1. KAUTIR (Kerala Agricultural University Theses Information and Retrieval)
Permanent URI for this communityhttp://localhost:4000/handle/123456789/1
Browse
14 results
Search Results
Item Genetic diversity analysis of gladiolus genotypes (Gladiolus grandiflorus L.) using molecular markers(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2022) Karthika Nair, A S; Beena ThomasCharacterization of plant genotypes based on crop genetic diversity is important for effective usage and conservation. This is generally achieved by either morphological tools or molecular tools or by using both. This study entitled “Genetic diversity analysis of gladiolus genotypes (Gladiolus grandiflorus L.) using molecular markers” was carried out in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, Thiruvananthapuram during 2020-2021 with an objective to analyse genetic diversity in the gladiolus genotypes using ISSR as well as morphological markers. Gladiolus (Gladiolus sp.) is a genus of perennial herbaceous cormous flowering plants in the family Iridaceae which is of high economic importance. Fifteen varieties of gladiolus including twelve varieties from IIHR, Bangalore and three varieties from NBRI, Lucknow were selected for this study. The study was divided into two parts- morphological characterization and molecular characterization. Morphological characterization was done by analysing both vegetative and floral characters. Different tools such as analysis of variance, co-variance, correlations, PCA and dendrogram were used for statistically analysing the recorded data. The dendrogram divided the genotypes into two principal clusters at a distance of 0.10. The major variables that contributed to the clustering of gladiolus genotypes were plant height, number of leaves per shoot, length of leaf, width of leaf, internode length, number of florets open at one time and number of florets per spike as revealed by PCA analysis. For molecular characterization using ISSR markers the genomic DNA was isolated using CTAB method of DNA isolation with little modifications. Ten ISSR primers were used for screening fifteen gladiolus genotypes. After the final PCR with selected primers, the amplicons were resolved in 2% agarose gel and polymorphic bands were obtained. Primers showed 94.56% polymorphism and the number of bands obtained ranged from 3(UBC 857) to 14 (UBC 890) with a mean value of 8.7 polymorphic bands per primer. A total of 87 polymorphic bands were obtained. The data analysed using NTSYS PC 2.02 program created a dendrogram, which grouped 113 the genotypes based on Jaccard’s similarity coefficient in which the fifteen genotypes were separated into two principal clusters. The first principal cluster consisted of most of the genotypes (12 genotypes). The second principal cluster comprised of ‘Arka Naveen’, ‘Archana’ and ‘Arka Gold’ with ‘Archana’ as an outlier. In molecular characterization, least similarity of 34% was observed between G3 (Arka Sapna) and G9 (Archana) whereas, maximum genetic similarity of 82% was observed between G6 (Arka Amar) and G10 (Arka Kumkum). The highest morphological similarity was also observed between G6 (Arka Amar) and G10 (Arka Kumkum) at a distance of 0.83 in UPGMA dendrogram based on Jaccard’s coefficient. Though some similarity in results existed between the morphological and molecular tools used for identifying the genetic relationships among selected gladiolus varieties in this study, it also revealed that the varieties were grouped as separate clusters based on morphological dendrogram. This may be due to the dependence of morphological expression on the physiological state of the individual plant along with environmental influence. Self-incompatibility, along with the outcrossing nature together might have contributed to the high variation observed among the gladiolus genotypes. Being a commercial cut flower crop, based on different floral parameters considered ‘Arka Sapna’, ‘Arka Nazrana’, Arka Darshan’, ‘Arka Amar’ and ‘Arka Poonam’ are recommended as the gladiolus genotypes that showed best performance in Kerala conditions. Tags from this library: No tags from this library for this title.Item Identification of microsatellite markers associated with root traits for drought tolerance in rice (Oryza sativa L.)(Department of Plant Technology, College of Agriculture, Vellayani, 2017) Rejeth, R; Beena, RItem Characterisation and systematic evaluation of genetic resources of the genus Vigna(Department of Plant Breeding and Genetics, College of Horticulture,Vellanikkara, 2010) Latha, M; Pushpalatha, K TVigna belonging to the family Leguminoseae is a large genus comprising of seven sub-genera and over 150 species. The two sub-genera Vigna and Ceratotropis contain the most important cultivated species. The taxonomical identification of many of these species is still confusing. The closely related wild species serve as a source of many desirable genes that can be utilised in the interspecific hybridisation programmes. This is possible only when the relationships among the different Vigna species are well understood. In this context, the present study “Characterisation and systematic evaluation