1. KAUTIR (Kerala Agricultural University Theses Information and Retrieval)
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Item Screening of Bitter gourd (Momordica charantia L.) genotypes for resistance to powdery mildew disease(Department of Vegetable Science, College of Agriculture, Vellanikkara, 2025-05-17) Chethan, S V; Prasanth, KItem Elucidating the morpho - physiological and molecular changes during seed priming in bitter gourd (Momordica charantia L.)(Department of Seed Science and Technology, College of Agriculture, Vellanikkara, 2024-05-24) Thabsheer, P.; Rose Mary FrancisA study to elucidate the morphological, biochemical and molecular changes leading to enhanced germination on priming in bitter gourd variety Preethi was conducted at College of Agriculture, Vellanikkara, Thrissur, during 2020-2023. Seeds of bitter gourd variety Preethi were dried to 6.84 per cent, and the quality parameters of the sourced seeds were assessed. They were divided into three equal sub-lots (5.0 kg each). Two sub-lots were packed in 700-gauge polyethylene bags and sealed airtight and one each stored for a period of six month under i). Ambient environment (Am.S), ii). Refrigerated storage (Rf.S) at ~18OC-20OC. The third sub-lot was packed in moisture-vapour proof aluminium foil bags seeds and stored for a period of six months at Medium term storage (MTS) facility at the ICAR-NBPGR (National Bureau of Plant Genetic Resources), Regional station, Vellanikkara. At the end of the storage period i.e., six months, the quality parameters of the stored seeds under the various storage environments were assessed following which they were further divided into 11 equal parts and subjected to 10 priming treatments (T1 to T10) following standard procedure, while unprimed seeds served as the control (T11). The priming treatments included T1: KNO3 0.3% for 2 h, T2: KNO3 150 ppm for 3 h, T3: KH2P04 10-2 M for 24 h, T4: PEG 6000 -1.5 MPa for 24 h, T5: GA3 100 ppm for 24 h, T6: Solid matrix priming with Perlite for 48 h, T7: Solid matrix priming with Cocopeat for 48 h, T8: Psuedomonas fluorescens 1x106 cfu.ml-1 for 24 h, T9: Hydration hot water at 50°C for 4 h and T10: Hydration cold water soaking for 24 h. Three separate studies as enumerated below were conducted to compare the effect of priming on germination of stored seeds under three different storage conditions viz. Ia) Effect of priming on seeds stored under ambient environment, Ib) Effect of priming on seeds stored in refrigerated storage (~18-20 OC) and Ic) Effect of priming on seeds stored in medium-term storage (~ 4-6 OC). Seed quality parameters viz., number of days to emergence, speed of germination, germination per cent, seedling root length (cm), seedling shoot length (cm), seedling dry weight (g), seed moisture content (%), germination capacity (GC), speed of germination, mean germination time (MGT), time taken for 50% germination (T50), energy of germination (GE), germination index (GI), coefficient velocity of germination (CVG) and seedling vigour indices were assessed after priming treatments in each experiment. In addition to the physiological observations, biochemical parameters viz., total soluble sugar (%), total protein (%), total oil content (%), H2O2 content (μmol g_1), lipid peroxidation (MDAμmol g_1), total dehydrogenase activity (OD), α–amylase activity (μmol g_1), catalase activity (μmol g_1), electrical conductivity of seed leachate (μScm-1), leakage of amino acid (μg leucine eqiv.ml -1) and leaching of sugar (μg glucose eqiv.ml -1) were recorded along with seed structural parameters viz., total seed coat thickness (μm), thickness of testa (μm), thickness of tegmen (μm), embryo length (μm), endosperm length (μm), endosperm breadth (μm), seed length (μm) and relative water absorption (RWA). Comparison of quality parameters before and after storage under the three environments pointed out that in addition to a reduction in germination, root length and shoot length the seedlings, the vigour indices I and II had also decreased in comparison to the estimates recorded prior to storage, while parameters like MGT and T50 of stored seeds were observed to have increased after six months of storage. The per cent reduction of speed of gemination, germination per cent, seedling