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Kerala Agricultural University, Thrissur
The history of agricultural education in Kerala can be traced back to the year 1896 when a scheme was evolved in the erstwhile Travancore State to train a few young men in scientific agriculture at the Demonstration Farm, Karamana, Thiruvananthapuram, presently, the Cropping Systems Research Centre under Kerala Agricultural University. Agriculture was introduced as an optional subject in the middle school classes in the State in 1922 when an Agricultural Middle School was started at Aluva, Ernakulam District. The popularity and usefulness of this school led to the starting of similar institutions at Kottarakkara and Konni in 1928 and 1931 respectively.
Agriculture was later introduced as an optional subject for Intermediate Course in 1953. In 1955, the erstwhile Government of Travancore-Cochin started the Agricultural College and Research Institute at Vellayani, Thiruvananthapuram and the College of Veterinary and Animal Sciences at Mannuthy, Thrissur for imparting higher education in agricultural and veterinary sciences, respectively. These institutions were brought under the direct administrative control of the Department of Agriculture and the Department of Animal Husbandry, respectively. With the formation of Kerala State in 1956, these two colleges were affiliated to the University of Kerala. The post-graduate programmes leading to M.Sc. (Ag), M.V.Sc. and Ph.D. degrees were started in 1961, 1962 and 1965 respectively.
On the recommendation of the Second National Education Commission (1964-66) headed by Dr. D.S. Kothari, the then Chairman of the University Grants Commission, one Agricultural University in each State was established. The State Agricultural Universities (SAUs) were established in India as an integral part of the National Agricultural Research System to give the much needed impetus to Agriculture Education and Research in the Country. As a result the Kerala Agricultural University (KAU) was established on 24th February 1971 by virtue of the Act 33 of 1971 and started functioning on 1st February 1972. The Kerala Agricultural University is the 15th in the series of the SAUs.
In accordance with the provisions of KAU Act of 1971, the Agricultural College and Research Institute at Vellayani, and the College of Veterinary and Animal Sciences, Mannuthy, were brought under the Kerala Agricultural University. In addition, twenty one agricultural and animal husbandry research stations were also transferred to the KAU for taking up research and extension programmes on various crops, animals, birds, etc.
During 2011, Kerala Agricultural University was trifurcated into Kerala Veterinary and Animal Sciences University (KVASU), Kerala University of Fisheries and Ocean Studies (KUFOS) and Kerala Agricultural University (KAU).
Now the University has seven colleges (four Agriculture, one Agricultural Engineering, one Forestry, one Co-operation Banking & Management), six RARSs, seven KVKs, 15 Research Stations and 16 Research and Extension Units under the faculties of Agriculture, Agricultural Engineering and Forestry. In addition, one Academy on Climate Change Adaptation and one Institute of Agricultural Technology offering M.Sc. (Integrated) Climate Change Adaptation and Diploma in Agricultural Sciences respectively are also functioning in Kerala Agricultural University.
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ThesisItem Open Access Characterization and identification of black pepper accessions (Piper nigrum L.) for stress tolerance and quality(Department of plant breeding, College of Horticulture, Vellanikkara, 2019) Prakash, K M; KAU; Jiji, JosephBlack pepper (Piper nigrum L.), often described as the ‘King of spices’ is the most important spice crop, grown for its berries in the world. Indian pepper is preferred across the globe due to its intrinsic qualities. Foot rot is a devastating disease of black pepper. In the changing climate, drought can be a major threat in black pepper production. Hence, the present study was taken up at College of Horticulture, Vellanikkara and ICAR-IISR, Kozhikode to characterise and to identify superior accessions of black pepper for yield, quality and tolerance to biotic and abiotic stresses. Fifty accessions of black pepper in the bearing stage maintained in the National Active Germplasm Site of ICAR-IISR, Kozhikode formed the base material for the study. The accessions were characterised for fifty qualitative and fifty quantitative characters following the descriptor developed by IPGRI (1995). Wide variability was observed among the accessions for ten qualitative characters. Quantitative characters of shoot, leaf, spike and fruit also showed wide variability. Field tolerance to foot rot disease and pollu beetle infestation was observed among the accessions. Twenty accessions were selected from the base collection based on superiority of yield (> 450g green berries/vine) , field tolerance to foot rot disease infection (biotic susceptibility score 1) and pollu beetle infestation (biotic susceptibility score 1-3). They were further evaluated for biochemical principles of quality, tolerance to foot rot disease under artificial inoculation and tolerance to drought by physiological and biochemical analyses. Piperine, essential oil and oleoresin ranged from 3.61 - 6.96 per cent, 3.00 - 5.87 per cent and 7.10 - 11.18 per cent, respectively, across the accessions. The accessions with high value of piperine, essential oil and oleoresin were identified as 7293, 7211 and 7289 respectively. The two accessions identified viz. 7293 and 7252 contained more piperine than the highest of Panniyur 2 (6.6 per cent) reported among the released varieties . Artificial inoculation of selected accessions using Phytophthora capsici culture for screening for foot rot disease resistance based on over all disease severity index of both stem and leaf lesions showed that accession 7259 was moderately resistant. The selected accessions did not exhibit significant variation for various physiological and biochemical parameters at field capacity. However higher value of photosynthesis, chlorophyll content, chlorophyll stability index, relative water content and membrane stability index and low leaf temperature were observed for