Open Access

Richa Sharma, V.K. Razdan, Vishal Gupta, Stanzin Dorjey, Kausar Fatima, P.K. Rai and Satish Sharma

Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Chatha – J&K, India.
J Pure Appl Microbiol. 2017;11(4):1973-1982
https://doi.org/10.22207/JPAM.11.4.39 | © The Author(s). 2017
Received: 09/11/2017 | Accepted: 20/12/2017 | Published: 31/12/2017
Abstract

Studies were carried out from 2013 to 2015 with the objectives to isolate and identify some indigenous rhizobacterial strains against major soil-borne pathogens of brinjal. Seventy thizobacterial isolates were characterized based on colony characters, morphological and biochemical test and identified as Pseudomonas flourescens (30), P. aeruginosa (13), P. aureofaciens (2) and Bacillus subtilis (25). In vitro bioassay of the seventy rhizobacterial isolates revealed that the isolate I-58 was the most effective rhizobacterial isolate, followed by I-30 and I-55 isolates in inhibiting the radial growth of test pathogens.

Keywords

Brinjal, soil-borne diseases, rhizobacteria, biochemical characterization.

Introduction

Brinjal or eggplant (Solanum melongena L.), of the family Solanaceae is quite popular as the poor man’s crop suffer from various soil-borne disease which emerged as major biotic stress in successful cultivation of this crop in India (Gupta et al., 2013). Soil borne plant pathogens responsible for the diseases due to their persistence nature in soil become more challenging and simultaneous infection by multiple soil-borne plant pathogens results in disease complex that further reduced the yield and quality of the brinjal crop (Koike et al., 2003). Currently, the management of soil- borne diseases has been done with the application of chemical fungicides. However, it may not be sustainable in the longer run as chemical fungicides are known to cause residual toxicity, toxicity to non-target organisms and other environmental hazards. In recent times much emphasis is being given to manage the soil borne diseases of the crop by employing the plant growth promoting rhizobacteria (Compant et al., 2005). The mechanisms of PGPR included regulating hormonal and nutritional balance, inducing resistance against plant pathogens and solubilizing nutrients for easy uptake by plants (Vejan et al., 2016). The use of PGPR inoculants as biofertilizers is due to the production of some plant growth promoting substances, production of enzymes and production of some antifungal and antibacterial secondary metabolites (Weller, 1988) and as antagonists of phytopathogens due to secretion of antibiotics provide a promising alternative to chemical fertilizers and pesticides (Apastambh et al. 2016; Glick, 2012). The objectives of this study is to isolates the plant growth promoting bacteria from the rhizosphere of brinjal crop, their bio-chemical characterization and bio-control potential against major soil – borne plant pathogens of brinjal crop.

Materials and Methods

Isolation of rhizobacterial isolates
Soil samples were collected from the rhizosphere of healthy brinjal plants was processed for isolation of rhizobacterial isolates by serial dilution method (Dhingra and Sinclair, 1995). Single colonies formed were picked and streaked separately on petriplates containing nutrient agar and King’s B media and purified by streak plate method (Koch, 1881).

Identification of isolated rhizobacteria
Isolates of rhizobacteria, isolated from the rhizosphere of healthy brinjal plants were identified based upon their colony characters, microscopic characteristics (Gram, 1884) and biochemical characters viz., starch hydrolysis (Iverson and Millis, 1974), catalase test (Schaad, 1992), oxidase test (Gordon and McLeod, 1928), spirit blue Agar, lipid hydrolysis (Cowan, 1974), levan production (Lelliot and Stead, 1987), casein hydrolysis (Kunz and Lonnerdal, 1990), arginine hydrolysis (Lelliot and Stead, 1987), gelatinase (Blazevic and Ederer, 1975), pectinase activity (Fogarty and Kelly, 1983), protease activity (Vieira, 1999) and endospore formation.

