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Phosphorus concentration and dry weight in phosphate solubilization ability in FePO4·(H2O)2 form

Phosphorus concentration and dry weight in phosphate solubilization ability in Ca3(PO4)2 form

Aspergillus sp. isolate 0/2 and I/4 cannot solubilize AlPO4

Biofertilizer development from the selected mineral solubilizing fungi for sweet basil (Ocimum basilicum) growth promotion

Laksana Nunan, Jaturong Kumla, Nakarin Suwannarach and Saisamorn Lumyong

Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand

INTRODUCTION

OBJECTIVES

MATERIALS AND METHODS

ReferencesJain et al. 2012. Effect of phosphate-solubilizing fungi Aspergillus awamori S29 on mungbean (Vigna radiata cv. RMG 492) growth. Folia Microbiologica,

57(6), 533–541.Qiao et al. 2019. The phosphate-solubilising ability of Penicilium guanacastense and its effects on the growth of Pinus massoniana in phosphate limiting

conditions. Biology open. 2019 Nov 15; 8(11): bio046797.Muraleedharan, H. Seshadri, S. and Perumal, K. 2012. Biofertilizer (Phosphobacteria). Shri A.M.M. Murugappa Chettiar Research Centre (MCRC). India

RESULTS

CONCLUSIONS

1. To vary and select the effective strains of phosphate mineral solubilizing fungi and siderophore production.2. To investigate phosphate solubilizing ability of fungi in different forms of phosphate minerals.3. To produce biofertilizer and evaluate sweet basil growth promoting ability of the biofertilizer from the selected fungi.

plants require mineral nutrients for their growth. However, the mineral in soil are usually in the insoluble form. A variety of mineral solubilizing microorganisms (MSMs) including bacterial and fungi, play an important role for solubilize an insoluble mineral in soil into available form for plant. MSMs use various mechanism to solubilize mineral such as acid production and secretion of siderophore. Moreover, MSMs can also increase the growth of plants.

1. Source of fungi Seven fungal isolates in genera Talaromyces (1 isolate) Penicillium (1 isolate)

and Aspergillus (5 isolates) were used in this study. All fungi isolated from the soil around the plant roots and identify species by SDBR laboratory.

Fungi on PDA: Talaromyces (isolate M1) Penicillium (isolate M4) and Aspergillus (isolates M3, PY, 0/2, I/1 and I/4)

2. Phosphate solubilizing

Pure culture

Inoculation

Pikovskaya’s (PKV) agar Clear zone on PKV agar

Incubation at 30 °c for 5 days Solubilization index (SI)

measurement

3. Siderophore production

Pure culture Chrome Azurol S (CAS) agar Color change zone on CAS agar

Diameter of color change zone measurement

4. Phosphate solubilization ability in different forms

Pure culture

Inoculation 3 pieces

Incubation on shaker at room temperature

PKV broth add Ca3(PO4)2, FePO4·(H2O)2 or AlPO4

Collect the sampleevery 24 hours for 5 days

Filtration out the fungal from media broth

Available phosphorus measurement bycolorimetric assay

Calculation of an available phosphorus content

5. Biofertilizer production

6. Evaluation of plant growth promoting

Incubation at 30°cfor 2 weeks

Spore suspension in water

vermiculite

perlitepeat moss

Compress and dry at 45°c for 48 hours

Spore concentration at 5x106 /g substrate

Cut fungal agar with Pasteur pipette

Bio-material

Transfer toplanting bag

seedling

Pla

nti

ng

Gro

wth

me

asu

rem

en

t

(6 treatment, 6 replication)

Leaves – leaves number, leaf area, fresh and dry weightStem – Height, stem diameter, fresh and dry weightRoot – length, fresh and dry weight

Treatment1 soil2 soil + Ca3(PO4)2

3 soil + biofertilizer Aspergillus sp. isolate 0/2

4 soil + biofertilizer Aspergillus sp. isolate I/4

5 soil + Ca3(PO4)2 + biofertilizer Aspergillus sp. isolate 0/2

6 soil + Ca3(PO4)2 + biofertilizer Aspergillus sp. isolate I/4

Table 1 Treatment details

Inoculation

Incubation at 30 °c for 3 days

Sweet basil seed

Plant for 60 days

Measure sweet basil growth

Fungal isolate SI value

Talaromyces sp. M1 1.09 ± 0.01 bc

Aspergillus sp. M3 1.11 ± 0.04 bc

Penicillium sp. M4 1.02 ± 0.02 c

Aspergillus sp. PY 1.05 ± 0.02 cd

Aspergillus sp. 0/2 1.12 ± 0.03 bc

Aspergillus sp. I/1 0.90 ± 0.0 d

Aspergillus sp. I/4 1.14 ± 0.03 a

1. Phosphate solubilizingTable 2 Solubilization index of each fungal isolate

Clear zone on PKV agar

Results are mean ± SD. The different letters in the same column indicated a significantly different at P < 0.05

