Table 1: Colony morphology and cultural characteristics of isolated strains after incubation at 28 o C for 72 h.

Transcription

1 Table 1: Colony morphology and cultural characteristics of isolated strains after incubation at 28 o C for 72 h. Bacterial Media used Colony morphology strains VR1 YEMA Small (2 mm), opaque, circular, highly EPS producing gummy colonies. VR2 YEMA 2.1 mm in size, translucent, round, highly gummy, convex type colonies. VR3 YEMA White coloured, circular, small, convex colonies. VR4 YEMA Small, translucent, granular, convex colonies. VR5 CrYEMA Translucent, round, highly LPS producing gummy colonies, small in size. VR6 CrYEMA Pinkish, translucent, medium sized, circular, gummy colonies. VR9 CrYEMA Small, rounded, white in colour, gummy, convex colonies VR10 CrYEMA Small, white, circular, less gummy, flat colonies VR11 BAM Circular, off-white in colour, very small, flat colonies. VR12 BAM Small, round, white coloured, opaque, convex colonies. VR13 BAM White coloured, small, flat, rounded, opaque colonies. VR14 BAM White coloured, opaque, round, convex colonies. VR15 KB medium Small, round, pale yellow colour, smooth convex colonies. VR16 KB medium Round, white coloured, convex colonies. VR19 KB medium Round, medium size, smooth, pale yellow in colour, convex colonies. VR20 KB medium Round, creamy white, opaque, small, flat colonies. YEMA= Yeast extract mannitol agar, CrYEMA= Congo red yeast extract mannitol agar, KB = King s B medium, BAM= Bacillus agar medium.

2 Table 2. Absorbance (at 610 nm) of isolates VR1 and VR1 after every 12 hours of intervals measured in UV-VIS spectrophotometer. Isolates Absorbance (610 nm) at every 12 h intervals * VR VR *, Values are mean of triplicate standard errors.

3 Table 3: Physiological and biochemical characteristics of different strains on YEMA/CrYEMA/ Bacillus agar/ King s B (KB) medium isolated from V. mungo. Isolates Biochemical characteristics VR1 - Rod N N N VR2 - Rod N N N VR3 - Rod N N N VR4 - Rod N N N VR5 - Rod N N N VR6 - Rod N N N VR9 - Rod N N N VR10 - Rod N N N VR11 + Rod N N N N VR12 + Rod N N N N VR13 + Rod N N N N VR14 + Rod N N N N VR15 - Rod N N VR16 - Rod N N VR19 - Rod N N VR20 - Rod N N Bradyrhizobium sp. NAIMCC-B Rod B. subtilis MTCC Rod Pseudomonas sp. MTCC Rod Gram reaction, 2- shape, 3- capsule, 4-motility, 5- endospore, 6- PHB accumulation, 7- catalase, 8- urease, 9- oxidase, 10- starch hydrolysis, 11- gelatin hydrolysis, 12- indole production, 13- growth on nutrient agar medium, 14- growth on YEMA, 15- growth on CrEMA, 16- esculin hydrolysis, 17- KNO 3 (8%) tolerance, 18- Voges Proskaur test, 19- methyl red, 20- ONPG, 21- citrate utlilization, 22- H 2 S prodyction, 23- nitrate reduction, 24- fluorescence, 25- pigment; N- not tested.

4 Table 4. Effect of different temperature on growth of bacterial isolates. Bacterial strains Temperature ( C) Cardinal Temperature Minimum Optimum Maximum VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR Bradyrhizo NAIMCC-B Bacillus MTCC441 Pseudomonas MTCC

5 Table 5. Growth of bacterial isolates on different ph values. Bacterial strains ph range Cardinal ph Minimum Optimum Maximum VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR NAIMCC-B Bacillus MTCC 441 Pseudomonas MTCC

6 Table 6. Effect of different salt concentrations (NaCl) (%) on growth of bacterial isolates. Bacterial strains Salt concentration (%) Cardinal salt concentration (%) Minimum Optimum Maximum VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR NAIMCC-B Bacillus MTCC Pseudomonas MTCC

