Isolation, maintenance and screening of symbiotic and non symbiotic diazotrophs BY Ariba Ajmal

2. Isolation, maintenance and screening of symbiotic and non symbiotic diazotrophs BY AribaAjmal 2010-ag-3582 From Soil Bacteriology Section Ayub Agriculture Research institute (AARI), Faisalabad Supervised By Muhammad AmjadQurashi

3. Research work • Case study; Growth Promotion Potential of Different PGPR on the Growth of Maize • Media preparation and isolations • Soil analysis

4. CASE STUDYGROWTH PROMOTION POTENTIAL OF DIFFERENT PGPR ON THE GROWTH OF MAIZE.

5. INTRODUCTION • Maize (Zeamays L.). • Most diffused crop world wide • 3rd most important crop after wheat and rice. • Use as both grain and fodder. • High economic and nutritional value • High starch content about 75% of weight • Maize kernels provide significant amount of proteins, involved in N storage

6. PGPR • Plant growth promoting rhizobacteria • potential tools for sustainable agriculture • Rhizobacteria are rhizosphere competent bacteria that aggressively colonize plant roots; they are able to multiply and colonize all the ecological niches. • Alternative source of mineral fertilizers

7. Objectives Keeping in view the above discussion, the present study was designed with the following objectives: • Isolation and screening of effective rhizobia and PGPR • Testing potential of selected rhizobia and PGPR strains alone as well as in combination (co-inoculation) for improving growth, yield and nodulation of maize. • Identification and characterization of selected rhizobia and PGPR that are beneficial for improving growth of maize.  

8. Rhizobium Chickpea • One of the most important dry land field and pulse crops. • Mainly grown in rainfed areas • Leguminous crop • Role play in BNF • Maximum N-fixation depends upon • cultivar • Number of nodules • Efficient strain of bacteria existing in their root nodules

9. Azotobacter • Free lioving • Aerobic organisms • Nitrogen fixer • Affect on seed growth • Bind atmosphric N and release ammonium ion into soil • Growth promoter

10. Azospirillum • General root colonizer • Spiral shaped • Produce hormones and vitamines • Fix the nitrogen • Provide nutritions to cereal crops

11. bacillus • Most abundant geneus in rhizosphere • Activity as PGPR known for many years • Release number of meatabolites • P-solublizers • N-fixer

12. Pseudomonas • Ubiquitous bacteria • Help in maintenance of soil health • P-solublizers • N-fixer

13. METHODOLOGY • Isolation of Az, As, Ps, Bc sp were isolated by standard dilution method. • Chick pea plants uprooted • Nodules separated and sterilized • Obtain suspension • Streaked on Congo red YMA medium • Colorless, purified colonies picked and re-streaked to obtain pure culture • Purified rhizobial culture, streaked on slants

14. YMA medium

15. Pot study • Complete Randomized Design (CRD) with six treatments and three replications. • T1=Control • T2=Azotobacter sp • T3=Azospirillum sp • T4 =Rhizobium sp (chick pea) • T5 = Pseudomonas sp • T6 = Bacillus sp

16. Results • Effect of treatments on root and shoot parameters of maize.

17. Effect of treatments on IAA equivalents and dry root /shoot mass of maize

18. conclusion • Clearly indicated that PGPR • Influenced the groeyh of maize considerably • Az show highest ABP and after it Rhizobium CP • Bacillus shows better growth of root, shoot parameters as compared to others

19. ISOLATION OF RHIZOBIUM Pink Healthy Nodules Washing Sterilization (95% Ehanol) Washing 0.1% HgCl2 Washing Crushing Streaking on YMA Petri Plate

20. ISOLATION OF FREE LIVINGS • Dilution Plate Technique

21. RESULTS • isolations of different microbes

22. SOIL ANALYSIS • Different soil samples were analyzed for: • Organic matter (walkley black method) • Phosphorus (Olsen and sommers method)

23. ResultsTable: Determination of available organic matter from different soil sample

24. ResultsTable: Determination of available Phosphorous from different soil sample

25. thanks