Sustainability Challenges in Oil Palm Mono-cultivation: Are Microbes the Solution?

Khim-Phin,Chong

Sustainable Palm Oil Research Unit (SPOR)

Universiti Malaysia Sabah

Sustainable Palm Oil Research Unit (SPOR)

Content of Talk:

Oil Palm Success and Challenges: In Brief

Fundamental of Microbial in Agriculture

Case Studies:

-Roles of Microbes in Oil Palm Improvement:

-Roles of Microbes in Enhancing Oil Palm Disease Resistant

-Microbial Diversity in Oil Palm Soil

Conclusions

Oil Palm

World Palm Oil production increase from 13 to 28% from 1990 to 2011, with increase of exports from 36 to 57%

Two key exporting countries: Malaysia (47%) and Indonesia (46%). Account 93% of world palm oil exports

Malaysia: 5 Million Ha of Oil Palm. Contributing over 11% of global supply of edible oils and fats with 0.1% of the total global agricultural land area

High Land Productivity Oil Palm vs Other Oil Seeds:

11x more than soyabean

10x more than sunflower

7x more than rapeseed

In 2011, Malaysia’s Export revenue of oil palm products reached a record high of RM80.4 billion, an increase of 34.5% against RM59.8 billion achieved in 2010.

Palm Oil Industry: At Crossroad or Under Real Threat? Workers

Socio-Environment Productivity

Cost of ProductionHuman Capital

Fundamental of Microbial in Agriculture

Soil Environments

• Surface and subsurface soils are typically nutrient-poor environments for microbes

• Rhizosphere is enriched in nutrients as a result of nearby plant activities

Rhizosphere

• Rhizoplane– soil in direct contact

with plant root

• Endophytes– microbes attached to

root surface

Decreasing moistureIncreasing organic C

Organic material in rhizosphere• Exudates

– low molecular weight compounds released from plant cells in a non-metabolic manner (leakage)

• Secretions– compounds metabolically released

from plant cells

• Lysates– compounds released from

moribund cells during autolysis

• Plant mucilage– plant polysacchrides

• Some bacteria and blue green algae are able to fix nitrogen from the atmosphere to enrich soil with nitrogen and increase its fertility.

• These microbes are commonly called biological nitrogen fixers.

Microbes Increase Soil Fertility

• Rhizobium bacteria is involved in the fixation of nitrogen in leguminous plants (pulses). Rhizobium lives in the root nodules of leguminous plants, such as beans and peas, with which it has a symbiotic relationship.

• Sometimes nitrogen gets fixed through the action of lightning.

Nitrogen Fixation

Nitrogen Fixation• Our atmosphere has 78% nitrogen gas. Nitrogen is one of the

essential constituents of all living organisms as part of proteins, chlorophyll, nucleic acids and vitamins. The atmospheric nitrogen cannot be taken directly by plants and animals.

• Certain bacteria and blue green algae present in the soil fix nitrogen from the atmosphere and convert into compounds of nitrogen.

• Once nitrogen is converted into these usable compounds, it can be utilized by plants from the soil through their root system.

Nitrogen Fixation

• Nitrogen is then used for the synthesis of plant proteins and other compounds. Animals feeding on plants get these proteins and other nitrogen compounds.

• When plants and animals die, bacteria and fungi present in the soil convert the nitrogenous wastes into nitrogenous compounds to be used by plants again.

• Certain other bacteria convert some part of them to nitrogen gas which goes back into the atmosphere. As a result, the percentage of nitrogen in the atmosphere remains more or less constant.

Beneficial root-microbe interactions• Atmosphere contains 1015 tons N2 gas

– Biological nitrogen fixation– Minimum of 70 million tons N fixed/year

Sources of Fixed Nitrogen

65%10%

25%

Biological Lightning Fertilizer

Disease-causing Microorganisms in Plants

Roles of Microbes inCrop Improvement:

Case Study

Vegetative Growth Improvement-Nursery

(Johor Estate)• Combinations of Bacillus spp,

Aspergillus spp & Pseudomonas spp

Feb 2010 : Seed sowing

May 2010 : Transplanting to main

nursery

June 2010 : 1st Application

Oct 2010 : 2nd Application

Dec 2010 : Field transplanting Courtesy of Agrinos

Microbes treated

Untreated

Observation (Avg) Treated Control

Frond Length (cm) 133.8 125.6

Leaflet Length (cm) 41.8 36.8

Number of Total Fronds 21.2 20.4

Girth (cm) 37.2 30.4

Root Length (cm) 122.8 111.4

Seedling Height (cm) 154.8 140.2

Results show the averages for each parameters measure for microorganisms treated oil palm seedlings compared to control (Kalimantan Nursery, Indonesia)

