820
0.8540.865
0.72
0.74
0.76
0.78
0.8
0.82
0.84
0.86
0.88
0 20 40 60 80 100 120
Abs
orba
nce
Temperature (oC)
Putra et al. JAAST 5(1): 38 –49 (2021)
42
The solubility value of Angkak pigments from this study was higher than the results of the
research of Fatimah, Suprihadi and Kusdiyantini (2014) where the lowest solubility average of
Monascus red pigment at 25oC was 0.28 while the highest solubility was at 100oC of 0.56.
according to Fatimah et al., (2014) Judging from the absorbance value obtained that the high
temperature is 100oC, fermentation powder has a color intensity that tends to be high compared to
low temperature, this shows that the color stability is influenced by temperature. this is in
accordance with the research results that have been obtained. 3.2. Effect of Temperature on the Stability of Angkak Pigment Powder
The stability test of Angkak pigment powder towards temperature aims to determine the
level of stability of pigment at high temperatures. This test uses different temperature levels, i.e.:
50oC, 100oC, 150oC and 200oC, and as a comparison is the initial pigment without heat treatment.
The results of the analysis of the effect of temperature on the stability of Angkak pigment powder
can be seen in Figure 2.
The results of the analysis showed a decrease in the intensity of angkak pigment as the
temperature increased. The intensity of the red pigment color in angkak pigment powder
absorbance values ranged from 0.307 to 0.886 for the best treatment of the type of solvent, namely
methanol solvent. The angkak pigment intensity at the beginning (without heat treatment) by
giving 50oC heat only slightly decreased the intensity of pigment. But after the heating temperature
reached 100oC, the intensity of angkak pigment showed a significant decrease until the temperature
of 200oC.
The results of the analysis of the stability of Angkak pigment powder on different heating
temperatures showed that there was a decrease in the intensity of Angkak pigment powder. Storage
of Angkak pigment powder at room temperature of 26oC for 1 hour does not cause changes in the
absorbance value of Angkak pigment, nor in heating at temperature 100oC Angkak pigment color
is still relatively stable, only slightly changes the absorbance value of Angkak pigment color. After
storage at 150oC for 1 hour there was a very significant change. This change in the form of a
decrease in the absorbance value of angkak pigment powder to a paler color. Similarly, heating at
200oC the intensity of angkak pigments decreases marked by the paler color produced.
This happens because at temperatures above 100oC, the red pigments, monascorubramin and
rubropunctamin, tend to become unstable due to heating. Reduction in red pigment color due to
heating due to red pigment damage to the pigment chromophore group. The damage is caused by
the release of functional groups or opening of functional groups that make up the red pigment
chromophore group. So there is a decrease in the intensity of red pigment along with an increase
in heating temperature (Asadayanti et al., 2010). Santoni, Darwis and Syahri (2013), added that
the main cause of the increasing of the heating temperature, the less the color intensity of the
Putra et al. JAAST 5(1): 38 –49 (2021)
43
anthocyanin extract solution, this is due to the degradation of the anthocyanin. Anthocyanin
degradation can be in the form of breaking of glycosidic bonds which causes unstable
anthocyanidins and changes in the structure of anthocyanidins to chalcone compounds.
Figure 2. Graph of pigment stability at different heating temperatures
This is in accordance with the statement of Tedjautama and Zubaidah (2014) the results of
the red pigment stability test against temperature showed a decrease in the intensity of the red
pigment after being incubated at a certain temperature for 1 hour. Angkak pigment stability
decreased at temperatures of 121 oC and 180 oC. It is suspected that the pigment was damaged due
to heat. According to Priatni (2015) monascus fermented rice pigment stable to at temperature
50oC for 30 minutes.
3.3. Effect of pH on Angkak Powder Solution
Angkak pigment powder stability testing at different pH conditions was carried out by
dissolving Angkak pigment powder in a solution with a pH condition of 3-9. Stability testing is
carried out for 6 hours where every 3 hours an analysis is performed. The results of the analysis
on the effect of different pH on the stability of Angkak pigment powder can be seen in Figure 3.
The analysis showed an increase in the intensity of angkak pigment along with the increase
in the pH value of the solution. Red pigment intensity, namely monascorubramin and
rubropunctamin on angkak powder ranged from 0.075 to 0.155 with 3 hours of coding and 0.052
to 0.140 with 6 hours of conditioning for the best treatment of the type of solvent, namely methanol
solvent.