root length, seedling shoot length, seedling dry weight, GC, GE, GI, CVG and vigour indices was however, the least in seeds stored in Am.S condition. Hence, it became evident that the per cent deterioration in seed stored at low temperatures is low, conversely, it reinforced the fact that storing bitter gourd seeds at low temperatures (MTs and Rf.S) would help maintain seed quality longer. The effect of seed priming treatments on germination (%) after six months of storage under i) S1: Ambient storage (Am.S), ii) S2: Refrigerated storage (Rf.S) and iii) S3: Medium-term storage (MTS) was anlaysed separately, each following a Completely randomized design with three replication and 11 priming treatments, in order to assess the best priming treatment under each storage environments studied. Germination of seeds stored under varying environmental conditions were found to be significantly influenced by the priming treatment they were subjected to. Germination of aged seeds from the three storage environments when subjected to Solid matrix priming with perlite (T6: Perlite for 48 h) cocopeat (T7: Cocopeat for 48 h) and hormonal priming with GA3 (T5: GA3 100 ppm for 24 h) were significantly superior over untreated seeds. Priming with PEG 6000 (T4: PEG 6000 -1.5 MPa for 24 h) was found to adversely affected germination irrespective of the storage environment the seeds were derived from. Although bio-priming seeds from Am.S with P. fluorescens (T8: P. f at 1x106 cfu.ml-1 for 24 h) and hydropriming (T9: Hydration hot water at 50°C for 4 h) was beneficial, it was not so in seeds derived from Rf.S and MTS. Similarly, hydration with hot water @ 50°C for 4 h was not found very beneficial in seeds stored under Rf.S and MTS as compared to Am.S. The impact of storage environment (S), priming treatment (T), and their interaction on physiological, biochemical and seed structural quality indices of seeds stored were analysed following a Completely randomized design with three replications and two factors viz., Factor I: Storage environments (S1, S2 and S3) and Factor II: Priming treatments (T1 to T11). Results indicated that, irrespective of the priming treatments, the quality indices of seed stored in MTS followed by Rf.S were found to be significantly superior to those stored under Am.S. It was evident that storing seeds at low temperatures (~ 18-20 0C in Rf.S, ~ 4-6 0C in MTS) was beneficial in bitter gourd. It resulted in higher germination as well as other physiological, biochemical and structural indices of the seed. Irrespective of the storage conditions, the quality indices of the aged seeds were found to be significantly influenced by the priming treatment they were subjected to. Solid matrix priming (SMP) with Perlite (T6: solid matrix priming with perlite for 48 h) invariably followed by solid matrix priming with cocopeat (T7: solid matrix priming with cocopeat for 48 h) and hormonal priming with GA3 (T5: 100 ppm for 24 h) were found to be significantly superior to other priming treatments as well as untreated seeds. Priming with PEG 6000 (T4: -1.5 MPa for 24 h) was found to be detrimental and registered the least seed indices. The interaction between seed priming treatment and storage conditions was found to significantly influence germination as well as other quality indices of the aged seeds. Results pointed out that germination and most seed indices studied were significantly superior in solid matrix primed seeds involving perlite (T6: solid matrix priming with perlite for 48 h) under both low temperature storage (MTS and Rf.S), as well as ambient storage (Am.S). Solid matrix priming with cocopeat (T7: solid matrix priming with cocopeat for 48 h) and hormonal priming with GA3 (T5: 100 ppm for 24 h), were found to be next best to SMP with perlite under low temperature storage environments viz., MTS and Rf.S. However, unlike in the low temperature storage environments, under ambient storage conditions, bio-priming with P. fluerescens (T8: P. f at 1x106 cfu.ml-1 for 24 h) was found to be next best to SMP with perlite and superior over SMP with cocopeat and hormonal priming with GA3. However, apart from untreated seeds, osmopriming seeds with PEG 6000 (T4: PEG 6000 -1.5 MPa for 24 h), hydration with cold water (T9: Hydration hot water at 50°C for 4 h) as well as with hot water (T10: Hydration cold water soaking for 24 h), were found to be detrimental under low temperature storage (MTS and Rf.S) and ambient storage as well. The results obtained thus point out that loss of germination, vigour and other quality parameters over storage is inevitable. However, the deterioration of seed indices in bitter gourd due to ageing can be slowed down to the maximum extent by storing them under Medium-Term storage (MTS). Storing seeds under refrigerated storage (Rf.S) is highly beneficial compared to storing them under ambient conditions (Am.S). Irrespective of the environment under which the seeds were stored, solid matrix priming of bitter gourd seeds with perlite (48 h) was the best priming treatment as it resulted in significantly superior germination, growth indices of seedlings, biochemical parameters and seed structural changes over untreated seeds. SMP with cocopeat (48 h) and hormonal priming with GA3 (100 ppm for 24 h) were next best to SMP with perlite in bitter gourd. Subjecting seeds of bitter gourd stored either in low temperature environment like medium term storage and refrigerated storage or under ambient environments to solid matrix priming with perlite would result in enhanced germination and seed quality indices. Solid matrix priming with cocopeat or hormonal priming of the aged seeds with GA3 (100 ppm for 24 h) were the next best priming options, if the seeds were stored at low temperature environments. However, in seed stored under ambient environments, bio-priming with P. fluorescens (1x106 cfu.ml-1 for 24 h) would be the best priming option, next to solid matrix priming with perlite.Item Breeding for downy mildew resistance in bittergourd (Momordica charantia L.)(Department of Vegetable Science, College of Agriculture, Vellayani, 2024-07-19) Syama, S Thampi; Sarada,S.The study entitled Breeding for downy mildew resistance in bittergourd (Momordica charantia L.) was carried out at department of vegetable science college of agriculture vellayani during 2021-2023.The objective of the study was to breed for resistance to downy mildew disease in bitter gourd through conventional method and molecular marker analysis.Item Development and evaluation of high yielding, mosaic tolerant backcross progenies in bitter gourd (Momordica charantia L.) variety Preethi using morphological, biochemical and molecular markers(Department of Genetics and Plant Breeding, College of Agriculture ,Vellayani, 2024-05-23) Ankitha, M O; KAU; Bindu, M RThe present research work entitled ‘Development and evaluation of high yielding, mosaic tolerant backcross progenies in bitter gourd (Momordica charantia L.) variety Preethi using morphological, biochemical and molecular markers’ was conducted in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani and Farming Systems Research Station (FSRS), Sadanandapuram during the year 2020-2023, with an objective to develop high yielding mosaic tolerant backcross progenies in bitter gourd using morphological, biochemical and molecular markers. Thirty three bitter gourd genotypes, including KAU released varieties (2 No’s), NBPGR accessions (13 No’s), and local collections from all over India were used for screening mosaic tolerance. Out of the 33 genotypes, 26 genotypes were Momordica charantia var. charantia and seven were Momordica charantia var. muricata. All these genotypes were artificially inoculated with the three viruses Cucumber Mosaic Virus (CMV), Tomato Leaf Curl New Delhi Virus (ToLCNDV) and Papaya Ringspot virus (PRSV) through wedge grafting. Wedge grafting was done using the infected plant shoots as scion and the collected genotypes as root stock and regrowth from the cotyledonary axis was examined for symptom expression. Out of the 33 genotypes screened, three were highly resistant, four were resistant, five were moderately resistant, six were moderately susceptible, ten were susceptible and five were highly susceptible. The genotypes Lodhi local, Udayagiri local and Therthali local recorded a lowest Vulnerability Index of zero. Molecular markers reported in Cucurbitaceae family were validated for bitter gourd mosaic resistance gene. SSR-11-1 marker for CMV resistance and CAPS marker for Potyvirus