Research Thrust Area

Research experiences: January 2009- Till date (13 Years)

Theme: Microbial Biotechnology for Agricultural and Environmental Sustainability

1. January 01, 2016 - Till date Assistant Professor at Eternal university 06 Years

2. October 03, 2009 -December 2015 SRF at ICAR-IARI, New Delhi 06 Years

3. January 01, 2009-May 31, 2009 M.Sc. Project work at ICAR-NBAIM, Mau, UP 5 Months

Current Research

1. Microbial biodiversity of crops growing in Himalayan regions

Objectives: Isolation, characterization, identification of plant microbes (Endophytic, rhizospheric and phyllopsheric) from cereal crops and soil microbiomes and their biotechnological applications in agriculture

Summary: The Himalayas are one of the most mystical, yet least studied terrains of the world. One of Earth’s greatest multifaceted and diverse montane ecosystems is also one of the thirty-four global biodiversity hotspots of the world. These are supposed to have been uplifted about 60-70 million years ago and support, distinct environments, physiography, a variety of orogeny and great biological diversity (plants, animals and microbes). Microbes are the pioneer colonizer of Himalayas that are involved in various bio-geological cycles and plays various significant roles. In the present study, total 588 plant associated microbial microbes were isolated from Amaranthus, barley, buckwheat, maize, millets, oats, wheat. All isolates were screened for multifarious plant growth promoting traits such as nitrogen fixation; phosphorus, potassium, zinc solubilization; production of siderophores, indole-3-acetic acid, ammonia, hydrogen cyanide, hydrolytic enzymes and 1-aminocyclopropane-1-carboxylate. The efficient strains were identified by 16S and 18S rRNA sequencing as Acinetobacter guillouiae, A. calcoaceticus, A. variabilis, Chryseobacterium arthrosphaerae, Citrobacter sp., Enterobacter ludwigii, Erwinia sp., E. persicina, E. rhapontici, Ewingella americana, Penicillium sp. Pantoea agglomerans, Pseudomonas sp., P. aeruginosa P. gessardii, P. panacis, P. libanensis, P. brenneri, P. extremorientalis, P. fluorescens, P. azotoformans, Rahnella sp., R. aquatilis, Stenotrophomonas rhizophila, Streptomyces laurentii, and Serratia marcescens. The efficient microbial strains when inoculated in wheat (under in vitro and in vivo conditions), foxtail millet sorghum (under in vitro conditions) and maize were improving the growth parameters such as shoot length, root length, fresh biomass and dry biomass of plants. The improvement in physiological parameters including accumulation of proline, glycine betaine, total soluble sugars, carotenoids, anthocyanin, increased chlorophyll content, e and Zn content, protein, and total protein content and antioxidant enzyme activities and decreased lipid peroxidation was also observed.

Publications: 9

2. Development of microbial consortium of NPK

Objectives: Isolation and enumeration of plant microbiomes from cereal crops growing in hilly regions. Screening of microbial isolates for NPK plant growth promoting attributes. Molecular characterization of selected potential plant growth promoting isolates. Bioformulations of liquid base microbial consortium. Validation of microbial consortium for different crops growing in hilly regions

Summary: Bioformulations of beneficial PGP microbes as biofertilizer and biopesticides are known from many years. Beneficial microbes exhibiting multifarious plant growth promoting traits as bioinoculants were used as singly or as consortium (mixture of two or more microbes). The majority of available bioformulations developed ever since was of single species. Particularly from the last decade, microbial bioformulations were developed containing two or more microbial species as combining the microbes gives combined benefits. In the present investigation, a total of 392 bacterial and 21 fungal isolates were obtained from wheat, oats, maize, millet, amaranth and buckwheat. All isolates were screened for plant growth promoting attributes such as nitrogen fixation and solubilization of P, and K and found 46 were nitrogen fixer, 92 and 58 were solubilizer of P and K, respectively. Out of these strains, 3 were having capability to fix nitrogen and solubilize P and K, whereas, 36 strains were able to solubilize both P and K mineral; 12 were fixing nitrogen and solubilizing P and 9 were fixing nitrogen and solubilize K. Potential microbe with nitrogen fixation, P and K solubilizing attributes were molecularly identified as Acinetobacter junii, Bacillus sp., B. mycoides, B. megaterium, Citrobacter sp. Erwinia sp., Chryseobacterium arthrosphaerae Pseudomonas brenneri, P. gessardii, P. fluorescens and Serratia marcescens,. The potential plant growth promoting microbe was selected for the liquid base bioformulations. The efficient microbial consortium of NPK when inoculated in wheat, pea, barley and pearl millet were improving the growth parameters as well as physiological parameters such as shoot length, root length, fresh biomass and dry biomass; content of carotenoids, chlorophyll, total soluble sugars, phenolics and flavonoids of plants.