In vitro evaluation of rhizobacterial isolates against the soil borne pathogens
The isolated rhizobacterial isolates were tested under in vitro conditions, against the major soil borne fungal pathogens viz., Fusarium oxysporum f. sp. melongenae, Rhizoctonia solani and Sclerotium rolfsii responsible for causing wilt, collar rot and root rot respectively in brinjal crop by using dual culture technique (Morton and Stroube, 1955). Observation regarding per cent inhibition of mycelial growth of pathogens was calculated (Wong et al., 2003)

RESULTS AND DISCUSSION

Isolation and identification rhizobacterial isolates
Seventy rhizobacterial isolates were isolated from the brinjal fields across the locations surveyed. The selected rhizobacterial strains were purified by streak plate method and characterized based on colony characters, morphological and biochemical characteristics. The isolated rhizobacterial isolates exhibited variations in their colony characteristics (Table 1). Out of 70 isolates, 23 produced light green, 24 green, 17 white and 6 cream coloured colonies. Of these, 25 colonies were round and 45 were irregular in shape. Forty five isolates produced non-spreading type of colonies, whereas, 25 produced colonies that were spreading in nature. Further, microscopic examinations of isolated rhizobacteria revealed that out of the 70 isolates, 41 isolates had long rod shaped cells and the remaining 29 had short rod shaped cells, of which 45 strains showed Gram negative reaction and remaining 25 showed Gram positive reactions.

Table (1):
Cultural and morphological characteristics of rhizo bacterial isolates.

Isolate Colony characteristics Microscopic characteristics
Colour Shape Nature Cell shape Gram reaction
I-1 White Round Non-spreading Long rods Gram+ve
I-2 Cream Round Non-spreading Long rods Gram+ve
I-3 Green Irregular Spreading Short rods Gram -ve
I-4 Light green Irregular Spreading Long rods Gram-ve
I-5 Green Irregular Spreading Short rods Gram -ve
I-6 Light green Irregular Spreading Short rods Gram -ve
I-7 White Round Non-spreading Long rods Gram +ve
I-8 Green Irregular Spreading Long rods Gram -ve
I-9 Light green Irregular Spreading Short rods Gram -ve
I-10 Green Irregular Spreading Long rods Gram -ve
I-11 Light green Irregular Spreading Short rods Gram -ve
I-12 White Round Non-spreading Short rods Gram +ve
I-13 Cream Round Non-spreading Long rods Gram +ve
I-14 Cream Round Non-spreading Long rods Gram +ve
I-15 Green Irregular Spreading Short rods Gram -ve
I-16 Light green Irregular Spreading Long rods Gram -ve
I-17 Green Irregular Spreading Long rods Gram -ve
I-18 White Round Non-spreading Long rods Gram +ve
I-19 White Round Non-spreading Short rods Gram +ve
I-20 Green Irregular Spreading Long rods Gram -ve
I-21 Green Irregular Spreading Long rods Gram -ve
I-22 Green Irregular Spreading Short rods Gram -ve
I-23 White Round Non-spreading Short rods Gram +ve
I-24 White Round Non-spreading Long rods Gram +ve
I-25 Light green Irregular Spreading Short rods Gram -ve
I-26 Light green Irregular Spreading Short rods Gram -ve
I-27 Green Irregular Spreading Long rods Gram -ve
I-28 Light green Irregular Spreading Long rods Gram -ve
I-29 White Round Non-spreading Short rods Gram +ve
I-30 Light green Irregular Spreading Long rods Gram -ve
I-31 Green Irregular Spreading Short rods Gram -ve
I-32 Light green Irregular Spreading Short rods Gram -ve
I-33 Cream Round Non-spreading Long rods Gram +ve
I-34 White Round Non-spreading Long rods Gram +ve
I-35 Green Irregular Spreading Short rods Gram -ve
I-36 Light green Irregular Spreading Long rods Gram -ve
I-37 Light green Irregular Spreading Long rods Gram -ve
I-38 Green Irregular Spreading Long rods Gram -ve
I-39 Light green Irregular Spreading Short rods Gram -ve
I-40 White Round Non-spreading Long rods Gram +ve
I-41 Green Irregular Spreading Long rods Gram -ve
I-42 Light green Irregular Spreading Short rods Gram -ve
I-43 Light green Irregular Spreading Short rods Gram -ve
I-44 White Round Non-spreading Long rods Gram +ve
I-45 White Round Non-spreading Short rods Gram +ve
I-46 Cream Round Non-spreading Short rods Gram +ve
I-47 Green Irregular Spreading Long rods Gram -ve
I-48 Green Irregular Spreading Long rods Gram -ve
I-49 Light green Irregular Spreading Short rods Gram -ve
I-50 Light green Irregular Spreading Long rods Gram -ve
I-51 White Round Non-spreading Short rods Gram +ve
I-52 Cream Round Non-spreading Short rods Gram +ve
I-53 Green Irregular Spreading Long rods Gram -ve
I-54 Green Irregular Spreading Long rods Gram -ve
I-55 Light green Irregular Spreading Short rods Gram -ve
I-56 White Round Non-spreading Long rods Gram +ve
I-57 White Round Non-spreading Long rods Gram +ve
I-58 Green Irregular Spreading Long rods Gram -ve
I-59 Green Irregular Spreading Short rods Gram -ve
I-60 Light green Irregular Spreading Short rods Gram -ve
I-61 Light green Irregular Spreading Long rods Gram –ve
I-62 Green Irregular Spreading Long rods Gram –ve
I-63 Light green Irregular Spreading Short rods Gram +ve
I-64 Green Irregular Spreading Long rods Gram +ve
I-65 Green Irregular Spreading Long rods Gram +ve
I-66 Light green Irregular Spreading Long rods Gram +ve
I-67 Green Irregular Spreading Short rods Gram +ve
I-68 White Round Non-spreading Long rods Gram –ve
I-69 White Round Non-spreading Long rods Gram +ve
I-70 Light green Irregular Spreading Long rods Gram -ve

