Papers by Jasdeep Padaria
Canadian Journal of Microbiology, 2015
Vegetative insecticidal proteins (Vip) represent the second generation of insecticidal proteins p... more Vegetative insecticidal proteins (Vip) represent the second generation of insecticidal proteins produced by Bacillus thuringiensis (Bt) during the vegetative growth stage of growth. Bt-based biopesticides are recognized as viable alternatives to chemical insecticides; the latter cause environmental pollution and lead to the emergence of pest resistance. To perform a systematic study of vip genes encoding toxic proteins, a total of 30 soil samples were collected from diverse locations of Kashmir valley, India, and characterized by molecular and analytical methods. Eighty-six colonies showing Bacillus-like morphology were selected. Scanning electron microscopy observations confirmed the presence of different crystal shapes, and PCR analysis of insecticidal genes revealed a predominance of the lepidopteran-specific vip3 (43.18%) gene followed by coleopteran-specific vip1 (22.72%) and vip2 (15.90%) genes in the isolates tested. Multi-alignment of the deduced amino acid sequences revealed that vip3 sequences were highly conserved, whereas vip1 and vip2 showed adequate differences in amino acid sequences compared with already reported sequences. Screening for toxicity against Helicoverpa armigera larvae was performed using partially purified soluble fractions containing Vip3A protein. The mortality levels observed ranged between 70% and 96.6% in the isolates. The LC50 values of 2 of the native isolates, JK37 and JK88, against H. armigera were found to be on par with that of Bt subsp. kurstaki HD1, suggesting that these isolates could be developed as effective biopesticides against H. armigera.
Physiology and Molecular Biology of Plants, 2015
Pearl millet (Pennisetum glaucum) is an important cereal of traditional farming systems that has ... more Pearl millet (Pennisetum glaucum) is an important cereal of traditional farming systems that has the natural ability to withstand various abiotic stresses. The present study aims at the identification and validation of major differentially expressed genes in response to drought stress in P. glaucum by Suppression Subtractive Hybridization (SSH) analysis. Twenty-two days old seedlings of P. glaucum cultivar PPMI741 were subjected to drought stress by treatment of 30 % Polyethylene glycol for different time periods 30 min (T1), 2 h (T2), 4 h (T3), 8 h (T4), 16 h (T5), 24 h (T6) and 48 h (T7) respectively, monitored by examining the RWC of seedlings. Total RNA was isolated to construct drought responsive subtractive cDNA library through SSH, sequenced to identify the differentially expressed genes in response to drought stress and validated by qRT-PCR.745 ESTs were assembled into a collection of 299 unigenes having 52 contigs and 247 singletons. All 745 ESTs were submitted to ENA-EMBL databases (Accession no. HG516611- HG517355). After analysis, 10 differentially expressed genes were validated namely Abscisic stress ripening protein, Ascorbate peroxidase, Inosine-5'-monophosphate dehydrogenase, Putative beta-1, 3-glucanase, Glyoxalase, Rab7, Aspartic proteinase Oryzasin, DnaJ-like protein and Calmodulin-like protein by qRT-PCR. The identified ESTs reveal a major portion of the stress responsive transcriptome that may prove to be a vent to unravel molecular basis underlying tolerance of pearl millet (Pennisetum glaucum) to drought stress. These genes could be utilized for transgenic breeding or transferred to crop plants through marker assisted selection for the development of better drought resistant cultivars having enhanced adaptability to survive harsh environmental conditions.
Pearl millet (Pennisetum glaucum L. R. Br.) is an important cereal crop grown mainly in the arid ... more Pearl millet (Pennisetum glaucum L. R. Br.) is an important cereal crop grown mainly in the arid and semi-arid regions
of India known to possess the natural ability to withstand thermal stress. To elucidate the molecular basis of high
temperature response in pearl millet, 12 days old seedlings of P. glaucum cv. 841A were subjected to heat stress at 46C for
different time durations ( 30 min, 2, 4, 8, 12 and 24 h) and a forward subtractive cDNA library was constructed from pooled
RNA of heat stressed seedlings. A total of 331 high quality Expressed Sequence Tags (ESTs) were obtained from randomly
selected 1050 clones. Sequences were assembled into 103 unique sequences consisting of 37 contigs and 66 singletons. Of
these, 92 unique sequences were submitted to NCBI dbEST database. Gene Ontology through RGAP data base and
BLASTx analysis revealed that about 18% of the ESTs showed homology to genes for “response to abiotic and biotic
stimulus”. About 2% of the ESTs showed no homology with genes in dbEST, indicating the presence of uncharacterized
candidate genes involved in heat stress response in P. glaucum. Differential expression of selected genes (hsp101 and CRT)
from the SSH library were validated by qRT-PCR analysis. The ESTs thus generated are a rich source of heat stress
responsive genes, which can be utilized in improving thermotolerance of other food crops.