Courtesy of Agrinos

Yield-Immature Plantings(Tawau Estate)

• Nitrogen based fertilizer reduction:

1st Yr Planting-0%

2nd Yr Planting-15%

Subsequent Yr-25%

• Yield Improvement (%)1st Yr Harvesting-80%

2nd Yr Harvesting-120%

3rd Yr Harvesting-49%

4th Yr Harvesting-17%

5th Yr Harvesting-11% Courtesy of Agrinos

3.57% bigger girth10% > LAI

Block Depth(cm) pH Exchg. K Available P (p.p.p.m)

Total Available

Organic C(%)

Organic N(%)

Opt 4.5-5.5

0.30 400 20.00 2.50 0.25

Treat-Blk 1

0-15 5.6 0.38 187 20.30 0.77 0.13

Treat-Blk 1

15-35 5.8 0.17 199 11.80 0.99 0.18

Cont-Blk 2

0-15 5.5 0.66 178 6.90 0.66 0.12

Cont-Blk 2

15-35 4.7 0.65 137 5.10 0.37 0.09

Soil Analysis

Courtesy of Agrinos

Yield-Mature Plantings(Sandakan Estate)

• 1st application of microbes: 10 years age

• Yield Improvement (%)1st Yr treatment-12%

2nd Yr treatment-15%

3rd Yr treatment-25%

4th Yr treatment-28%

*Salinity and sandy problem

Courtesy of Agrinos

Years of treatment

Roles of Microbes inEnhancing Disease Resistance:

Case Study

Microbes Ergosterol (Average)

Infection percentage

GSM percentage

Bacillus spp. and Trichoderma spp.

0.19 µg/mL-1 30 % 30%

Lactobacillus, Nattobacillus and Yeasts

0.19 µg/mL-1 60 % 60 %

Bacillus spp, Aspergillus spp. and Pseudomonas spp.

Control (Infect without microbes)

Control (Healthy)

0.23 µg/mL-1

1.52 µg/mL-1

0 µg/mL-1

100 %

100%

0%

100 %

100%

0%

Percentage of Ganoderma infection after two months based on ergosterol (fungal sterol) content and growth on GSM for oil palm seedlings pre-treated with various combination of microbes

Microbes Ergosterol (Average)

Infection percentage

GSM percentage

Bacillus spp. and Trichoderma spp.

0.22 µg/mL-1 60 % 60%

Lactobacillus, Nattobacillus and Yeasts

0.34 µg/mL-1 50 % 50%

Bacillus spp, Aspergillus spp. and Pseudomonas spp.

Control (Infect without microbes)

Control (Healthy)

0.22 µg/mL-1

2.35 µg/mL-1

0 µg/mL-1

60 %

100%

0%

60 %

100%

0%

Percentage of Ganoderma infection after four months based on ergosterol (fungal sterol) content and growth on GSM for oil palm seedlings pre-treated with various combination of microbes

Ergosterol

Ergosterol

HPLC Chromatograms of ergosterol, peak was detected at RT 7-8 min. (A) Ergosterol standard (B) Healthy palm (C) Infected palm. Ergosterol peaks are arrowed.

Oil palm roots cultured on Ganoderma Selective Media (GSM) after 5 days of incubation. A: Infected oil palm roots, indicated by Ganoderma growth. B: Uninfected oil palm roots.

A

A

C

B

B

Ganoderma-inoculated oil palm seedling roots treated with different combination of microbes. A: Bacillus sp and Trichoderma sp. B: Bacillus sp., Aspergillus sp. and Pseudomonas sp. C: Lactobacillus, Nattobacillus and Yeasts , D: Control. Bar= 3 cm

Oil palm estate soil inoculated with Ganoderma boninense and treated with combinations of Bacillus spp, Aspergillus spp and Pseudomonas spp

1st App

2nd App

Blue line: controlGreen line: Microbes treated

Courtesy of Agrinos & UPM

Hyphal extension of G. boninense in sterile and non sterile soil and frond debris (FD). Colonised wheat grains were used as inoculum source. a: Mycelial grown in FD after 4 days. b: Extension in FD after 10 days. c: Growth in soil after 4 days. d: Hyphal extension in soil after 10 days

2nd App

Cooper, R. (2011) in Sustainable Agriculture: An Insight into Ganoderma

Assessment on field application is currently on going. Data collection due mid Nov 2012

2nd App

Courtesy of Martin Kong, One Good Earth

Microbial Diversity in Oil Palm Soil

Population density of microorganisms in all soil samples collected from oil palm plantation of Sapi Nangoh, Sandakan.