Biochemical characterization
Data in the (table 2) indicated that the isolated rhizobacterial strains were tested for their biochemical characters. In case of starch hydrolysis test, isolate 1, 2, 7, 12, 13, 14, 18, 19, 23, 24, 29, 33, 34, 40, 44, 45, 46, 51, 52, 56, 57, 61, 62, 68 and 69 formed a transparent zone around the rhizobacterial colonies that indicated positive test for Bacillus spp., whereas, remaining forty five isolates showed negative test, whereas, in case of catalase test, all the 70 rhizobacterial isolates produced gas bubbles when drops of hydrogen peroxide were flooded on them revealing that all the isolates were strict aerobes as well as facultative anaerobes. For oxidase test, the colony colour of 45 isolates viz., 3, 4, 5, 6, 8, 9, 10, 11, 15, 16, 17, 20, 21, 22, 25, 26, 27, 28, 30, 31, 32, 35, 36, 37, 38, 39, 41, 42, 43, 47, 48, 49, 50, 53, 54, 55, 58, 59, 60, 63, 64, 65, 66, 67 and 70 changed to maroon, indicating that the rhizobacterial isolates belonged to the genus Pseudomonas, whereas, remaining 25 isolates gave negative result for the test. In case of spirit blue agar test, 25 isolates viz., 1, 2, 7, 12, 13, 14, 18, 19, 23, 24, 29, 33, 34, 40, 44, 45, 46, 51, 52, 56, 57, 61, 62, 68 and 69 formed a clear halo around the rhizobacterial colonies that indicated positive test for Bacillus spp., whereas, remaining forty five isolates showed negative test. In lipid hydrolysis test, isolate, 3, 4, 5, 6, 8, 9, 10, 11, 15, 16, 17, 20, 21, 22, 25, 26, 27, 28, 30, 31, 32, 35, 36, 37, 38, 39, 41, 42, 43, 47, 48, 49, 50, 53, 54, 55, 58, 59, 60, 63, 64, 65, 66, 67 and 70 formed a transparent zone around the rhizobacterial colonies indicating positive test for Pseudomonas spp., remaining 25 isolates gave negative result for the test. In case of levan test, large white, domed, mucoid colonies was formed in forty five isolates viz., 3, 4, 5, 6, 8, 9, 10, 11, 15, 16, 17, 20, 21, 22, 25, 26, 27, 28, 30, 31, 32, 35, 36, 37, 38, 39, 41, 42, 43, 47, 48, 49, 50, 53, 54, 55, 58, 59, 60, 63, 64, 65, 66, 67 and 70 indicating that these rhizobacterial isolates belonged to Pseudomonas spp. and remaining twenty five isolates showed negative test. In case of casein hydrolysis, isolate 1, 2, 7, 12, 13, 14, 18, 19, 23, 24, 29, 33, 34, 40, 44, 45, 46, 51, 52, 56, 57, 61, 62, 68 and 69 formed a halo zone around the rhizobacterial colonies indicating positive test for Bacillus spp., whereas, remaining forty five isolates showed negative test. In arginine hydrolysis, isolate 3, 4, 5, 6, 8, 9, 10, 11, 15, 16, 17, 20, 21, 22, 25, 26, 27, 28, 30, 31, 32, 35, 36, 37, 38, 39, 41, 42, 43, 47, 48, 49, 50, 53, 54, 55, 58, 59, 60, 63, 64, 65, 66, 67 and 70 formed a clear zone around the rhizobacterial colonies indicated positive test for Pseudomonas spp. and remaining twenty five isolates showed negative test. In gelatinous test, 25 isolates viz., 1, 2, 7, 12, 13, 14, 18, 19, 23, 24, 29, 33, 34, 40, 44, 45, 46, 51, 52, 56, 57, 61, 62, 68 and 69 the liquid medium failed to solidify in gelatin tubes upon refrigeration and a clear halo appeared around the colonies of rhizobacterial isolates indicating positive test for Bacillus spp., whereas, remaining forty five isolates showed negative test. For pectinase test, isolate 1, 2, 7, 12, 13, 14, 18, 19, 23, 24, 29, 33, 34, 40, 44, 45, 46, 51, 52, 56, 57, 61, 62, 68 and 69 formed a clear halo around rhizobacterial colonies indicating positive for Bacillus spp., whereas, the remaining forty five isolates gave negative result against this test. In protease activity, isolate 1, 2, 7, 12, 13, 14, 18, 19, 23, 24, 29, 33, 34, 40, 44, 45, 46, 51, 52, 56, 57, 61, 62, 68 and 69 formed a clear zone around the rhizobacterial colonies that indicated positive test for Bacillus spp., whereas, remaining forty five isolates showed negative test. In case of endospore formation test isolate 1, 2, 7, 12, 13, 14, 18, 19, 23, 24, 29, 33, 34, 40, 44, 45, 46, 51, 52, 56, 57, 61, 62, 68 and 69 indicated positive test, therefore confirming the rhizobacteria being Bacillus spp., as by adopting differential staining technique the endospores retained the primary dye i.e., malachite green, whereas, the vegetative cells lost the stain. Based on the studies regarding the morphological, biochemical and physiological characteristics and the capability of the isolates to grow at different temperatures, 30 isolates were identified as Pseudomonas flourescens, 13 were identified as P. aeruginosa, 2 as P. aureofaciens and 25 as Bacillus subtilis (Table 3). Our results are in confirmatory with the findings of earlier workers who also characterize the rhizobacteria on the basis of phenotypic and bio-chemical test (Rhodes, 1959; Palleroni, 1975; Fritze, 2002). Bacillus and Pseudomonas are the most frequently reported genera of PGPR (Zahid et al., 2015)
Table (2):
Biochemical characteristics of rhizo bacterial strains.

