Browsing by Author "Kumar, Rajesh"
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ThesisItem Open Access Agro morphological and molecular characterization of local rice (Oryza sativa L.) landraces(Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur (Bihar), 2019) Sinha, Naincy; Kumar, RajeshThe present investigation was carried out at Rice Research Farm, Dr. Rajendra Prasad Central Agricultural University Pusa during kharif 2018 and molecular work in plant breeding and genetics laboratory in the month of Feb –April. It was done to assess the Agro-morphological characterization, genetic variability, Correlation, Path Coefficient analysis, genetic divergence (Cluster analysis), and molecular characterisation. The experimental material consisted of 23 rice landraces including check Sugandha. Analysis of variance indicated that the mean sum of squares due to treatments were significant for all fifteen quantitative trait. High estimates of PCV were observed for traits grain yield per plant, days to50% flowering, plant height, panicle length of main axis, panicle number per plant, length of leaf blade, width of leaf blade, 1000 grain weight, grain width, decorticated grain length, decorticated grain width, root volume. In present set of materials high heritability with high genetic advance as per cent of mean was recorded for panicle length of main axis, panicle number per plant, length of leaf blade, root volume, grain yield per plant. The highest magnitude of coefficient of variation was noted for grain yield per plant followed by panicle number per plant, panicle length of main axis, width of leaf blade, decorticated grain width, stem thickness. In the present investigation simple correlation coefficients among fifteen quantitative characters revealed that grain yield per plant had significant positive correlation with panicle number per plant. Path analysis revealed very high positive direct effect by days to 50% flowering, length of leaf blade,1000 grain weight, grain length, root volume and decorticated grain length and very high negative direct effect by days to maturity, plant height, stem thickness, panicle length of main axis, decorticated grain length on grain yield per plant. Hence, selection based on these characters would be more effective for yield improvement. The landraces in cluster D and cluster E due to maximum inter cluster distance between them, exhibited high degree of genetic diversity and thus may be utilized in future breeding programme for getting high yielding recombinants. Cluster D showed higher mean performance for most of the yield attributing characters therefore, selection of parents from this cluster would be effective. Grain yield per plant followed by 1000 grain weight, root volume, days to 50% flowering, length of leaf blade, grain width were major characters contributing towards genetic divergence. Scoring was done for 31 morphological DUS descriptors on rice as per the IIRR (formerly known as DRR) guidelines. Out of 31 characteristics, 6 were monomorphic, partially monomorphic was found in single trait, 6 were dimorphic and 18 were polymorphic. . The dimorphic and polymorphic characters showed their potential for genotype characterization. The landrace sathi-1 was observed to be more distinct, with the rarest morphological feature like no panicle exertion, black lemma and palea colour, early flowering and early maturity were observed. The landrace Parwa Pankhi with unique feature i.e. longest sterile lemma. Character decorticated grain aroma divided 23 landraces into scented and non- scented landraces. Scented landraces are Lalmati, Rajbhog, kankirbi, sugandha, Kasturi, Basmati-570, Marcha-1, Marcha-2, Tarori Basmati, CSR-30. The descriptors offering the most discrimination were lemma and palea colour, flag leaf: attitude at early and late observation. A panel of fifteen primers, namely RM 555, RM 5791, RM 566, RM28166, RM 70, RM 286, RM 319, RM 321, RM 324, RM 416, RM431, RM520, RM521, MRG 2805, and MRG2894 generated amplified products Altogether 81 allelic variants were detected among the twenty-three rice genotypes with an average of 3.52 alleles per primer. A total of 74 shared and 7 unique allelic variants were generated in the form of amplified products by using 15 primer pairs. Considerably the greater number of alleles had been detected in primers RM520, RM321, RM70, MRG2894, and MRG2805. RM566, RM520, RM324, RM321, RM70, MRG2894 generated considerably unique alleles rest the primer pairs generated shared alleles. The PIC values revealing allele diversity and frequency among the landraces varied from 0.32 in the case of RM286 to 0.87 in the case of RM520 and RM521 with an average of 0.725 per