of genetic resources of the genus Vigna” was undertaken in the Department of Plant Breeding and Genetics, College of Horticulture at Vellanikkara. Investigations were undertaken to characterise the accessions of Vigna germplasm available at National Bureau of Plant Genetic Resources (NBPGR) Regional Station, Vellanikkara, Thrissur using morphological markers and to confirm the results using biochemical and molecular markers in distinct variants belonging to different taxa as well as to prepare a key for the identification of different Vigna taxa. The 150 accessions available at NBPGR Regional Station, Vellanikkara were subjected to morphological, biochemical and molecular characterisation. For morphological characterisation 48 qualitative and 24 quantitative characters were taken. The biochemical characterisation of the selected distinct variants from each taxa was done by isozymes, peroxidase and poly-phenol oxidase. The molecular characterisation was done with Inter Sequence Repeat Analysis using 10 different primers. The clustering patterns based on all three characterisation were compared and key for identification of different taxa of Vigna was developed. Among the qualitative characters evaluated, type of seed germination, nature of attachment of primary leaves, size of stipule, shape of stipule, presence of ligule, shape of bracteole, nature of pod attachment to peduncle, curvature of pod, shape of seed and shape of hilum were distinct for each taxa. Variability was observed in size and shape of stipules and bracteoles. Based on the qualitative characters the 150 accessions were reclassified into 22 taxa. One accession originally classified as V.radiata var.sublobata was found to be distinct taxa of Vigna and hence regrouped as distinct taxa. All the 24 quantitative characters studied exhibited wide range of variability. The keel pocket was present in all taxa except V.unguiculata, V.marina and V.pilosa. The length of keel pocket also varied from taxa to taxa. Cluster analysis based on qualitative, quantitative, biochemical and molecular characters resulted in 10, 5, 4 and 12 clusters respectively. A statistical methodology was worked out to compare the parallelism among the different clustering patterns. The result showed that there existed a similarity between clusters formed based on quantitative and qualitative characters, with majority of accessions of each taxa in a qualitative cluster falling in the same quantitative cluster. The accessions taken for isozyme and molecular study were distinct. Accessions of same taxa which fell in same clusters based on isozyme and molecular markers fell in different clusters based on quantitative characters and vice-versa, indicating the differences and similarities among these accessions at isozyme and molecular level. Key quantitative characters were also identified for each taxa based on weighted average. Based on morphological, biochemical and molecular characters, a dichotomous key was developed to identify different taxa. The key that is now proposed is different from the existing one which is based on floral and fruit characters alone.Item Identification of the population genetic structure of Carcharhinus longimanus (oceanic white tip shark or brown Milbert's shark) using mitochondrial DNA markers(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2019) Sreelekshmi, S; Sandhya SukumaranEven though sharks are the largest fishes in the world with their size varying size and behaviour, they were over exploited and most of them were at the fear of extinction. Among these Carcharhinus longimanus, an epipelagic bottomless shark considered as at the point of extinction were IUCN Red list points out this shark as a “vulnerable” species at global level. In order to implement the management measures for these species which require the information regarding its population in interoceanic regions. Population genetics can be characterized as the study of how hereditary variance is dispersed among the species and population on a very basic level (Hansen, 2003). Assessment of genetic makeup and variability of fish stock is important for scientific management of fishery, conservation and rejuvenation of endangered species. Mitochondrial DNA (mtDNA), which in general possess a five to ten times greater variability than single copy nuclear genes hence, served as a powerful tool for elucidating population structures studies. Among the 150 specimens of C. longimanus sequenced, we obtained sequences ranging from 720 base pairs were obtained 12 polymorphic sites yielding 13 haplotypes. Genetic differentiation among the populations of C. longimanus from Indian Ocean was revealed as a non-significant statistical analysis. Vital insights were gained from this study indicating lack of significant substructuring and its capability to migrate across large expanses of Open Ocean. The capability to migrate may provide it with some buffering against habitat loss and climate change, but excessive fishing is a danger to its populations. Globally sharks are in danger due to their inherent vulnerabilities like long gestation time and reduced number of offsprings coupled with over fishing. Our study also corroborated the findings of shark decline, as decline in genetic diversity is an indicator of decrease in resilience capacity. The present study calls for restrictions on its fishery so that populations will get sufficient time to replenish and consequently their resilience is ensured in the face of changing oceans.Item Breeding cowpea (Vigna unguiculata (L.) Walp.) for resistance to spotted pod borer (Maruca vitrata Fab.)