7 Table 7. Utilization of various carbon sources by bacterial strain isolated from Vigna mungo. Carbon sources VR1 VR2 VR3 VR4 VR5 VR6 VR9 VR10 VR11 VR12 VR13 VR14 VR15 VR16 VR19 VR20 NAIMCC -B MTCC 441 MTCC 129 Monosaccharides pentoses Xylose L-Arabinose Ribose Rhamnose D-Arabinose Monosaccharides Hexoses Fructose Dextrose Galactose Mannose Sorbose Disaccharides Lactose Maltose Trehalose Melibiose Sucrose Cellobiose Trisaccharide Raffinose Melezitose Polysaccharides Inulin Organic compounds Sodium gluconate Glycerol Salicin Organic compounds Glucosamine α-methyl-dglucoside α-methyl-dmannoside ONPG Esculin Citrate Sugar alcohol Dulcitol Inositol Sorbitol Mannitol Adonitol

8 Table 8: Plant growth promoting (PGP) activities found in different bacterial isolates from V. mungo. Bacterial strains IAA production HCN production Phosphate solubilization Siderophore production Chitinase activity ACC deaminase Antagonism against M. phaseolina VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR Bradyrhizobium sp. NAIMCC-B Bacillus subtilis MTCC 441 Pseudomonas MTCC

9 Table 9. Quantitative estimation of siderophore by isolates VR1, VR2, VR11 and VR13. Incubation time (h) Siderophore ( g/ml) produced by different Bradyrhizobium isolates * VR1 VR2 VR11 VR v * Values are mean of three replicates standard error.

10 Table 10: Antagonistic effect of Bradyrhizobiun isolates VR1 VR6 against M. phaseolina in vitro. Isolates Growth inhibition of M. phaseolina colony (%) * Dual culture Cell-free culture VR VR VR VR VR VR Bradyrhizobium NAIMCC-B * = Values are mean of three replicates standard error.

11 Table 11: Antagonistic effect of Bacillus isolates VR11 VR14 against M. phaseolina in vitro. Isolates Growth inhibition of M. phaseolina colony (%) * Dual culture Cell-free culture VR VR VR VR Bacillus subtilis MTCC * Values are mean of three replicates standard error;

12 Table 12: Effect of cell-free culture filtrates (CFCF) of Bradyrhizobium isolates on mycelial yield of M. phaseolina. Bacterial strains Mycelial dry weight (mg)# Concentration of CFCF (%, v/v) 15% 30% 45% Control VR VR2 VR3 VR4 VR5 VR6 NAIMCC-B (50.9) (58.4) (41.9) (36.9) (33.9) (39.6) (48.1) (96.8) (92.5) (55.9) (38.7) (26.3) (32.7) (68.0) (100) (53.8) (61.3) (54.5) (56.7) (91.4) SEM CD at 1% CD at 5% # Values are mean of three replicates, = standard errors; values in parentheses represent inhibition in mycelia yield (%); LSD at P > 0.1 in comparison with control values.

13 Table 13: Effect of cell-free culture filtrates (CFCF) of Bacillus isolates on mycelial yield of M. phaseolina. Bacterial strains Mycelial dry weight (mg)# Concentration of CFCF (%, v/v) Control VR11 VR12 VR13 VR14 Bacillus MTCC (43.6) (28.3) (42.6) (31.6) (39.0) (72.4) (37.1) ((58.8)) (51.8) (41.9) 0 (100) (80.7) 0 (100) (84.1) (92.8) SEM CD at 1% CD at 5% #, Values are mean of three replicates, = standard errors; values in parentheses represent inhibition in mycelia yield (%); LSD at P > 0.1 in comparison with control values.