accessions viz. 7215, 7240, P 5 and 7241 after five days and ten days of moisture stress induction following field capacity compared to other accessions. Higher values of proline, SOD, catalase and peroxidase were also observed for these accessions. The visual scoring showed that accessions with higher values for most of physiological and biochemical parameters of drought tolerance viz. 7215, 7240, P5, and 7241 had lesser number of fallen leaves and more number of leaves retained at permanent wilting point (PWP). The accessions 7215 and 7240 took twenty days to reach PWP compared to eleven accessions which took only 16 days to reach PWP. Foliar nutrition with sulphate of potash, IISR - Power mix and Pink Pigmented Facultative Methylotrophs (PPFM) had positive effect on drought tolerance for the accessions (7215, 7240, P5 and 7241) having natural tolerance. The identified accessions with high yield , quality and tolerance to biotic or abiotic stress can be used for further breeding programme.ThesisItem Open Access Development of Cytoplasmic male sterile line in an identified rice variety of Kerala through marker assisted back crossing(Department of Plant Breeding & Genetics, College of Agriculture, Vellayani, 2019) Tejashree Shivaputra, Lachyan; KAU; Jayalekshmy, V GProduction and productivity of rice, the staple food for over half of the world's population, is insufficient to meet the demand of the increasing population in the rice consuming nations. One of the readily available and economically viable technological options for meeting the projected global demand of rice is exploitation of heterosis through large-scale cultivation of rice hybrids. The role of hybrids in enhancing the productivity is widely acknowledged. The commercial rice hybrids are currently based on cytoplasmic genic male sterility (CGMS) system. The availability of stable cytoplasmic male sterility and fertility restoring system is vital for commercial exploitation of heterosis in rice. The cytoplasmic male sterile (CMS) lines introduced from China were found unsuitable to use as such in developing hybrid rice in India. Limited resources of CMS and low variation of CMS lines causes genetic vulnerability. Therefore, to diversify CMS lines, breeders can transfer this character to the existing putative maintainer lines through repeated backcrosses. So the present study entitled “Development of cytoplasmic male sterile line in an identified rice variety of Kerala through marker assisted back crossing” was undertaken as an initial step for the development of hybrid rice for Kerala with the objective of transferring the cytoplasmic male sterility from the CMS line to the Kerala rice variety, identified as maintainer for CGMS system through marker assisted back crossing. This study was conducted from December 2016 to August 2019. In the present study, four Kerala rice varieties were used, which were reported as potential maintainers of CRMS31A line in the research work conducted by Das (2017). For identification of the best maintainer among the four potential maintainers under study, these lines were crossed with CMS line CRMS31A. The pollen fertility per cent and spikelet fertility percent of hybrids was lowest in the cross CRMS 31A x Jyothi and highest in the cross CRMS 31A x Aruna. The highest pollen and spikelet fertility percent was observed in cross with Aruna with 60% seed set. Kanchana and Bharathy on crossing with CRMS31A showed 91% and 57% pollen sterility which lead to 99.11% and 74.83% of spikelet sterility respectively. Among 30 F1’s obtained from the cross CRMS31A x Jyothi, two F1 plants showed 99.99% pollen sterility and 100% spikelet sterility. Pollen and spikelet sterility of the hybrids indicated that Kanchana, Bharathy and Aruna were partial maintainers for CMS line. Rice variety Jyothi, alone was identified as maintainer for cytoplasmic male sterile line CRMS31A. The transfer of ms gene was confirmed at molecular level using mitochondrial WA-CMS-specific marker “drrcms”. Foreground selection was carried out in 30 F1 plants and it showed homozygous band similar to male sterile line. F1 plants with 99.99% pollen sterility and 100% spikelet sterility, were backcrossed with maintainer Jyothi to obtain BC1F1 seeds.. These BC1F1plants were screened with cytoplasmic male sterility specific marker and all the plants were identified with gene of interest i.e. cms gene. Selection of polymorphic markers was done for background selection in the identified plants with 100% spikelet sterility. 170 genome wide SSR primers were used for screening of the maintainer and CMS line to locate 31 SSR markers specific to the maintainer. The background selection was carried out on 17 BC1F1 plants using 31 loci polymorphic in the parents. The BC1F1 plants with highest percentage of recurrent parent genome recovery were selected based on the background markers. The maximum percentage of recovery of recurrent parent genome was found to be 68.13% in BC1F1 generation. Pollen sterility in these plants was also studied by screening for fertility restoration gene (Rf). Out of 10 Rf markers used for screening only four markers for Rf4 Rf6 and Rf7 gene were present in the maintainer line Jyothi. The identified markers were then used for screening of BC1F1 plants. It was observed that the BC1F1plants had only Rf7 gene. Pollen and spikelet sterility percent of these plants could not be recorded due to absence of flowering even on the 240th day after sowing. The plants were under vegetative phase upto 260-270 days, after which they started wilting. Due to absence of flowering, these plants were screened with marker specific to flowering time gene Hd3a. On gel documentation the band was obtained revealing the presence of Hd3a gene at 179bp. To confirm the possible reason for absence of flowering even in the presence of Hd3a gene, the expression of Hd3a gene was studied. Expression was studied at different stages of plant growth i.e. 15, 30, 45, 60, 75 and 110 days after sowing in Jyothi variety and 240 days old BC1F1 plants. It was observed that the expression of target gene in the variety Jyothi was reduced at 30 days after sowing. But in later stages, gradual increase in the expression level was observed. A drastic increase in expression was recorded at flowering stage (75DAS) of the Jyothi