(Department of Plant Breeding and Genetics, Vellanikkara, 2018) Ambavane Ajinkya Rajendra; Jiji JosephItem Introgression of mosaic resistance in popular short duration cassava varieties of Kerala through marker assisted selection(Deparment of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2018) Darshan, S; Arya, KThe present study entitled “Introgression of mosaic disease resistance in to popular short duration cassava varieties of Kerala through marker assisted breeding”was conducted in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, Kerala Agricultural University and Division of Crop Improvement, ICAR- Central Tuber Crops research Institute, Sreekariyam, Thiruvananthapuram, Kerala during the period 2014 - 2017 with the core objective of introgression of cassava mosaic disease (CMD) resistance to short duration varieties of cassava through marker assisted selection (MAS) and to study the inheritance of early bulking nature. The research work was carried out as four experiments. In the first experiment, Five early bulking high yielding lines viz, Sree Jaya, Sree Vijaya, Vellayani Hraswa, CI 889 and 9S 75 and three testers viz, CR 54A3, IMS2-5 and CI 273 with resistance to cassava mosaic disease were selected and planted in a pollination block and crossed in Line x Tester (LxT) design to produce hybrid seeds of 15 F1 combinations. Experiment II was conducted in two parts. Screening of F1 seedlings for CMD resistance and early bulking nature was carried out in the first part of experiment II, where hybrids along with the parents were evaluated. Analysis of variance revealed significant differences among the genotypes for all the traits studied. All the agronomic traits were recorded and inheritance of early bulking and its correlation with other traits were studied. The CMD incidence expressed significant and negative correlation with tuber yield per plant where as significant and positive correlation for all other traits with tuber yield per plant was observed among the F1’s. As a part of experiment II (b), seedlings without the CMD visual symptoms were subjected to multiplex PCR and the results revealed that among the parents Sree Jaya, Sree Vijaya, Vellayani Hraswa expressed presence of Srilankan Cassava mosaic Virus (SLCMV) and Vellayani Hraswa expressed the presence of both SLCMV and Indian Cassava mosaic Virus (ICMV). Among the crosses, Sree Jaya x CR54 A3 (L1x T1), Sree Jaya x IMS2-5 (L1xT2), Vellayani Hraswa x IMS2-5 (L3xT2), CI 889 x CI 273 (L4xT3) expressed the presence of SLCMV. Real time PCR (qPCR) assay for seedlings identified CI 889 (L4), 9S 75(L5), CR 54A3 (T1), IMS2-5 (T2) and CI 273(T3) among the parents and Sree Jaya x CR54 A3 (L1x T1), Sree Jaya x IMS2-5 (L1xT2), Sree Jaya x CI 273(L1x T3) and 9S 75 x CR54 A3 (L5x T1), 9S 75 x IMS2-5 (L5xT2) and 9S 75 x CI 273 (L5x T3) among the crosses as highly resistant, based on viral load present in the DNA sample. Based on the previous report ten CMD resistance linked markers were screened through BSA and five of which SSRY 28, SSRY 44 SSRY 40, SSRY 106 and SSRY 235 were selected. Among the CMD linked SSR markers studied, the maximum polymorphism was elucidated by SSRY 28, SSRY 44 and followed by SSRY 235. SSRY 28 is a strongly linked marker to CMD2 which is a dominant gene conferring resistance among the clones of combinations (L1xT1, L2xT2, L3xT1 and L3xT3) three of five markers revealed alleles associated with CMD2 gene In the third experiment to evaluate the early bulking clones, field was laid out in randomized block design (RBD) with three replications consisting of CMD resistant clones along with parental clones using miniset technique. Analysis of variance revealed significant differences among the genotypes for all the traits. Measurement of heterosis was carried out considering parent Vellayani Hraswa (L3) as check and results revealed that standard heterosis was positive and significant in the combinations Sree Jaya x CR 54A3 (L1xT1) and Sree Jaya x CI 273 (L1xT3) for all the yield contributing traits. The crosses Sree Jaya x CR54 A3 (L1x T1) and Sree Jaya x CI 273 (L1xT3) exhibited negative standard heterosis for CMD. Combining ability analysis showed significant gca, sca variances and gca, sca effects for all the traits. Moreover gca/sca variance ratio indicated preponderance of dominance / non-additive gene action for the inheritance of all traits. Among the lines, Sree Jaya (L1) exhibited positive and significant gcaeffect for tuber yield and yield contributing traits. Among the testers, IMS2-5 (T2) exhibited negative and significant gca effect for CMD. Among the crosses Sree Jaya x CR54 A3 (L1x T1) exhibited positive and significant scaeffect for girth of tuber and stem