The results of the analysis of the influence of pH on the stability of Angkak powder pigments
showed that there was a change in the absorbance value at each difference in the pH value. In
acidic atmosphere, the Angkak powder pigment stability is lower than neutral and basic
atmosphere. According to Fabre et al., (1993) red pigments are most sensitive to acidic pH and are
0.886 0.875
0.791
0.4030.307
00.10.20.30.40.50.60.70.80.9
1
0 50 100 150 200 250
Abs
orba
nce
Temperature (oC)
Putra et al. JAAST 5(1): 38 –49 (2021)
44
more stable under alkaline conditions. Radiastuti (2005) research results, also reported the same
results that angkak pigments are less stable in acidic conditions and more stable in neutral or basic
conditions. The color stability of pigments produced by microorganisms including fungi is
generally more stable to heat, light and pH compared to pigments from plants (Malik et al., 2012).
However, pigments from microbes are unstable at too acidic pH and high temperatures. Carvalho
et al. (2005), in the analysis of the stability of the red biopigment produced by Monascus fungi on
the treatment of pH and temperature reported that biopigment treatment at pH 4-8 obtained
information that the smaller the pH, the more significant the pigment damage was. This red
pigment from Monascus is stable at pH 7.
Figure 3. Graph of pigment stability at different pH
Natural pigments that are sensitive to acidic pH are present in the form of flavilium cations
which are red in color and lack of electrons. With increasing pH the flavilium cation group gets an
electron donor and turns into a colorless chalcone. This is indicated by a decrease in absorbance
in sample testing (Jenie et al., 1997). According to Radiastuti (2005), the mechanism of damage
to angkak pigments due to changes in pH is unclear, but it can be suspected that the damage also
occurs due to changes in donor or electron acceptor in the cation group, causing the structure of
the compound to change.
The results of Tedjautama and Zubaidah (2014) research on the stability of the red pigment
against pH show that the red pigment is more stable at neutral pH (pH 7) than at acidic pH (pH 3).
this is in accordance with the results of the research that has been done. at acidic pH the stability
of the pigment in Angkak decreased. It is thought that the pigment has been damaged by the H+
ions in the acid. According to Priatni (2015) stability of MFR pigment was decreased under acidic
conditions and stable at at pH value 5-6. Kaur et al. (2009) Angkak pigment stability is stable at
pH 6-8 and more stable at neutral pH conditions.
0.052
0.055
0.105
0.1270.140
8, 0.1260.078
0.0750.079
50.106
0.1410.155
0.135
0.096
00.020.040.060.080.1
0.120.140.160.18
0 2 4 6 8 10
Abs
orba
nce
pH
6 Hours3 Hours
Putra et al. JAAST 5(1): 38 –49 (2021)
45
3.4. Effect of Heating Time on the Stability of Angkak Pigment Powder
Testing the stability of pigment powder Angkak pigment against heating time carried out for
3 hours using an oven with a temperature of 100oC. Every hour the sample of pigment intensity
was measured using a spectrophotometer. The results of the analysis of the effect of heating
duration can be seen in Figure 4.
Figure 4. Graphic stability of pigment on heating time.
The analysis showed that Angkak pigment powder which was heated for 3 hours at 100oC
showed a decrease in pigment intensity due to this treatment. The intensity of red pigment in
angkak powder absorbance value ranges from 0.333 - 0.887. The highest pigment intensity in the
treatment without heating with an absorbance value of 0.887 but not different from the treatment
with heating for one hour with an absorbance value of 0.851. The decrease in color intensity of
Angkak pigment powder is caused by damage to the chromophore group of the dyestuff, namely
changes in bonding or functional groups. This occurs marked by a decrease in the absorbance
value at every one hour heating for 3 hours. In addition, the effect of long heating in a long time
will cause kinetic energy that is thought to be the cause of the damage. Damage to the chromophore
group will cause the dyeing of the dye but if seen visually there is no significant change in color
(Sutrisno, 1987). The results of research conducted by Radiastuti (2005) stated that Monacus
pigment heated for 4 hours at 100 ° C showed a decrease in the intensity of the red pigment due to
treatment.
3.5. The Effect of Irradiation Time on the Stability of Angkak Pigment Powder
Testing stability of Angkak pigment powder pigments against irradiation time is done by
using Philips 40 watt incandescent lamp. Where the sample is conditioned in a closed container or
box and given an incandescent lamp for 6 hours and every 3 hours analysis of angkak pigments is
analyzed. The results of the analysis of the stability of pigment powder Angkak pigment against
the long exposure can be seen in Figure 5.
0.8870.851
0.474
0.333
00.10.20.30.40.50.60.70.80.9
1
0 0.5 1 1.5 2 2.5 3 3.5
Abs
orba
nce
Time of Heating (Hours)
Putra et al. JAAST 5(1): 38 –49 (2021)
46
Figure 5. Graph of pigment stability at exposure time.