Bacteria capable of degrading the sulfonated azo dye Red HE7B were isolated from textile mill eff... more Bacteria capable of degrading the sulfonated azo dye Red HE7B were isolated from textile mill effluent contaminated soil. The most efficient isolate was identified as Bacillus sp. Azo1 and the isolate could successfully decolorize up to 89 % of the dye. The decolorized cultural extract analyzed by HPLC confirmed degradation. Enzymatic analysis showed twofold and fourfold increase in the activity of azoreductase and laccase enzymes, respectively, indicating involvement of both reductive and oxidative enzymes in biodegradation of Red HE7B. Degraded products which were identified by GC/MS analysis included various metabolites like 8-nitroso 1-naphthol, 2-diazonium naphthalene. Mono azo dye intermediate was initially generated from the parent molecule. This mono azo dye was further degraded by the organism, into additional products, depending on the site of cleavage of R-N=N-R molecule. Based on the degradation products identified, three different pathways have been proposed. The mechanism of degradation in two of these pathways is different from that of the previously reported pathway for azo dye degradation. This is the first report of a microbial isolate following multiple pathways for azo dye degradation. Azo dye Red HE7B was observed to be phytotoxic, leading to decrease in root development, shoot length and seedling fresh weight. However, after biotreatment the resulting degradation products were non-phytotoxic.
Journal of environmental biology / Academy of Environmental Biology, India, 2014
Bio-fuel produced from ethanol is economically and environmentally advantageous in context of cha... more Bio-fuel produced from ethanol is economically and environmentally advantageous in context of changing global climate. A large number of microorganisms are capable of cellulase production but most of them cannot be utilized commercially due to their low activity. In the present study, an effiecient cellulose degrading strain of Bacillus pumilus was obtained after thorough screening for the production of extracellular cellulases. Out of a total of 144 microbes isolated from soils of Darjeeling hills of India, nineteen were found to be cellulose degrader under in vitro conditions as observed by clearing zone on CMC - agar plates. Isolate #35 had high cellulolytic activity as observed by a clearing zone of 26.83 mm diameter formed on CMC - agar plate. The isolate was characterized and identified as Bacillus pumilus. The isolate was submitted to National Agriculturally Important Microbial Culture Collection (NAIMCC), NBAIM, Mau with Accession number NAIMCC-B-01415. Transposon (Tn5) muta...
Journal of environmental biology / Academy of Environmental Biology, India, 2013
Transposon Tn5 induced, four Mesorhizobium ciceri auxotroph were isolated and characterized. Unli... more Transposon Tn5 induced, four Mesorhizobium ciceri auxotroph were isolated and characterized. Unlike its wild type parent (TL 620), all four mutants were found defective for amino acid and pyramiding biosynthesis. The auxotroph mutants were characterized and found TL130 as cytosine and uracil, TL 196 for guanine, cytocine, uracil and riboflavin, TL 141 as serine and TL 38 as argentine defective. Symbiotic characterization of these mutants revealed phenotypic deformities and deficiencies in biological nitrogen fixation. All the four auxotrophic mutants were characterized as nod+/fix+ nature with reduced nitrogenase activity of 42.2, 26.3 and 17.13% respectively as compared to the wild type which is further supported by sub cellular examination of the nodules section by TEM study.
