Subramaniam Gopalakrishnan

Evaluation of Streptomyces strains isolated from herbal vermicompost for their plant growth-promotion traits in rice.

Six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140, CAI-155 and KAI-180, isolated from six different herbal vermi-composts were characterized for in vitro plant growth-promoting (PGP) properties and further evaluated in the field for PGP activity in rice. Of the six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140 and CAI-155 produced siderophores; CAI-13, CAI-93, CAI-155 and KAI-180 produced chitinase; CAI-13, CAI-140, CAI-155 and KAI-180 produced lipase; CAI-13, CAI-93, CAI-155 and KAI-180 produced protease; and CAI-13, CAI-85, CAI-140 and CAI-155 produced ß-1-3-glucanase whereas all the six actinomycetes produced cellulase, hydrocyanic acid and indole acetic acid (IAA). The actinomycetes were able to grow in NaCl concentrations of up to 8%, at pH values between 7 and 11, temperatures between 20 and 40 °C and compatible with fungicide bavistin at field application levels. In the rice field, the actinomycetes significantly enhanced tiller numbers, panicle numbers, filled grain numbers and weight, stover yield, grain yield, total dry matter, root length, volume and dry weight over the un-inoculated control. In the rhizosphere, the actinomycetes also significantly enhanced total nitrogen, available phosphorous, % organic carbon, microbial biomass carbon and nitrogen and dehydrogenase activity over the un-inoculated control. Sequences of 16S rDNA gene of the actinomycetes matched with different Streptomyces species in BLAST analysis. Of the six actinomycetes, CAI-85 and CAI-93 were found superior over other actinomycetes in terms of PGP properties, root development and crop productivity. qRT-PCR analysis on selected plant growth promoting genes of actinomycetes revealed the up-regulation of IAA genes only in CAI-85 and CAI-93.

Plant growth promoting rhizobia: challenges and opportunities

Modern agriculture faces challenges, such as loss of soil fertility, fluctuating climatic factors and increasing pathogen and pest attacks. Sustainability and environmental safety of agricultural production relies on eco-friendly approaches like biofertilizers, biopesticides and crop residue return. The multiplicity of beneficial effects of microbial inoculants, particularly plant growth promoters (PGP), emphasizes the need for further strengthening the research and their use in modern agriculture. PGP inhabit the rhizosphere for nutrients from plant root exudates. By reaction, they help in (1) increased plant growth through soil nutrient enrichment by nitrogen fixation, phosphate solubilization, siderophore production and phytohormones production (2) increased plant protection by influencing cellulase, protease, lipase and β-1,3 glucanase productions and enhance plant defense by triggering induced systemic resistance through lipopolysaccharides, flagella, homoserine lactones, acetoin and butanediol against pests and pathogens. In addition, the PGP microbes contain useful variation for tolerating abiotic stresses like extremes of temperature, pH, salinity and drought; heavy metal and pesticide pollution. Seeking such tolerant PGP microbes is expected to offer enhanced plant growth and yield even under a combination of stresses. This review summarizes the PGP related research and its benefits, and highlights the benefits of PGP rhizobia belonging to the family Rhizobiaceae, Phyllobacteriaceae and Bradyrhizobiaceae.

Evaluation of bacteria isolated from rice rhizosphere for biological control of charcoal rot of sorghum caused by Macrophomina phaseolina (Tassi) Goid.

A total of 360 bacteria, isolated from the rhizospheres of a system of rice intensification (SRI) fields, were characterized for the production of siderophore, fluorescence, indole acetic acid (IAA), hydrocyanic acid (HCN) and solubilization of phosphorus. Of them, seven most promising isolates (SRI-156, -158, -178, -211, -229, -305 and -360) were screened for their antagonistic potential against Macrophomina phaseolina (causes charcoal rot in sorghum) by dual culture assay, blotter paper assay and in greenhouse. All the seven isolates inhibited M. phaseolina in dual culture assay, whereas six isolates solubilized phosphorous (except SRI-360), all seven produced siderophore, four produced fluorescence (except SRI-178, -229 and -305), six produced IAA (except SRI-305) and five produced HCN (except SRI-158 and -305). In the blotter paper assay, no charcoal rot infection was observed in SRI-156-treated sorghum roots, indicating complete inhibition of the pathogen, while the roots treated with the other isolates showed 49–76% lesser charcoal rot infection compared to the control. In the antifungal activity test (in green house on sorghum), all the isolates increased shoot dry mass by 15–23% and root dry mass by 15–20% (except SRI-158 and -360), over the control. In order to confirm the plant growth-promoting (PGP) traits of the isolates, the green house experiment was repeated but, in the absence of M. phaseolina. The results further confirmed the PGP traits of the isolates as evidenced by increases in shoot and root dry mass, 22–100% and 5–20%, respectively, over the control. The sequences of 16S rDNA gene of the isolates SRI-156, -158, -178, -211, -229, -305 and -360 were matched with Pseudomonasplecoglossicida, Brevibacterium antiquum, Bacillus altitudinis, Enterobacter ludwigii, E. ludwigii, Acinetobacter tandoii and P. monteilii, respectively in BLAST analysis. This study indicates that the selected bacterial isolates have the potential for PGP and control of charcoal rot disease in sorghum.