primer. The pair-wise estimates of similarity coefficients ranged from 0.062 to 0.733. The magnitude of Dice similarity coefficient between Dihawan and Moti; Sugapankhi and Lalka Dhan (0.7333) was found to be the maximum.By drawing the phenon line at 25 similarity units. A perusal of the dendrogram clearly indicates that the entries were basically divided into four groups. Cluster I, II, III, IV consisting 13, 2, 7, 1 landraces respectively. Analysis of divergence pattern based on morphological and molecular characterization having 15 quantitative traits and fifteen SSR markers allowed differentiation and classification of twenty three rice landraces into five and four clusters, respectively revealed that genotype Marcha-1, Marcha-2; Jadhan, Rajbhog; Karahani Dudha Ladu, Moti, Parwa Pankhi, Lalka Dhan belong to same group in both clusters showed high similarity between genotypes present in same clusters.ThesisItem Open Access An Analysis of the role of institutional Credit int eh Marketing of Agricultural Produce in Punjab -A Case Study in Ludhiana District(Punjab Agricultural University, Ludhiana, 1974) Kumar, Rajesh; Johl, S. SThesisItem Open Access Application of starter cultures for the production of dry sausages(College of Agriculture Chaudhary Charan Singh Haryana Agricultural University Hisar, 1994) Kumar, Rajesh; Berwal, J. SThesisItem Open Access Buffalo milk lactoperoxidase : Isolation, purification and characterization.(NDRI, Karnal, 1994) Kumar, Rajesh; Bhatia, K.L.ThesisItem Open Access Certain clinicopathological studies on experimental candidiasis in Japanese quail(College of Veterinary Sciences Chaudhary Charan Singh Haryana Agricultural University Hisar, 1991) Kumar, Rajesh; Gupta, R. K. PaulThesisItem Open Access Clinicoimmunological studies on Experimental nephropathy in calves(College of Veterinary Sciences Chaudhary Charan Singh Haryana Agricultural University Hisar, 1995) Kuamr, Lokesh; Kumar, RajeshThesisItem Open Access Cloning and characterization of RNA Binding Protein (ALL 4377) from Anabaena PCC7120.(Department of Genetics and Plant Breeding Institute of Agricultural Science BHU, 2021) Rai, Tushita; Rai, L.C.; Kumar, RajeshThe presence of prevalent nitrogen-fixing, photosynthetic cyanobacteria is a common inhabitant of the tropical rice fields. Their early emergence on earth and survival in extreme conditions through biotic and a-biotic stresses allied with their plant-like habits and phylogenetic relationship with plant chloroplasts makes them the most appropriate model system for studying plant response to different biotic along with abiotic environmental stresses. In terms of the magnitude of damage and global distribution, cold stress emerges as the principal determinant of crop productivity among all the stress. Acclimation of cyanobacteria in cold stress involves induction of the expression of several families of genes. In response to low temperature, several biological and physicochemical changes occur in cellular physiology so as (i) decrease in membrane fluidity, (ii) stabilization of secondary structures of nucleic acids leading to reduced efficiency of mRNA translation and transcription, (iii) inefficient folding of some proteins, and (iv) hampered ribosome function. The present study shows the importance of Cold-shock response and adaptation towards cold stress, which has been studied extensively in Anabaena PCC 7120. Many RNA Binding Proteins have been produced to counteract these harmful effects of temperature downshift. Cold stress enhances the level of RNA binding protein (rbp) in the cell and acts as an essential player in RNA metabolism5. RNA-binding proteins play a significant role in regulating transcription and termination and generally accumulate after a fall in temperature. The following study intended to determine the effect of cold stress on the expression of RNA-binding proteins by using In-silico and wet lab approaches. The transcript levels of genes All 4377 were monitored in Anabaena sp. PCC 7120 cultures. Steady-state transcript levels of gene all 4377 increased transiently in response to a temperature shift from 30 to 20‡C under both the circumstances of nitrogen regimes. Recombinant plasmid transformed in E. coli showed better survival under low temperatures. Therefore, this study concluded that low temperature modulates the stress-responsive regulation of RNA-binding proteins in cyanobacteria, providing a coherent mechanism to integrate environmental and molecular signals.ThesisItem Open Access Cloning and Characterization of the Dengue Virus Envelope Gene from Clinical Sample.