variety and then the expression was gradually reduced towards maturity stage. Whereas the Hd3a gene was not expressed in BC1F1 plants i.e. the gene was silenced. The reason for non expression of Hd3a gene in the background of sterile cytoplasm and ms gene requires further investigation. Advanced generation of the cross between another CMS line, UPRI95-17A and maintainer line Kanchana, which was developed earlier under a project for transferring male sterility to Kanchana variety was further studied. Sterility was studied by pollen test and spikelet test. Three of the plants showed 99.09% - 100% pollen sterility and 100% spikelet sterility. All the plants were screened with 28 SSR markers. Foreground selection was carried out using mitochondrial WA-CMS-specific marker “drrcms”. Band obtained in the advanced generation plants were similar to the band of male sterile parent. Recovery of the recurrent parent genome was analysed using 28 SSR markers. One plant of this advanced generation, recovered 92.86% of recurrent parent genome. Observations were taken on eight morphological traits viz. plant length, leaf length, leaf breadth, total number of tillers, panicle length, days for flowering, number of spikelets per panicle and pollen fertility percentage. It was observed that out of eight quantitative traits, four traits i.e. plant length, leaf length, total number of tillers and panicle length were almost similar to the maintainer line Kanchana. The number of spikelets per panicle and leaf breadth was less than Kanchana. In the present study, Kerala rice varieties, Jyothi and Kanchana showed 100% pollen and spikelet sterility when crossed with CMS line CRMS 31A and UPRI95-17A respectively. On the basis of pollen and spikelet sterilirty Jyothi and Kancjana were identified as maintainers for WA-CMS line CRMS 31A and UPRI95-17A. The reason for non expression of Hd3a gene in the background of sterile cytoplasm i.e. backcrossed plants of Jyothi variety needs to be further investigated. The identified plants from the backcross population of UPRI95-17A and Kanchana, with higher recurrent parent genome (RPG) recovery percentage can be backcrossed for another generation to get RPG of 99%. Later it can be used for commercial hybrid seed production with compatible restorers for UPRI95-17A. This will be the first CMS line in the background of Kerala rice variety.ThesisItem Open Access 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; KAU; Jiji JosephCowpea [Vigna unguiculata (L.) Walp.] is important pulse crop rich in nutrients, especially proteins. This crop is cultivated in the tropics of Asia, Africa and other parts of the world. Nevertheless, the production of cowpea is unable to achieve its summit. One of the prime reasons for this is the infestation of a notorious pest, the spotted pod borer, (Maruca vitrata Fab.; Lepidoptera: Crambidae). Spotted pod borer is one of the most important post-flowering pests of cowpea in the tropics. It is a major lepidopteran pest and damage cause to cowpea by the pest almost always crosses economic threshold level. Hence, the present investigation was conducted in the Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, Kerala Agricultural University, Thrissur during 2015 to 2018 with the objective of identification and incorporation of resistance against spotted pod borer in high yielding varieties of cowpea and assessment of parental polymorphism at the molecular level Thirty genotypes of cowpea formed the material for the study. These genotypes were subjected to field screening against spotted pod borer. These genotypes were also evaluated for morphological and biochemical basis of resistance. Five selected genotypes from experiment 1 then hybridised with four high yielding genotypes viz., Geethika, Vellayani Jyothika, Lola and Kashi Kanchan following Line × Tester mating design. Twenty F 1 hybrids evaluated for field resistance and the morphological basis of resistance. Progenies of selected F 1 hybrids grown as F 2 populations and evaluated for same parameters as like F 1 s. Thirty genotypes were also subjected to molecular screening by 40 SSR primers. Wide variation was observed in terms of different damage parameters. Ten genotypes viz., Hridya, Palakkadan thandan payar, EC 300039, EC 98668, EC 101216, IC 52110, IC 39945, IC 2918, IC 39922 and IC 39916 recorded total damage below five per cent. Among them, IC 39922 observed to have no flower bud and flower damage, EC 300039, EC 98668, IC 52110, IC 39945, IC 2918 and IC 39916 recorded no flower damage, whereas, Palakkadan thandan payar, IC 39945, IC 2918 and IC 39947 were free from pod damage. The highest damage was recorded in the variety Bhagyalakshmy (48.46 %) followed by variety Lola (30.04 %). Analysis of the morphological basis of resistance to spotted pod borer revealed the negative correlation of trichome density and length on flower bud, trichome density on the pod and pod wall thickness with respective damage parameters. With respect to the biochemical basis of resistance to spotted pod borer, total sugar content, reducing sugar content and non-reducing sugar content of flower bud and pod showed a positive correlation with damage parameters, but the correlation was not significant. However, the total protein content of pod showed a strong and positive correlation with pod damage. Total phenol content of flower bud showed strong negative correlation with damage parameters. Polyphenol oxidase activity in flower bud and pod exhibited a strong negative correlation with damage parameters. The crude fibre content of pod also showed a strong negative correlation with pod damage. In experiment 3, three SSR primers viz., CLM0061, CLM0295 and CLM0300 recorded high polymorphic information content (0.70, 0.71 and 0.76, respectively). Primer CLM0190 observed to have high amplicon size (307.03-415.73 bp). Jaccard’s similarity coefficient was highest between IC 52118 and IC 39916 (0.643) and was lowest between Kashi Kanchan and TVX-944 (0.022). Cluster analysis of SSR data grouped 30 genotypes in 22 clusters, and the 21 st cluster was observed to have more members (3 genotypes). Most resistant genotype, IC 2918, grouped in a separate cluster which proved its diverse nature from other genotypes. Principal component analysis of SSR data placed three yard-long bean