R. C. Dubey

Bacillus strains isolated from rhizosphere showed plant growth promoting and antagonistic activity against phytopathogens

Seven bacterial isolates screened from rhizosphere of common bean growing at Uttarakhand Himalaya showed potential plant growth promoting (PGP) and antagonistic activities. Based on 16S rRNA gene sequence the isolate BPR7 was identified as Bacillus sp. BPR7. The strain BPR7 produced IAA, siderophore, phytase, organic acid, ACC deaminase, cyanogens, lytic enzymes, oxalate oxidase, and solubilized various sources of organic and inorganic phosphates as well as potassium and zinc. Strain BPR7 strongly inhibited the growth of several phytopathogens such as Macrophomina phaseolina, Fusarium oxysporum, F. solani, Sclerotinia sclerotiorum, Rhizoctonia solani and Colletotricum sp. in vitro. Cell-free culture filtrate of strain BPR7 also caused colony growth inhibition of all test pathogens. PGP and antifungal activities of Bacillus sp. BPR7 suggest that it may be exploited as a potential bioinoculant agent for P. vulgaris.

Chitinase-mediated destructive antagonistic potential of Pseudomonas aeruginosa GRC1 against Sclerotinia sclerotiorum causing stem rot of peanut

Pseudomonas aeruginosa GRC1 exhibited strong antagonistic activity against Sclerotinia sclerotiorum, in vitro and in vivo. Scanning electron microscopic (SEM) studies showed morphological abnormalities such as perforation, lysis and fragmentation of hyphae of S. sclerotiorum caused by P. aeruginosa GRC1. This strain produced extracellular chitinase enzyme, the role of which was clearly demonstrated through Tn5 mutagenesis. Bacterization of peanut seeds with GRC1 resulted in increased seed germination and reduced stem-rot of peanut in S. sclerotiorum-infested soil by 97%. Other vegetative and yield plant parameters such as nodules per plant, pods and grain yield per plant were enhanced with a statistical significance in comparison to control. Neomycin resistant (GRC1neo+) bacterium was a good root colonizer and frequently isolated from rhizosphere of peanut plants. These findings showed P. aeruginosa GRC1 as a potential biocontrol agent against S. sclerotiorum.

Antagonistic interactions between fungal pathogens and phylloplane fungi of guava

Dominant phylloplane fungi of guava (Psidium guajava L.) were screened for their antagonistic activities against the two pathogens,Colletotrichum gloeosporioides andPestalotia psidii, bothin vitro andin vivo. Culture filtrates ofAspergillus niger, Fusarium oxysporum andPenicillium citrinum caused more than 50% growth inhibition ofC. gloeosporioides. Filtrates ofCephalosporium roseo-griseum andF. oxysporum were most effective in reducing the growth ofP. psidii. Volatiles produced from the cultures ofA. niger, F. oxysporum, P. citrinum andP. oxalicum inhibited the growth ofC. gloeosporioides, whereas volatiles fromC. roseo-griseum, F. oxysporum andTrichoderma harzianum inhibited the growth ofP. psidii. The inhibitory effect of volatiles decreased with increase in incubation time. In general, the maximum effect of volatiles was noticed after 48 h incubation. Different grades of colony interactions in dual cultures were recognised between the two pathogens and the phylloplane fungi examined. Maximum inhibition ofC. gloeosporioides was caused byAureobasidium pullulans, Cladosporium cladosporioides, epicoccum purpurascens, F. oxysporum andMyrothecium roridum, whereasAspergillus terreus, C. roseo-griseum andP. oxalicum significantly reduced the growth ofP. psidii. Application of a spore suspension of each test fungus inhibited lesion development of guava leaves caused by the test pathogensin vitro. Inhibition was more pronounced when the spore concentration was increased.A. pullulans, C. cladosporioides, E. purpurascens, F. oxysporum, andT. harzianum were found to be strongly antagonistic toC. gloeosporioides. A. niger, A. terreus, C. roseo-grisem andT. harzianum were strongly antagonistic toP. psidii.

Effect of environmental conditions and inoculum density on infection of guava fruits by Colletotrichum glososporioides.

