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Kalia R.K.,Center for Plant Biotechnology | Kalia R.K.,University of California at Riverside | Rai M.K.,Center for Plant Biotechnology | Kalia S.,NRC on Plant Biotechnology | And 2 more authors.
Euphytica | Year: 2011

In recent years, molecular markers have been utilized for a variety of applications including examination of genetic relationships between individuals, mapping of useful genes, construction of linkage maps, marker assisted selections and backcrosses, population genetics and phylogenetic studies. Among the available molecular markers, microsatellites or simple sequence repeats (SSRs) which are tandem repeats of one to six nucleotide long DNA motifs, have gained considerable importance in plant genetics and breeding owing to many desirable genetic attributes including hypervariability, multiallelic nature, codominant inheritance, reproducibility, relative abundance, extensive genome coverage including organellar genomes, chromosome specific location and amenability to automation and high throughput genotyping. High degree of allelic variation revealed by microsatellite markers results from variation in number of repeat-motifs at a locus caused by replication slippage and/or unequal crossing-over during meiosis. In spite of limited understanding of the functions of the SSR motifs within the plant genes, SSRs are being widely utilized in plant genome analysis. Microsatellites can be developed directly from genomic DNA libraries or from libraries enriched for specific microsatellites. Alternatively, microsatellites can also be found by searching public databases such as GenBank and EMBL or through cross-species transferability. At present, EST databases are an important source of candidate genes, as these can generate markers directly associated with a trait of interest and may be transferable in close relative genera. A large number of SSR based techniques have been developed and a quantum of literature has accumulated regarding the applicability of SSRs in plant genetics and genomics. In this review we discuss the recent developments (last 4-5 years) made in plant genetics using SSR markers. © 2010 Springer Science+Business Media B.V. Source


Mittal A.,Kurukshetra University | Nagar S.,Kurukshetra University | Kansal R.,NRC on Plant Biotechnology | Gupta V.K.,Kurukshetra University
Indian Journal of Agricultural Biochemistry | Year: 2013

This work investigates the factors affecting the extraction of trypsin inhibitor from kidney bean (Phaseolus vulgaris) seeds. Trypsin inhibitor extracted with 0.1% NaCl rendered a higher recovery than other solvents. The extraction was also affected by pH and extraction time. It took 5 h for complete recovery of trypsin inhibitor activity. Response surface methodology was used to study the effect of temperature and pH with respect to time on the extraction of trypsin inhibitor. Statistical optimization studies revealed that trypsin inhibitor activity from kidney bean flour was stable to 60 °C for 52 min at neutral and acidic pH and labile under strong alkaline conditions. © 2013, Indian Society of Agricultural Biochemists. All rights reserved. Source


Lochab S.,Guru Gobind Singh Indraprastha University | Kumar P.A.,NRC on Plant Biotechnology | Raghuram N.,Guru Gobind Singh Indraprastha University
Archives of Microbiology | Year: 2014

The nitrate assimilation pathway and its regulation in the high-protein neutraceutical cyanobacterium, Arthrospira (Spirulina), were studied. A complete characterization of the genes of the nitrate uptake and assimilatory pathway in Arthrospira platensis strain PCC 7345 was done including cloning, sequencing, phylogenetic analysis and expression studies. Genomic localization studies revealed that their clustering is different from the operons known in other cyanobacteria; only nrtP and narB are organized together, while nirA, glnA and gltS exist in separate genomic locations. The presence of both types of nitrate transporters (nrtP/ABC types) in A. platensis is rare, as their occurrence is usually specific to marine and freshwater microorganisms, respectively. The positive effect of nitrate on transcript accumulation of narB, nirA and nrtP genes in N-depleted and N-restored cultures confirmed nitrate induction, which is abolished by the addition of ammonium ions into the medium. Gene expression studies in response to nitrate, nitrite, ammonium and glutamine provided the first evidence of differential regulation of multiple genes of nitrate assimilatory pathway in Arthrospira. © 2014 Springer-Verlag. Source


Mittal A.,Kurukshetra University | Kansal R.,NRC on Plant Biotechnology | Kalia V.,Indian Agricultural Research Institute | Tripathi M.,Indian Agricultural Research Institute | Gupta V.K.,Kurukshetra University
Acta Physiologiae Plantarum | Year: 2014

In the present study, trypsin inhibitor extracts of ten kidney bean seed (Phaseolus vulgaris) varieties exhibiting trypsin and gut trypsin-like protease inhibitor activity were tested on Helicoverpa armigera and Spodoptera litura. Trypsin inhibitor protein was isolated and purified using multi-step strategy with a recovery of ~15 % and purification fold by ~39.4. SDS-PAGE revealed a single band corresponding to molecular mass of ~15 kDa and inhibitory activity was confirmed by reverse zymogram analyses. The inhibitor retained its inhibitory activity over a broad range of pH (3-11), temperature (40-60 °C) and thermostability was promoted by casein, CaCl2, BSA and sucrose. The purified inhibitor inhibited bovine trypsin in 1:1 molar ratio. Kinetic studies showed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 1.85 μM. The purified trypsin inhibitor protein was further incorporated in the artificial diet and fed to second instar larvae. A maximum of 91.7 % inhibition was obtained in H. armigera, while it was moderate in S. litura (29 %) with slight varietal differences. The insect bioassay showed 40 and 22 % decrease in larval growth followed by 3 and 2 days delay in pupation of H. armigera and S. litura, respectively. Some of the adults emerged were deformed and not fully formed. Trypsin inhibitor protein was more effective against H. armigera as it showed 46.7 % mortality during larval growth period compared to S. litura (13.3 %). © 2013 Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków. Source


Durgesh K.,Indian Grassland And Fodder Research Institute | Durgesh K.,Indian Agricultural Research Institute | Raje R.S.,Indian Agricultural Research Institute | Singh A.K.,Indian Agricultural Research Institute | And 3 more authors.
Indian Journal of Genetics and Plant Breeding | Year: 2015

In the present study, genetic diversity among 73 genotypes comprising of cultivars and stable inter-specific derivatives from pigeonpea (Cajanus cajan x C. scarabaeoides) was assessed based on thirteen agro-morphological and quality traits and 42 genic microsatellite markers. Cluster analysis based on Tocher’s methods of pooled fielddata of agromorphological traits from all three environment and two locations grouped pigeonpea genotypes into nine clusters. Genetic divergence was mainly contributed by days to 50% flowering (30.14%) followed by100 seed weight (20.78%) and protein content (12.56%). The range of polymorphic information content (PIC) for microsatellites was 0.055- 0.695. A total of 104 alleles were amplified with 42 SSR markers with an average of 2.48 alleles per locus.Ten alleles were unique to inter-specific derivatives. Based on Shannon’s information index (I), ASSR 17 was the most informative locus for all the genotypes. Two loci, ASSR114 and ASSR317 were polymorphic only among the interspecific derivatives. Cophenetic correlations were statistically significant for phenograms based on Euclidean distances and average Manhattan distance analysis.The first three principal coordinates explained 60.17% and 57.22% of the accumulated variation in case of agromorphological traits and microsatellite data respectively. The information accrued is potential for broadening the genetic base in pigeonpea. © 2015, Indian Society of Genetics and Plant Breeding. All rights reserved. Source

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