(Department of Genetics and Plant Breeding, Banaras Hindu University, Varanasi., 2022) Singh, Deepa; Kumar, RajeshDengue feveris one of the most significant arboviral infections that affect people. It is principally transmitted by the mosquitoes Aedes aegypti and rarely by Aedes albopictus. It is brought on by the dengue virus, a member of the flavivirus genus that belongs to the Flaviviridae family. Dengue virus is enveloped,single-stranded RNA genomes of positive polarity. The genome is ~11kb in length, capped at 5’end but lacks poly-A tail at 3’ end. The ORF region of the genome is flanked by the two untranslated regions (UTR’s) at the 5’end and 3’ end.The genomic organization consists of three structural proteins (capsid, C; membrane, M and envelope, E) and seven non-structural proteins (NS1, NS2, NS3, NS4, NS4A, NS4B, and NS5). The dengue virus exists as four antigenically distinct serotypes,DENV1, DENV2, DENV3, and DENV4 which diverge ~30% in their genomic similarity.The E protein present on the outer surface is the first point of attachment for the viral entry and contains the epitope-specific for dengue serotypes.The pathogenicity of viruses can be affected by mutations in this protein. Thus, this study targeted the E gene of dengue for the characterization of the dengue virus.Reverse transcription-polymerase chain reaction (RT-PCR) was used to establish the diagnosis of dengue from the viral RNA recovered from the suspected dengue clinical sample. DENV-specific gene primers amplified the 1700 bp of the targeted E-gene. With the help of restriction enzyme analysis and nucleotide sequencing of genomic fragments, the desired amplified product was further examined for serotype identity. The restriction enzyme Eco RI was used to carry out the digestion. A T/A cloning vector was used to ligate the digested product. The ligated product was cloned into competent cells DH5α of E.coli plated on an LB-AIX plate. The study based on cloning the gene fragments of DENV1 and DENV2 is useful to diagnose dengue fever and study the structure and function of E-genes in detail. In addition, the selection of the E coding sequence as a sequencing target provides a sufficient phylogenetic signal to distinguish DENV from other flaviviruses and between serotypes.ThesisItem Open Access Comparative analysis of genetic diversity in Bottle gourd [Lagenaria siceraria (Mol.) Standl.] using RAPD and ISSR markers for developing crop improvement strategies(Department of Horticulture (Olericulture), BAU, Sabour, 2019-05) Kumar, Rajesh; Kumar, RandhirAmong the all cultivated cucurbitaceous vegetables, bottle gourd (Lagenaria siceraria (Mol.) Standl.) with chromosome no. 2n = 22, is an annual monoecious crop synonymously called as calabash gourd. For the above investigation twenty-four genotypes collected from various states adjoining Bihar including breeding lines developed, collected and maintained at department. The genotypes of bottle gourd laid out in statistical design named Randomized Block Design with 3 replications in spring summer season of 2014 and 2015, respectively to examine the magnitude of variability and genetic diversity for several traits horticultural importance. In morphological study genotypes performed significantly in all traits studied, however BRBG-65 (591.10 q/ha) was proved to be high yielder with supporting traits like yield of marketable fruits/plant (15.19 kg) and no. of fruits/vine (14.85) in pooled analysis of both season than compare to the all three check varieties used in past study. Traits like fruit length, fruit circumference, no. of fruits per vine, yield of marketable fruits/vine, no. of seeds/fruit and fruit yield/hectare exhibited high GCV and PCV. Almost all the traits under study shown high percentage of heritability with maximum genetic advancement percentage for the traits like fruit length, fruit circumference, average fruit weight, no. of fruits/vine, yield of marketable fruits/vine, no. of seeds/fruit and yield per hectare. Traits like average fruit weight, no. of fruits plant and yield of marketable fruits/vine shown strong character association with fruit yield/hectare. Based on D2 values, twenty-four genotypes were classified amongst five clusters which indicated a large genetic diversity. Cluster II produced maximum no. (11) of genotypes followed closely by cluster I (8), whereas cluster V was proved to be mono-genotypic. Fruit length has given maximum contribution for total divergence. In the molecular study, average polymorphism percentage was obtained by RAPD pattern was studied by using 10 random primers. Total 58 reproducible amplicons were generated by 10 RAPD primers, of an average around 5.8 amplicon per primer. The value corresponding average polymorphic information content was 0.58 and ranged from 0.43 to 0.81. The