girth, 9S 75 x CI 273 (L5xT3) exhibited positive and significant scaeffect for tuber yield per plant, CI 889 x CR 54A3 (L4xT1) exhibited negative and significant scaeffect for CMD. In the last experiment, through bulk segregants analysis using 5 SSR markers linked to early bulking in cassava were selected out of 9 SSR markers selected. Among 5 SSR markers of CMD and early bulking nature two SSR markers (SSRY 28 and SSRY 106) associated with resistance to CMD and One SSR marker, ESTs (SSRY) 292 associated to early bulking nature has been identified. Among the crosses, clones from Sree Jaya x CR54 A3 (L1xT1), Sree Jaya x CI 273 (L1x T3) and 9S 75 x CR 54A3 (L5xT1) are being confirmed with CMD resistance as well as early bulking nature.Item Molecular characterization and in vitro conservation of taro (Colocasia esculenta (L.)Schott)(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2017) Sreevidya, M R; Asha Devi, AItem Tagging of phytophthor pod rot disease resistance gene in cocoa (Theobroma cacao L.) using ISSR markers(Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2017) Jeughale Kishor, Pundlik; Minimol, J SCocoa (Theobroma cacao L.) known as ‘Chocolate tree’, is a major cash crop in tropical countries. Cocoa production is seriously affected by pod rot diseases caused by many Phytophthora species. Among these, the pod rot caused by Phytophthora palmivora has been reported in India. Yearly losses to the cocoa growers around the world from Phytophthora diseases were assessed at 30 per cent of the total yield loss. Disease resistance can be scored using a number of morphological and physiological characters. However, the morpho-physiological characters greatly depend upon the environment which ultimately affect the experimental data. Hence, confirmation of transfer of genes by tagging with the help of a strong tool is of utmost importance in crop breeding. Molecular markers such as Inter simple sequence repeats (ISSRs) have already proven to be a good tool to detect and tag the genes of interest and will help to reduce the breeding cycle. In this context, the present study was taken up with an objective to develop a strategy to tag gene(s) for Phytophthora pod rot (PPR) resistance in cocoa using ISSR markers. Morphological characterization of 28 hybrid progenies of SVI 1.26 × PII 12.11 was carried out by recording five pod and bean characters. High variability was observed for characters viz., pod weight, pod length and breadth, wet bean weight per pod and single dry bean weight among the progeny of the same cross. Detached pod inoculation technique was adopted to classify the hybrids into resistant and susceptible ones. The wide variability was also recorded for disease reaction among the progenies. Based on the resistance score, three resistant and three susceptible hybrids were selected from the segregating progeny. The eight accessions were screened with fifty ISSR and 15 SSR primers to observe polymorphism between resistance and susceptible genotypes. Polymorphism was observed in 11 ISSR primers and from these, six primers viz., UBC 810, UBC 826, UBC 827, UBC 857, Oligo ISSR 04 and Oligo ISSR 08 were eluted and cloned. Plasmid DNA was isolated from clones and sequenced. Though various SSR primer sets screened were found to yield polymorphism, none of them was successful to give a clear distinction among the resistant and susceptible hybrids. This may be due to the fact that, Quantitative trait loci (QTLs) associated with these reported SSR primers may be absent in the genotypes considered for the study. BLASTn analysis specific to plants was done for all six sequences. Upon analysis, Oligo ISSR 04561 had shown 98 per cent identity with Predicted: T. cacao histidine-containing phosphotransfer protein 1 (HPt). HPts play an important role in propagating cytokinin signal transduction. Cytokinins are instrumental in mediating disease resistance by generating a green island around the infection zones, exhibiting delayed leaf senescence and upregulating the expression of the pathogenesis related (PR) gene/s. In addition to this, the auxin-cytokinin antagonism that occurs as part of a complex hormonal interplay, exerts a critical influence on the core SA-JA/ET plant immunity pathways. The BLASTn analysis of marker UBC 810877 resulted in 99 per cent sequence identity with Predicted: T. cacao phospholipid: diacylglycerol acyltransferase (PDAT) 1 mRNA. This protein regulates the synthesis of triacylglycerol, which is a building component of oils in the plant. Accumulation of oil content in plant cells could impart resistance against the pathogen. UBC 827571 had shown 73 per cent sequence identity with T. cacao clone TCC_BA049P20 complete sequence and it is reported to be QTL rich region associated with different traits of T. cacao. Moreover, ISSR markers UBC 810877, UBC 826535 and UBC 857839 are located on chromosome nine, six and four respectively as inferred from NCBI Genome Data Viewer tool through BLASTn annotations. These markers are found to be located in PPR resistance regions rich in defense associated genes. Further validation and exploitation of polymorphic amplicons or markers in response to PPR would be required. The linkage of Oligo ISSR 04561 and UBC 810877 with HPts and PDAT correspondingly have to be validated to elucidate the association and role of cytokinin and triacylglycerol with PPR disease resistance. If validated, UBC 810877, UBC 826535 and UBC 857839 and Oligo ISSR 04561 could be employed as a marker in PPR resistance breeding programmes in cocoa.Item Development of near isogenic lines of rice variety 'uma' for blast resistance genes through molecular marker assisted backcross breeding(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2017) Harikrishnan, P J; Jayalekshmy, V GItem Genetic diversity and combining ability in cucumber (Cucumis sativus L.)