genotypes viz., Geethika, Vellayani Jyothika and Lola near to each other in a two-dimensional score plot. The same analysis also formed two clusters with more number of genotypes which placed resistant and susceptible genotypes separately. In field screening of F 1 hybrids, Hybrid 20 observed to have total damage below 5 per cent. Eight hybrids recorded total damage in the range of 5 to 10 per cent. These hybrids were selected for next experiment. Line × Tester analysis of F 1 s revealed Kashi Kanchan, EC 98668 and IC 2918 as a good combiner for resistance against spotted pod borer. Hybrid 5, Hybrid 6, Hybrid 10, Hybrid 11, Hybrid 13, Hybrid 15, Hybrid 16, Hybrid 17, Hybrid 18, Hybrid 19 and Hybrid 20 observed to have desired negative heterosis (mid-parent) for total damage. In F 2 plant screening, Hybrid 1 population recorded low mean for total damage (6.76 %), whereas, the population of Hybrid 3 recorded high mean (12.17 %). Around 100 plants of F 2 generation recorded total damage below ten per cent. Out of them, around 38 plants also registered good yield. These plants should be further evaluated to isolate high yielding resistant segregants.ThesisItem Open Access Diversity analysis in landraces of rice(oryza sativa L.) in Wayanad through morphological and molecular polymorphism study(Department of Plant Breeding and Genetics,College of Horticulture, Vellanikkara, Thrissur, 2018) Manjunatha, G A; KAU; Elsy, C RThe present investigation entitled “Diversity analysis in landraces of rice (Oryza sativa L.) in Wayanad through morphological and molecular polymorphism study” was conducted at the Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, Thrissur and Regional Agricultural Research Station, Ambalavayal, Wayanad during the period 2015- 2018. The study aimed to collect and characterize the rice landraces of Wayanad at morphological level and also to characterize the popular aromatic genotypes of the area at molecular level. DUS characterization revealed that, all the landraces under the study exhibited presence of leaf collar, absence of anthocyanin colouration of leaf collar, presence of leaf ligule, split shape of leaf ligule, absence of anthocyanin colouration for plant parts like lemma (keel), area below apex of lemma, stem nodes and presence of secondary branching in panicle. Variability was exhibited for majority of characters namely coleoptile colour, basal leaf sheath colour, intensity of green colour of leaf, anthocyanin colouration of plant parts like leaf, leaf sheath, auricles, ligules, lemma apex, area below apex of lemma, distribution of anthocyanin colouration in leaf, pubescence of leaf blade surface, leaf auricles, culm attitude, density of pubescence of lemma, colour of stigma, attitude of flag leaf blade, curvature of main axis of panicle, lemma and palea colour, presence and colour of awns, distribution of awns in panicle, secondary branching in panicle, attitude of branches in panicle, panicle exsertion, leaf senescence, sterile lemma colour, shape, colour and aroma of decorticated grain, gelatinization temperature and lodging nature.Thavalakannan, Mullan puncha and Sugandhamathi exhibited distinct morphological characters. Among the 60 landraces, Kanni kayama, Ambalavayal-1, Kothandan, Onamottan, Thondi-2, Chenthadi, Kannali and Thondi-1 exhibited significantly high grain yield per plant, which was on par with that of Uma and Aathira (high yielding check varieties), indicating the possibility of commercial cultivation of these cultivars. High PCV value than GCV for all the characters under study, indicated the influences of environmental factors on the characters. High heritability and high genetic gain was exhibited by the characters like length of leaf blade, width of leaf blade, stem thickness, stem length, number of tillers per plant, number of panicles per plant, length of panicle main axis, number of spikelets per panicle, number of grains per panicle, seed setting (per cent), 1000 grain weight, grain length, grain L/B ratio, decorticated grain L/B ratio and grain yield per plant, indicating that these characters could be used in crop improvement programmes. Character association studies including correlation and path-analysis revealed that, five characters viz., width of leaf blade, stem length, number of panicles per plant, weight of 1000 grains and decorticated grain width showed both positive correlation and positive direct effect on grain yield per plant. Cluster analysis based on qualitative characters categorized 60 landraces into 11 clusters at 74 percent similarity level. Based on D2 statistics analysis, the landraces were grouped into seven clusters with variable number of landraces in each cluster, indicating wide genetic diversity in the Wayanad landraces. All the inter-cluster distances were higher than the intra-cluster distance, indicating the wider genetic diversity among landraces of different clusters. Molecular characterization of aromatic rice genotypes of Wayanad by SSR (RM) profiling revealed high level of genetic polymorphism among the genotypes studied. Out of 86 SSR markers used for molecular characterization, 44 markers were polymorphic and remaining 42 were monomorphic. Maximum number of amplicons was exhibited by RM247 with five amplicons, followed by RM85, RM251, RM248 and RM493 with four amplicons each. The highest PIC value was exhibited by RM247 (0.90), followed by RM85, RM251 and RM493 with 0.88 PIC value each. Out of 86 SSR (RM) markers, 21 markers distinguished Basmati from traditional aromatic landraces of Wayanad viz., Gandhakasala and Jeerakasala. Seven SSR markers distinguished Gandhakasala from Jeerakasala, whereas 23 markers distinguished Basmati from Jeerakasala. Twenty-two markers distinguished Basmati from Gandhakasala and 23 markers distinguished aromatic group from non-aromatic group. Cluster analysis for molecular characterization revealed maximum similarity coefficient (1.00) within all the Jeerakasala morphotypes and all the Gandhakasala morphotypes. Cluster analysis effectively differentiated Basmati, Jeerakasala, Gandhakasala, Uma and Aathira from each other. Among the five clusters formed, cluster III was the largest one comprising all the 12 Gandhakasala morphotypes, followed by cluster IV with all Jeerakasala morphotypes. Cluster I, Cluster II and