major allele frequency was ranged from 0.33 to 0.73 with the average of 0.57. The major genetic diversity was ranged from 0.30 to 0.59 with the average of 0.41. ISSR pattern was studied by using 7 primers. Total 48 reproducible amplicons were generated by 7 ISSR primers of an average around 6.85 amplicon per primer. The value of average polymorphic information content was 0.56 and ranged from 0.29 to 0.76. The major allele frequency was ranged from 0.39 to 0.80 with the average of 0.59. The major gene diversity was ranged from 0.20 to 0.71 with the average of 0.44. Wide range of value regarding Jaccard’s similarity coefficient in RAPD markers (0.46 to 0.86), ISSR markers (0.37 to 0.910) and in combination (0.50 to 0.80). The dendrogram concluded relative magnitude of resemblance also amongst different clusters formed. Dendrogram and similarity matrix produced from both the markers RAPD and ISSR data then compared and concluded similar but not genetically identical phylo-genetic relationships, when RAPD, ISSR, and RAPD combination with ISSR derived dendrograms were compared, the discrimination among genotypes within these clusters was more effective with the combined analysis. Most genotypes studied in the present experiment differed with their geographic origin. The promising lines collected from different source were classified amongst different clusters and sub clusters. The genotype BRBG 65 was extremely diverse from other genotypes and the genotype BRBG 11-1 was genetically similar to BRBG 52 and BRBG 12-3. The investigation highlights the potential utilization of these germplasm for future breeding programmes.The genotypes of more divergent cluster may be utilized in breeding programme for developing high yielding varieties and F1 hybrids with desirable quantitative traits. The most promising genotype BRBG 65 may be recommended for commercial cultivation.ThesisItem Open Access Comparative analysis of genetic diversity in Bottle gourd [Lagenaria siceraria (Mol.) Standl.] using RAPD and ISSR markers for developing crop improvement strategies(Department of Horticulture (Olericulture), BAU, Sabour, 2019-05-21) Kumar, Rajesh; Kumar, RandhirAmong the all cultivated cucurbitaceous vegetables, bottle gourd (Lagenaria siceraria (Mol.) Standl.) with chromosome no. 2n = 22, is an annual monoecious crop synonymously called as calabash gourd. For the above investigation twenty-four genotypes collected from various states adjoining Bihar including breeding lines developed, collected and maintained at department. The genotypes of bottle gourd laid out in statistical design named Randomized Block Design with 3 replications in spring summer season of 2014 and 2015, respectively to examine the magnitude of variability and genetic diversity for several traits horticultural importance. In morphological study genotypes performed significantly in all traits studied, however BRBG-65 (591.10 q/ha) was proved to be high yielder with supporting traits like yield of marketable fruits/plant (15.19 kg) and no. of fruits/vine (14.85) in pooled analysis of both season than compare to the all three check varieties used in past study. Traits like fruit length, fruit circumference, no. of fruits per vine, yield of marketable fruits/vine, no. of seeds/fruit and fruit yield/hectare exhibited high GCV and PCV. Almost all the traits under study shown high percentage of heritability with maximum genetic advancement percentage for the traits like fruit length, fruit circumference, average fruit weight, no. of fruits/vine, yield of marketable fruits/vine, no. of seeds/fruit and yield per hectare. Traits like average fruit weight, no. of fruits plant and yield of marketable fruits/vine shown strong character association with fruit yield/hectare. Based on D2 values, twenty-four genotypes were classified amongst five clusters which indicated a large genetic diversity. Cluster II produced maximum no. (11) of genotypes followed closely by cluster I (8), whereas cluster V was proved to be mono-genotypic. Fruit length has given maximum contribution for total divergence. In the molecular study, average polymorphism percentage was obtained by RAPD pattern was studied by using 10 random primers. Total 58 reproducible amplicons were generated by 10 RAPD primers, of an average around 5.8 amplicon per primer. The value corresponding average polymorphic information content was 0.58 and ranged from 0.43 to 0.81. The major allele frequency was ranged from 0.33 to 0.73 with the average of 0.57. The major genetic diversity was ranged from 0.30 to 0.59 with the average of 0.41. ISSR pattern was studied by using 7 primers. Total 48 reproducible amplicons were generated by 7 ISSR primers of an