resistance gene were used, but no amplification was obtained. Double Antibody Sandwich ELISA (DAS-ELISA) was performed to confirm the resistance reaction of three highly resistant genotypes identified in seedling screening. Optical density (OD) value of the genotypes for the three viruses, CMV, ToLCNDV and PRSV were less than twice the OD value of the un-inoculated healthy plant which confirmed highly resistant disease reaction of genotypes. Molecular confirmation was done by using coat protein primer (Deng primer) specific to the Begomovirus group. Deng primer amplifies coat protein gene of ToLCNDV (520 bp), so that band will be present in only susceptible genotypes and will be absent in resistant ones. Plant defense related enzymes such as peroxidase, polyphenol oxidase and phenyl alanine ammonialyase was estimated and there was increased rate of synthesis of these enzymes in the identified resistant genotypes. So the identified resistant genotypes, Lodhi local, Udayagiri local and Therthali local were used as the donor parent for imparting mosaic resistance into the bitter gourd variety Preethi. Lodhi local is M. charantia var. charantia genotype where as both Udayagiri local and Therthali local are M. charantia var. muricata genotypes. High yielding variety released from KAU viz., ‘Preethi’ was selected as the recurrent parent in the study. Preethi was crossed with the three donor parents and F1s were produced. The F1s were morphologically evaluated with the parents for seventeen characters and it was observed that all the characters of F1 were approximately the average of two parents. All the F1s were backcrossed with Preethi to produce BC1F1 segregants. In the backcross progeny of the cross involving Preethi and Lodhi local, a total of 176 BC1F1 lines were developed. BC1F1 lines were artificially inoculated for their disease reaction. Among the 176 BC1F1 lines, 22 were found to be highly resistant to mosaic disease, 30 were resistant, 30 were moderately resistant, 26 were moderately susceptible, 35 were susceptible and 33 were highly susceptible. Confirmation of resistance was done using DAS- ELISA, Deng primers and estimation of defense enzymes. All the 17 biometrical characters were recorded and the Euclidean distance of the highly resistant BC1F1 lines from the recurrent parent Preethi was calculated using proximity dissimilarity matrix analysis. The 14 BC1F1 lines with high phenotypic similarity to Preethi was backcrossed to develop BC2F1 lines. In the backcross progeny of the cross involving Preethi and Udayagiri local, a total of 170 BC1F1 lines were produced. Among them 15 BC1F1 lines were highly resistant. Resistant reaction of identified 15 BC1F1 was confirmed by DAS-ELISA, molecular screening and biochemical analysis. Euclidean distance of the highly resistant 15 BC1F1 lines from the recurrent parent revealed that eight lines showed similarity with Preethi and they were backcrossed to get BC2F1 lines. A total 147 BC1F1 lines of the cross involving Preethi and Therthali local were screened at seedling stage. Out of the 147 lines, 16 BC1F1 lines were highly resistant. DAS-ELISA, molecular screening using aforementioned Deng primer confirmed the resistant reaction of these lines. Euclidean distance using biometric characters found that, out of 16 highly resistant BC1F1 lines eight lines had close proximity with Preethi. These lines were used to produce BC2F1 lines. The 190 BC2F1 lines of the cross involving Preethi and Lodhi local were screened at seedling stage and in 24 BC2F1 lines, there was absence of virus coat protein band which confirmed the highly resistant disease reaction of the aforementioned lines. The 12 BC2F1 lines with the shortest Euclidean distance and high phenotypic similarity with Preethi were selfed to generate BC2F2 seeds. In the 134 BC2F1 lines of the cross involving Preethi and Udayairi local, seedling screening recorded 17 highly resistant lines. After molecular confirmation of mosaic resistance four BC2F1 lines with close proximity to Preethi were selfed to get BC2F2 seeds. Out of the 143 BC2F1 lines of the cross involving Preethi and Therthali local, 20 BC2F1 lines were highly resistant. The molecular analysis of the 20 BC2F1 lines also confirmed the highly resistant reaction. Four