Cluster V exhibited one genotype each namely Aathira, Uma and Basmati, indicating their genetic distinctness.ThesisItem Open Access Distant hybridization and compatibility studies in wild orchids(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2018) Seeja, G; KAU; Arya, KThe present investigation entitled “Distant hybridization and compatibility studies in wild orchids” was carried out with the objective of studying the cross compatibility through distant hybridization in wild orchids and molecular characterization of hybrid seedlings using RAPD/ISSR- markers. The study was carried out at Saraswathy Thangavelu Extension Centre of KSCSTE- JNTBGRI, Puthenthope, Thiruvananthapuram and at College of Agriculture, Vellayan, Thiruvananthapuram. The research work included experiment I, II, III and IV. Experiment I was carried out in two parts as evaluation of parents and hybridization, experiment II was cross compatibility studies of selected species, experiment III was in vitro culture of resultant hybrids and evaluation of seedlings and experiment IV was molecular characterization of hybrids. In experiment I, wild types belonging to different genera/species namely Vanda (Vanda tessellata, Vanda spathulata, Vanda sp.), Rhynchostylis retusa, Spathoglottis (Spathiglottis albida, Spathoglottis unguiculata, Spathoglottis gracilis, Spathoglottis plicata), Aerides ringens and Arundina graminifolia available at the orchid conservatory of Saraswathy Thangavelu Extension Centre of JNTBGRI, Puthenthope, Thiruvananthapuram were selected as parents based on variability in floral characteristics and these were evaluated for various quantitative and qualitative characters. Statistical analysis was carried out in Completely Randomized Design and analysis of variance revealed highly significant differences for all the biometrical characters included in the evaluation. Estimation of components of variance revealed that Phenotypic Coefficient of Variation (PCV) was higher than both Genotypic Coefficient of Variation (GCV) and Environmental Coefficient of Variation (ECV) for all the traits studied. PCV (143.93) and GCV (128.52) were the highest for number of spikes plant-1. Heritability was the highest for flower width (99.06 %) and it was high for all thirteen traits. Genetic advance as percent of mean was high for all the traits studied. It was the highest for number of spikes plant-1 (239.80). Estimated phenotypic correlation coefficient revealed highly significant and positive association between flower length and flower width (0.9519). The highly significant and negative association was obtained between internodal length and days for inflorescence emergence to first flower opening (-0.6896). Similarly genotypic correlation coefficient exhibited highly significant and positive association between flower length and flower width (0.9545) and highly significant and negative association between plant spread and number of leaves plant-1 (-0.9903). Based on this, parents were selected and were crossed in all possible combinations. In Vanda a hybrid evolved from the wild species was available at the centre and it was also crossed with the wild parents to study the cross compatibility of wild species with hybrids. In experiment II, cross compatibility of 152 cross combinations made was studied based on successful capsule setting. Capsule characteristics were recorded. The genus Spathoglottis took only four days for attaining pod setting whereas for the genus Rhynchostylis it was ten days for pod setting. Capsule attained maturity much earlier (30 days) in the genus Spathoglottis but the species Vanda spathulata took more days for attaining capsule maturity i.e., harvesting stage (150-154 days). In experiment III, capsules developed among thirty one cross combinations were subjected to seed culture for rapid germination under in vitro condition and seedling development. Observations regarding seed germination were recorded and seeds of ten crosses exhibited seed greening and initiation of protocorm development. Seeds of seven cross namely one intergeneric hybrid and six interspecific hybrids were developed into seedlings. Evaluation of hybrid seedlings was made based on variability in morphological characters at monthly intervals. The six interspecific hybrids were Spathoglottis unguiculata x Spathoglottis albida (Culture 622), Spathoglottis gracilis Spathoglottis unguiculata (Culture 652), Spathoglottis albida x Spathoglottis unguiculata (Culture 778), Spathoglottis albida x Spathoglottis unguiculata (Culture 1368), Spathoglottis plicata x Spathoglottis unguiculata (Culture 1386), Spathoglottis unguiculata x Spathoglottis plicata (Culture 1388) and intergeneric hybrid was Spathoglottis unguiculata x Vanda sp. (Culture 679). In experiment IV, molecular characterisation of seedlings evolved through the crossing was done based on ISSR analysis to distinguish F1 hybrid seedlings from selfed seedlings and their parents. Percentage of polymorphism between hybrids and parents ranged from 11.99 to 32.97 and these results confirmed that all the seven crossed seedlings tested were F1 hybrids and these were distinct from their relevant parents. The above study revealed that through distant hybridization in wild orchids, one intergeneric hybrid Spathoglottis unguiculata x Vanda sp. and six interspecific hybrids viz., Spathoglottis unguiculata x Spathoglottis albida, Spathoglottis gracilis x Spathoglottis unguiculata, Spathoglottis albida x Spathoglottis unguiculata, Spathoglottis albida x Spathoglottis unguiculata, Spathoglottis plicata x Spathoglottis unguiculata, Spathoglottis unguiculata x Spathoglottis plicata could be obtained and molecular characterization confirmed the hybridity of the intergeneric and interspecific hybrids. Therefore, the intergeneric and six interspecific F1 hybrids evolved through distant hybridization can be carried forward for the development of varieties or can be utilised for genetic improvement programme in future.ThesisItem Open Access Characterization and taxonomic evaluation of landraces of capsicum spp. in Kerala(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2018) Asish, I Edakkalathur; KAU; Minimol, J SCapsicum is a unique genus coming under the family Solanaceae. This genus includes nearly 40 species among which five