Applied & Translational Genomics, 2014
A set of thermotolerant strains isolated from hot springs of Manikaran and Bakreshwar (India) wer... more A set of thermotolerant strains isolated from hot springs of Manikaran and Bakreshwar (India) were selected with an aim to isolate dnak gene which encodes DnaK protein. The gene dnaK along with its flanking region was successfully amplified from 5 different strains (4 from Bakreshwar and one from Manikaran). Restriction fragment length polymorphism (RFLP) revealed that amplicons were almost identical in sequence. The dnak gene from one representative, Bacillus pumilus strain B3 isolated from Bakreshwar hot springs was successfully cloned and sequenced. The dnaK gene was flanked by gene grpE on one side. The dnaK gene was 1842 bp in length encoding a polypeptide of 613 amino acid residues. Calculated molecular weight and pI of the protein were 66,128.36 Da and 4.72 respectively. The deduced amino acid sequence of this gene shared high sequence homology with other DnaK proteins and its homologue Hsp 70 from other microorganisms, but possessed 36 substitutions and two insertions, as compared to DnaK protein of Bacillus subtilis. The dnaK gene of B. pumilus was successfully expressed in Escherichia coli BL 21 (DE3) using pET expression systems. Heterologous expression of dnaK of B. pumilus in E. coli BL 21 (DE3) allowed for the growth of E. coli up to 50°C and survival up to 60°C for 16 h, suggesting that dnak from B. pumilus imparts tolerance to host cells under high temperature. This novel gene can be an important component for possible utilization in abiotic stress management of plants.
The diversity of culturable, aerobic and heterotrophic
Bacillus and Bacillus-derived genera (BBD... more The diversity of culturable, aerobic and heterotrophic
Bacillus and Bacillus-derived genera (BBDG) was investigated
in various extreme environments (including thermal
springs, cold deserts, mangroves, salt lakes, arid regions, salt
pans and acidic soils) of India. Heat treatment followed by
enrichment in different media led to a total of 893 bacterial
isolates. Amplified ribosomal DNA restriction analysis
(ARDRA) using three restriction enzymes AluI, MspI and
HaeIII led to the clustering of these isolates into 12–74 groups
for the different sites at 75 % similarity index, adding up to
559 groups. Phylogenetic analysis based on 16S rRNA gene
sequencing led to the identification of 392 bacilli, grouped in
two families, Bacillaceae (89.03 %) and Paenibacillaceae
(10.97 %), and included 13 different genera with 75 distinct
species. It was found that among the thirteen genera, nine
(Bacillus, Halobacillus, Lysinibacillus, Oceanobacillus, Pontibacillus, Salinibaci l lus, Sediminibacillus,
Thalassobacillus and Virgibacillus) belonged to Bacillaceae
and four (Ammoniphilus, Aneurinibacillus, Brevibacillus and
Paenibacillus) belonged to Paenibacillaceae. Novel isolates
tolerant to low and high pH and temperature, salt and low
moisture were identified. The major outcome of the present
investigation was the identification of niche-specific species
and also the ubiquitous presence of selected species of BBDG,
which illustrate the diversity and pervasive nature of BBDG in
extreme environments.
Environmental Deterioration and Human Health, 2013
BMC research notes, 2014
Heat stress leads to accelerated production of reactive oxygen species (ROS) which causes a huge ... more Heat stress leads to accelerated production of reactive oxygen species (ROS) which causes a huge amount of oxidative damage to the cellular components of plants. A large number of heat stress related genes as HSPs, catalases, peroxidases are overexpressed at the time of stress. A potent stress responsive gene peroxisomal ascorbate peroxidase (TapAPX) obtained from heat stress (42 °C) responsive subtractive cDNA library from a thermo tolerant wheat cv. Raj3765 at anthesis stage was cloned, characterized and its role was validated under heat stress by proteomics and in-silico studies. In the present study we report the characterization at molecular and in-silico level of peroxisomal TapAPX gene isolated from heat tolerant wheat cultivar of India. qPCR studies of TapAPX gene displayed up to 203 fold level of expression at 42 °C heat stress exposure. A full length cDNA of 876 bp obtained by RACE deduced a protein of 292 amino acid residues which gives a complete 3D structure of pAPX by ...