Based on the graph above the reduction in the intensity of angkak pigment along with the
longer irradiation. The intensity of the red pigment in angkak powder absorbance value ranges
from 0.721 - 0.886. In the first 3 hours irradiation the absorbance value of the sample was 0.823
and not much different from the initial sample with an absorbance value of 0.886, but after
irradiation for 6 hours there was a considerable decrease with an absorbance value of 0.721.
Based on the analysis of the effect of irradiation time it was found that the angkak pigment
powder was relatively stable in irradiation for 3 hours, this was marked by a slight change in the
absorbance value of the sample. But after irradiation for 6 hours has seen a decrease in the intensity
value of angkak pigment powder which is marked by the decreasing value of the absorbance of
the sample. This decrease in absorbance value is because during the irradiation process the
temperature around the box increases to 45oC, the longer the irradiation the longer the sample will
be exposed to heat from the incandescent lamp used.
Light rays can affect the stability of angkak pigments, Smith (1975) states that source of
light are emits energy and some of this energy is converted into visible light. This is reinforced by
the opinion of Markakis (1982) that the pigment has a strong tendency to absorb visible light and
the radiation energy of the light causes photochemical reactions in the visible spectrum and results
in discoloration. Bilyk et al. (1981) conducted a study of the effect of light conditions on the
stability of red beet pigments resulting in color loss of up to 50% -60%. According to Kaur et al.
(2009) Angkak pigments are stable during irradiation for up to 24 hours. Color reduction in angkak
pigment occurs after irradiation for more than 24 hours.
3.6. Brine Shrimps Method of Angkak Pigment Powder Toxicity Test
The toxicity test of Angkak pigment powder uses the Brine Shrimps Lethality Test (BSLT)
method, which is a screening method to determine the toxicity of a compound or extract of natural
0.886 0.8230.721
00.10.20.30.40.50.60.70.80.9
1
0 1 2 3 4 5 6 7
Abs
orba
nce
Time (Hours)
Putra et al. JAAST 5(1): 38 –49 (2021)
47
substances that are cytotoxically acutely by using experimental animals Artemia salina Leach
larvae. To calculate the LC50 value, a curve that states the log concentration of the test extract that
causes death of 50% A. salina larvae as the x-axis, and mortality of A. salina 50% larvae after a
24-hour incubation period as the y-axis, is plotted in the linear regression equation y = a + bx.
Using this formula, values a and b are obtained based on data from the 10 concentration points
used. LC50 value is determined through probit analysis of the data obtained. Data on the results of
the toxicity testing of angkak products against A. salina larvae can be seen in Table 1.
Table 1. Angkak toxicity against Artemia salina Larvae Treatment LC50 (ppm)
Angkak pigment powder 2897.05
Information: LC50 <1000 ppm: Toxic LC50 >1000 ppm: Non toxic Data obtained from the average results of each test
Toxicity test based on the Table 1 it is known that the toxicity value of Angkak pigment
powder is 2,897.05 ppm. From the results of the toxicity analysis it can be concluded that the
product of the Angkak Pigment Powder is not dangerous (toxic) to humans if applied to food
products, because the value of LC50 is obtained more greater than 1000 ppm. An extract is declared
to have acute toxicity if it has an LC50 value of less than 1000 µg / mL. Lethal Concentration 50
(LC50) is a concentration of substances that cause the death of 50% of experimental animals,
namely larvae Artemia salina Leach (Rolliana, 2010).
Angkak pigment powder does not contain toxic compounds due to the process of extracting
pigment from Angkak with methanol as a solvent. Furthermore, concentration of angkak pigment
extract was carried out with a rotary vacum evaporator until no solvent was left. Furthermore, it is
encapsulated using fillers that can be consumed and dried with a freeze dryer, so that the final
results of Angkak pigment in powder form. According to Triana and Nurhidayat (2009), angkak
pigments are also not carcinogenic, besides the presence of lovastatin in angkak can reduce
cholesterol and triglycerides. Asben and Permata (2018) in their research the effect of sago hampas
particle size in angkak pigmen production using Monascus purpureus, it was found that angkak's
toxicity value was 1719.86 ppm which indicates that angkak from sago pulp is not toxic.
4. Conclusion
The results showed that the pigment powder Angkak will increasingly dissolve at higher
temperatures. Stability of angkak pigments tends to decrease with longer heating, the higher the
heating temperature and the longer the irradiation. Angkak pigment powder is more stable at
neutral and basic pH compared to acidic pH. Angkak pigment residue pigment powder is not toxic
to experimental animals with LC50 value (toxicity value) Angkak pigment powder of 2897.05 ppm.
Putra et al. JAAST 5(1): 38 –49 (2021)
48
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Tedjautama, E., and Zubaidah, E. (2014). Increased Production of Red Pigment Angkak High Lovastatin Using Co-cultures of Monascus purpureus and Saccharomyces cerevisiae. Jurnal Pangan dan Agroindustri, 2(4), 78-88.
