Hari S. Gupta

Isolation and Characterization of Nonrhizobial Plant Growth Promoting Bacteria from Nodules of Kudzu (Pueraria thunbergiana) and Their Effect on Wheat Seedling Growth

The leguminous vine Kudzu (Pueraria thunbergiana) is an introduction into the N. W. Himalayan region of India. Despite its value as a fodder and cover crop, little is known about the nature of the nodule microflora. In an attempt to study the nodule bacteria, we isolated and characterized three nonrhizobial plant growth promoting bacteria from surface sterilized nodules of Kudzu. Based on the sequencing of the 16 S r RNA gene, the isolates were designated as Bacillusthuringiensis KR-1, Enterobacter asburiae KR-3, and Serratia marcescens KR-4. Crystalline bodies were detected in the isolate KR-1, confirming its identity as B. thuringiensis. Under in vitro conditions, all three isolates were found to produce indole acetic acid. Other plant growth promotion attributes such as P solubilization, hydrogen cyanide production, and ammonia production varied among the isolates. All of the three isolates promoted growth and positively influenced nutrient uptake parameters of wheat seedlings.

Phosphate solubilization and growth promotion by Pseudomonas fragi CS11RH1 (MTCC 8984), a psychrotolerant bacterium isolated from a high altitude Himalayan rhizosphere

Phosphate solubilization and growth promotion by Pseudomonas fragi CS11RH1 (MTCC 8984), a psychrotolerant bacterium isolated from a high altitude garlic rhizosphere from the Indian Himalayas, are reported here. The identity of the isolate was arrived on the basis of its biochemical features and sequencing of the 16S rRNA gene. The isolate grew and solubilized phosphate at temperatures ranging from 4 to 30°C. Besides solubilizing P it produced indole acetic acid (IAA) and hydrogen cyanide (HCN). Seed bacterization with the isolate significantly increased the percent germination, rate of germination, plant biomass and nutrient uptake of wheat seedlings. While Pseudomonas fragi is normally associated with the spoilage of dairy products stored at cold temperatures, this is an early report on the plant growth promoting ability of the bacterium.

Characterization and phylogenetic analysis of an entomopathogenic Bacillus cereus strain WGPSB-2 (MTCC 7182) isolated from white grub, Anomala dimidiata (Coleoptera: Scarabaeidae)

Abstract White grubs (Coleoptera: Scarabaeidae) are cosmopolitan and polyphagous insect pests of agricultural crops, forests and pastures around the world. The lack of an environmentally sound approach for white grub management has prompted the exploration and detection of a novel microbial biocontrol agent against these sub-terranean insect pests. In this study we describe the isolation, establishment of pathogenesis, biochemical characterization and phylogenetic analysis of an entomopathogenic Bacillus cereus strain WGPSB-2 (MTCC 7182), isolated from an atrophied pupa of Anomala dimidiata, collected from the N.W. Indian Himalayas. The sequencing and subsequent comparison of the 16S rDNA revealed that the strain has100% similarity with Bacillus cereus sequences. Phylogenetic analysis of the 16S rDNA sequence revealed that the isolate is closely related to Bacillus thuringiensis and Bacillus sphaericus. In vitro bioassays showed that the isolate was able to infect and cause 92 and 67% mortality in second instar larvae of Anomala dimidiata and Holotrichia seticollis, respectively. The infected larvae exhibited bacterial septicemia like symptoms and mortality occurred between the third and ninth weeks after inoculation. The culture has been granted the accession number MTCC 7182 by the Microbial Type Culture Collection and Gene Bank, Institute of Microbial Technology, Chandigarh, India.

Effect of organic manures and integrated nutrient management on yield potential of bell pepper (Capsicum annuum) varieties and on soil properties

A field experiment was conducted during the summer season of 2005 and 2006 at Almora (Indian Himalayas) to evaluate the comparative performance of five varieties of bell pepper (Capsicum annuum L.) and changes in soil properties under organic and integrated nutrient management (INM) systems. The number of fruits plant−1 and fruit yield was significantly higher under INM compared with organic nutrient supply. The yield reduction in the latter was 22.1% in 2005 and 21.7% in 2006 compared with INM. Among the bell pepper varieties, ‘US Agri 181’ produced significantly higher fruit yield (25.42 mg ha−1 in 2005 and 28.82 mg ha−1 in 2006) compared with other varieties. The soil pH, organic carbon, and microbial activities in terms of urease, dehydrogenase and alkaline phosphatase were higher in the plots treated with organic manures alone compared to INM. The latter, however, recorded higher N, P, and K contents in soil. The gross margin and benefit:cost (B:C) ratio were higher under INM compared with organic nutrient supply. ‘US Agri 181’ gave the highest net returns followed by ‘Aishwarya’ and ‘California Wonder’. However, the latter recorded the highest B:C ratio (9.2) compared with other varieties. We conclude that at least 25–50% price premium for organic bell pepper may be required to offset the higher cost of cultivation and low yields under organic production system. Among the bell pepper varieties, ‘US Agri 181’ was the best variety as it yielded highest followed by ‘Aishwarya’.

