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 for sustainable food security Muhammad Manjurul Karim, PhD Department of Microbiology, University of Dhaka, Bangladesh 2

3

WAY OUT for a sustainable development? Bioinnovation and Bioeconomy  Development of salt-resistant crop varieties  Application of salt-tolerant Plant growth promoting rhizobacteria (PGPR) Prologue 4

Direct effect Indirect effect Roles of PGPR Antimicrobials N 2 fixation production Hydrolytic Phosphate enzyme solubilization Completely Green Induced approach Siderophores system production resistance K EPS production Phytohormone production Why PGPR?

Bacillus aryabhattai MS3 Sampling site Plant Growth Promoting abilities A Venn diagram to isolate potential bacteria Background

Pot Experiments under saline and non-saline condition Materials Source of collection Seed Laboratory of Plant Biotechnology, ( Oryza sativa BR-28) Department of BMB, DU A salt-sensitive variety Agriculture field soil Gazipur Earthen pot Local market In vivo experiments 7

In v n vivo e expe periment Timeline of in vivo experiment (PGPR a applicatio ion as b biofe iofert rtil ilizer on on ric rice p plant) t) Day No. 1 Seeds incubated at 56⁰C seeds incubated at 56⁰C (2 days) 3 seeds transferred to a Seeds transferred to a petriplate containing petriplate containing whatman whatman filter paper (2 days) filter paper 5 germinated seeds transferred Germinated seeds transferred to to hypotonic solution hydroponic solution (8 days) (8 days) Cultivation on pot under normal condition 13 cultivation on pot under non- (45 days) saline condition (45 days) Application of salt (for 25 days) 58 application of salt 83 Physiological, phenotypic and bio- molecular analyses of plants Methods 8

Cultivated on earthen pots Day 15 Day 11 Day 10 Day 7 Day 1 Picture Profile 9

Preparation of bio-fertilizer • Bio-fertilizer was formulated according to the Bureau of Indian Standards (BIS) guidelines. – Charcoal powder : Calcium carbonate : Gum acacia: Microbial culture = 7: 1: 0.2: 1 × 10 9 Biofertilizer Biofertilizer preparation 10

Survival Survivability of MS3 applied 87% 84 12 plant plants Live plants number Dead plants number Survivability of controled 25 7 78% plants MS3-applied plants Control plants E.coli DH5 α -appl plants Results 11

Obse servatio ion a n after s salt a applica icatio ion

After 25 days 46% Survivability of MS3 applied plants 39 45 8% Survivability of controled plants 2 23 MS3 Applied plants Control plants E.coli DH5 α -appl plants 13 Results

Observing ng physi siological a and nd p pheno notypic sta state te of plants nts

Plant length in cm 60 50 40 Length in cm Comparative length 30 (cm) of plant stem 20 and leaves under normal and saline 10 conditions 0 normal condition saline condition normal condition saline condition stem leaf Plant dry weight in normal and saline conditions 25 None (control) 20 Plant dry weight in gm E. coli DH5 α (control) 15 B. aryabhattai MS3 10 5 0 normal condition saline condition Results: Plants’ phenotypic state 15

12 30 10 Carbohydrate conc. (µM/g) IAA concentration((µg/g) 25 8 20 6 15 4 10 2 5 0 0 normal condition saline condition normal condition saline condition normal condition saline condition normal condition saline condition stem leaf stem leaf IAA conc. of plants (stem and leaf) under normal and Carbohydrate conc. of plants (stem and leaf) under normal saline conditions and saline conditions 3 2.5 chlorophyl conc.(µg/g) 2 1.5 None (control) 1 E. coli DH5 α (control) 0.5 0 B. aryabhattai MS3 normal condition saline condition normal condition saline condition stem leaf Chlorophyll conc. of plants (stem and leaf) under normal and saline conditions Results: Plants’ biochemical state Carbohydrate conc. of plants (stem and leaf) under saline and normal conditions

16 14 MDA Melandealdehyde High MDA content indicates 12 conc.(µM/g) membrane lipid 10 peroxidation, sustainability 8 of plant growth under high salinity is associated with 6 reduced MDA formation 4 2 0 None normal condition saline condition normal condition saline condition E.coli DH5 α stem leaf MS3 350 Proline conc. (µM/g) 300 Proline 250 Mainly produced by 200 plants as a compatible solute under saline 150 condition to overcome 100 the oxidative damage of tissues. 50 0 Results: Plants’ biochemical state

MOLECULAR MECHANISM OF PLANT’S SALT RESISTANCE BY PGPR Results 18

Plant Gene Expression Analysis 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 osmotic signaling Ref: C.S. Nautiyal et al. / Plant Physiology and Biochemistry 66 (2013) Results

Transcriptomic analysis Semi quantitative RT- PCR Total mRNA Collect tissue RT PCR DNA pol dNTP + buffer cDNA Specific amplicon pool of target cDNAs Results

Analysis of plant gene expression upon salt stress C C+S B B+S C C+S B B+S C C+S B B+S 3000 1500 1000 700 BZ8 600 GIG 200 NHX1 100 bp Keys: Results C = Control, S= salinity with 200 mM, and B= Bacillus aryabhattai

Analysis of plant gene expression upon salt stress Results

PGP GPR & & Plant t • Nitrogen fixation NH NHX1 • Phosphate solubilization • Siderophore production • Phytohormone production Plant cell Model

Conclusion  Application of MS3 could induce plant growth even under salinity stress conditions, as a result of plant-microbe interaction by  increasing availability of nutrients (Fe, P)  decreasing reduction of IAA and chlorophyll content  enhancing proline accumulation  avoiding MDA formation • We propose stimulation by Bacillus arybhattai MS3 as a mechanism of inducing salt tolerance in rice by modulating differential transcription in a set of salt-tolerant genes. Conclusion

Research Experience Sea Level Acts to Rise Climate Achieve SDG Goal Change Salinity 13 ‘Climate Intrusion Action” Endophytic PGPR Bioinoculants Global Warming Substantial Failure to Achieve Crop Yield SDG 2030’s Initial Goal ‘No Poverty’ Achieved Crop Loss and Food Threatened Food Security Poverty Security 2 nd SDG Goal ‘Zero Food Food Hunger’ Price ↑ Price ↓ Concluding remarks 25

Final Words The suitable application of PGPR can bring the coastal agriculture in to sustainable crop production 12-Jan-19 26

Acknowledgements : 1. Dr Sirajul Hoq Dept of Soil, Water and Environment University of Dhaka 2. Dr Zeba I Seraj Dept of Biochemistry & Molecular Biology University of Dhaka Thank you My students: 1. Shahnaz Sultana 2. Sumonto C Paul 3. Samia Rahman 4. Bushra Zannat and 5. Naziza Rahaman Funding body: Ministry of Education Government of the People’s Republic of Bangladesh 27