Triana, E. & Nurhidayat, N. (2009). Pengaruh Saccharomyces cerevisiae terhadap Kadar Lovastatin dalam Angkak yang Dihasilkan dari Fermentasi Beras oleh Monascus purpureus JMBA. Berkala Peneliti Hayati, 14, 203-207. Retrieved from http://berkalahayati.org/index.php/jurnal/article/view/342
Yuliani, L. A. (2014). Pengaruh Konsentrasi Inokulum Monascus purpureus terhadap Produksi Pigmen pada Substrat Tepung Biji Nangka (Artocarpus heterophyllus) (Thesis). Retrieved from http://repository.upi.edu/3294/
Journal of Applied Agricultural Science and Technology E-ISSN: 2621-2528 5(1): 50-61 (2021) ISSN: 2621-4709
Received February 23, 2021; Accepted March 9, 2021; Published March 10, 2021 https://doi.org/10.32530/jaast.v5i1.12 This is an open access article under the CC BY-SA 4.0 license https://creativecommons.org/licenses/by-sa/4.0
50
THE DESIGN AND BUILDING OF MEDIUM CAPACITY DRYING HOUSE FOR BOKAR
Sri Aulia Novita*,1, Hendra 2, Perdana Putera2,3, Fithra Herdian1, Muhammad Makky4, Khandra Fahmi4
1Department of Agricultural Mechanisation Technology, Politeknik Pertanian Negeri Payakumbuh, 50 Kota,
Indonesia 2Department of Computer Engineering Technology, Politeknik Pertanian Negeri Payakumbuh, 50 Kota,
Indonesia 3Department of Electrical and Electronics Engineering, University of Nottingham, Nottingham, United
Kingdom 1Department of Agricultural Technology, Andalas University, Padang, Indonesia
*Corresponding author
Email: [email protected]
Abstract. Bokar, processed rubber material, is latex obtained from rubber trees of community plantation, and it can be formed into various kinds of rubber products such as: unsmoked sheet/ USS, slab and lump. Good quality of garden latex can be identified from the white bokar. It is not hard and odorless. Bokar that has been made is then ground using a rubber mill to produce milled rubber in accordance with Indonesian National Standard of Bokar no. 06-2047-2002. The purpose of this research was to design a simple rubber sheet drying house, to examine the quality of dry rubber in accordance with Indonesian National Standard, to determine the level of dry rubber produced, to find out the temperature and time constant for drying crumb rubber to get the quality of SIR 20 rubber, and to examine the odor of dry rubber. The results from this research showed that Bokars are processed using a liquid smoke coagulant with a concentration of 10-15% and it obtained a clean white rubber, odorless and slightly smelly. Bokar was ground to gain the thickness of the unsmoked sheet/ USS around 3-5 mm, then it was dried in a drying house. The components of the drying house include the drying room, heating room, heater, thermometer, blower, plenum chamber, ventilation, drying room door and electric motor. The capacity of the drying house is 200 kg of rubber sheet, with a drying temperature of 35 - 46 0C and a drying time for 6 hours. The rubber produced has good quality with average dry rubber content (DRC) was 73.75%. Keyword: bokar, drying house, dry rubber content
1. Introduction
Natural rubber in Indonesia is one of the important plantation commodities taking the role as a
source of foreign exchange income, job opportunity, and a trigger for economic growth in new centers
in rubber plantation areas. Indonesia is the second largest producer of natural rubber in the world after
Thailand. The total export of Indonesian natural rubber in 2014 shared about 2.62 million tones with
a value reaching 4.74 billion USD (Ditjenbun, 2015). Indonesian natural rubber export products are
mostly in the form of crumb rubber of the SIR 20 quality, which reaches 92% from the total
production. The level of demand and production of rubber has always increased significantly. It is
shown by the high demand of companies – rubber products supplying companies in complying of
Novita et al. JAAST 5(1): 50 –61 (2021)
51
production’s need. Based on the results of studies by experts, it shows that the trade prospects of
rubber products (Bokar) is very good.
The process of rubber farming by smallholder produces natural rubber materials such as latex,
lumps, unsmoked sheet/USS, crepe and slab. Smallholder rubber is usually processed by using
traditional methods. This kind of processing produces low quality rubber, so that the selling price of
Bokar received by farmers is relatively very cheap. The process of rubber coagulant usually uses acid
ant which produces smelly and dull color rubber. The good quality of Bokar is reflected by the dried
rubber content (DRC) and the high level of cleanliness. Effort to improve the quality of Bokar must
begin from latex to the final processing stage Solichin & Anwar (2003). The quality of Bokar must be
in accordance with Indonesian National Standard - SNI Bokar No. 06-2047-2002 (Badan
Standardisasi Nasional (BSN), 2002), which include the following conditions: it is not added with
non-rubber materials, frozen with formic acid / other freezer with proper dosage, stored in a shady
place and unsubmerged in water. The quality of this Bokar greatly determines the competitiveness of
Indonesian natural rubber in international market.