Publications: 7

3. Development of microbial consortium of mineral solubilizing microbes

Objectives: Isolation and characterization of microbes associated with crops growing in hilly regions, Screening of isolated microbes for mineral solubilizing attributes, Molecular characterization of selected mineral solubilizing microbes, Development of microbial consortium of mineral solubilizing microbes for plant growth and enhanced crop production, Evaluation and validation of mineral solubilizing microbes for plant growth promotion under controlled and natural conditions

Summary: Minerals solubilizing microbes could use for stimulating growth and productivity of plant crop and could be used as a bioinoculants. Total 10 samples of plant samples from cereal crops (wheat, oats, maize, millet, amaranth and buckwheat) were collected from experimental farms of Eternal University, Baru Sahib and epiphytic, endophytic and rhizospheric microbes were isolated using eleven different selective and complex growth media. A total of 431 bacterial isolates were obtained from wheat, oats, maize, millet, amaranth and buckwheat. Isolated microbes were characterized for their morphological and cultural characteristics. All isolates were screened for mineral solubilizing microbes such as phosphorus, potassium, zinc and selenium and other plant growth promoting attributes like fixation of nitrogen, and production of siderophores, ammonia, indole-3- acetic acid, 1-aminocyclopropane-1-carboxylic acid deaminase activity and hydrogen cyanide. The efficient mineral solubilizing microbes were identified by 16S rRNA gene sequencing as Acinetobacter guillouiae, Bacillus sp., B. mycoides, B. horikoshii, Brevibacillus brevis, Erwinia rhapontici, E. persicina, Klebsiella pneumoniae, Pseudomonas gessardi, P. extremorientalis, P. trivalis, Rahnella aquatilis Serratia marcescens, and Stenotrophomonas rhizophila,. The efficient mineral solubilizing microbial consortium when inoculated in wheat, oats, maize, Amaranthus (under in vitro and in vivo conditions), capsicum, garlic and chilli (under in vitro conditions) were improving the growth parameters such as shoot length, root length, fresh weight and dry biomass of plants. The physiological parameters such as chlorophyll, carotenoids, total sugar, phenolics and flavonoids contents were also improved.

Publications: 3

Previous Reseach

Title: Studies of methylotrophic community from the phyllosphere and rhizosphere of tropical crop plants

M.Sc. (Microbial Biotechnology)- [January-May, 2009].

Summary: The diversity analysis of methylotrophic bacteria from tropical crop plants was investigated from different location at Mau U.P, in India using structural and functional approaches were considered. A total of 93 methylotrophic bacteria were isolated using ammonium minerals salt medium and all the isolated were screen for diverse plant growth promoting attributes. The selected methylotrophs were identified using molecular and biochemical analysis. The functional gene, coding methanol dehydrogenase (key enzyme of methanol oxidizing bacteria) coded by maxF locus has been used as genus specific locus for restricting the sub population of these Methylotrophs. The PGP activities shown by culture were HCN production, ammonia production, siderophores production and phosphorus solubilization. The methylotrophic bacterial communities could be utilize for plant growth promotion.

Publications: 3 (Research- 1; Review-1; Book chapter-1)

Title: “Bacterial diversity of cold deserts and mining of genes for low temperature tolerance”

Ph.D. (Microbial Biotechnology)- 2010-2016.

Summary: Cold deserts represent ice covered regions which are habitats of psychrophilic or psychrotrophic microbes. Indian Himalayas are hot spots for psychrophilic microorganisms whose biological attributes are not well characterized. In present investigation a total 325 psychrotrophic bacteria from Leh Ladakh and 232 bacteria from Rohtang Pass region were isolated. All the 557 bacteria from ten different sites were characterized employing 16S rDNA-Amplified Ribosomal DNA Restriction Analysis with three restriction endonucleases Alu I, Msp I and Hae III, which led to formation of different groups for the different sites at >95 % similarity index, adding up to 175 groups for Leh Ladakh and 169 groups for Rohtang Pass regions. Phylogenetic analysis based on 16S rRNA gene sequencing led to the identification of 175 bacteria belongs to phyla, Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes. Based on their optimal temperature for growth, bacteria were grouped as psychrophilic, psychrotrophic and psychrotolerant. The representative strains from each cluster were screened for cold active enzyme activities Amylase, β-glucosidase, pectinase, protease, cellulase, xylanase, β-galactosidase, laccase, chitinase and lipase activity. The isolates from each cluster were screened for their plant growth promoting attributes included production of ammonia, HCN, gibberellic acid, IAA and siderophore; solubilization of phosphorus and activity of ACC deaminase. In vitro antifungal activity assay was performed against Rhizoctonia solani and Macrophomina phaseolina. The selected isolates were screened for anti-freezing compounds productions. Cold adapted microorganisms may serve as inoculants for crops growing under cold climatic conditions.