(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2017) Suma, A; Elsy, C RAssessment of genetic diversity is the key tool in any crop improvement and germplasm management programme. Evaluation of genetic variation will help to provide valuable information about new sources of genes. The studies on combining ability and heterosis can support utilization of promising lines in further crop improvement programmes. Cucumber (Cucumis sativus L. 2n= 2x= 14) is an indigenous vegetable crop of India. Even though rich diversity for cucumber is available in India, studies on genetic diversity of this crop are scanty. Therefore, the present project was proposed to explore genetic diversity in cucumber using morphological and molecular markers and to study combining ability and heterosis in selected genotypes. Morphological characterization of 50 accessions of cucumber revealed presence of significant difference among accessions for majority of vegetative, floral and fruit characters. Mean days to first male and female flower opening was 36 and 43 days respectively. Majority of the accessions possessed elliptical elongated fruits with light green skin colour and white flesh colour. Sixteen accessions exhibited significantly higher fruit length than AAUC-2, the standard check, the maximum being exhibited by IC613472 (20.85 cm). Accessions with oblong ellipsoid fruits possessed higher fruit diameter. Mean fruit weight showed high variability among accessions with a range of 33 g to 343 g. Fourteen accessions exhibited significantly high yield than AAUC-2. Number of fruits per plant, yield per plant, loss of weight during storage and sex ratio showed high values for all the genetic parameters studied. IC613481 was the promising genotype identified in morphological characterization, followed by IC613480. Cluster analysis grouped accessions into seven distinct clusters based on the level of similarity in quantitative characters. Random grouping of accessions into various clusters indicated absence of parallelism between genetic diversity and geographical diversity. Cluster II and III were the largest clusters, with 14 accessions each and Cluster V and VI, the smallest ones with single accession each. Results of Principal component analysis revealed that first three principal components, with Eigen values more than unity accounted for 85.80 per cent of cumulative variance, contributed by fruit weight, fruit length, fruit diameter and days to first harvest. The diversity analysis of the accessions was done using DIVA-GIS by generating grid maps. The results of the study indicated that highly diverse accessions with respect to the selected characters were sourced from Mizoram, Tripura and West Bengal. Molecular characterization revealed high level of genetic distinctness between genotypes. SSR11742 and AF202378 were found to be highly polymorphic markers, with high polymorphism information content and number of polymorphic bands. In-depth evaluation of selected 22 genotypes revealed significant difference for all fruit characters except days to last harvest and harvest duration and further revealed the superiority of IC613480. Evaluation of 15 hybrid combinations developed through half diallel mating design and their parents indicated presence of significant difference among parents and hybrids for various characters studied. Among the parental genotypes, IC613480, exhibited significantly high GCA effects for fruit length, number of fruits per plant and yield per plant whereas IC595508A, for fruit weight and loss of weight during storage, and IC613485 for fruit diameter, thus proving to be promising parents for accumulating genes for these characters. The hybrids, IC613480 x IC595508A and IC613480 x IC613471 showed significant SCA effects for yield per plant and sex ratio. IC613480 x IC613471, IC613480 x IC595508A and IC613471 x IC595508A were exhibiting significantly high relative heterosis, heterobeltiosis and standard heterosis for number of fruits per plant and yield per plant. IC613480 and IC613485 were the most promising genotypes identified from the study whereas IC613480 x IC613471 was the most promising hybrid based on SCA effects, heterosis, per se performance on yield contributing characters and organoleptic qualities. This hybrid showed high fruit length (17.01 cm), yield per plant (2163.45 g), number of fruits per plant (11.43) and sex ratio (0.11). IC613480 x IC613476 and IC613485 x IC595508A were the other promising hybrids.