Bokar material is obtained from Limapuluh Kota regency. This latex will be mixed with liquid
smoke coagulant from coconut shell. The use of liquid smoke in processing rubber is proven to be
able to produce good quality of rubber in accordance with the requirements of the Indonesian National
Standard (SNI) 06-1903-2000 about Standard Indonesian Rubber (SIR) (BSN, 2000). Liquid smoke
is commonly used as a latex coagulant and and as odor remover. The results of the research by
Tedjaputra et al. (2007) showed that liquid smoke can be applied in rubber processing as a coagulation
material (clotting) and as odor control (malodor).
The ability of liquid smoke to agglomerate latex is due to the acetic acid content in liquid smoke.
Meanwhile, the control of stink odors by liquid smoke is more due to the content of phenols and other
aromatic compounds. The ability of phenol as an anti-microbial can prevent the growth of
microorganisms on rubber blankets (Karseno & Rahayu, 2002).
Bokars that have been made is then ground using a rubber grinder to produce milled rubber with
a thickness of 3-5 mm and flat (in accordance with Bokar of Indonesian National Standard). The
grinding capacity of this rubber is 37.40 kg / hour. Its capacity is quite high. The rubber grinding
process is carried out to separate most of the water contained in Bokar, so it is easier for the drying
process. Then, the milled rubber is dried by being aerated for ± 10-14 days so that Bokar is drier which
is called an unsmoked sheet.
To lessen the effort and speed up the drying process of the unsmoked sheet, the rubber drying
house was designed. There were two treatments implemented in this study which include the provision
Novita et al. JAAST 5(1): 50 –61 (2021)
52
of liquid smoke coagulant and the drying process to analyze the effect of drying on the quality of
Bokar, especially unsmoked sheet. The equipment used in the drying process is a drying house that
works automatically. The temperature and the drying time must be controlled so that the expected
level of production quality can be achieved.
During the rainy season, 320 kg of the sheet can be evenly dried to the required 0.5% moisture
content in 5 days. Compared to conventional smokehouses, a better design of the house can greatly
reduced fuelwood consumption, from 1.0–1.5 kg per kg to 0.3 kg (Breymayer et al., 1993). A of
rubber sheets heat storage with height of 100 cm has drying time 78 hours with a thermal efficiency
of the drying system of 6.71% (Tekasakul et al., 2017). The drying efficiency of the mixed-mode
dryer is 15.4% higher than the indirect solar dryer (13.3%). The moisture content of the rubber sheet
decreased from 32.3 to 2.0% and 29.4 to 8.0% on a wet basis (Dejchanchaiwong et al., 2016). Ortiz-
Rodríguez et al. (2021) have compared two direct and indirect solar drying technologies. The moisture
content of the rubber sheet decreased from 45.8 to 0.59% and from 49.7 to 0.33% on a dry basis. The
temperature of the rubber dryer greenhouse with an additional area panel (PGEP) was 5 ° C hotter
than the rubber dryer greenhouse without an additional area panel (PGEP) (Jitjack et al., 2016). Drying
the rubber sheet using the Sandwich model greenhouse is 15 °C and 5 °C higher than the ambient
temperature during the day and night and the rubber sheet moisture content drops from 36.4% to 2.8%
in less than 2 days (Tanwanichkul et al., 2013). Drying at 50 °C provides good rubber properties with
high PRI values, good mechanical properties, and thermal properties within the normal range (Pianroj
et al., 2018).
The design of the drying house should emphasize on the ease of the application. It is also
economical and environmentally friendly. Therefore, this research will be carried out by making a
simple rubber drying house and providing liquid smoke coagulant to coagulate rubber.
The purpose of this research was to design a simple rubber sheet drying house, to examine the
quality of drying rubber in accordance with Bokar of Indonesian National Standard 06-2047-2002
(BSN, 2002), to determine the level of dry rubber produced, to determine the temperature and time
constant for drying crumb rubber in order to obtain quality of rubber SIR 20, and to examine the odor
of drying rubber produced.
2. Methods
The equipment used in this study were workshop equipment, heaters, furnaces and laboratory
analysis equipment, while the materials were building materials of drying house, Bokar, liquid smoke
and etc. The design of the rubber drying house can be seen in Figure 1.