The influence of environmental factors (temperature and humidity), inoculum density on infection by Colletotrichum glososporioides and development of anthracnose lesions were determined on uninjured, sand-injured and punctured fruits. The optical temperature for severe infection was 30 °C, whereas the disease incidence was less at 20 and 35 °C. Inoculated guavas that received 1–60 h of continuous free moisture developed lesions, but the disease was minimal (0–7%) after 1–6 h free moisture. Infection rates of uninjured, sand-injured and punctured fruits receiving 60 h of free moisture were 34, 70 and 100%, respectively. Disease incidence increased as inoculum density increased from 101 to 106 conidia/ml. In field conditions, the development of anthracnose lesions was greater on punctured guavas than on uninjured or sand-injured ones, in both rainy and winter seasons. In general, the number of lesions was highest in sand-injured fruits, followed by punctured and uninjured fruits. In rainy season the number of lesions on injured and uninjured fruits was greater than similarly treated guavas in winter.

Combined effect of chemical fertilisers and rhizosphere-competent Bacillus subtilis BSK17 on yield of Cicer arietinum

A chemical fertiliser-adaptive variant, Bacillus subtilis BSK17, showed induction in growth at 0.32 M of Urea, 0.05 M of DAP, 0.04 M of MoP and 0.08 of gypsum. In addition, B. subtilis BSK17 produced various plant growth-promoting substances and showed higher colony growth inhibition of Fusarium oxysporum that increased with increase in incubation time and reached the maximum by 78% at 120 h. In field, antibiotic-resistant marker strains of B. subtilis BSK17ery+ and B. subtilis BSK17tet+ showed more improvement in seed yield (90% than the control and 24% than full dose of chemical fertilisers) of Cicer arietinum when applied with half dose of chemical fertilisers (N5+5P15+15K15S10+10+10). Root length, shoot length, fresh and dry weight of root and shoot of plants were enhanced after 120 days in comparison to control; all values were significant at 1% CD. The strain significantly colonised the rhizosphere of C. arietinum by 6.64 log cfu after 120 days.

Growth enhancement of Sesamum indicum L. by rhizosphere-competent Azotobacter chroococcum AZO2 and its antagonistic activity against Macrophomina phaseolina

Indigenous strains isolated from rhizosphere may contain highly competent genotypes to enhance the plant growth and often perform better than the introduced isolates. The present study deals with the characterisation of plant growth-promoting (PGP) attributes and antagonistic activity of Azotobacter chroococcum AZO2 against Macrophomina phaseolina causing charcoal rot disease and their effect on the growth of sesame (Sesamum indicum L.). Eight strains of Azotobacter were isolated from sesame rhizosphere on nitrogen-free medium, which exhibited significant PGP parameters such as phosphate solubilisation, indole acetic acid and siderophore production. The strain A. chroococcum AZO2 (EU274299) was characterised by 16S rDNA gene sequencing. Amplification of 781 bp nif H gene confirms nitrogenase activity of all the strains. A. chroococcum AZO2 exhibited strong antagonistic activities against M. phaseolina causing 81% colony growth inhibition and resulted in hyphal perforations, empty cell (halo) formation, hyphal twisting, shrinking and lysis of fungal mycelia along with degeneration of sclerotia. A. chroococcum AZO2 produced chitinase that caused degradation and digestion of the cell wall component of M. phaseolina. Different vegetative and reproductive parameters of sesame were found to be enhanced significantly upon application of A. chroococcum AZO2 + half doses of chemical fertilisers. A. chroococcum AZO2 was also found to be an effective root coloniser, plant growth promoter and potential antagonistic bacterium. It can be concluded that A. chroococcum AZO2 strain bears the characteristics of technological applications for inoculant preparation and growth enhancement of sesame besides being utilised as a better PGP bacterium as well as an effective agent for biocontrol of M. phaseolina.

Evaluation of diversity of Bacilli from chickpea rhizosphere by 16S ARDRA and assessment of their plant-growth- promoting attributes

A total of eight motile, aerobic, Gram-positive and straight rod-shaped, endospore forming Bacillus spp. were isolated from rhizosphere of chickpea plants collected from different agricultural fields. Phylogeny of isolates was studied by partial sequencing of 16S rDNA and comparative analysis of the sequence data confirmed that the isolates belong to distinct phylogenetic lineage corresponding to Bacillus. Phenotyping clusters correlate with ARDRA pattern and showed resemblance to partial 16S rDNA sequencing. Bacillus spp. BSK5 and Bacillus subtilis BSK17 were the most potent strains for having plant-growth-promoting attributes. These two strains solubilised inorganic phosphate, produced Indole acetic acid, siderophore, Hydrocyanic acid and secreted extracellular chitinase and mild β-1,3-glucanase which antagonised and caused mycelial deformities in two phytopathogens Macrophomina phaseolina and Fusarium oxysporum in dual culture and by culture filtrate.