Prem Lal Kashyap

Myconanotechnology in agriculture: a perspective

Myconanotechnology is an emerging field, where fungi can be harnessed for the synthesis of nanomaterials or nanostructures with desirable shape and size. Though myconanotechnology is in its infancy, potential applications provide exciting waves of transformation in agriculture and fascinate microbiologists and other researchers to contribute in providing incremental solutions through green chemistry approaches for advancing food security. In this article, we provide a brief overview of the research efforts on the mycogenic synthesis of nanoparticles with particular emphasis on mechanisms and potential applications in agriculture and allied sectors.

Diversity and antagonistic potential of Bacillus spp. associated to the rhizosphere of tomato for the management of Rhizoctonia solani

Abstract Bacillus spp. has emerged as the most effective alternative to synthetic chemical fungicides. To get a better insight in the antagonistic potential of Bacillus strains, rhizospheric soil samples of healthy tomato plants from Indo-gangetic plain regions of India were analysed. A total of 108 Bacillus strains were obtained from preliminary screening. Potent strains identified on the basis of in vitro antagonistic and biochemical assays were subjected to diversity analysis using 16S-rDNA, BOX and ERIC-PCR. Furthermore, the four best performing antagonistic Bacillus strains under in vitro plant growth promotion and antagonistic assay were selected for pot experiment. In field study, Bacillus amyloliquefaciens MB101 and Bacillus subtilis MB14 showed drastic reduction in disease index by 55.7 and 41.74% with significant elevation in fruit yield up to 220 and 184 qha–1, respectively. The present study was successful in selecting effective Bacillus strains by performing phenotypic and genotypic characterisation of Bacillus strains that can be used as an integral component of integrated disease management of tomato root rot and damping-off.

Bacterial xylanases: biology to biotechnology

In this review, a comprehensive discussion exclusively on bacterial xylanases; their gene organization; different factors and conditions affecting enzyme yield and activity; and their commercial application have been deliberated in the light of recent research findings and extensive information mining. Improved understanding of biological properties and genetics of bacterial xylanase will enable exploitation of these enzymes for many more ingenious biotechnological and industrial applications.

Optimization of media components for chitinase production by chickpea rhizosphere associated Lysinibacillus fusiformis B‐CM18

Chitinase producing strain B‐CM18 was isolated from chickpea rhizosphere and identified as Lysinibacillus fusiformis B‐CM18. It showed in vitro antifungal activity against a wide range of fungal plant pathogens and was found to produce several PGPR activities. Further, a multivariate response surface methodology was used to evaluate the effects of different factors on chitinolytic activity and optimizing enzyme production. A central composite design was employed to achieve the highest chitinase production at optimum values of the process variables, viz., temperature (20–45 °C), sodium chloride (2–7%), starch (0.1–1%) and yeast extract (0.1–1%), added in the minimal medium supplemented with colloidal chitin (1–10%; w:w). The fit of the model (R2 = 0.5692) was found to be significant. The production medium to achieve the highest chitinase production (101 U ml−1) was composed of the minimal medium composed of chitin (6.09%), NaCl (4.5%), starch (0.55%) and yeast extract (0.55%) with temperature (32.5 °C). The results show that the optimization strategy led to an increase in chitinase production by 56.1‐fold. The molecular mass of the chitinase was estimated to be 20 kDa by anion exchange and gel filtration chromatography. Further, purified chitinase showed strong antifungal activity against test pathogens. Overall, these results may serve as a base line data for enhancing the chitinolytic potential of bacterial antagonists for bio‐management of chickpea pathogens.

Multifarious plant growth promoting characteristics of chickpea rhizosphere associated Bacilli help to suppress soil-borne pathogens

Wilt and root rot are the major constraints in chickpea production and very difficult to manage through agrochemicals. Hence, for an ecofriendly and biological management, 240 strains of Bacillus and Bacillus derived genera were isolated from chickpea rhizosphere, further narrowed down to 14 strains on the basis of in vitro production of indole acetic acid, siderophore, phosphate solubilization, hydrolytic enzymes and were evaluated for antagonism against chickpea pathogens (Fusarium oxysporum f. sp. ciceri race 1, F. solani and Macrophomina phaseolina). The strains were identified on the basis of physiological characters and 16S RNA gene sequencing. The genotypic comparisons of strains were determined by BOX-polymerase chain reaction profiles and amplified rDNA restriction analysis. These isolates were evaluated in greenhouse assay in which B. subtilis (B-CM191, B-CV235, B-CL-122) proved to be effective in reducing wilt incidence and significant enhancement in growth (root and shoot length) and dry matter of chickpea plants. PCR amplification of bacillomycin (bmyB) and β-glucanase genes suggests that amplified genes from the Bacillus could have a role to further define the diversity, ecology, and biocontrol activities in the suppression of soil-borne pathogens.

Isolation and characterization of siderophore producing antagonistic rhizobacteria against Rhizoctonia solani

Plant protection through siderophore producing rhizobacteria (SPR) has emerged as a sustainable approach for crop health management. In present study, 220 bacteria isolated from tomato rhizosphere were screened for in vitro antagonistic activity against Rhizoctonia solani AG‐4. Nine potent antagonistic strains viz., Alcaligenes sp. (MUN1, MB21, and MPF37), Enterobacter sp. (MPM1), Pseudomonas sp. (M10A and MB65), P. aeruginosa (MPF14 and MB123) and P. fluorescens (MPF47) were identified on the basis of physiological characters and 16S rDNA sequencing. These strains were able to produce hydrolytic enzymes, hydrogen cyanide, indole acetic acid, although, only few strains were able to solubilize phosphate. Two strains (MB123 and MPF47) showed significant disease reduction in glasshouse conditions were further evaluated under field conditions using three different application methods. Application of P. fluorescens (MPF47) in nursery as soil mix + seedling root treatments prior to transplantation resulted in significant disease reduction compared to control. Total chlorophyll and available iron were significantly higher in the MPF47 treated plants in contrast to infected control. In conclusion, siderophore producing bacteria MPF47 have strong biocontrol abilities and its application as soil mix + seedling root treatments provided strong shield to plant roots against R. solani and could be used for effective bio‐management of pathogen.