Isolate
Starch Hydrolysis
Catalase
Oxidase
Spirit blue Agar
Lipid hydrolysis
Levan
Casein hydrolysis
Arginine hydrolysis
Gelatinous activity
Pectinase Activity
Protease activity
Endospore formation
I-1
+
+
+
+
+
+
+
+
I-2
+
+
+
+
+
+
+
+
I-3
+
+
+
+
+
I-4
+
+
+
+
+
I-5
+
+
+
+
+
I-6
+
+
+
+
+
I-7
+
+
+
+
+
+
+
+
I-8
+
+
+
+
+
I-9
+
+
+
+
+
I-10
+
+
+
+
+
I-11
+
+
+
+
+
I-12
+
+
+
+
+
+
+
I-13
+
+
+
+
+
+
+
+
I-14
+
+
+
+
+
+
+
+
I-15
+
+
+
+
+
I-16
+
+
+
+
+
I-17
+
+
+
+
+
I-18
+
+
+
+
+
+
+
+
I-19
+
+
+
+
+
+
+
+
I-20
+
+
+
+
+
I-21
+
+
+
+
+
I-22
+
+
+
+
+
I-23
+
+
+
+
+
+
+
I-24
+
+
+
+
+
+
+
+
I-25
+
+
+
+
+
I-26
+
+
+
+
+
I-27
+
+
+
+
+
I-28
+
+
+
+
+
I-29
+
+
+
+
+
+
+
+
I-30
+
+
+
+
+
I-31
+
+
+
+
+
I-32
+
+
+
+
+
I-33
+
+
+
+
+
+
+
+
I-34
+
+
+
+
+
+
+
I-35
+
+
+
+
+
I-36
+
+
+
+
+
I-37
+
+
+
+
+
I-38
+
+
+
+
+
I-39
+
+
+
+
+
I-40
+
+
+
+
+
+
+
+
I-41
+
+
+
+
+
I-42
+
+
+
+
+
I-43
+
+
+
+
+
I-44
+
+
+
+
+
+
+
+
I-45
+
+
+
+
+
+
+
+
I-46
+
+
+
+
+
+
+
+
I-47
+
+
+
+
+
I-48
+
+
+
+
+
I-49
+
+
+
+
+
I-50
+
+
+
+
+
I-51
+
+
+
+
+
+
+
+
I-52
+
+
+
+
+
+
+
+
I-53
+
+
+
+
+
I-54
+
+
+
+
+
I-55
+
+
+
+
+
I-56
+
+
+
+
+
+
+
I-57
+
+
+
+
+
+
+
+
I-58
+
+
+
+
+
I-59
+
+
+
+
+
I-60
+
+
+
+
+
I-61
+
+
+
+
+
+
+
+
I-62
+
+
+
+
+
+
+
I-63
+
+
+
+
+
I-64
+
+
+
+
+
I-65
+
+
+
+
+
I-66
+
+
+
+
+
I-67
+
+
+
+
+
I-68
+
+
+
+
+
+
+
+
I-69
+
+
+
+
+
+
+
+
I-70
+
+
+
+
+