Novita et al. JAAST 5(1): 50 –61 (2021)
53
2.1. Functional Approach
Tool design or drying house has parts that function as:
1. Drying Room is a dry rubber area
2. Blower functions to drain dry air
3. The heater serves to heat the drying chamber
4. Plenum Chamber is a place to distribute hot air
5. Burning Furnace functions to produce hot air by burning bricks with liquefied petroleum gas (LPG)
6. The thermometer measures the temperature in the drying chamber
Figure 1. Design of drying house
2.2. The Process of Making Liquid Smoke and Bokar
Liquid smoke is made from coconut shells through a pyrolysis process using high
temperatures. The formed liquid smoke will be left alone for one week to be separated from the tar,
so that it can be used as a natural latex coagulant. The latex will be given liquid smoke around 10 -
15 ml / kg of material. Giving liquid smoke is expected to eliminate the smell of latex. After the
latex coagulates, Bokar will be milled with a rubber grinder to reduce the high water content, so it
eases the process of drying rubber sheet.
2.3. The Process of Drying of Bokar
Bokar (unsmoked sheet) that has been given a liquid smoke coagulant will be dried in the
drying chamber. Iin this drying process the temperature of the drying air, the drying time, the dry
rubber content, the level of cleanliness and the quality of rubber sheet will be determined.
Determination of Dry Rubber Content of Unsmoked Sheet
1
2
3
4 5
6
Novita et al. JAAST 5(1): 50 –61 (2021)
54
Determination of dry rubber content
𝐾 = 𝑊𝑊𝑡% × 100% (1)
Note: K is the dry rubber content; Wt is the weight of latex; W is the weight of the crepe from the
clumping of latex.
Average K = (K1 + K2 + …… + Kn) / n with K1… .Kn = dry rubber content for each sample
3. Result and Discussion
3.1. Bokar (Community Processed Rubber Material)
Bokar is latex obtained from rubber trees (Indonesian National Standard of Bokar 06-2047-
2002) (BSN, 2002). The rubber processed material can be formed into various kinds of rubber
products such as unsmoked sheet, slab and lump. Good quality of latex is the main requirement to
produce good Bokar. To get high quality of Bokar, a clean work environment is the most important
requirement that must be considered including the cleanliness of equipment used during working
and the possibility of latex contamination by any dirt. To prevent the acceleration of latex clumping,
the following ways should be considered: equipment for tapping and transportating must always be
clean and stainless, the latex must be transfered smoothly to the processing area without frequent
shakes, latex should not be exposed to direct sunlight, it can use anti-coagulants such as ammonia
(NH3) or sodium sulfite (Na2SO3) (Budiman, 2012).
Deliberate coagulation which is commonly done today is by adding the acids such as formic
and acetic acids to reduce the pH of the latex. Meanwhile, latex can coagulate naturally due to the
formation of acidic compounds as a result of overhauling the carbohydrates and lipids contained in
latex by microorganisms.
To make unsmoked sheet, the raw material used in this research is fresh latex which has not
been added with coagulant material which is obtained from several areas in Limapuluh Kota
regency. The latex must be free from any dirt such as; crumbs, leaves and other debris. Then, latex
is filtered and put into a rectangular container, and mixed with 10 ml of liquid smoke, stirred and
left alone for ± 4 hours, so that the latex will agglomerated.
After that, the processed latex material will be washed to clean the dirt and then grinding the
rubber to reduce water content material to ease the drying process. Liquid smoke grade 3 contains
acid, carbon and phenol which can be used as a natural latex coagulant.
The acetic acid in liquid smoke can be used as a coagulation for latex (Solichin & Anwar,
Novita et al. JAAST 5(1): 50 –61 (2021)
55
2003), while the phenolic compounds are proven as anti-bacterial, anti-oxidant and anti-fungal
(Novita, 2011). The addition of liquid smoke can prevent fungal growth and smelly odor in Bokar
which has the impact like the rubber smoking process. Proper treatment in rubber processing will
produce high-quality rubber, so that the price of smallholder rubber farming can be increased. Liquid
latex is added with liquid smoke at a concentration around 10-15%. The process of latex with liquid
smoke coagulants can be seen in Figure 2.
Figure 2. Garden Latex Added liquid smoke
Bokar is left for several hours, until the latex is completely coagulated. The white color
showed that it has very good quality as seen in Figure 3. After this process, reducing the amount
of air content was done in the grinder.
Figure 3. Coagulated latex
3.2. Rubber Grinding Process
Bokar will be ground to reduce the water content in Bokar. This rubber grinding process uses
a rubber grinding tool that uses three rollers to flatten Bokar. The grinding process must be done
after 4-5 hours of coagulation of the latex. If the time is too long, the rubber becomes harder so
that it will be difficult to be ground. In the grinding process, the grinding is carried out 4-5 times
to get a grinding thickness about 3-5 mm. The thickness of the unsmoked sheet that is obtained is
in accordance with the Indonesian National Standard of Bokar. From the performance of the
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56
experiment of Bokar, it was found that the average working capacity of the tool was 37.40 kg /
hour.