viz. C. annuum, C.chinense, C. frutescens, C. baccatum and C. pubescens are cultivated. Large number of types, the transitory nature of these types, introgressive hybridization and environmental influence make species arrangement in Capsicum a confusing task. Even though Kerala is rich in the diversity of Capsicum, a comprehensive study of landraces available has not yet been carried out. In this background, the present project was carried out with the objective of characterizing the landraces of Capsicum species in Kerala using morphological markers and validating the key for the identification of different taxa using molecular and biochemical markers. Ripened fruit samples of 262 accessions were collected from the different districts of Kerala during December, 2013 to August, 2014. These accessions were planted in augmented block design along with released varieties from Kerala Agricultural University as checks at National Bureau of Plant Genetic Resources, Regional Station, Thrissur. Morphological characterization was done by considering both qualitative and quantitative characters. Numerical Taxonomy and Multivariate Analysis System (NTSYS) software version 2.10 z was used for construction of phenogram based on forty four qualitative characters available in the Capsicum descriptor (IPGRI, 1995). Accessions belonging to three species viz., C. annuum, C.chinense and C. frutescens formed species specific clusters. C. baccatum accessions clustered with C. frutescens. C. chinense accessions were clearly separated and distantly placed from C. annuum accessions revealing genetic dissimilarity between the above two species.. Out of 42 clusters formed at 70 per cent similarity level, 39 are mono-specific clusters. Morphological characterization based on qualitative unit characters aligned 95 per cent of the accessions into species specific clusters. Diversity relationship accessed on the basis of 32 quantitative characters grouped the accessions into 14 clusters. C. annuum accessions formed mono- specific clusters where as C. chinense and C. frutescens accessions grouped together in three different clusters revealing that C. frutescens and C. chinense are inseparably related. C. annuum accessions were found standing as a separated group. C. baccatum accessions clustered along with C. frutescens - C. chinense group. Representative accessions selected from each qualitative cluster along with eight released varieties were analysed using 17 RAPD primers. Dendrogram consisted of eleven mono-specific clusters indicating the ability of RAPD markers to align the genotypes into correct taxa. Clustering of C. annuum accession with C. frutescens (two clusters) and C. chinense (two clusters) as well as clustering between C. frutescens and C. chinense (three clusters) accessions points to the genetic similarity within the annuum complex. C. baccatum accession expressed more similarity with C. frutescens. The genetic background of the annuum complex was further scrutinized with 18 SSR primers among 19 genotypes comprising of eight C. annuum, seven C. chinense and four of C. frutescens accessions. Out of five clusters formed, two were mono-specific C. annuum clusters. Other three clusters composed of both C. frutescens and C. chinense accessions in each clusters. Thus SSR characterization clearly revealed the close genetic similarity between C. frutescens and C. chinense species and this group is distantly placed from C. annuum species. Clustering pattern based on RAPD primers for the above 19 genotypes strongly supported the result of SSR analysis with formation of two major clusters viz., one composed of C. annuum type only and the other major cluster comprising both species viz., C. frutescens and C. chinense. Dendrogram constructed based on RAPD and SSR banding pattern also substantiated above result by forming mono-specific C. annuum clusters and clusters comprising accessions from both species viz., C. frutescens and C. chinese. Both RAPD and SSR analysis points to the close similarity among the released varieties. All the released varieties of C. annuum type are grouped in to the same cluster. Combined analysis based on RAPD and SSR data resulted in the formation of two major clusters viz., first cluster comprising of only released varieties and second of landraces. Compared to separate RAPD or SSR characterization, combined analysis based on RAPD and SSR data more precisely assigned accessions into corresponding species specific clusters and formed mono-specific clusters. At 70 per cent similarity level, combined RAPD-SSR analysis expressed 79 per cent accuracy for allotting accessions to species specific clusters. More number of molecular markers improve the accuracy of species alignment and molecular markers serve a complementary role to assist phenetics. Quantity of ascorbic acid ranges from 81.40 to 327.58 mg per 100g of ripe chilly. The upper limit of capsaicin and oleoresin were 0.87 and 23.5 per cent, respectively. Accessions superior in performance to the check varieties for economic and biochemical characters were identified through the agronomic characterization. Majority of the yield contributing characters exhibited high GCV coupled with moderate to high heritability and high genetic gain revealing wide variability contributed by genetic factors and indicating scope for selection. DNA fingerprinting profile of released varieties and elite accessions were developed using SSR primers. Species specific fingerprinting of C. annuum, C. chinense and C. frutescens species were also made for differentiation of species with SSR primers. Validation of taxonomic key carried out integrating characters viz., spot colour at the throat of corolla, annular constriction at calyx, number of flowers per axil, corolla colour, position of flower at anthesis, leaf shape, margin of calyx at fruiting, fruit to pedicel length ratio and level of main stem bifurcation. SSR primers viz., CAMS-101, CAMS-806 and CAMS-864 as well as RAPD primers viz., OPB-10,OPG-02, OPG-03, OPB-08 and OPA-03 identified as promising in identification and differentiation of cultivated species of Capsicum.ThesisItem Open Access 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; KAU; 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.ThesisItem Open Access Stability analysis and molecular characterization of F1 hybrids in brinjal (Solanum melongena L.)