Table (3):
Identification of the isolated rhizo bacterial isolates.

Isolate
Identification
I-1, I-2, I-7, I-13, I-14, I-18, I-19, I-23, I-24, I-30, I-33, I-34, I-40, I-45, I-46, I-51, I-52, I-56, I-57, I-61, I-62, I-68 and I-69
Bacillus subtilis(25)
I-3, I-4, I-5, I-6, I-9, I-10, I-16, I-17, I-20, I-22, I-25, I-26, I-27, I-31, I-32, I-37, I-38, I-39, I-42, I-43, I-48, I-49, I-50, I-53, I-54, I-58, I-60, I-63, I-67 and I-70
Pseudomonas fluorescens(30)
I-8, I-11, I-15, I-21, I-28, I-29, I-35, I-41, I-47, I-55, I-64, I-65 and I-66
Pseudomonas aeruginosa(13)
I-36 and I-59
Pseudomonas aureofaciens(2)

Evaluation of rhizobacterial isolates against the test pathogens
A perusal of the data presented in Table 4 revealed out of the 70 rhizobacterial isolates, the efficacy of Pseudomonas flourescens isolate I-58, Bacillus subtilis isolate I-30 and Pseudomonas fluorescens isolate I-55, was superior to other isolates with regard to inhibiting the growth of the test pathogens. The rhizobacterial isolate I-58 (P. flourescens) shown significant reduction in the radial growth of the test pathogens i.e., F. oxysporum f. sp. melongenae, R. solani and S. rofsii. In case of F. oxysporum f. sp. melongenae the minimum radial growth of 13.06 mm was recorded thereby effecting 85.48 per cent reduction in radial growth over control. In case of R. solani the isolate I-58 effected 82.90 per cent reduction over control (15.33 mm radial growth). With the isolate I-58 minimum radial growth of 14.00 mm with 84.44 per cent inhibition over control was recorded against S. rofsii. The rhizobacterial isolate I-58 was statistically at par with P. flourescens isolates I-55, I-43, I-50, I-67 and I-70 and Bacillus subtilis isolates I-30, I-33 and I-51. As per the effectivity of the rhizobacterial isolates, I-58 was followed by isolate I-30. influenced radial growth of 14.50 mm resulting in growth inhibition of 83.88 per cent over control in case of F. oxysporum f. sp. melongenae, 16.66 mm growth with 81.48 per cent growth inhibition over control in case of R. solani and 14.66 mm growth with 83.71 per cent reduction in growth over control in case of S. rofsii. The isolate I-30 was found to be at par with P. flourescens isolates I-55 and I-67 and Pseudomonas aeruginosa isolate I-41. The data further exhibited that isolate I-55 resulted in the radial growth of 15.00 mm with 83.33 per cent reduction in growth over control in case of F. oxysporum f. sp. melongenae. It also resulted in radial growth of 15.66 mm with 82.60 per cent reduction in growth over control in case of R. solani. Isolate I-55 caused 81.11 per cent inhibition of mycelial growth of S. rofsii (17.00 mm radial growth). However, I-55 was at par with the P. flourescens isolates I-32 and I-70. Tennakoon (2007) reported the inhibition of the mycelial growth of one or the other pathogens (Pestolotia thea, Fusarium oxysporum f. sp. carthami and Rhizoctonia bataticola) by volatile metabolites produced by 7 out of 11 isolates of fluorescent pseudomonads and two out of three Bacillus isolates. Biswas and Singh (2008) used Bacillus subtilis, P. fluorescens and T. viridae against wilt of tomato and observed that P. fluorescens was effective in minimizing the disease. B. subtilis inhibited the growth of F. oxysporum and and recommended to purify antifungal compounds (Khan et al., 2011). As reported by Nandakumar et al. (2001) and Asha et al. (2011) inhibition of mycelial growth of R. solani was due to nutritional competition, colonization of fungal hyphae production of inhibitory compounds by the rhizobacteria.
Table (4):
In vitro evaluation of rhizo bacterial isolates against the soil borne pathogens obtained from brinjal plants.