average around 6.85 amplicon per primer. The value of average polymorphic information content was 0.56 and ranged from 0.29 to 0.76. The major allele frequency was ranged from 0.39 to 0.80 with the average of 0.59. The major gene diversity was ranged from 0.20 to 0.71 with the average of 0.44. Wide range of value regarding Jaccard’s similarity coefficient in RAPD markers (0.46 to 0.86), ISSR markers (0.37 to 0.910) and in combination (0.50 to 0.80). The dendrogram concluded relative magnitude of resemblance also amongst different clusters formed. Dendrogram and similarity matrix produced from both the markers RAPD and ISSR data then compared and concluded similar but not genetically identical phylo-genetic relationships, when RAPD, ISSR, and RAPD combination with ISSR derived dendrograms were compared, the discrimination among genotypes within these clusters was more effective with the combined analysis. Most genotypes studied in the present experiment differed with their geographic origin. The promising lines collected from different source were classified amongst different clusters and sub clusters. The genotype BRBG 65 was extremely diverse from other genotypes and the genotype BRBG 11-1 was genetically similar to BRBG 52 and BRBG 12-3. The investigation highlights the potential utilization of these germplasm for future breeding programmes.The genotypes of more divergent cluster may be utilized in breeding programme for developing high yielding varieties and F1 hybrids with desirable quantitative traits. The most promising genotype BRBG 65 may be recommended for commercial cultivation.ThesisItem Open Access ThesisItem Open Access Denovo transcriptomic profiling of roots of Carica papaya under drought stress condition.(Department of Genetics and Plant Breeding Institute of Agricultural Sciences Banaras Hindu University Varanasi, 2022) Prakash, Chhavi; Kumar, Rajesh“Papaya”(Carica papaya L) is an incredibly healthy tropical fruit, first domesticated in Central America but now grows worldwide in tropical and subtropical regions.A soft, fleshy cerise-orange coloured juicy fruit, loaded with lots of vitamins and minerals,&well known for its nutritional benefits and medicinal properties.Papaya plant have been regarded as relatively resistant to drought, responding through a desiccation delay mechanism. However, the transcriptomic evaluation for the elucidation of papaya plant molecular responses to drought stress still remains scarce.Despite the complexity of its genome, Carica papaya's genomic data sources are still deficient, but advancements in Next Generation Sequencing have made it possible to assemble transcriptomes of organisms without the availability of their entire genome sequences using De novo transcriptome profiling.In the present study, we employed de novo transcriptomic profiling on the RNA-seq raw reads data of the “roots ofCaricapapayaunder well-watered (control) condition and after 10 days of drought stress condition”, for the assembly, validation, and reporting of the transcriptome with their biological identities. The raw data were obtained from the NCBI’s sequenced library of SRA (Sequence Read Archive) experiment. The sample sequences were projected to De novo transcriptome analysis and further filtration had be done.Andfinallywe obtained a total of 13 unique true transcript ids and their corresponding sequences from this. These sequences were searched against the Arabidopsis thalianaas well as Brassica oleraceacDNA sequences using BLASTn, to determine the sequence similarity of these significant transcripts based on their homology and to provide the biological identities. The complete names of homologous sequences were found out using DAVID annotation tool.ThesisItem Open Access Designing of Insecticides against Bemisia tabaci using computational approaches(Department of Genetics and Plant Breeding Institute of Agricultural Sciences Banaras Hindu University Varanasi, 2022) Indu; Kumar, RajeshBemisia tabaci is a major destructive pest that destroys more than 600 crop species worldwide. Its feeding behaviour on phloem sap and secretion of honeydew on leaves leads to the accumulation of mold that impairs photosynthesis and fruit quality. Additionally, it is responsible for transferring more than 100 viruses in plants which interferes with plant growth by becoming a limiting growth factor. This pest is responsible for a huge number of economic losses. This research aims to find novel natural compounds from available databases using molecular docking (MD), and molecular dynamics simulation (MDS) approaches. In this work, I have targeted the ecdysone receptor of Bemisia tabaci as it is involved in metamorphosis, cell differentiation