Botany - Mphil Thesis
Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/30
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Item Development and chacterization of genomic microsatellite markers in rhdodendron arboreum(Central University of Punjab, 2014) Choudhary, Shruti; Bhardwaj, PankajRhododendron arboreum, a key species inhabiting Indian Himalayas, has ecological and economic importance. Global climate change and anthropogenic activities can pose a threat to the biodiversity richness. Population genetics characteristics are the fundamentals of conservation and management practices. However, no genetic background for the species is presently available. To provide an insight into the population structure of the species, an effective marker resource was developed. A genomic library enriched for microsatellites was constructed using biotin- streptavidin hybridization technique, followed by sequencing of positive clones. With 54% enrichment rate, 41 primers were designed from SSR clones, consisting of perfect or interrupted repeats of AG/CT, AC/GT motifs. 38 loci showed successful amplification on genotypes of three populations with number of alleles ranging from 2 to 14, observed and expected heterozygosity of 0.167 to 0.933 and 0.422 to 0.917, respectively and average polymorphic information content of 0.104 to 0.811. A high gene flow (Nm= 5.436) demonstrated high genetic diversity within the populations. 19 loci displayed significant deviations from Hardy Weinberg equilibrium. Overall no linkage disequilibrium and bottleneck was detected among sampled populations. DARwin and STRUCTURE analysis grouped the populations into two original clusters and admixed genetic stock irrespective of their geographical locations. These novel loci will support further genotyping studies in R. arboreum.Item Development and charaterization of UGMS markers for genetic diversity analysis in Rhododendron arboreum(Central University of Punjab, 2013) Sharma, Gagan; Bhardwaj, PankajRhododendron arboreum is an ecologically important species growing in Himalayan regions. It is widely popular due to its medicinal potential, edible, sacred & economic value, however very little is known about its genetic diversity. The development and characterization of UGMS markers will provide a major impact on genetic diversity analysis. Total 792 (615 singletons and 177 contigs) unigenes were predicted from 1,241 publicly available EST database from NCBI (http://www.ncbi.nlm.nih.gov/) site in R.catawbiense by clustering of 2 - 33 random EST sequences. From 50 SSR contained Unigenes, 36 UGMS primers were designed for the cross amplification approach and six pairs of microsatellite primers were produced clear PCR amplification. The cross related species/genera transferability rate was 16.66% from R.catawbiense to R.arboreum. The polymorphism information content (PIC) ranged from 0.2756 to 0.9212 with an average of 0.5765.The average observed heterozygosity (Ho) was 0.8666 i.e. greater than average expected heterozygosity (He) was 0.6792. The genetic diversity was found with an average of 0.6222 within population of R.arboreum and genetic diversity found with an average of 0.3436 among the population.This study revealed the insight of abundance & distribution of microsatellite in the expressed region of the Rhododendron arboreum genome.