Isolate Radial growth (mm) Inhibition over control
F R S F R S
I-1 34.16 23.33 30.00 62.04 74.07 66.66
I-2 34.00 26.16 25.00 62.22 70.93 72.22
I-3 43.50 36.00 36.00 51.66 60.00 60.00
I-4 34.66 32.66 26.66 61.48 63.71 70.37
I-5 35.00 32.00 30.00 61.11 64.44 66.66
I-6 40.66 45.33 35.33 54.82 49.63 60.74
I-7 20.66 25.33 22.00 77.04 71.85 75.55
I-8 31.50 29.66 26.66 65.00 67.04 70.37
I-9 39.33 40.00 36.33 56.30 55.55 59.63
I-10 38.83 31.50 28.66 56.85 65.00 68.15
I-11 22.33 24.33 21.33 75.18 72.96 76.30
I-12 36.16 35.66 33.33 59.82 60.37 62.96
I-13 36.33 26.66 32.66 59.63 70.37 63.71
I-14 32.16 24.33 28.00 64.26 72.96 68.88
I-15 34.00 24.00 32.16 62.22 73.33 64.26
I-16 31.33 34.66 26.83 65.18 61.48 70.18
I-17 22.83 23.00 20.00 74.63 74.44 77.77
I-18 25.76 24.83 24.00 71.37 72.41 73.33
I-19 25.00 25.00 30.00 72.22 72.22 66.66
I-20 30.33 29.00 30.16 66.30 67.77 66.48
I-21 35.83 33.66 31.66 60.18 62.60 64.82
I-22 24.33 22.66 24.00 72.96 74.82 73.33
I-23 35.33 27.33 34.00 60.74 69.63 62.22
I-24 39.16 35.33 36.50 56.48 60.74 59.44
I-25 39.16 33.66 35.00 56.48 62.60 61.11
I-26 43.50 41.00 41.00 51.66 54.44 54.44
I-27 35.00 29.16 29.00 61.11 67.60 67.77
I-28 34.66 27.33 24.66 61.48 69.63 72.60
I-29 25.16 25.66 24.83 72.04 71.48 72.41
I-30 14.50 16.66 14.66 83.88 81.48 83.71
I-31 23.33 26.33 23.00 74.07 70.74 74.44
I-32 21.33 29.00 18.00 76.30 67.77 80.00
I-33 19.16 27.66 20.66 78.71 69.26 77.04
I-34 42.33 45.00 36.33 52.96 50.00 59.63
I-35 36.83 32.66 35.83 59.07 63.71 60.18
I-36 25.66 22.33 24.66 71.48 75.18 72.60
I-37 26.83 26.33 24.33 70.18 70.74 72.96
I-38 36.00 36.66 27.33 60.00 59.26 69.63
I-39 28.83 32.33 27.50 67.96 64.07 69.44
I-40 32.00 22.66 26.00 64.44 74.82 71.11
I-41 40.66 16.66 34.00 54.82 81.48 62.22
I-42 35.66 27.16 32.00 60.37 69.82 64.44
I-43 15.50 25.33 17.33 83.71 71.85 82.22
I-44 29.50 29.50 31.00 67.22 67.22 65.55
I-45 27.00 24.66 27.00 70.00 72.60 70.00
I-46 33.85 30.00 30.33 62.38 66.66 66.30
I-47 43.00 34.00 40.00 52.22 62.22 55.55
I-48 25.00 24.66 25.33 72.22 72.60 71.85
I-49 46.00 38.00 40.00 48.88 57.77 55.55
I-50 16.00 20.00 16.66 82.22 77.77 80.74
I-51 17.83 22.33 18.33 80.18 75.18 79.63
I-52 28.33 27.66 22.00 68.52 69.26 75.55
I-53 30.33 29.33 29.00 66.30 67.41 67.77
I-54 24.00 25.66 24.66 73.33 71.48 72.60
I-55 15.00 17.00 15.66 83.33 81.11 82.60
I-56 29.33 28.33 32.66 67.41 68.52 63.71
I-57 32.16 30.66 31.66 64.26 65.93 64.82
I-58 13.06 15.33 14.00 85.48 82.96 84.44
I-59 21.33 24.66 22.00 76.30 72.60 75.55
I-60 21.76 24.00 22.00 75.82 73.33 75.55
I-61 21.40 24.00 21.33 76.22 73.33 76.30
I-62 34.66 30.00 33.33 61.48 66.66 62.96
I-63 30.66 35.00 31.66 65.93 61.11 64.82
I-64 34.00 28.00 31.33 62.22 68.88 65.18
I-65 39.16 33.66 31.00 56.48 62.60 65.55
I-66 30.83 26.33 25.00 65.74 70.74 72.22