BC2F1 lines with the shortest Euclidean distance was selected and selfed to produce BC2F2 seeds. Although there were BC2F2 seeds of three different crosses, only the BC2F2 seeds of the cross involving Preethi and Lodhi local was carried forward for further backcrossing. This is due to the low yield potential of the backcross progenies of the crosses involving M. charantia var. muricata genotypes. So 206 BC2F2 lines of the cross involving Preethi and Lodhi local were artificially screened at seedling stage for mosaic incidence. Out of the 206 BC2F2 lines, 42 plants were highly resistant to bitter gourd mosaic viruses. The 42 mosaic tolerant backcross inbred lines developed in the study can be carried forward for the development of a mosaic tolerant essentially derived variety (EDV) in the background of high yielding variety Preethi. The backcross progenies obtained in the crosses involving muricata genotypes can be further evaluated for its nutraceutical values.Item Development and evaluation of high yielding, mosaic tolerant backcross progenies in bitter gourd (Momordica charantia L.) variety Preethi using morphological, biochemical and molecular markers(Department of Genetics and Plant Breeding, College of Agriculture, Vellayani, 2024-05-23) Ankitha, M O; KAU; Bindu, M RThe present research work entitled ‘Development and evaluation of high yielding, mosaic tolerant backcross progenies in bitter gourd (Momordica charantia L.) variety Preethi using morphological, biochemical and molecular markers’ was conducted in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani and Farming Systems Research Station (FSRS), Sadanandapuram during the year 2020-2023, with an objective to develop high yielding mosaic tolerant backcross progenies in bitter gourd using morphological, biochemical and molecular markers. Thirty three bitter gourd genotypes, including KAU released varieties (2 No’s), NBPGR accessions (13 No’s), and local collections from all over India were used for screening mosaic tolerance. Out of the 33 genotypes, 26 genotypes were Momordica charantia var. charantia and seven were Momordica charantia var. muricata. All these genotypes were artificially inoculated with the three viruses Cucumber Mosaic Virus (CMV), Tomato Leaf Curl New Delhi Virus (ToLCNDV) and Papaya Ringspot virus (PRSV) through wedge grafting. Wedge grafting was done using the infected plant shoots as scion and the collected genotypes as root stock and regrowth from the cotyledonary axis was examined for symptom expression. Out of the 33 genotypes screened, three were highly resistant, four were resistant, five were moderately resistant, six were moderately susceptible, ten were susceptible and five were highly susceptible. The genotypes Lodhi local, Udayagiri local and Therthali local recorded a lowest Vulnerability Index of zero. Molecular markers reported in Cucurbitaceae family were validated for bitter gourd mosaic resistance gene. SSR-11-1 marker for CMV resistance and CAPS marker for Potyvirus resistance gene were used, but no amplification was obtained. Double Antibody Sandwich ELISA (DAS-ELISA) was performed to confirm the resistance reaction of three highly resistant genotypes identified in seedling screening. Optical density (OD) value of the genotypes for the three viruses, CMV, ToLCNDV and PRSV were less than twice the OD value of the un-inoculated healthy plant which confirmed highly resistant disease reaction of genotypes. Molecular confirmation was done by using coat protein primer (Deng primer) specific to the Begomovirus group. Deng primer amplifies coat protein gene of ToLCNDV (520 bp), so that band will be present in only susceptible genotypes and will be absent in resistant ones. Plant defense related enzymes such as peroxidase, polyphenol oxidase and phenyl alanine ammonialyase was estimated and there was increased rate of synthesis of these enzymes in the identified resistant genotypes. So the identified resistant genotypes, Lodhi local, Udayagiri local and Therthali local were used as the donor parent for imparting mosaic resistance into the bitter gourd variety Preethi. Lodhi local is M. charantia var. charantia genotype where as both Udayagiri local and Therthali local are M. charantia var. muricata genotypes. High