(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2018) Kavishetti Vinay, Vishwanath; KAU; Lekha Rani, CStability analysis helps in assessing genotype × environment interaction in order to identify stable genotypes in large multi-environment trials. Therefore the present study entitled “Stability analysis and molecular characterization of F1 hybrids in brinjal (Solanum melongena L.)” was carried out to evaluate ten hybrids along with one check across four locations. The locations selected for trials were College of Agriculture, Vellayani and farmer’s fields at Thiruvalla, Sadanandapuram and Kayamkulam in Kerala. The trial seasons were kharif (2015- 16) and summer (2016-17). The objective was to study the performance of superior hybrids over different locations and seasons from heterotic crosses of brinjal and to confirm the hybridity using SSR markers. Randomized block design with four replications was employed. Stability and adaptability of yield and yield attributing characters of hybrids were analysed by Eberhart-Russell model (1966). Pooled analysis of variance revealed significant differences among the genotypes, environments and genotype × environment interaction for all the characters studied. The indication was that the hybrids responded differently to changes in the environment. Promising hybrids were identified on the basis of stability parameters viz., overall mean, regression coefficient (bi) and deviation from regression (S2di). In kharif season, the hybrid Wardha local × Palakurthi local was observed to be stable and widely adapted to all environments for days to first flowering, number of fruits plant-1, fruit weight, fruit length, fruit girth, calyx length, yield plant-1, yield plot-1 and plant height. The hybrid Wardha local × Swetha was identified as stable with regard to fruit weight, yield plant-1 and yield plot-1. The hybrid Wardha local × Vellayani local was identified as stable for favourable environments with regard to fruit length, yield plant-1 and yield plot-1. The hybrid Surya × Vellayani local was found stable for poor environments with regard to days to first flowering. Stability analysis for the summer season crop also revealed the hybrid Wardha local × Palakurthi local as stable across all environments with respect to days to first flowering, days to first harvest, number of fruits plant-1, fruit weight, fruit length and yield plant-1. Swetha × Vellayani local hybrid was stable across all environments with regard to days to first harvest, fruit weight, fruit length, calyx length, yield plant-1, yield plot-1 and plant height. The hybrid Surya × NBR-38 was found stable with respect to yield plant-1, yield plot-1 and plant height for favourable environments. Qualitative characters viz., total phenols, total sugars and vitamin C content and biotic stress traits viz., shoot and fruit borer infestation and bacterial wilt incidence were recorded for the eleven hybrids in kharif and summer season in all four locations. On the basis of overall mean performance of hybrids, results illustrated that the hybrid Swetha × Vellayani local recorded the highest values for total phenols and the hybrid NBR-38 × Vellayani local recorded the lowest values. Total sugar content was highest in Wardha local × Swetha and NBR-38 × Selection Pooja and lowest in Surya × NBR-38. The hybrid Neelima and Swetha × Vellayani local recorded higher quantity of vitamin C and NBR-38 × Vellayani local and NBR-38 × Selection Pooja recorded the lowest values. The hybrids Wardha local × Palakurthi local, Swetha × Vellayani local and Neelima recorded minimum infestation of shoot and fruit borer and was found resistant to bacterial wilt also. Hybrids and their parental lines were characterized using SSR markers. Among the four markers studied, three markers viz., emb01M15, eme08D09 and CSM31 were found to be polymorphic among the parental lines of respective hybrids viz., Surya × NBR-38, Surya × Vellayani local, NBR-38 × Selection Pooja, Swetha × Vellayani local, Wardha local × Swetha, Wardha local × Palakurthi local, Palakurthi local × Vellayani local and NBR-38 × Vellayani local which could be used for ensuring the genetic purity of respective parental lines and hybrids. F2 families viz., Wardha local × Palakurthi local, Wardha local × Swetha, Wardha local × Vellayani local and Swetha × Vellayani local were selected on the basis of yield performance from F1 for further evaluation. F2 populations revealed that family Wardha local × Palakurthi local and Wardha local × Vellayani local were superior in yield performance and yield attributing characters. The present investigation revealed that the hybrids Wardha local × Palakurthi local and Swetha × Vellayani local were stable and widely adapted over different locations and seasons and the hybridity was confirmed with the SSR markers, emb01M15 and eme08D09.ThesisItem Open Access Induced mutagenesis for delayed flowering and high tillering in guinea grass (Panicum maximum Jacq.)(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2018) Sudrik Bibhishan, Popat; KAU; Mareen AbrahamThe present investigation entitled “Induced mutagenesis for delayed flowering and high tillering in guinea grass (Panicum maximum Jacq.)”, was carried out at College of Agriculture, Vellayani during the period 2014-2017. The major objective of the study was to develop high yielding guinea grass types with delayed flowering and high tillering through induced mutagenesis. The first experiment consisted of evaluation and characterization of germplasm accessions of guinea grass. For assessing genetic variability and diversity, thirty seven germplasm accessions were studied with respect to fourteen morphological and three biochemical characters. ANOVA revealed highly significant differences among the genotypes for all the traits analysed except length of internode. High estimates of phenotypic coefficient of variation (PCV) and genotypic coefficient of variation (GCV) were obtained for green fodder yield, dry fodder yield, leaf stem ratio, weight of seeds hill-1 and gibberellic acid content and high PCV and moderate GCV were obtained for leaf area index, number of tillers hill-1, number of leaves hill-1 and number of panicles hill-1. Moderate values of PCV and GCV were obtained