14 Table 14: Effect of cell-free culture filtrates of Bradyrhizobium isolates VR1-VR6 on germination of M. phaseolina sclerotia. Bacterial isolates Concentration (%, v/v) *Values are mean of five replicates, = standard errors Sclerotia germination (%)* Incubation (hours) Control VR VR VR VR VR VR Brady. NAIMCC- B

15 Table 15: Effect of culture filtrates of Bradyrhizobium isolates on hyphal development during sclerotia germination of M. phaseolina after 48 h incubation. Bacterial isolates Concentration (%, v/v) Sclerotia germination (%)* No. of hypha produced /sclerotium >7 Total Control VR1 VR2 VR VR VR VR Bradyrhizobium sp. NAIMCC-B *Values are mean of five replicates, 1 standard error; 0 = No germination

16 Table 16: Effect of cell-free culture filtrates of isolates of Bacillus on germination of M. phaseolina sclerotia. Bacterial isolates Concentration (%, v/v) Sclerotia germination (%)* Incubation time (hours) Control VR VR VR VR Bacilus MTCC *Values are mean of five replicates, standard errors

17 Table 17: Effect of culture filtrates of isolates of Bacillus on hyphal development during sclerotia germination of M. phaseolina after 48 h incubation. Bacterial Concentration Sclerotia germination (%)* isolates (%), v/v No. of hypha produced /sclerotium > 7 Total Control VR11 VR12 VR13 VR14 Bacillus MTCC % % % % % % % % % % % % % % % *Values are mean of five replicates; standard error.

18 Table 18. In vitro interaction between bacterial isolates. Isolated VR1 VR2 VR3 VR4 VR5 VR6 VR9 VR10 VR11 VR12 VR13 VR14 VR15 VR16 VR19 VR20 strains VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR VR1 + +, Synersism, -, Antagonism.

19 Table 19. Effect of cell-free filtrates of Bradyrhizobium sp. VR2 on growth of Bacillus sp. VR11. Bacillus sp. VR11 Optical density at 660 nm # Incubation time (hours) Control VR ns ** ** * SEM CD at 1% CD at 5% #, Values are mean of three replicates, standard error; * and **, LSD at P > 0.1 and P > 0.5, respectively in comparison with control.

20 Table 20: Intrinsic antibiotic activity shown by bacterial isolates. Antibiotics Antibiotic disk (mcg) Zone of resistance (mm) Zone of sensitivity (mm) VR1 VR2 VR11 Chloramphenicol C (S) 26 (S) 22 (S) Nalidixic acid Na (S) 09 (R) 10 (R) Furazolidone Fr (R) 20 (R) 23 (S) Norfloxacine Nx (S) 21 (S) 11 (R) Oxytetracycline O (S) 24 (S) 21 (S) Cephotaxime Ce (R) 06 (R) 13 (R) Cephalexin Cp (R) 06 (R) 09 (R) Co-trimoxazol Co (S) 06 (R) 24 (S) Ceftazidime Ca (S) 24 (S) 31 (S) Ciprofloxacin Cf (S) 23 (S) 28 (S) Nitrofurantoin Nf (R) 08 (R) 21 (S) Norfloxacin Nx (S) 28 (S) 23 (S) Netillin Nt (S) 19 (S) 21 (S) Ofloxacin Of (S) 24 (S) 21 (S)

21 Table 21. Effect of seed bacterisation with bacterial isolates on seed germination and seedling growth of V. mungo 30 days after sowing. Treatments Seed germination (%) Root Shoot Nodule No. length (cm) Weight (g) length (cm) Weight (g) Control T1 62 ** 4.4 ** ns 17.1 ** ns 4 * T2 86 ** ** 25.0 ** ns 11 ** T3 80 * 7.8 ** * 23.4 ** ns 10 ** T4 82 ** 8.0 ** * 24.2 ** ns 11 ** T5 88 * 8.9 ** ** 26.7 ** * 12 ** T6 77 ** 7.4 * ns 21.2 ** * 8 ** T7 76 ** 7.2 ** ns 20.0 ** * 8 * T8 78 ** 7.8 ** ** 23.9 ** ** 9 ** SEM CD at 1% CD at 5% Abbreviations: * and **, LSD at P > 0.1 and > 0.5, respectively in comparison with control; T1 M. phaseolina, T2- B. japonicum VR2 nal+, T3- VR1 fur+, T4- VR11 nor+, T5- VR1 fur+ +VR2 nal+ + VR11 nor+, T-6- M. phaseolina + VR1 fur+ +VR2 nal+, T7- M. phaseolina + VR1 fur+ + VR11 nor+, T8- M. phaseolina + VR2 nal+ + VR11 nor+.