Current Microbiology, 2014
Bacteria capable of degrading the sulfonated azo dye Red HE7B were isolated from textile mill eff... more Bacteria capable of degrading the sulfonated azo dye Red HE7B were isolated from textile mill effluent contaminated soil. The most efficient isolate was identified as Bacillus sp. Azo1 and the isolate could successfully decolorize up to 89% of the dye. The decolorized cultural extract analyzed by HPLC confirmed degradation. Enzymatic analysis showed twofold and fourfold increase in the activity of azoreductase and laccase enzymes, respectively, indicating involvement of both reductive and oxidative enzymes in biodegradation of Red HE7B. Degraded products which were identified by GC/MS analysis included various metabolites like 8-nitroso 1-naphthol, 2-diazonium naphthalene. Mono azo dye intermediate was initially generated from the parent molecule. This mono azo dye was further degraded by the organism, into additional products, depending on the site of cleavage of R-N=N-R molecule. Based on the degradation products identified, three different pathways have been proposed. The mechanism of degradation in two of these pathways is different from that of the previously reported pathway for azo dye degradation. This is the first report of a microbial isolate following multiple pathways for azo dye degradation. Azo dye Red HE7B was observed to be phytotoxic, leading to decrease in root development, shoot length and seedling fresh weight. However, after biotreatment the resulting degradation products were non-phytotoxic.
SUMMARY
High temperature adversely affects the growth, yield, and quality of crops. Prosopis cine... more SUMMARY
High temperature adversely affects the growth, yield, and quality of crops. Prosopis cineraria, an indigenous plant in
India, is highly tolerant of environmental stresses. In the present study, an expressed sequence tag (EST) library was
constructed using pooled RNAs from 1-month-old P. cineraria seedlings subjected to heat stress at 48ºC for between
15 min to 48 h. A total of 84 unigenes (20 contigs and 64 singletons) were generated from the cluster assembly of 132
good ESTs. Of these, nine unigenes did not show any homology to sequences in the NCBI database. A BLASTX
comparison of the remaining 75 unigenes revealed significant similarities to known genes.The 84 ESTs were classified
into 14 functional categories, of which 13.75% were in the stress-response category, including approx. 10% that were
for heat-shock proteins. The heat stress-induced ESTs reported here are the first for P. cineraria. A phylogenetic
analysis based on the complete coding sequence of the Pchsp17.9 gene showed a close relationship to hsp17.9 of
Acacia mangium. Levels of expression of genes related to hsp chaperones were analysed by reverse transcriptionquantitative
PCR (RT-qPCR) in 1-month-old heat-stressed seedlings of P. cineraria and were found to be highly upregulated
Annals of Plant Protection Sciences 20 (1), 195-200
Plant Omics 6 (2), 150, 2013
Journal of Applied Horticulture 1 (1), 38-40, 1999
PRECISION FARMING IN HORTICULTURE
Page 248. GENETIC ENGINEERING: A STRATEGIC APPROACH FOR HI-TECH HORTICULTURE Jasdeep Chatrath Pad... more Page 248. GENETIC ENGINEERING: A STRATEGIC APPROACH FOR HI-TECH HORTICULTURE Jasdeep Chatrath Padaria1 and Ramesh Chandra2 Horticultural crops constitute a significant component of total agricultural produce in India. ...
Management of Microbial Resources in the …, 2013
ABSTRACT Microbes are known to play an important role in numerous metabolic processes like nutrie... more ABSTRACT Microbes are known to play an important role in numerous metabolic processes like nutrient cycling, environmental detoxification, production of antibiotics, vitamins, industrial enzymes etc. Therefore it is important to efficiently harness and utilize the biologically important properties of microbes and their products to tackle the ever growing challenges of food security, healthcare and environmental pollution. A complete knowledge of these microbes with respect to the role played by them in ecosystem function is essential to fully exploit them for the benefit of mankind. Unfortunately only 4–5% of the microbes have been explored so far whereas the rest ≈95% is are still un-culturable. A number of uncertainties still exist with respect to the microbial diversity as knowledge regarding the number of species of microorganisms that exist, their distribution, stability in the environment and the important roles played by them are lacking to a greater extent. Microbial diversity is an unseen global resource that deserves to be conserved and utilized judiciously. Microbial resource centers play an important role in this regard as they act as living libraries holding microorganisms. The primary function of these centers is to collect, maintain and distribute microbial strains and/or their products to researchers and industrialists all over the world. The role of microbial Culture Collections with respect to conservation and propagation of microbial resources and the difficulties and uncertainties of conservation faced are discussed.
