“Characterization of Cotton (Gossypium Spp.) Cultivars through DNA Fingerprinting, Isozymes and Protein Profiling”
| dc.contributor.advisor | Dr. D. N. Vakharia | |
| dc.contributor.author | Mr. Kahodariya J. H. | |
| dc.date.accessioned | 2017-06-07T06:18:25Z | |
| dc.date.available | 2017-06-07T06:18:25Z | |
| dc.date.issued | 2010-06 | |
| dc.description.abstract | The present investigation on “Characterization of Cotton (Gossypium Spp.) Cultivars through DNA Fingerprinting, Isozymes and Protein Profiling” was planned to conduct with two main objectives, (1) DNA fingerprinting through RAPD, ISSR, SSR, STMS primers and development of locus specific SCAR markers, (2) Biochemical analysis through four isoenzymes Viz. catalase, esterase, peroxidase and polyphenol oxidase, and protein profiling using Native-PAGE. Twenty one RAPD primers generated a total of 245 bands/alleles with the 97.88 % polymorphism with an average of 11.66 bands per primer. The dendrogram generated two main clusters that consists all the cultivars of three species grouped together in their respective sub-cluster. Among the screened primers OPA-05, OPA-10, OPA-15, and OPB-11 showed cultivar specific markers while OPA-09, OPA-15, OPA-16, OPA-19 and OPB-11 produced species specific DNA fragments. Nine ISSR primers engendered 85 bands/alleles with 95.69 % polymorphism with an average of 9.44 bands per primer. The cluster analysis revealed the two main clusters. The clustering of G. hirsutum cultivars found to be the same as RAPD. Cultivar and species specific markers were also observed with the ISSR primers Viz. IS-4, IS-7, ISSR-6, ISSR-7, ISSR-10 and ISSR-12. Ten SSR primers out of fifteen and four STMS primers generated total 19 and 14 bands respectively out of which 12 and 14 bands were polymorphic. In case of SSR, 58.33 % polymorphism was recorded while STMS markers were 100 % polymorphic. The character specific band was obtained with STMS primer CM43 that was linked to the leaf red colour of cotton leaves present only in two cultivars Viz. G.Cot-19 and G.Cot MDH-11. The microsatellite primers Viz. BNL1053, BNL3408 and CM43 also produced cultivar and species specific markers in studied cotton cultivars. Fifteen cotton cultivars were grouped into two main clusters and the grouping of cultivars were somewhat similar to the RAPD analysis. Among the studied technique SSR and STMS seems to be more effective than other techniques. The cluster analysis revealed grouping of cultivars according to species. The average similarity of 15 cotton cultivars was 25 % in RAPD and ISSR that is not possible in nature among the cultivars of the same genus, while was 47 % in SSR and STMS. The locus specific SCAR marker was developed by cloning and sequencing the band of around 900 bp present in all G. arboreum cultivars. The sequenced fragment generated a novel sequence of 914 bp. The 20-mer primer was designed from the sequence to develop 297 bp SCAR and was submitted in NCBI genebank with Accession No. 490146. Thus by developing one SCAR marker specific to each cultivars could eliminate the use of all other technique of DNA fingerprinting i.e RAPD, ISSR, SSR and STMS and biochemical markers. Isoenzymes and protein were used for the characterization of cotton cultivars. The isoenzyme analysis was carried out at 4, 8 and 12 DAG and protein analysis was done from the seeds as well as from the seedlings of 4, 8 and 12 DAG. There was no isoenzyme variation found in catalase. The maximum numbers of 7 bands at 8 DAG, 5 bands at 4 DAG and 8 bands at 4 DAG were visualized on Native-PAGE by esterase, peroxidase and polyphenol oxidase isoenzymes respectively. The esterase, peroxidase and polyphenol oxidase isoenzymes generated dissimilar clusters of 15 cotton cultivars from each other. However somewhat similarity was observed in cluster-I of all the dendrogram generated from these three isoenzymes i.e. consists only G. hirsutum cultivars. The polyphenol oxidase isoenzyme was found to be superior in clustering of cotton cultivars according to species. The average similarity was 86 %, 21 %, 38% and 24 % for catalase, esterase, peroxidase and polyphenol oxidase isoenzymes respectively. The protein profile generated the highest number of 10 bands at 12 DAG. First time in the present investigation the cultivars of G. arboreum Viz. G.cot-15, G.cot-19 and G.Cot MDH-11 was found in a separate cluster than G. herbaceum. The pooled study of all molecular and biochemical markers revealed a dendrogram consisted of two clusters. First cluster consisted only of G. hirsutum cultivars while cluster-II consisted of G. herbaceum and G. arboreum cultivars both in different sub-clusters. The cultivars G.Cot hyb-8 and G. cot hyb-10, and G.cot-15 G.cot-19 were found to be similar. The cultivars of G. herbaceum Viz. G.Cot-21, G.Cot-23 and V797 shared a same position in most of the dendrogram generated using various techniques. The results pointed out that biochemical techniques were not able to distinguish cotton cultivars precisely as compared to the DNA fingerprinting techniques. The cultivar identification through molecular and biochemical markers resulted in developing highly diversified map of 15 cotton cultivars belonging to three different species of Gossypium. The results revealed that molecular techniques are more accurate than biochemical markers, and can be used for characterization of cotton cultivars and seed purity study. | en_US |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810018483 | |
| dc.language.iso | en | en_US |
| dc.pages | 263 | en_US |
| dc.publisher | jau,junagadh | en_US |
| dc.sub | Agricultural Biotechnology | en_US |
| dc.subject | biotechnology | en_US |
| dc.theme | “Characterization of Cotton (Gossypium Spp.) Cultivars through DNA Fingerprinting, Isozymes and Protein Profiling” | en_US |
| dc.these.type | M.Sc | en_US |
| dc.title | “Characterization of Cotton (Gossypium Spp.) Cultivars through DNA Fingerprinting, Isozymes and Protein Profiling” | en_US |
| dc.type | Thesis | en_US |
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