Differential toxicity of Bacillus thuringiensis strains and their crystal toxins against high-altitude Himalayan populations of diamondback moth, Plutella xylostella L.

BACKGROUND Diamondback moth, Plutella xylostella (L.), is a major insect pest of crucifers in the biodiversity-rich north-western Indian Himalayan hills. The present investigation was aimed at determining the susceptibility pattern of P. xylostella populations collected from different locations of this region to autochthonous and standard Bacillus thuringiensis strains. RESULTS Among the reference as well as indigenous B. thuringiensis strains tested, sub spp. kurstaki HD-1, kurstaki HD-73, galleriae HD-8, local galleriae/colmeri strain BtOa1 and some of their Cry1 class toxins were found to be highly toxic. Surprisingly, the sub sp. tolworthi HD-125, local tolworthi strain BtHa1 and Cry9 class toxins were found to be non-toxic. Midgut homogenate from fourth-instar larvae was found to activate 130 kDa protoxin from the local tolworthi strain BtHa1 into 68 kDa toxin, but failed to exert any larval mortality, probably owing to lack of receptor binding. CONCLUSION The present study provides valuable baseline susceptibility data for the deployment of B. thuringiensis-based control methods, as well as for future monitoring of development of resistance in P. xylostella to B. thuringiensis in this ecologically sensitive region.

Brevibacterium frigoritolerans a novel entomopathogen of Anomala dimidiata and Holotrichia longipennis (Scarabaeidae: Coleoptera)

We describe the isolation, biochemical characterization, phylogenetic analysis, and pathogenecity of a novel entomopathogenic bacterium Brevibacterium frigoritolerans to first instar larvae of Anomala dimidiata and Holotrichia longipennis. The almost full length 16S rRNA sequence of the bacterium has 99% identity with the type strain of B. frigoritolerans, while phylogenetic analysis revealed that the isolate formed a tightly linked branch with the type strain of B. frigoritolerans. Under in vitro bioassay conditions, the isolate infected and caused 89±5.4 and 74±7.7% mortality, in first instar larvae of A. dimidiata and H. longipennis, respectively. The infected larvae exhibited bacteremia like symptoms and mortality occurred between the second and fifth weeks after inoculation. This is an early report on the entomopathogenic potential of the hitherto lesser-known bacterium Brevibacterium frigoritolerans.

Genomewide Expression and Functional Interactions of Genes under Drought Stress in Maize

A genomewide transcriptome assay of two subtropical genotypes of maize was used to observe the expression of genes at seedling stage of drought stress. The number of genes expressed differentially was greater in HKI1532 (a drought tolerant genotype) than in PC3 (a drought sensitive genotype), indicating primary differences at the transcriptional level in stress tolerance. The global coexpression networks of the two genotypes differed significantly with respect to the number of modules and the coexpression pattern within the modules. A total of 174 drought-responsive genes were selected from HKI1532, and their coexpression network revealed key correlations between different adaptive pathways, each cluster of the network representing a specific biological function. Transcription factors related to ABA-dependent stomatal closure, signalling, and phosphoprotein cascades work in concert to compensate for reduced photosynthesis. Under stress, water balance was maintained by coexpression of the genes involved in osmotic adjustments and transporter proteins. Metabolism was maintained by the coexpression of genes involved in cell wall modification and protein and lipid metabolism. The interaction of genes involved in crucial biological functions during stress was identified and the results will be useful in targeting important gene interactions to understand drought tolerance in greater detail.

‘Vivek Matar 11’: A New Garden Pea Cultivar

Garden pea (Pisum sativum L. var. hortense) is cultivated during winter in subtropical and early spring in temperate zones. The major constraint to its cultivation, especially in low- and mid-hills, is infection by the causal organism of powdery mildew. Varietal diversification is needed to develop early- and medium-duration resistance to powdery mildew. Breeding work was initiated to develop a high-yielding garden pea cultivar with medium maturity, dark green pods, and resistance to powdery mildew. ‘Vivek Matar 11’ is a new garden pea cultivar developed by hybridization between ‘Azad Pea 1’ × ‘PRS-18-6-4-5-1’. It is a medium maturity cultivar having attractive long green curved pods with 8–10 sweet seed/pod and is highly resistant to powdery mildew. Average green pod yield is 10–11 Mt·ha−1. It is suitable for cultivation under organic and inorganic conditions. This cultivar may be adapted to other temperate regions of the world.