Muhammad Manjurul Karim Department of Microbiology University of - - PowerPoint PPT Presentation
Muhammad Manjurul Karim Department of Microbiology University of Dhaka Greetings from University of Dhaka, Bangladesh Salinity intrusion and coastal agriculture: Adaptation strategies using salt-tolerant plant-growth promoting rhizobacteria
Greetings from University of Dhaka, Bangladesh Muhammad Manjurul Karim Department of Microbiology University of Dhaka
Muhammad Manjurul Karim, PhD Department of Microbiology, University of Dhaka, Bangladesh
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Prologue
Why PGPR?
Direct effect N2 fixation Phosphate
solubilization Siderophores
production K production
Phytohormone
production Indirect effect
Antimicrobials
production Hydrolytic enzyme Induced system resistance EPS
Completely Green approach
Background
Sampling site Plant Growth Promoting abilities A Venn diagram to isolate potential bacteria Bacillus aryabhattaiMS3
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In vivo experiments Materials Source of collection
Seed (Oryza sativa BR-28) A salt-sensitive variety Laboratory of Plant Biotechnology, Department of BMB, DU Agriculture field soil Gazipur Earthen pot Local market
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Methods
seeds incubated at 56⁰C seeds transferred to a petriplate containing whatman filter paper germinated seeds transferred to hypotonic solution cultivation on pot under non- saline condition application of salt (8 days) (45 days) Seeds incubated at 56⁰C (2 days) Seeds transferred to a petriplate containing whatman filter paper (2 days) Germinated seeds transferred to hydroponic solution (8 days) Cultivation on pot under normal condition (45 days) Application of salt (for 25 days)
In v n vivo e expe periment (PGPR a applicatio ion as b biofe iofert rtil ilizer on
rice p plant) t)
1 3 5 13 58 83 Day No.
Physiological, phenotypic and bio- molecular analyses of plants
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Cultivated on earthen pots Day 1 Day 7 Day 10 Day 11 Day 15
Picture Profile
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Biofertilizer preparation
Indian Standards (BIS) guidelines.
– Charcoal powder : Calcium carbonate : Gum acacia: Microbial culture = 7: 1: 0.2: 1×109
Biofertilizer
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Results
Control plants MS3-applied plants E.coli DH5α-appl plants 25 84 7 12 78% 87% Survivability of controled plants Survivability of MS3 applied plants Live plants number Dead plants number Survival plant
Obse servatio ion a n after s salt a applica icatio ion
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Results
Control plants MS3 Applied plants E.coli DH5α-appl plants After 25 days 2 39 23 45 8% 46% Survivability of controled plants Survivability of MS3 applied plants
Observing ng physi siological a and nd p pheno notypic sta state te of plants nts
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Results: Plants’ phenotypic state
Comparative length (cm) of plant stem and leaves under normal and saline conditions
Plant length in cm Plant dry weight in normal and saline conditions
10 20 30 40 50 60 normal condition saline condition normal condition saline condition stem leaf Length in cm 5 10 15 20 25 normal condition saline condition Plant dry weight in gm
None (control)
IAA conc. of plants (stem and leaf) under normal and saline conditions Carbohydrate conc. of plants (stem and leaf) under normal and saline conditions Carbohydrate conc. of plants (stem and leaf) under saline and normal conditions
None (control)
Results: Plants’ biochemical state
2 4 6 8 10 12 normal condition saline condition normal condition saline condition stem leaf IAA concentration((µg/g) 5 10 15 20 25 30 normal condition saline condition normal condition saline condition stem leaf Carbohydrate conc. (µM/g) 0.5 1 1.5 2 2.5 3 normal condition saline condition normal condition saline condition stem leaf chlorophyl conc.(µg/g)
Chlorophyll conc. of plants (stem and leaf) under normal and saline conditions
2 4 6 8 10 12 14 16 normal condition saline condition normal condition saline condition stem leaf
Melandealdehyde conc.(µM/g) 50 100 150 200 250 300 350 Proline conc. (µM/g)
Proline Mainly produced by plants as a compatible solute under saline condition to overcome the oxidative damage of tissues. MDA High MDA content indicates membrane lipid peroxidation, sustainability
salinity is associated with reduced MDA formation
None E.coli DH5α MS3
Results: Plants’ biochemical state
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Results
To observe if the PGPR can confer salt tolerance ability in rice plant by modulating in cellular transcription level. 3 salt responsive genes were selected to be analyzed. Gene name Function NHX1 Sodium proton exchanger; reduces Na+ concentration in cytosol during salt stress. GIG Negative regulation of cellular protein translation. BZ8 Regulate different transcriptional pathway through
Ref: C.S. Nautiyal et al. / Plant Physiology and Biochemistry 66 (2013)
Results
Transcriptomic analysis
Collect tissue Total mRNA
RT PCR
cDNA DNA pol dNTP + buffer Specific amplicon pool of target cDNAs
Semi quantitative RT-PCR
Results
Results
BZ8 GIG NHX1 700 600 200 100 bp 3000 1500 1000 C C+S B B+S C C+S B B+S C C+S B B+S
Keys: C = Control, S= salinity with 200 mM, and B= Bacillus aryabhattai
Results
Plant cell
Model
PGP GPR & & Plant t NH NHX1
Conclusion
salinity stress conditions, as a result of plant-microbe interaction by
mechanism of inducing salt tolerance in rice by modulating differential transcription in a set of salt-tolerant genes.
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Global Warming Sea Level Rise Salinity Intrusion Climate Change Crop Loss and Threatened Food Security Food Price ↑ Poverty Failure to Achieve SDG 2030’s Initial Goal ‘No Poverty’ Endophytic PGPR Bioinoculants Substantial Crop Yield Acts to Achieve SDG Goal 13 ‘Climate Action” Achieved Food Security Food Price ↓ 2nd SDG Goal ‘Zero Hunger’
Concluding remarks
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12-Jan-19
Funding body: Ministry of Education Government of the People’s Republic of Bangladesh
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Acknowledgements :
Dept of Soil, Water and Environment University of Dhaka
Dept of Biochemistry & Molecular Biology University of Dhaka My students: 1. Shahnaz Sultana 2. Sumonto C Paul 3. Samia Rahman 4. Bushra Zannat and 5. Naziza Rahaman