2 * Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28 2 87/2/27 : 87/2/3 :. -- - () ( 68) Sinorhizobium meliloti 33. ( 58) R. leguminosarum bv. phaseoli ( 44) Rhizobium leguminosarum bv. viciae ( 6) Mesorhizobium ciceri. 28/2. 8/6 R. leguminosarum bv. Viciae 3/6 S. meliloti 46/5 M. ciceri 6/7 R. leguminosarum bv. Phaseoli.. 8-6-8 4-6 2-4 (HCN) (IAA)... 24 2 (IAA). 2/7 CAS... : bahar@nigeb.ac.ir : 24485353 : *. IAA (Plant PGPR Growth Promoting Rhizobacteria) 66
Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28 (99).(8 7) Pseudomonas.(23) 6 (999) (Rhizosphere).(5). - PGPR.. deaminase. Sinorhizobium meliloti 4 35 deaminase.(7).. 4 PGPR.(5) Azotobacter Acetobacter Azospirillum. Pseudomonas. (994). Rhizobium leguminosarum bv. trifoli.(2) -.()..(). 25 /25.(4) - PGPR deaminase -- - -) ( -. 662
Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28 (Plant infection test).. (Gram). ) 33.( (YeastMannitol Agar ) YMA.. - -7.. - 68 75 Sinorhizobium meliloti 44 44 Rhizobium leguminosarum bv. viciae 58 7 Rhizobium leguminosarum bv. phaseoli 6 Mesorhizobium ciceri 33 4. (Rhizobium Minimal Medium) 2 ІІІ ІІ ІІ І І. ІІІ. /25 5 ІІІ. 9-2 ; YMA /2 ;NaCl /; MgSO 4.7H 2 O /5 K 2 HPO 4 5 /5 ;Yeast extract ;Mannitol. ph (23).(2).(5). 663
Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28.(3) HCN.(24) (IAA) 53.(2) CAS-Agar. 25 (Chrome Azurol S). YMA.(2). 3 28/2. 46/5 Sinorhizobium meliloti Mesorhizobium ciceri 3. 6/7 (23).(6) K 2 HPO 4 2.5 gr KH 2 PO 4.45 gr ** (І Distilled water 2 ml NaCl.5gr CaCl 2. gr MgSO 4.7H 2 O.5 gr Mannitol. gr ** (ІІ Microelements solution *.5 ml Distilled water 8 ml Agar 5 gr Pantathonic acid mg.ml - Biotin mg.ml - (ІІІ Thiamine mg.ml - : * /5 7 ІІ І ph **. 2 ( ). 2.. /45 /45 () NH 4 Cl NH 4 Cl. ( ) (HCN).. YMA... HCN 28 HCN. 664
27.(6) deaminase 233 (28) 3 Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28-3 46/5 3/6 8/6 6/7 28/2 78 6 5 4 93 68 44 58 6 33.. 8- % 2/4 4 /2 4 Sinorhizobium meliloti Rhizobium leguminosarum bv. viciae Rhizobium leguminosarum bv. phaseoli Mesorhizobium ciceri ( ) 6-8 % 8/3 4 4/6 2 4/9 6 4-6 % 6 /7 3/3 2-4 % 29/8 5 9/ 4 6/9 4 6/7 8/8 4 62-4 ) 8-6-8 4-6 2.(4 % 53/5 9 86/4 38 9/4 53 93/3 7/8 56 237-4 Sinorhizobium meliloti Rhizobium leguminosarum bv. viciae Rhizobium leguminosarum bv. phaseoli Mesorhizobium ciceri S. meliloti. S. 4 meliloti. 665
Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28 (22) (8) (26).(7) 35-4.(25 ) Sino3 Sino44 Sino27 Sino45 Sino43 S. meliloti YMA 5.. (9) S. meliloti -5 Sino66 Sino97 Sino74 Sino5 Sino37 Sino4 Sino7 Sino49 Sino64 Sino58 Sino5 Sino6 Sino62 Sino72 Sino95 deaminase ( ) 8- (IAA) 6 IAA IAA ** 2/7 /6 /3 /7 * /2 /2 2/2 /8 3/7 7/3 /7 6/7 Sino6 Sino62 Sino64 Sino7 Sino72 Sino74 Sino97 Sino95 Sino3 Sino66 ** /7 /4 2/2 6-8 4-6 2-4 * 6 5/5 /5 3/2 /3 7/3 2/8 2 2/8 4 Sino5 Sino5 Sino27 Sino37 Sino4 Sino43 Sino44 Sino45 Sino49 Sino58 CAS ** 24 * 666
Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28... ph 6 Tm 8-8/5 (3 7). - PGPR. Sinorhizobium meliloti.. S. meliloti. PGPR. 6 CAS.. -.().(9) (6 ) (PGPR).. HCN.. ATP HCN.(4). 667
Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28.. 2. Alexander, D.B. and Zuberer, D.A. 99. Use of chrome azurol S reagents to evaluate siderophore production by rhizosphere bacteria. Biology and Fertility of Soils, 2:39-45. 3. Alström, S. and Burns, R.G. 989. Cyanide production by rhizobacteria as a possible mechanism of plant growth inhibition. Biology and Fertility of Soils, 7:232-238. 4. Baker, A.W. and Schippers, B. 986. Microbial cyanide production in the rhizosphere in relation to potato yield reduction and pseudomonas spp.- mediated plant growth-stimulation. Soil Biology and Biochemistry, 9: 45-457. 5. Cattelan, A.J. ; Hartel, P.G. and Fuhrmann, J.J. 999. Screening for Plant growth promoting rhizobacteria to promote early soybean growth. Soil Science Society of America Journal, 63:67-68. 6. Duan, J.; Müller, K.M. ; Charles, T.C. ; Vesely, S. and Glick B.R. 28. -aminocyclopropane- -carboxylate () deaminase genes in Rhizobia from Southern Saskatchewan. Microbial Ecology, DOI.7/s248-8- 947-6. 7. Glick, B.R. 995. The enhancement of plant growth by free-living bacteria. Canadian Journal of Microbiology, 4:9 7. 8. Glick, B.R. ; Penrose, D.M, and Li., J. 998. A model for the lowering of plant ethylene concentrations by plant growth-promoting bacteria. Journal of Theoretical Biology, 9: 63 68. 9. Grichko, V.P. and Glick, B.R. 2. Amelioration of flooding stress by deaminase-containing plant growth-promoting bacteria. Plant Physiology and Biochemistry, 39:-7.. Grichko, V.P.; Filby, B. and Glick, B.R. 2. Increased ability of transgenic plants expressing...376... 72 the bacterial enzyme to accumulate Cd, Co, Cu, Ni, Pb and Zn. Journal of Biotechnology, 8: 45-53.. Guerinot, M.L. 99. Iron uptake and metabolism in the rhizobia/legume symbiosis. Plant and Soil, 3:99-29. 2. Hoflich, G. ; Wiehle, W. and Kuhn, G. 994. Plant growth stimulation by inoculation with symbiotic and associative microorganisms. Experientia, 5: 897-95. 3. Hontzeas, N. ; Hontzeas, C.E. and B. R. Glick. 26. Reaction mechanisms of the bacterial enzyme -aminocyclopropane--carboxylate deaminase. Biochemistry advances, 24:42-426. 4. Klassen, S. and Bugbi, B. 2. Differential sensitivity of crops to ethylene and interactions with elevated CO 2. Life support and biosphere science, 7: 23-83. 5. Lemieux, J. 24. Présence de l enzyme désaminase chez les bactéries de la famille des Rhizobiacées. Mémoire de maitrise, Université Laval, Quebec,Canada, 9 pages. 6. Ma, W.; Sebestianova, S.; Sebestian, J.; Burd, G.I.; Guinel, F. and Glick, B.R. 23. Prevalence of -aminocyclopropaqne-- carboxylate in deaminase in Rhizobia spp. Antonie Van Leeuwenhoek, 83: 285-29 7. Ma, W. ; Charles, T.C. and Glick, B.R. 24. Expression of an exogenous - aminocyclopropane--aarboxylate deaminase gene in Sinorhizobium meliloti increases its ability to nodulate alfalfa. Applied and Environmental Microbiology, 7: 589 5897. 8. Mayak, S. ; Tirosh, T. and Glick, B.R. 24. Plant growth-promoting bacteria that confer resistance to water stress in tomatoes and peppers. Plant Science, 66: 525-53. 9. Nordustrum, A.C. and Eliasson, L. 984. Regulation of root formation by auxine-ethylene 668
Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28 interaction in pea stems cuttings. Physiologia Plantarum, 6:298-32. 2. Penrose, D.M. and Glick, B.R. 23. Methods for isolating and characterizing deaminase-containing plant growth-promoting rhizobacteria. Physiologia Plantarum, 8:- 5. 2. Rubio, M.G.T. ; Plata, S.A. ; Castillo, J.B. ; and. Nieto, P.M. 2. Isolation of Enterobacteria, Azotobacter and Pseudomonas producers of Indole acetic acid and siderophores from Colombian rice rhizosphere. Revista Latinoamrican de Microbiologia, 5: 7-76. 22. Saravanakumar, D. and Samiyappan, R. 27. from Pseudomonas fluorescens mediated saline resistance in groundnut (Arachis hypogea) plants. Journal of Applied Microbiology, 2(5): 283-292. 23. Sheehy, R. E.; Yamada, M.H.; Sasaki, T.; Martineau, B. and Hiatt, W.R. 99. Isolation, sequence, and expression in Escherichia coli of the Pseudomonas sp. strain ACP gene encoding -aminocyclopropane--carboxylate deaminase. Journal of Bacteriology, 73: 526-5265. 24. Sperber, J.I. 958. Solution of apatite by soil microorganisms producing organic acids. Australian Journal of Agricultural Research, 9:782-787. 25. Stearns, J.C. ; Shah, S. ;. Greenberg, B.M. ; Dixon, D.G. and Glick, B.R. 25. Tolerance of transgenic canola expressing - aminocyclopropane--carboxylic acid deaminase to growth inhibition by nickel. Plant Physiology and Biochemistry, 43: 7-78. 26. Wang, C. ; Knill, E. ; Glick, B.R. and Défago, G. 2. Effect of transferring - aminocyclopropane--carboxylic acid () deaminase genes into Pseudomonas fluorescens strain CHA and its gaca derivative CHA96 on their growth-promoting and disease-suppressive capacities. Canadian Journal of Microbiology, 46:898-97. 669
Potential Evaluation of Some Native Rhizobia as Plant growth Promoting Bacteria and their Role in Decreasing of Stress Ethylene Khosravi H., Yakhchali. B. 2 and Alikhani H.A. Soil Science Dep., Faculty of Agriculture, Tehran University, Tehran, I.R. of IRAN Downloaded from ibs.org.ir at 3:32 +33 on Sunday December 3th 28 2 National Institute of Genetic Engineering and Biotechnology, Tehran, I.R. of IRAN Abstract The amount of ethylene increases in stress conditions in plants. Some of plant growth rhizobacteria contain enzyme that is capable to convert - aminocyclopropane--carboxylate, the immediate precursor of ethylene in higher plants to ammonia and α-ketobutyrate that causes decrease in stress ethylene level. In this study 33 strains of indigenous rhizobia including Sinorhizobium meliloti (68 strains), Rhizobium leguminosarum bv. viciae (44 strains), R. leguminosarum bv. phaseoli (58 strains) and Mesorhizobium ciceri (6 strains) native to different soils of Iran were evaluated for production. The results showed that 28.2% of strains synthesized enzyme. The most number was belonged to the Sinorhizobium meliloti (46.5%) and the least number to the Mesorhizobium ciceri (6.7%). The strains with enzyme were categorized in five groups of, 2-4, 4-6, 6-8 and 8- mm based on the colony diameter as compared to control. Four isolates of each group were selected for future investigation. The selected isolates did not show phosphate solubilizing properties and HCN production as well. The selected strains synthesize auxine (IAA) between -2 µgr.ml -.24h -. In these strains, the ability to produce siderophores based on the ratio of corona to colony diameter on CAS agar medium was zero up to 2.7. The future work on these strains would be the development of biofertilizers with properties. Keywords: Rhizobium,, Ethylene, Biofertilizers. 67