Deciphering Diversity of Salt-Tolerant Bacilli from Saline Soils of Eastern Indo-gangetic Plains of India

The intensive use of chemical fertilizers, monoculture and irrigation with surface saline water has resulted in the deterioration of soil health by enhancing the level of salinity in the Eastern Indo-Gangetic Plains of India. Therefore, diversity of halotolerant bacteria adapted to that environment and possessed the ability to produce plant growth hormones was explored, that could be used for salt stress amelioration. The 16S rRNA gene sequencing and fatty acid methyl ester (FAME) were used for diversity analysis of salt-tolerant bacilli. Among the 95 isolates, 55 strains showed plant growth promotion traits, production of industrially important enzymes (amylase, protease and cellulase) and tolerance to more than 4% NaCl. Using partial 16S rRNA sequences and FAME comparisons, 21 different species of Bacillus and Bacillus-derived genera were identified, viz. Bacillus megaterium, B. subtilis, B. licheniformis, B. firmus, B. horikoshii, B. pumilus, Bacillus sp., B. safensis, B. thuringiensis, B. simplex, B. agri, B. flexus, B. oceanisediminis, B. cereus, B. arsenicus, Paenibacillus dendritiformis, Lysinibacillus sp., L. sphaericus, B. marisflavi, Terribacillus sp., and B. mycoides. These isolates possess the ability to tolerate high salt, form endospores, withstand harsh environments, and also have the potential for plant growth promotion, which could be useful in formulation of new inoculants to enhance the availability of nutrients for crop growth under saline conditions.

Trichoderma for climate resilient agriculture

Climate change is one of the biggest challenges of the twenty-first century for sustainable agricultural production. Several reports highlighted the need for better agricultural practices and use of eco-friendly methods for sustainable crop production under such situations. In this context, Trichoderma species could be a model fungus to sustain crop productivity. Currently, these are widely used as inoculants for biocontrol, biofertilization, and phytostimulation. They are reported to improve photosynthetic efficiency, enhance nutrient uptake and increase nitrogen use efficiency in crops. Moreover, they can be used to produce bio-energy, facilitate plants for adaptation and mitigate adverse effect of climate change. The technological advancement in high throughput DNA sequencing and biotechnology provided deep insight into the complex and diverse biotic interactions established in nature by Trichoderma spp. and efforts are being made to translate this knowledge to enhance crop growth, resistance to disease and tolerance to abiotic stresses under field conditions. The discovery of several traits and genes that are involved in the beneficial effects of Trichoderma spp. has resulted in better understanding of the performance of bioinoculants in the field, and will lead to more efficient use of these strains and possibly to their improvement by genetic modification. The present mini-review is an effort to elucidate the molecular basis of plant growth promotion and defence activation by Trichoderma spp. to garner broad perspectives regarding their functioning and applicability for climate resilient agriculture.

Characterization of antagonistic‐potential of two Bacillus strains and their biocontrol activity against Rhizoctonia solani in tomato

To investigate the biocontrol mechanism of two antagonistic Bacillus strains (Bacillus subtilis MB14 and Bacillus amyloliquefaciens MB101), three in vitro antagonism assays were screened and the results were concluded that both strains inhibited Rhizoctonia solani growth in a similar manner by dual culture assay, but the maximum percent of inhibition only resulted with MB101 by volatile and diffusible metabolite assays. Moreover, cell free supernatant (CFS) of MB101 also showed significant (p > 0.05) growth inhibition as compared to MB14, when 10 and 20% CFS mix with the growth medium of R. solani. After in vitro-validation, both strains were evaluated under greenhouse and the results concluded that strain MB101 had significant biocontrol potential as compared to MB14. Strain MB101 was enhanced the plant height, biomass and chlorophyll content of tomato plant through a higher degree of root colonization. In field trials, strain MB101 showed higher lessening in root rot symptoms with significant fruit yield as compare to strain MB14 and infected control. Next to the field study, the presence of four antibiotic genes (srfAA, fenD, ituC, and bmyB) also concluded the antifungal nature of both Bacillus strains. Phylogenetic analysis of protein sequences revealed a close relatedness of three genes (srfAA, fenD, and ituC) with earlier reported sequences of B. subtilis and B. amyloliquefaciens. However, bmyB showed heterogeneity in among both strains (MB14 and MB101) and it may be concluded that higher degree of antagonism, root colonization and different antibiotic producing genes may play an important role in biocontrol mechanism of strain MB101.

Nanodiagnostics for plant pathogens

Rapid detection technologies with high sensitivity and selectivity for plant pathogens are essential to prevent disease spread and minimize losses to assure optimal productivity and food security. Traditional laboratory techniques such as microscopy and culture are time-consuming, labour intensive and require complex sample handling. Immunological and molecular techniques have advanced but have some issues related to rapidity, signal strength and instrumentation. The integration of immunological and molecular diagnostics with nanotechnology systems offers an option where all detection steps can be accommodated on a portable miniaturized device for rapid and accurate detection of plant pathogens. The sensitive nature of functionalized nanoparticles can be used to design phytopathogen detection devices with smart sensing capabilities for field use. This review summarizes the current status and future prospects of nanotechnology for detection and diagnosis of plant pathogens.