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Papers by Jasdeep Padaria
of India known to possess the natural ability to withstand thermal stress. To elucidate the molecular basis of high
temperature response in pearl millet, 12 days old seedlings of P. glaucum cv. 841A were subjected to heat stress at 46C for
different time durations ( 30 min, 2, 4, 8, 12 and 24 h) and a forward subtractive cDNA library was constructed from pooled
RNA of heat stressed seedlings. A total of 331 high quality Expressed Sequence Tags (ESTs) were obtained from randomly
selected 1050 clones. Sequences were assembled into 103 unique sequences consisting of 37 contigs and 66 singletons. Of
these, 92 unique sequences were submitted to NCBI dbEST database. Gene Ontology through RGAP data base and
BLASTx analysis revealed that about 18% of the ESTs showed homology to genes for “response to abiotic and biotic
stimulus”. About 2% of the ESTs showed no homology with genes in dbEST, indicating the presence of uncharacterized
candidate genes involved in heat stress response in P. glaucum. Differential expression of selected genes (hsp101 and CRT)
from the SSH library were validated by qRT-PCR analysis. The ESTs thus generated are a rich source of heat stress
responsive genes, which can be utilized in improving thermotolerance of other food crops.
Bacillus and Bacillus-derived genera (BBDG) was investigated
in various extreme environments (including thermal
springs, cold deserts, mangroves, salt lakes, arid regions, salt
pans and acidic soils) of India. Heat treatment followed by
enrichment in different media led to a total of 893 bacterial
isolates. Amplified ribosomal DNA restriction analysis
(ARDRA) using three restriction enzymes AluI, MspI and
HaeIII led to the clustering of these isolates into 12–74 groups
for the different sites at 75 % similarity index, adding up to
559 groups. Phylogenetic analysis based on 16S rRNA gene
sequencing led to the identification of 392 bacilli, grouped in
two families, Bacillaceae (89.03 %) and Paenibacillaceae
(10.97 %), and included 13 different genera with 75 distinct
species. It was found that among the thirteen genera, nine
(Bacillus, Halobacillus, Lysinibacillus, Oceanobacillus, Pontibacillus, Salinibaci l lus, Sediminibacillus,
Thalassobacillus and Virgibacillus) belonged to Bacillaceae
and four (Ammoniphilus, Aneurinibacillus, Brevibacillus and
Paenibacillus) belonged to Paenibacillaceae. Novel isolates
tolerant to low and high pH and temperature, salt and low
moisture were identified. The major outcome of the present
investigation was the identification of niche-specific species
and also the ubiquitous presence of selected species of BBDG,
which illustrate the diversity and pervasive nature of BBDG in
extreme environments.
High temperature adversely affects the growth, yield, and quality of crops. Prosopis cineraria, an indigenous plant in
India, is highly tolerant of environmental stresses. In the present study, an expressed sequence tag (EST) library was
constructed using pooled RNAs from 1-month-old P. cineraria seedlings subjected to heat stress at 48ºC for between
15 min to 48 h. A total of 84 unigenes (20 contigs and 64 singletons) were generated from the cluster assembly of 132
good ESTs. Of these, nine unigenes did not show any homology to sequences in the NCBI database. A BLASTX
comparison of the remaining 75 unigenes revealed significant similarities to known genes.The 84 ESTs were classified
into 14 functional categories, of which 13.75% were in the stress-response category, including approx. 10% that were
for heat-shock proteins. The heat stress-induced ESTs reported here are the first for P. cineraria. A phylogenetic
analysis based on the complete coding sequence of the Pchsp17.9 gene showed a close relationship to hsp17.9 of
Acacia mangium. Levels of expression of genes related to hsp chaperones were analysed by reverse transcriptionquantitative
PCR (RT-qPCR) in 1-month-old heat-stressed seedlings of P. cineraria and were found to be highly upregulated
of India known to possess the natural ability to withstand thermal stress. To elucidate the molecular basis of high
temperature response in pearl millet, 12 days old seedlings of P. glaucum cv. 841A were subjected to heat stress at 46C for
different time durations ( 30 min, 2, 4, 8, 12 and 24 h) and a forward subtractive cDNA library was constructed from pooled
RNA of heat stressed seedlings. A total of 331 high quality Expressed Sequence Tags (ESTs) were obtained from randomly
selected 1050 clones. Sequences were assembled into 103 unique sequences consisting of 37 contigs and 66 singletons. Of
these, 92 unique sequences were submitted to NCBI dbEST database. Gene Ontology through RGAP data base and
BLASTx analysis revealed that about 18% of the ESTs showed homology to genes for “response to abiotic and biotic
stimulus”. About 2% of the ESTs showed no homology with genes in dbEST, indicating the presence of uncharacterized
candidate genes involved in heat stress response in P. glaucum. Differential expression of selected genes (hsp101 and CRT)
from the SSH library were validated by qRT-PCR analysis. The ESTs thus generated are a rich source of heat stress
responsive genes, which can be utilized in improving thermotolerance of other food crops.