2. Siderophore production

Fungal isolate Color change zone (cm)

Talaromyces sp. M1 2.35 ± 0.07 d

Aspergillus sp. M3 6.85 ± 0.07 b

Penicillium sp. M4 3.90 ± 0.42 c

Aspergillus sp. PY 7.15 ± 0.35 c

Aspergillus sp. 0/2 8.15 ± 0.07 a

Aspergillus sp. I/1 6.75 ± 0.21 b

Aspergillus sp. I/4 8.35 ± 0.07 a

Table 3 Diameter of color change zone of each fungal isolate

Results are mean ± SD. The different letters in the same column indicated a significantly different at P < 0.05

Color change zone on CAS agar

3. Phosphate solubilization ability in different forms

1. All fungal isolates can solubilize phosphate and produce siderophore. The high effective isolate were Aspergillus sp. isolate 0/2 and I/4.2. Aspergillus sp. isolate 0/2 and I/4 can solubilize Ca3(PO4)2 and FePO4·(H2O)2

but can’t solubilize AlPO4

3. Biofertilizer from Aspergillus sp. isolate 0/2 and I/4 could promote growth of sweet basil.

Acknowledgments

Microbiology Division, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand

The Sustainable Development of Biological Resources Laboratory (SDBR), Chiang Mai, ThailandCenter of Excellence in Microbial Diversity and Sustainable Utilization, Chiang Mai, Thailand

Treatment Leaves number Leaf area (cm2)Weight (g)

Fresh Dry

1 82.00 ± 9.20 cd 503.53 ± 190.92 b 15.22 ± 2.13 c 1.58 ± 0.26 c

2 73.75 ± 14.20 d 602.00 ± 83.70 ab 17.53 ± 3.84 bc 1.69 ± 0.46 b

3 89.00 ± 6.00 bc 710.18 ± 38.50 ab 18.18 ± 1.94 bc 1.90 ± 0.17 ab

4 80.50 ± 5.97 cd 783.20 ± 215.84 a 20.34 ± 0.89 b 2.06 ± 0.20 ab

5 104.00 ± 8.16 a 793.62 ± 105.29 a 24.20 ± 3.43 a 2.26 ± 0.13 a

6 97.25 ± 10.78 ab 801.70 ± 128.38 a 20.17 ± 0.95 b 1.87 ± 0.13 ab

6. Evaluation of plant growth promotingTable 4 Sweet basil growth after 60 days: Leaves

Treatment Height (cm) Diameter (mm)Weight (g)

Fresh Dry

1 43.2 ± 1.50 ab 1.15 ± 0.26 b 12.32 ± 1.39 a 2.61 ± 0.44 a

2 41.75 ± 3.68 b 1.30 ± 0.22 ab 11.27 ± 3.18 a 2.20 ± 0.69 a

3 45.75 ± 1.89 a 1.30 ± 0.25 ab 10.72 ± 0.59 a 2.54 ± 0.81 a

4 46.50 ± 1.19 a 1.22 ± 0.22 b 12.02 ± 3.54 a 2.54 ± 0.82 a

5 46.50 ± 1.29 a 1.58 ± 0.10 a 13.20 ± 1.26 a 2.67 ± 0.15 a

6 46.00 ± 2.94 a 1.45 ± 0.13 ab 10.54 ± 1.63 a 2.17 ± 0.21 a

Treatment Length (mm)Weight (g)

Fresh Dry

1 143.75 ± 20.56 b 1.23 ± 0.23 b 0.34 ± 0.53 b

2 145.00 ± 41.83 b 1.79 ± 0.47 ab 0.39 ± 0.14 b

3 147.50 ± 15.00 b 1.84 ± 0.39 ab 0.42 ± 0.06 ab

4 158.75 ± 31.19 ab 1.89 ± 0.15 ab 0.45 ± 0.05 ab

5 212.50 ± 51.23 a 2.18 ± 0.40 a 0.52 ± 0.04 a

6 192.50 ± 40.31 ab 1.92 ± 0.74 ab 0.46 ± 0.06 ab

Table 5 Sweet basil growth after 60 days: Stem

Table 6 Sweet basil growth after 60 days: Root

Sweet basil growth measurement of each treatment

Results are mean ± SD. The different letters in the same column indicated a significantly different at P < 0.05Results are mean ± SD. The different letters in the same column indicated a significantly different at P < 0.05

Results are mean ± SD. The different letters in the same column indicated a significantly different at P < 0.05