I-67 15.80 19.66 16.00 82.77 78.15 81.48
I-68 25.83 26.00 24.00 71.30 71.11 73.33
I-69 26.00 27.00 25.00 71.11 70.00 72.22
I-70 17.00 24.00 16.33 81.11 73.33 81.85
Control 90.00 90.00 90.00 0.00 0.00 0.00
S.Em(±) 2.37 1.35 1.54
CD (P=0.05) 6.64 3.73 4.32
References
  1. Apastambh, A. R, Tanveer, K. and M. M. V. Baig. Isolation and Characterization of Plant Growth Promoting Rhizobacteria from Banana Rhizosphere. Int. J. Curr. Microbiol. App. Sci., 2016; 5: 59-65.
  2. Asha, B. B., Chandra, N. S., Shankar, U. A. C., Srinivas, C. and Niranjana, S. R. Biological control of Fusarium oxysporum f. sp. lycopersici causing wilt of tomato by Pseudomonas fluorescens. International J. Microbio. Res., 2011; 3: 79-84.
  3. Biswas, S. and Singh, N. P. Integrated management of wilt of tomato caused by Ralstonia solanacearum. J. Myco. and Plnt Path., 2008; 38: 18-20.
  4. Blazevic, D. J. and Ederer, G. M. Principles of Biochemical Tests in Diagnostic Microbiology. Wiley and Company, New York, USA, 1975; pp 13 – 45.
  5. Compant, S., Duffy, B., Nowak, J., Clement, C., and Barka, E. A. Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Appl. and Envir. Microbio., 2005; 71: 4951-4959.
  6. Cowan, S. T. Cowan and Steel’s Manual for the identification of medical bacteria, 2nd edition, Cambridge University, UK, 1974; pp 331.
  7. Dhingra, O. D. and Sinclair, J. B. Basic Plant Pathology Methods. 2nd Edition, Lewis Publishers, USA, 1995; pp 434.
  8. Fogarty, M. V. and Kelly, C. T. In: Microbial Enzymes and Biotechnology. Fogarty, M.W. (ed.) London and NewYork: Elsevier Applied Science Publishers, 1983; pp 131-182.
  9. Fritze D. Bacillusidentification- traditional approaches. In: Berkeley R., Heyndrickx M., Logan N., De Vos P., editors. Applications and systematics of Bacillus and relatives. Blackwell Science Ltd.; 2002.
  10. Glick, B. R. Plant Growth-Promoting Bacteria: Mechanisms and Applications. Hindawi Publishing Corporation, Scientifica, 2012.
  11. Gordon, J. and Mcleod, J. W. The practical application of the direct oxidase reaction in bacteriology. J. Path. and Bacterio., 1928; 31: 185.
  12. Gram, C. 1884. The differential staining of Schizomycetes in tissue sections and in dried preparations. Fortschr chitte der Medi., 1884; 2: 185-189.
  13. Gupta, V., Razdan, V. K., Kumar, D. and Gupta, V. 2013. Status of soil borne diseases of brinjal and the frequency of associated pathogens. Vegetos, 26: 27-35.
  14. Iverson, W. G. and Millis, F. N. 1974. A method for the detection of starch hydrolysis by bacteria. J. Appl. Microbio., 1974; 37: 443-446.
  15. Khan, A. A., Naseem, L. R. and Prathibha, B. Screening and potency evaluation of antifungal from soil isolates of B. subtilis on selected fungi. Advanced Biotech., 2011; 10: 35-37.
