sustainability challenges in oil palm mono-cultivation: are microbes the solution? khim-phin,chong...
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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