Plant growth-promoting activities of Streptomyces spp. in sorghum and rice

Five strains of Streptomyces (CAI-24, CAI-121, CAI-127, KAI-32 and KAI-90) were earlier reported by us as biological control agents against Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceri (FOC). In the present study, the Streptomyces were characterized for enzymatic activities, physiological traits and further evaluated in greenhouse and field for their plant growth promotion (PGP) of sorghum and rice. All the Streptomyces produced lipase, β-1-3-glucanase and chitinase (except CAI-121 and CAI-127), grew in NaCl concentrations of up to 6%, at pH values between 5 and 13 and temperatures between 20 and 40°C and were highly sensitive to Thiram, Benlate, Captan, Benomyl and Radonil at field application level. When the Streptomyces were evaluated in the greenhouse on sorghum all the isolates significantly enhanced all the agronomic traits over the control. In the field, on rice, the Streptomyces significantly enhanced stover yield (up to 25%; except CAI-24), grain yield (up to 10%), total dry matter (up to 18%; except CAI-24) and root length, volume and dry weight (up to 15%, 36% and 55%, respectively, except CAI-24) over the control. In the rhizosphere soil, the Streptomyces significantly enhanced microbial biomass carbon (except CAI-24), nitrogen, dehydrogenase (except CAI-24), total N, available P and organic carbon (up to 41%, 52%, 75%, 122%, 53% and 13%, respectively) over the control. This study demonstrates that the selected Streptomyces which were antagonistic to FOC also have PGP properties.

Plant growth-promoting traits of biocontrol potential bacteria isolated from rice rhizosphere

Seven isolates of bacteria (SRI-156, SRI-158, SRI-178, SRI-211, SRI-229, SRI-305 and SRI-360) were earlier reported by us as having potential for biocontrol of charcoal rot of sorghum and plant growth promotion (PGP) of the plant. In the present study, the seven isolates were characterized for their physiological traits (tolerance to salinity, pH, temperature and resistance to antibiotics and fungicides) and further evaluated in the field for their PGP of rice. All the seven isolates were able to grow at pH values between 5 and 13, in NaCl concentrations of up to 8% (except SRI-156 and SRI-360), temperatures between 20 and 40°C and were resistant to ampicillin (>100 ppm; except SRI-158 and SRI-178) but sensitive (<10 ppm) to chloramphenicol, kanamycin, nalidixic acid, streptomycin (except SRI-156 and SRI-211) and tetracycline. They were tolerant to fungicides benlate and captan, except SRI-158 and SRI-178, bavistin and sensitive to thiram (except SRI-156 and SRI-211) at field application level. In the field, four of the seven isolates (SRI-158, SRI-211, SRI-229 and SRI-360) significantly enhanced the tiller numbers, stover and grain yields, total dry matter, root length, volume and dry weight over the un-inoculated control. In the rhizosphere soil at harvest, all the isolates significantly enhanced microbial biomass carbon (except SRI-156), microbial biomass nitrogen and dehydrogenase activity (up to 33%, 36% and 39%, respectively) and total N, available P and% organic carbon (up to 10%, 38% and 10%, respectively) compared to the control. This investigation further confirms that the SRI isolates have PGP properties.