SampleGrowth medium

NA(Bacteria) PDA (Fungi) MEA (Yeast)

S1 3.2 x 103 5.0 x 102 2.8 x 102

S2 5.1 x 103 1.0 x 102 1.3 x 104

S3 7.0 x 102 - 1.5 x 104

S4 7.2 x 104 - 4.7 x 104

S5 3.8 x 103 3.0 x 102 7.4 x 103

S6 - 3.0 x 102 5.3 x 103

S7 3.0 x 106 - 1.5 x 103

S8 4.0 x 103 - 4.8 x 103

S9 2.4 x 107 9.0 x 102 1.1 x 103

S10 1.6 x 103 1.0 x 102 2.0 x 106

S11 3.6 x 103 - 3.6 x 106

S12 7.0 x 102 1.0 x 102 4.2 x 104

S13 5.0 x 106 - 1.9 x 103

S14 1.2 x 106 - 7.9 x 103

S15 4.6 x 103 - 6.4 x 104

S16 6.7 x 105 - 7.2 x 104

S17 1.7 x 103 1.0 x 102 6.3 x 102

S18 5.7 x 103 - 4.7 x 104

S19 3.2 x 103 - 3.1 x 103

S20 3.2 x 103 1.0 x 102 4.7 x 103

Cont…..

SampleGrowth medium

NA(Bacteria) PDA (Fungi) MEA (Yeast)

S21 2.0 x 103 1.0 x 102 1.8 x 103

S22 2.0 x 105 3.0 x 102 9.6 x 103

S23 1.3 x 103 1.0 x 102 1.6 x 103

S24 3.2 x 103 3.0 x 102 1.1 x 103

S25 2.2 x 104 1.0 x 102 8.0 x 102

S26 3.1 x 104 1.0 x 102 1.6 x 103

S27 3.3 x 104 - 7.6 x 103

S28 3.1 x 104 - 2.4 x 103

S29 1.9 x 104 - 4.8 x 103

S30 3.6 x 103 - 2.0 x 103

S31 1.6 x 103 - 2.5 x 103

S32 7.3 x 103 8.0 x 102 2.7 x 103

S33 5.6 x 104 2.0 x 102 4.7 x 103

S34 1.5 x 103 - 4.0 x 102

S35 1.2 x 103 - 4.9 x 103

Microrganisms Genus SpeciesBacteria Bacillus B. thuringiensis

B. pumilusB. humiB. lichenformisB. albusB. pseudomycoidesB. amyloliquefaciens

Arthobacter A. woluwensisA. globiformis

Kytococous K. sedentariusRalstonia R. picketii

Actinomycetes Corynebacterium C. borisC. mycetoides

Brevibacterium B. epidermisRhodotococcus R. fascians

Yeast Rhodotorula R. minutaR. graminisR. pustula

Guilliermondella G. selenosporaFilobasidiella F. neoformansbacillisporusFellomyces F. fuzhouensisSporidiobolus S. johnsoniiBulleromyces B. albusCryptococcus C. albidus

Fungi Penicilium P. solitumwestlingP. neoechinulatum

Cladosporium C. herbarumFusarium F. tricinctumAcremonium A. kiliense

Identification using Biolog technique

GEN III microplate (left) showed purple colour -change pattern due to the carbon utilization by the bacteria after 24 hr of incubation. Based on the colour changes, bacteria was identified as Bacillus pumilus (squared in red) with the probability of 1.0 and 0.764 similarity.

Microbes Identification based on Biolog technique

Bacterial culture (left) isolated from soil sample which identified as B.pumilus based on Biolog identification technique. Observation of B. pumilus (right) under light microscope using 40x magnification power. Bar=100µm

Bacterial population: 102 to 107

cfu/g of soil

No growth was observed from soil sample of S6 on NA.

Fungal population: only 102cfu/g of soil.

No growth was observed on PDA from soil of S3, S4, S7, S8, S13, S14, S15, S16, S18, S19, S27, S28, S29, S30, S31, S34 and S35.

All soil samples (S1-S35) showed the presence of yeasts

Yeasts population: 102 to 106

cfu/g of soil

Oil PalmBacterial populations: 105 to 108 cfu/g of soil

Fungal population: 105 to 106

cfu/g of soil

Yeasts population: 103 cfu/g of soil

VS

Forest

Conclusion

• Soil microbial ecosystem is complex but combinations of microbes have the ability to enhance Oil Palm nutrient uptake, increase availability of nutrient to Oil Palm and suppress pathogens attack especially Ganoderma

• Ganoderma is weak competitor with many other microorganisms. We are the one who provides suitable soil condition for them to become the champion!

• Is extensive use of agrochemicals lead to depletion of microbial population in oil palm soil? Need further investigation

• More funding is required to accelerate research related to potential indigenous microbes for the betterment/enrichment of agriculture/plantation soil for the future.

Thank You

One Good Earth (M) Sdn Bhd All my Postgraduates & Co-

researchers