With the rubber grinding, it will ease subsequent drying rubber process, because the water
content in the rubber has been reduced. If the grinding is uneven, the cooking process will decrease
the value of the accelerated storage hardening test (ASHT) after the drying process is carried out.
The process of grinding Bokar can be seen in Figure 4.
Figure 4. Rubber grinding process
3.3. The Manufacture of Drying House
The drying house is a building designed for the drying process of the unsmoked sheet. This
drying house consists of a drying chamber (Figure 6), drying shelf, ventilation, roof of the drying
house, heater, plenum chamber, blower, combustion chamber (Figure 5), bricks and liquid
petroleum gas (LPG).
Figure 5. Burning room
Figure 6. Drying room
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57
The drying house space is allocated mostly to the heating room as an area to produce hot air
by burning the bricks with liquid petroleum gas to dry the unsmoked rubber sheet and the drying
chamber as an area where the ground rubber is dried by stringing it. Other main components of
the house consist of the heater, blower, plenum chamber and ventilation. These components have
function to convert electrical to heat energy to produce hot air and distribute the hot air.
The drying house can dry Bokar or unsmoked sheet well, so that the quality of Bokar can
meet the Indonesian National Standard of Bokar. With proper drying, it will facilitate the rubber
storage process for a long time.
3.4. The Performance of Drying House Bokar
In order to produce high quality pocessed rubber material that is based on the standard of
crumb rubber, SIR 20, proper treatment during the process is required. One of the crumb rubber
processing processes that determines the quality of a rubber product is the drying process which
aims to maintain the ASHT value of the processed rubber product. Although the ASHT value for
SIR 20 is not attached in the Standard Indonesian Rubber (SIR), it is very important to maintain
the quality of the crumb rubber produced.
In the SIR 20 crumb rubber process, there are other factors that influence including the type
of raw material, the grinding process, and the age of raw material. Considering how important the
drying process is to reduce the ASHT value, it is necessary to handle it properly. Therefore,
temperature and drying time must be maintained so that the expected level of production quality,
which is Standard Indonesian Rubber (SIR) 20, can be met.
Table 1. Data dry rubber content
No Initial weight (Kg)
After drying weight (Kg)
Water content (Kg) Drying content (%)
1 1.3 0.98 0.32 75.38 2 1.3 0.95 0.35 73.07 3 1.1 0.8 0.3 72.73 4 1.1 0.79 0.31 71.82 5 0.9 0.66 0.24 73.33 6 0.9 0.65 0.25 72.22 7 0.75 0.55 0.2 73.33 8 0.75 0.56 0.19 74.67 9 0.55 0.42 0.13 76.36 10 0.55 0.41 0.14 74.54
Average of rubber drying content 73.75
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58
Based on the discussion above, it is clear that the temperature and drying time need to be
kept constant, because this affects the quality level of rubber production. Data from the
performance of the rubber grinding house to determine the dry rubber content can be seen in Table
1.
The thickness of the unsmoked sheet of Bokar is in the range of 3 - 5 mm, in accordance
with the thickness of unsmoked sheet of Indonesian National Standard of Bokar. The average of
dry rubber content obtained from the research data is 73.75%. It shows that the processed rubber
has good quality, so that the rubber farmers can increase the price of rubber.
The high amount of dry rubber content that was hanged was due to the low water content of
the rubber sheet which was caused by the evaporation process during hanging. In the coagulation
of rubber sheet, besides rubber particles, it also contains other materials such as water (Jayanthy
& Sankaranarayanan, 2005). Dry rubber content is the percentage of natural rubber (poliisoprena)
particles contained in Bokar, which is rubber sheet. Dry rubber content is a common term used in
the natural rubber processing industry (Kumar et al., 2007).
The advantage of having a high dry rubber content on blankets is that the drying process can
be done faster because there is less amount of water that has to be evaporated. Drying is an
important process to reduce the water content of materials and to ensure the consistency of product
quality (Ng et al., 2015) This fast drying process will increase the efficiency of production costs.
Tham et al. (2014) and Ekphon et al. (2013) stated that the drying process requires a large amount
of energy in the natural rubber industry. The lower water content, the shorter the drying time, so
that the energy required is lower or more efficient.
The drying temperature using a heat source from gas bricks is in the range of 35 - 46 0C for
6 hours, and temperature control is done by adjusting the gas output in the 3 kg liquid petroleum
gas (LPG). Temperature measurement is carried out by using a thermometer. High drying
temperatures can increase or decrease the ASHT value of the rubber depending on the drying time.