Evaluation of Streptomyces spp. for their plant-growth-promotion traits in rice

Five strains of Streptomyces (CAI-17, CAI-68, CAI-78, KAI-26, and KAI-27) were previously reported to have potential for charcoal rot control and plant growth promotion (PGP) in sorghum. In this study, those 5 Streptomyces strains were characterized for their enzymatic activities and evaluated for their PGP capabilities on rice. All the Streptomyces strains were able to produce lipase and β-1,3-glucanase; grew in NaCl (up to 8%), at pH 5-13, and at temperatures 20-40 °C; and were resistant to ampicillin, sensitive to nalidixic acid, and highly sensitive to chloramphenicol, kanamycin, streptomycin, and tetracycline. They were highly tolerant to the fungicide bavistin but were highly sensitive to benlate, benomyl, and radonil. When evaluated on rice in the field, Streptomyces significantly enhanced tiller and panicle numbers, stover and grain yields, dry matter, root length, volume and dry weight, compared with the control. In the rhizosphere at harvest, microbial biomass carbon and nitrogen, dehydrogenase activity, total nitrogen, available phosphorus, and % organic carbon were also found significantly higher in Streptomyces-treated plots than in the control plots. This study further confirms that the selected Streptomyces have PGP activities.

Plant growth-promoting traits of Streptomyces with biocontrol potential isolated from herbal vermicompost

Abstract Three strains of Streptomyces (CAI-21, CAI-26 and MMA-32) were earlier reported by us as having potential for biocontrol of charcoal rot of sorghum, caused by Macrophomina phaseolina (Tassi) Goid., and plant growth promotion (PGP) of the plant. In the present investigation, the three Streptomyces were characterised for their physiological traits (tolerance of salinity, temperature, pH and resistance to antibiotics) and further evaluated in the field for their PGP of rice, grown by a system of rice intensification methods. All three Streptomyces were able to grow in NaCl concentrations of up to 12% (except MMA-32), at pH values between 5 and 13 and temperatures between 20 and 40°C. They were highly resistant to ampicillin and trimethoprim (>800 ppm), sensitive to chloramphenicol, kanamycin and nalidixic acid (50–100 ppm) and highly sensitive to streptomycin and tetracycline (5–25 ppm). When evaluated for their PGP activity on seedlings of rice, % germination and shoot and root lengths were significantly enhanced over the control. In the field, the Streptomyces strains significantly enhanced the panicle length, filled grain numbers and weight, panicle weight, 1000 seed weight, tiller numbers, total dry matter, root length (39–65%), root volume (13–30%), root dry weight (16–24%), grain yield (9–11%) and stover yield (11–22%) over the control. In the rhizosphere soil (0–15 cm from root) at harvest, the population of actinomycetes was significantly enhanced as was microbial biomass carbon (27–83%) and nitrogen (24–43%), dehydrogenase activity (34–152%), available P (13–34%) and N (30–53%) and % organic carbon (26–28%). This study further confirms that the selected Streptomyces have PGP properties.

Efficacy of botanical extracts and entomopathogens on control of Helicoverpa armigera and Spodoptera litura

Interest in biological control of insect-pests of economically important plants has been stimulated in recent years by trends in agriculture towards greater sustainability and public concern about the use of hazardous pesticides. Botanicals and microorganisms have the capability to synthesize biologically active secondary metabolites such as antibiotics, herbicides and pesticides. In this investigation, washings of herbal vermicompost (called biowash; viz. Annona, Chrysanthemum, Datura, Jatropha, Neem, Parthenium, Pongamia, Tridax and Vitax) and plant growth promoting (PGP) bacteria [viz. Bacillus subtilis (BCB-19), Bacillus megaterium (SB-9), Serratia mercescens (HIB-28) and Pseudomonas spp. (SB-21)] and fungus (Metarhizium anisopliae ) were evaluated for their efficacy against Helicoverpa armigera and Spodoptera litura. When the feed was treated with crude biowash for healthy larvae (4-day old), 42 and 86% mortality and 32 and 71% weight reduction over control was reported for H. armigera, while in the case of S. litura , it was between 46 and 74% larval mortality and 47 and 77% weight reduction over control. When healthy larvae were treated with PGP bacteria and fungus, the mortality rate varied between 59 and 73%, with 55 and 92% weight reduction over control on H. armigera, while for S. litura, 54 and 72% larval mortality and 44 and 79% weight reduction over control was reported. The results of the compatibility studies (entomopathogenic potential biowash of the botanicals with PGP bacteria and fungus) indicate that there was no definite sign of suppression of any of the botanicals on the PGP bacteria and fungus, except Datura with B. subtilis BCB-19, whereas, there was definite sign of enhanced growth of B. megaterium SB-9 with all the botanicals. Compatibility studies between PGP bacteria and fungus showed that all bacteria are compatible with each other except M. anisopliae . Crude biowash of the promising botanicals (Annona, Datura, Jatropha, Neem, Parthenium and Pongamia) were further fractionated on C18 solid phase extraction cartridge (SPE) and the resultant adsorbed and non-adsorbed fractions were tested against H. armigera. Results indicate that both adsorbed as well as non-adsorbed fractions showed significant mortality on H. armigera . Adsorbed fractions of all the six biowash showed mortality between 81 and 93% (64 and 73% for nonadsorbed fraction) over control and the weight reduction of the larvae was found between 73 and 91% (80 and 97% for non-adsorbed fraction) over control. It was therefore concluded that the aforementioned six botanicals and five entomopathogens has great potential in the management of H. armigera and S. litura. Key words: Botanicals, entomopathogens, biowash, secondary metabolites, Helicoverpa armigera, Spodoptera litura.