22 Table 22: Effect of seed bacterization with B. japonicum VR1, Bradyrhizobium sp. (Vigna) VR2 and VR11 on plant height and vigour index of V. mungo after 30 DAS in pots. Treatments Plant length Vigour index (cm) Control T ** T ** T ** T ** T ** T ** T ** T ** SEM CD at 5% CD at 1% Abbreviations: * and **, LSD at P > 0.1 and P > 0.5, respectively in comparison with control; T1 M. phaseolina, T2- B. japonicum VR2 nal+, T3- VR1 fur+, T4- VR11 nor+, T5- VR1 fur+ +VR2 nal+ + VR11 nor+, T-6- M. phaseolina + VR1 fur+ +VR2 nal+, T7- M. phaseolina + VR1 fur+ + VR11 nor+, T8- M. phaseolina + VR2 nal+ + VR11 nor+.

23 Table 23. Effect of seed bacterisation with bacterial isolates on growth of V. mungo 60 days after sowing. Treatment Root Shoot Length (cm) Weight (g) length (cm) Weight (g) Nodule No. Control T1 9.3 * ns ** * 5 * T ** ** ** ** 13 ** T ** ** 38.3 ** ** 10 ** T ** ** 39.7 ** ** 12 ** T ** ** ** ** 14 ** T ** * ** ** 10 ** T * ** ** ** 10 ** T ** ** ** ** 11 ** SEM CD at 1% CD at 5% Abbreviations: * and **, LSD at P > 0.1 and P > 0.5, respectively in comparison with control; T1 M. phaseolina, T2- B. japonicum VR2 nal+, T3- VR1 fur+, T4- VR11 nor+, T5- VR1 fur+ +VR2 nal+ + VR11 nor+, T- 6- M. phaseolina + VR1 fur+ +VR2 nal+, T7- M. phaseolina + VR1 fur+ + VR11 nor+, T8- M. phaseolina + VR2 nal+ + VR11 nor+.

24 Table 24. Effect of seed bacterization with twin/tree consortia of Bradyrhizobium sp. (Vigna) isolate VR1 fur+ and VR2 nal+, and Bacillus sp. isolate VR11 nor+ on disease reduction (%). Treatment Number of plants showing disease symptoms # 30 days 60 days T T5 2.0 (66.7%)* 2.5 (68.7%) T6 4.0 (33.4%) 5.3 (33.7%) T7 4.8 (20.0%) 6.2 (22.5%) T8 3.2 (46.7%) 3.9 (51.2%) #, Values are mean of three replicates of 10 plants of V. mungo. *, Values in parentheses are disease reduction (%). T1 M. phaseolina, T5- VR1 fur+ +VR2 nal+ + VR11 nor+, T-6- M. phaseolina + VR1 fur+ +VR2 nal+, T7- M. phaseolina + VR1 fur+ + VR11 nor+, T8- M. phaseolina + VR2 nal+ + VR11 nor+

25 Table 25. Root colonization by B. japonicum VR1 fur+, Bradyrhizobium sp. (Vigna) VR2 nal+ and Bacillus sp. VR11 nor+ in the presence of M. phaseolina after 30 and 60 days of sowing (2011). Treatments Log CFU g -1 root segments # Marker strain 30 days 60 days T2 VR2 nal T3 VR1 fur T4 VR11 nor T5 (Consortium A) T6 (Consortium B) T7 (Consortium C) T8 (Consortium D) VR1 fur VR2 nal VR11 nor VR1 fur VR2 nal VR1 fur VR11 nor VR2 nal VR11 nor ##, Values are mean Log cfu of 10 plants; = standard error. T2- Bradyrhizobium sp. VR2 nal+, T3- B. japonicum VR1 fur+, T4- VR11 nor+, T5- VR1 fur+ +VR2 nal+ + VR11 nor+, T6- M. phaseolina + VR1 fur+ +VR2 nal+, T7- M. phaseolina + VR1 fur+ + VR11 nor+, T8- M. phaseolina + VR2 nal+ + VR11 nor+