The duration of the drying process in the drying room is influenced by the sheet internal
factors such as the thickness of the sheet layer (Figure 7), pattern form and softness of the sheet
layer. Whilst the external factor are the hanging treatment, way and the tightness of the hanging,
airflow control, heat setting , insulation of drying room, fuel type and weather condition.
Novita et al. JAAST 5(1): 50 –61 (2021)
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Figure 7. Dried Ribbed Smoked Sit The result of the unsmoked sheet obtained from drying by using a drying house shows a very
good quality. This can be seen from the good color of the unsmoked sheet. It does not have a lot
of dirt and odorless or slightly smelly due to the addition of liquid smoke coagulants.
4. ConclusionBased on the report data above, it can be concluded processing of Bokars is in accordance
with Indonesian National Standard of Bokar. To produce high-quality Bokar, a natural coagulant
which is liquid smoke is used so Bokar is cleaner, odorless and has good quality. Latex will
coagulate for 4 - 10 hours after filtration. Bokar which has agglomerated is then being ground
using a rubber grinder so that it is called rubber milled (unsmoked sheet), which has a thickness
of 3-5 mm. The unsmoked sheet/ USS will be dried in the drying house with two heaters, namely
bricks and gas heater and a heater. The drying process using bricks and gas with the drying
temperature of 35 -46oC, and with a drying time of 6 hours resulting in dry rubber content as much
as 73.75%, to produce unsmoked sheets of good quality. It does not have a lot of dirt and odorless
or slightly smelly due to the addition of liquid smoke coagulants.
AcknowledgementWe would like to thank the Ministry of Research, Technology and Higher Education and
Centre of Research and Community Development Payakumbuh State Agricultural Polytechnic
who have been willing to fund this research.
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Journal of Applied Agricultural Science and Technology E-ISSN: 2621-2528 5(1): 62-63 (2021) ISSN: 2621-4709
https://doi.org/10.32530/jaast.v5i1.19 This is an open access article under the CC BY-SA 4.0 license https://creativecommons.org/licenses/by-sa/4.0
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THE PROSPECT OF KNOWLEDGE GROWING SYSTEM (KGS) FOR PLANT DISEASE EARLY DETECTION SYSTEM
Ika Noer Syamsiana*
Department of Electrical Engineering, Politeknik Negeri Malang, Malang, Indonesia
*Corresponding author
Email: [email protected]
Pests and diseases significantly contribute to crop production losses. The primary diseases
found are fungi, bacteria, and viruses. These diseases affect the growth and development of the plant
and lead to harvest loss or failure. The microscopic test is one of the conventional technology that is
still used for detecting plant disease. This test is very accurate in delivering the results but the test
process is very expensive.
Pattern recognition combined with image processing offers a low-cost approach to identify
plant disease and many research articles proposed Machine Learning and Deep Learning to deliver
very high accuracy in determining and classifying healthy plants. However, most of the methods need
a huge dataset and require significant time to make the annotations.
Knowledge Growing System (KGS), one of the methods in Cognitive Artificial Intelligence
(CAI). KGS uses a different approach from Machine Learning and Deep Learning, It does not learn
from the prepared annotated data but it learns directly from the phenomenon or object and grows its
own knowledge during interactions.
Figure 1. The model of KGS mechanism in obtaining new knowledge.
Basically, KGS is a system that is able to grow its own knowledge as the accretion of
information as the time elapsed. This concept derived from the observation of the mechanism that
Syamsiana JAAST 5(1): 62 –63 (2021)
63
occurs within human brain when performing information inferencing fusion to generate new
knowledge. KGS is developed by referring to Human Inference System (HIS). The process in
obtaining new knowledge is carried out in three simple steps after receiving data or information
regarding the phenomenon or object being observed, namely information fusion, information
inferencing, and information inferencing fusion as depicted in Figure 1. New knowledge is obtained
by applying a mechanism called as Degree of Certainty (DoC), which means the value that determines
the knowledge obtained by KGS after making interactions with the phenomenon or object.
KGS has already been applied in various fields such as bioinformatics, decision making,
cybersecurity, health, early warning system, Covid-19 detection system. and others including
cognitive processor. KGS is CAI technology invented by two Indonesian researchers, namely Colonel
Assist. Prof. Dr. Ir. Arwin Datumaya Wahyudi Sumari, S.T., M.T., IPM, ASEAN Eng., ACPE and
Prof. Dr. Ir. Adang Suwandi Ahmad, DEA, IPU. It is an original product of Indonesia and has already
gotten registered patent number P00201902101 dated March 12, 2019. In the near future, KGS is very
prospective to be applied as a plant disease early detection system.
Ika Noer Syamsiana, ST, MT Member of Editorial Board
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