Publications: 12 (Research-05, Review-02, Chapter-05)

Title: Diversity analysis of Bacillus and other predominant genera in extreme environments and its utilization in Agriculture

I have work as Senior Research Fellow (SRF) in NAIP project on “Diversity analysis of Bacillus and other predominant genera in extreme environments and its utilization in Agriculture” (2009-2012) with PI-Dr. A.K. Saxena, FNAAS, FIAMS, Head Division of Microbiology, IARI, New Delhi-110012

Summary:

Microbes are an essential component of biological diversity, without which there can be no sustainable ecosystems. They make up most of the biodiversity on earth, and it is estimated that fifty to ninety percent of the living biomass on the earth and ocean is microbial consortia and also provide a major source of genetic information for molecular biology and biotechnology. Microorganisms perform several processes like cycling of nutrients and degradation of various compounds. Since long, microbes have been used as a biofertilizers to stimulate plant growth, bio-control agent to manage insect pest and soil-borne pathogens and as a natural accelerator for decomposition of organic matter and rapid nutrient mobilization to plants. From past few decades, it is realized that the amount and quality of the world’s natural habitat harboring millions of microorganisms are disappearing rapidly and only the deepest parts of the ocean appear to have maintained some degree of pristine quality. Bacillus species predominate in most of the environments and are used as plant growth promoting rhizobacteria as well as potent biocontrol agents. The nutritional versatility of Bacillus allows them to use a range of carbon sources. They are extremely competitive organisms with remarkable resistance to desiccation and starvation. However, species isolated from extreme conditions have developed acclimation proteins allowing them to sustain life under extreme conditions of salinity, drought, high or low temperatures and acidity. Identification of new osmolytes and the relevant genes can be a boon to Indian agriculture as these genes could be utilized to develop transgenics tolerant to abiotic stress. Advances in biotechnology have produced improved prospects for developing new Bt insecticides and an ability to place Bt toxins within crop plants in a variety of ways. Indigenous strains of B. thuringiensis have not been exploited for allele mining and to develop variants of cry genes that codes for more virulent protein. The native strains of Bt needs to be screened for cry genes, sequenced and to clone them in suitable vectors for their efficient utilisation in transgenic programme of the country. Since the diversity of Bacillus and Bacillus derived genera in the country is not well known, there is an ample opportunity to explore or screen Bacillus strains for insecticidal properties. The work in this project deals with the screening of different isolates that may lead to identification of new species or strains that have insecticidal attribute.

Title: Role of Archaebacteria in Alleviation of Salinity and Moisture Stress in Plants

I have work as Senior Research Fellow (SRF) in NFBSFARA project on “Role of Archaebacteria in Alleviation of Salinity and Moisture Stress in Plants” (2012-2015) with PI-Dr. A.K. Saxena, FNAAS, FIAMS, Head Division of Microbiology, IARI, New Delhi-110012

Summary:

Archaebacteria (also referred to as archaea), known to inhabit extreme environments; have never been studied with perspectives to understand their interactions with eubacteria and to sustain vegetation in extremes of salinity, moisture stress and temperature. This is probably because culturing of archaea till early 2000 was a tedious task. The achievements of the project will contribute to the scientific advancement and/or technological advancement in the following ways:

• The database of archaea isolated from the rhizosphere of plants growing in extreme environments of the country does not exist. The project would lead to the development of database and baseline information on archaebacteria isolated from extreme environments of the country.

• The archaebacteria, eubacteria and fungi isolated in different seasons would lead to selection of microbes tolerant to fluctuating salinity and moisture regimes and could be a resource to be utilized for alleviation of salinity and moisture stress in plants.

• The role of archaea as plant growth promoting bacteria or nutrient provider would be understood.

• Development of technologies to sustain plants in extreme saline and moisture deficit regimes through rhizospheric engineering with archaebacteria, eubacteria and/or fungi.

• Demonstration of archaebacteria, eubacteria and fungi as nutrient providers and mobilizers to sustain vegetation in extreme saline and moisture deficit regimes.

• Application of microbial bio-inoculants for various crop plants grown in saline soils to alleviate salinity stress.

• The potential archaeal strains can be exploited for allele mining and identification of genes for developing crop plants tolerant to abiotic stress.