Plant growth-promotion and biofortification of chickpea and pigeonpea through inoculation of biocontrol potential bacteria, isolated from organic soils

Seven strains of bacteria [Pseudomonas plecoglossicida SRI-156, Brevibacterium antiquum SRI-158, Bacillus altitudinis SRI-178, Enterobacter ludwigii SRI-211, E. ludwigii SRI-229, Acinetobacter tandoii SRI-305 and Pseudomonas monteilii SRI-360; demonstrated previously for control of charcoal rot disease in sorghum and plant growth-promotion (PGP) in rice] were evaluated for their PGP and biofortification traits in chickpea and pigeonpea under field conditions. When treated on seed, the seven selected bacteria significantly enhanced the shoot height and root length of both chickpea and pigeonpea over the un-inoculated control. Under field conditions, in both chickpea and pigeonpea, the plots inoculated with test bacteria enhanced the nodule number, nodule weight, root and shoot weights, pod number, pod weight, leaf weight, leaf area and grain yield over the un-inoculated control plots. Among the seven bacteria, SRI-229 was found to significantly and consistently enhance all the studied PGP and yield traits including nodule number (24 and 36%), nodule weight (11 and 44%), shoot weight (22 and 20%), root weight (23 and 16%) and grain yield (19 and 26%) for both chickpea and pigeonpea, respectively. When the harvested grains were evaluated for their mineral contents, iron (up to 18 and 12%), zinc (up to 23 and 5%), copper (up to 19 and 8%), manganese (up to 2 and 39%) and calcium (up to 22 and 11%) contents in chickpea and pigeonpea, respectively, were found enhanced in test bacteria inoculated plots over the un-inoculated control plots. This study further confirms that the selected bacterial isolates not only have the potential for PGP in cereals and legumes but also have the potential for biofortification of mineral nutrients.

Biological activity of entomopathogenic actinomycetes against lepidopteran insects (Noctuidae: Lepidoptera)

Vijayabharathi, R., Kumari, B. R., Sathya, A., Srinivas, V., Abhishek, R., Sharma, H. C. and Gopalakrishnan, S. 2014. Biological activity of entomopathogenic actinomycetes against lepidopteran insects (Noctuidae: Lepidoptera). Can. J. Plant Sci. 94: 759-769. The aim of the present study was to identify an efficient broad-spectrum bio-pesticide for the control of lepidopteran insects from microbes in various ecological niches. A total of 111 microbes isolated from various herbal vermi-composts and organically cultivated fields were evaluated for their intracellular metabolites (ICM), extracellular metabolites (ECM) and whole culture (WC) against early instars of lepidopteran insects. Fifteen actinomycete isolates which showed insecticidal activity against 2nd instar Helicoverpa armigera were selected and further screened against Spodoptera litura and Chilo partellus. A significant broad spectrum insecticidal activity was found in the order ECM>ICM>WC against all the insects under laboratory conditions. All these actinomycete isolates also registered significant activity under greenhouse conditions on 2nd instar H. armigera. The actinomycete isolates were identified by 16S rDNA sequencing and matched with Streptomyces species using BLAST search. Among all the 15 isolates, SAI-25 (S. griseoplanus), CAI-155 (S. bacillaris) and BCA-698 (S. albolongus) showed consistent entomopathogenic activity against all the three insects suggesting their potential as broad-spectrum biocontrol agents against other lepidopterans.