yielding variety released from KAU viz., ‘Preethi’ was selected as the recurrent parent in the study. Preethi was crossed with the three donor parents and F1s were produced. The F1s were morphologically evaluated with the parents for seventeen characters and it was observed that all the characters of F1 were approximately the average of two parents. All the F1s were backcrossed with Preethi to produce BC1F1 segregants. In the backcross progeny of the cross involving Preethi and Lodhi local, a total of 176 BC1F1 lines were developed. BC1F1 lines were artificially inoculated for their disease reaction. Among the 176 BC1F1 lines, 22 were found to be highly resistant to mosaic disease, 30 were resistant, 30 were moderately resistant, 26 were moderately susceptible, 35 were susceptible and 33 were highly susceptible. Confirmation of resistance was done using DAS- ELISA, Deng primers and estimation of defense enzymes. All the 17 biometrical characters were recorded and the Euclidean distance of the highly resistant BC1F1 lines from the recurrent parent Preethi was calculated using proximity dissimilarity matrix analysis. The 14 BC1F1 lines with high phenotypic similarity to Preethi was backcrossed to develop BC2F1 lines. In the backcross progeny of the cross involving Preethi and Udayagiri local, a total of 170 BC1F1 lines were produced. Among them 15 BC1F1 lines were highly resistant. Resistant reaction of identified 15 BC1F1 was confirmed by DAS-ELISA, molecular screening and biochemical analysis. Euclidean distance of the highly resistant 15 BC1F1 lines from the recurrent parent revealed that eight lines showed similarity with Preethi and they were backcrossed to get BC2F1 lines. A total 147 BC1F1 lines of the cross involving Preethi and Therthali local were screened at seedling stage. Out of the 147 lines, 16 BC1F1 lines were highly resistant. DAS-ELISA, molecular screening using aforementioned Deng primer confirmed the resistant reaction of these lines. Euclidean distance using biometric characters found that, out of 16 highly resistant BC1F1 lines eight lines had close proximity with Preethi. These lines were used to produce BC2F1 lines. The 190 BC2F1 lines of the cross involving Preethi and Lodhi local were screened at seedling stage and in 24 BC2F1 lines, there was absence of virus coat protein band which confirmed the highly resistant disease reaction of the aforementioned lines. The 12 BC2F1 lines with the shortest Euclidean distance and high phenotypic similarity with Preethi were selfed to generate BC2F2 seeds. In the 134 BC2F1 lines of the cross involving Preethi and Udayairi local, seedling screening recorded 17 highly resistant lines. After molecular confirmation of mosaic resistance four BC2F1 lines with close proximity to Preethi were selfed to get BC2F2 seeds. Out of the 143 BC2F1 lines of the cross involving Preethi and Therthali local, 20 BC2F1 lines were highly resistant. The molecular analysis of the 20 BC2F1 lines also confirmed the highly resistant reaction. Four BC2F1 lines with the shortest Euclidean distance was selected and selfed to produce BC2F2 seeds. Although there were BC2F2 seeds of three different crosses, only the BC2F2 seeds of the cross involving Preethi and Lodhi local was carried forward for further backcrossing. This is due to the low yield potential of the backcross progenies of the crosses involving M. charantia var. muricata genotypes. So 206 BC2F2 lines of the cross involving Preethi and Lodhi local were artificially screened at seedling stage for mosaic incidence. Out of the 206 BC2F2 lines, 42 plants were highly resistant to bitter gourd mosaic viruses. The 42 mosaic tolerant backcross inbred lines developed in the study can be carried forward for the development of a mosaic tolerant essentially derived variety (EDV) in the background of high yielding variety Preethi. The backcross progenies obtained in the crosses involving muricata genotypes can be further evaluated for its nutraceutical values.Item Relative susceptibility of population of Amrasca biquttula biquttula (Ishida) infesting bittergourd Momordica charantia L. collected from different locations to insecticides(Department of Agricultural Entomology, College of Horticulture, Vellanikkara, 1992) Sabitha, R; Sosamma Jacob