and reproduction processes. No similar receptor is present in mammals which makes it an ideal target. The sequence of EcR has retrieved from the NCBI database and then BLAST (Sayers et al., 2021) against the UniProt database (The UniProt Consortium et al., 2021). Multiple sequence alignment by MUSCLE web-server (Edgar, 2004) has been performed. Further BLAST against PDB database to find out available 3D structures. Only one ligand-binding domain structure was available; hence I have modelled the full-length protein using alphafold 2.2.0 (Jumper et al., 2021). I have checked for the disordered regions of the protein by using two IDP predictor software, i.e. DEPICTER (Barik et al., 2020) and PrDOS (Ishida & Kinoshita, 2007). Further, molecular dynamics simulation was performed to equilibrate the best model predicted by alphafold in an aqueous and ionic environment to check it’s the stability and flexibility. From the simulation, it has been found out though the IDR part is highly flexible, but the ligand-binding domain is very stable and compact. Therefore, I considered the last conformation from the simulation for further studies. Next, 32,552 bacterial and fungal secondary metabolites were retrieved from the npatlas 2.0 database (van Santen et al., 2022) and docked each metabolite with the simulation obtained last conformation of EcR protein using idock 2.2.3 software (Li et al., 2012). I have chosen cut-off -11kcal/mol binding energy and found 37 metabolites. I have redock these 37 metabolites again with another well- known docking software, i.e. Autodock vina 1.1.2 (Trott & Olson, 2009), to validate idock 2.2.3 results and found an almost similar result with minor deviations. These dockings were compared with 20E, a natural hormone binding with EcR protein. The top five consistent compounds were selected, and protein-ligand interactions were studied using ligplots (Wallace et al., 1995). Finally, I concluded these 37 molecules might be possible hits for EcR protein, and the top five selected compounds are needed priority for experimental validations.ArticleItem Open Access Development and assessment of green synthesis of hydrazides(2004) Saha, Ajoy; Kumar, Rajesh; Kumar, Rajendra; Devakumar, C.ThesisItem Open Access Development of a mechanised process for continous drainage & pressing of paneer(ICAR-NDRI, Karnal, 1984) Kumar, Rajesh; Kohli, R.K.ThesisItem Open Access Development of bioprocess for generation of antimicrobial and antioxidative peptides from milk proteins.(NDRI, Karnal, 2014) Rana, Seema; Kumar, RajeshThesisItem Open Access Differential time utilization pattern of farm women in crop and dairy farming(NDRI, KARNAL, 1992) Kumar, Rajesh; Fulzele, R MThesisItem Open Access Diversity and Management of sucking insect pests of Brinjal(MPUAT, Udaipur, 2017) Kumar, Rajesh; Mahla, M. K.The investigation on “Diversity and Management of Sucking Insect pests of brinjal” was carried out at the Horticulture Farm, Rajasthan College of Agriculture, Udaipur (Rajasthan) with the objectives of:- i. To study the influence of abiotic environmental factors on the incidence of sucking insect pests of brinjal. ii. To establish the diversity of insect pests and natural enemies in brinjal. iii. To evaluate the relative efficacy of newer insecticides against sucking insect pests of brinjal. The major sap sucking insect pests including aphid commenced in the last week of September and attained its peak during third week of October (43rd SMW). Jassid commenced in the last week of September and attained its peak during last week of October (44rd SMW) and whiteflies in the last week of September attained its peak during last week of October (44rd SMW). The pests showed a positive significant correlation with temperature and negative and with humidity. Among the major pests infesting brinjal, as per relative density and mean density values for brinjal shoot and fruit borer were (13.49, 10.53), Epilachna beetle (6.15, 4.80), Lace wing bug, (9.05, 7.06) aphid, (22.55, 17.59) jassid (22.98, 17.93) and whitefly were (25.78, 20.11) respectively. The natural enemies relative density and mean density values for Lady bird beetle (42.91, 15.69) Syrphids (29.01, 10.63) and Spiders (28.08, 10.29) respectively. On the basis of first and second sprays overall efficacy of pesticidal treatments at, three, five and seven, days after each spray against aphid, jassid and whitefly revealed that imidacloprid 0.5 ml /lit as most effective The treatment of azadiractin 3000 ppm proved least effective followed by NSKE (5.0%). The fruit yield was maximum (28.69 kg /plot) in the treatment of imidacloprid. The fruit yield drastically reduced in untreated check plots (15.90 kg /plot).