for crude protein content and crude fibre content. High heritability coupled with high genetic advance as percent of mean was observed for number of tillers hill - 1 , leaf stem ratio, weight of seeds hill-1, crude protein content, crude fibre content and gibberellic acid content. High heritability coupled with moderate genetic advance as percent of mean was observed for days to flowering and length of panicle. Moderate heritability coupled with high genetic advance as percent of mean was observed for number of leaves hill-1, number of panicles hill-1 and green fodder yield. Green fodder yield showed significant positive correlation with dry fodder yield and weight of seeds hill -1 at phenotypic level. Green fodder yield showed significant positive correlation with dry fodder yield, weight of seeds hill-1 and crude fibre content at genotypic level. Path coefficient analysis revealed that green fodder yield showed high positive direct effect on dry fodder yield followed by length of panicle, weight of seeds hill-1, plant height and leaf stem ratio. Based on observations four varieties viz. two early flowering (FP-553, MS-4690) and two late flowering (PGG-208, MS- 4600) were selected for further studies. The second experiment included induced mutagenesis using physical mutagens on seeds and slips. The physical mutagen used was gamma rays from 60Co source and the initial trial was for fixing LD50 value. For this, seeds of four varieties viz., FP-553, MS-4600, MS- 4690 and PGG-208 were treated with six doses of gamma rays ranging from 60 to 140 Gy along with control. The effects of gamma rays were studied on different parameters such as germination percent, seedling height and survival in M1 generation. Reduction of germination, seeding height and survival were studied. Accordingly LD50 doses were fixed and three doses 60, 80 and 100 Gy were administered to the seeds of four different varieties and field planting of M1 was done. M1 was evaluated for fifteen morphological characters. Statistical analysis of quantitative characters showed that for an increase in gamma ray doses, a proportionate significant increase for various plant growth parameters was evident. Occurrence of major viable mutants from seedling to adult growth stages varied viz., broad and narrow leaves, paired panicle, early and late maturity, dwarf and tall plants, low and high number of tillers and leaves hill-1, short and long internode. Mutagenesis by gamma ray irradiation was employed to treat slips of four genotypes viz., FP-553, MS-4600, MS-4690 and PGG-208. They were treated with six doses of gamma ray ranging from 20 to 100 Gy along with control for fixing LD50 value. The effects of the irradiation doses were studied on regeneration, survival of shoots and height of plants. Based on LD50 value, three doses 20, 40 and 60 Gy were administered along with control to the different varieties and field planting of M1V1 was done. M1V1 were evaluated for fifteen morphological characters. Most of quantitative characters showed an increase in mean values with increase in doses of mutagen. Analysis of data showed that all characters studied were significantly affected by irradiation at 60 Gy. The third experiment was the study of frequency and spectrum of chlorophyll and viable mutants, mutagenic efficiency and effectiveness in M2 generation while employing the three doses of gamma rays and one control. In general the frequencies of chlorophyll mutations increased in linear fashion at low (60 Gy) and medium (80 Gy) doses and the frequency was erratic at higher (100 Gy) doses. The spectrum of chlorophyll mutants included albina and xantha. The highest frequency of viable mutants was produced at 60 Gy dose. There were variations in the occurrence of major viable mutants during the crop growth period stages viz., broad and narrow leaves, paired panicle, days to maturity, dwarf and tall plants, high and low tillering with variations in number of leaves hill-1, short and long internode, plant height, protein content, fibre content and gibberellic acid content. In the M1V2 generation, the chlorophyll mutation frequency increased in a linear fashion at low (20 Gy) and medium (40 Gy) doses and was erratic at higher doses (60 Gy). The chlorophyll mutant spectrum included albino and xantha. The frequency of these mutants varied with treatments. Superior plants with high green fodder yield as a result of increased number of tillers ranging from 37-42 were observed in 60 Gy populations of MS-4600 and PGG-208 when compared to the control (30-32 tillers). Based on single plant performance, plants which recorded high biomass were identified and M1V2 slips were collected and studied in further generations. The mutagenic treatments were effective in inducing various types of chlorophyll and morphological mutants, such as those with significant change in days to flowering, plant height, leaf mutations (narrow, broad, wrinkled); panicle mutations (twin panicle, compact, spread) and biochemical variations in protein content, fibre content and gibberellic acid content. The fourth experiment was the study of M3 and M1V3 generation. The altered morphological traits were stable as shown by their growth performance in different seasons. The mutagenic treatments were effective in inducing both qualitative and quantitative characters. Tall mutants with delayed flowering, high green fodder and dry fodder yield displayed two fold increase in number of tillers and leaves hill-1 compared to control at 60 Gy irradiation. Considerable variation was observed with regard to leaf stem ratio at 60 Gy from slips. To conclude genetic variability was found to be high among the genotypes of guinea grass. Gibberellic acid content was found to vary with genotypes. Genotypes with late flowering habit had low gibberellic acid content. Gamma rays induced variations in both seeds and slips for plant height, days to flowering, number of tillers hill-1, number of leaves hill-1, green fodder yield, crude protein content and gibberellic acid content at dose of 60 Gy. The study yielded mutants with delayed flowering and high tillering which were found to be stable in growth performance in M3 and M1V3. These stable genotypes could be carried forward to develop better varieties.
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