Bacillus and Bacillus-derived genera (BBDG) was investigated
in various extreme environments (including thermal
springs, cold deserts, mangroves, salt lakes, arid regions, salt
pans and acidic soils) of India. Heat treatment followed by
enrichment in different media led to a total of 893 bacterial
isolates. Amplified ribosomal DNA restriction analysis
(ARDRA) using three restriction enzymes AluI, MspI and
HaeIII led to the clustering of these isolates into 12–74 groups
for the different sites at 75 % similarity index, adding up to
559 groups. Phylogenetic analysis based on 16S rRNA gene
sequencing led to the identification of 392 bacilli, grouped in
two families, Bacillaceae (89.03 %) and Paenibacillaceae
(10.97 %), and included 13 different genera with 75 distinct
species. It was found that among the thirteen genera, nine
(Bacillus, Halobacillus, Lysinibacillus, Oceanobacillus, Pontibacillus, Salinibaci l lus, Sediminibacillus,
Thalassobacillus and Virgibacillus) belonged to Bacillaceae
and four (Ammoniphilus, Aneurinibacillus, Brevibacillus and
Paenibacillus) belonged to Paenibacillaceae. Novel isolates
tolerant to low and high pH and temperature, salt and low
moisture were identified. The major outcome of the present
investigation was the identification of niche-specific species
and also the ubiquitous presence of selected species of BBDG,
which illustrate the diversity and pervasive nature of BBDG in
extreme environments.
High temperature adversely affects the growth, yield, and quality of crops. Prosopis cineraria, an indigenous plant in
India, is highly tolerant of environmental stresses. In the present study, an expressed sequence tag (EST) library was
constructed using pooled RNAs from 1-month-old P. cineraria seedlings subjected to heat stress at 48ºC for between
15 min to 48 h. A total of 84 unigenes (20 contigs and 64 singletons) were generated from the cluster assembly of 132
good ESTs. Of these, nine unigenes did not show any homology to sequences in the NCBI database. A BLASTX
comparison of the remaining 75 unigenes revealed significant similarities to known genes.The 84 ESTs were classified
into 14 functional categories, of which 13.75% were in the stress-response category, including approx. 10% that were
for heat-shock proteins. The heat stress-induced ESTs reported here are the first for P. cineraria. A phylogenetic
analysis based on the complete coding sequence of the Pchsp17.9 gene showed a close relationship to hsp17.9 of
Acacia mangium. Levels of expression of genes related to hsp chaperones were analysed by reverse transcriptionquantitative
PCR (RT-qPCR) in 1-month-old heat-stressed seedlings of P. cineraria and were found to be highly upregulated
With the advancement of recombinant DNA technology, the gene of interest for useful trait has been successfully incorporated into the genome of important crop plants or by altering the crop’s own genetic code, the function of the products coded by genes or change the way genes are expressed by switching on or off. But, Conflicting assessments and incomplete substantiation of the benefits, risks and limitations of transgenic crops have added to existing controversies. A major roadblock in the rapid adoption of transgenic crops is the prolonged process of regulatory approval. There is an urgent need for appropriate science based cost-effective and time-effective regulatory systems that are responsible and rigorous but not onerous for small and poor developing countries.