  16. Koch, R. Zur Untersuchung von pathogenen Organismen. Mitthdungen uus dem Kaiserlichen Gesundbeitsamte, 1881; 1: 1-48.
  17. Koike, S. T., Gordon, T. R. and B. J. Aegerter. Root and Basal Rot of Leek Caused by Fusarium culmorum in California. Plant Dis., 2003; 87: 601.
  18. Kunz, C. and Lonnerdal, B. Casein and casein subunits in preterm milk, colostrums and mature human milk. J. Pedi. Gastoentero. and Nutri., 1990; 14: 454-461.
  19. Lelliott, R. A. and Stead, D. E. Methods for the diagnosis of bacterial diseases of plants. Blackwell Scientific Publications, Oxford. 1987; pp 216.
  20. Morton, D. J. and Stroube, W. H. 1955. Antagonistic and stimulatory effect of soil microorganism upon Sclerotium rolfsii. Phytopath., 1955; 45: 417-420.
  21. Nandakumar, R., Viswanathan, B. S., Sheela, J., Raguchander, T. and Samiyappan, R. A new formulation containing plant growth promoting rhizobacterial mixture for the management of sheath blight and enhanced grain yield in rice. Biocntrl., 2001; 46: 493-510.
  22. Palleroni, N. J. General properties and taxonomy of the genus Pseudomonas. In: Genetics and Biochemistry of Pseudomonas, Clarke, P. H. and Richmond, M. H., (eds.), John Wiley and Sons, London. 1975; pp 36.
  23. Rhodes, M.E., The characterization of Pseudomonas fluorescens. J Gen. Microbiol. 1959. 21: 221-225.
  24. Schaad, N.W. Laboratory guide for identification of plant pathogenic bacteria. In: The American Psychopatho. Soc., Schaad, N. W. (ed.), Minneapolis, USA, 1992; pp 89-94.
  25. Tennakoon, P. L. Studies on plant growth promoting rhizo-microorganisms of Tea (Camellia sinensis) plants. M.Sc. Thesis, University of Agricultural Sciences, Dharwad, India. 2007; pp 103.
  26. Vejan, P., Abdullah, R. R., Khadiran, T., Ismail, S. and Nasrulhaq, B. A. Role of Plant Growth Promoting Rhizobacteria in Agricultural Sustainability. Molecules, 2016; 21: 573.
  27. Vieira, J. D. G. Purificacao e caracterizacao de uma á-amilase de Streptomyces sp. PhD. Thesis, Universidade de Sao Paulo, Brazil. 1999; pp 286.
  28. Weller, D. M. Biological control of soilborne plant pathogens in the rhizosphere with bacteria. Annual Rev. Phytopath., 1988; 26: 379-407.
  29. Wong, M. Y. Fusarium oxysporum f. sp. lycopersici (Sacc.). In: Soil Borne Plant Pathogens, Snyder, W.C. and Hans, H. N. (eds.), Kansas State University. 2003; pp 728.
  30. Zahid M., Abbasi M. K., Hameed S., Rahim N. 2015. Isolation and identification of indigenous plant growth promoting rhizobacteria from Himalayan region of Kashmir and their effect on improving growth and nutrient contents of maize (Zea mays L.). Front. Microbiol., 2015; 6: 207.

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