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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 Heterosis breeding in sesame (Sesamum indicum L.).(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2011) Gayathri, G; KAU; Dijee, BastianThe study entitled ‘Heterosis breeding in sesame (Sesamum indicum L.)’ was undertaken at the Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara. The objectives of the study were to collect and evaluate different genotypes of sesame for morphological traits and yield attributes, to identify useful parents producing heterotic crosses and developing hybrids in sesame. The study also intended to develop male sterile lines in sesame through interspecific hybridization with Sesamum malabaricum. Sesamum indicum and Sesamum malabaricum accessions were collected from Kerala and Tamil Nadu and evaluated for their morphological traits. Wide range of variation was noticed for characters like plant height, number of days to flowering and seed yield per plant which contributed maximum to genetic divergence. The genotypes studied were grouped into six clusters. High genotypic coefficient of variation (GCV) was recorded for number of capsules per plant, plant height, seed yield per plant and number of branches per plant. High heritability with high genetic advance as per cent of mean was recorded for number of days to flowering, plant height, number of branches per plant, number of capsules per plant and seed yield per plant. This indicates that the characters are governed by additive gene effects and selection for these traits will be effective. Association analysis revealed that seed yield per plant was correlated to plant height, number of capsules per plant and number of days to flowering. Path coefficient analysis indicated maximum positive direct effect by number of capsules per plant, capsule length, plant height and 1000 seed weight on seed yield per plant. In order to develop hybrids, fourteen parents were selected based on the per se performance of the genotypes. They were crossed in line X tester mating design. Forty eight hybrid combinations obtained were raised in the field along with the parents and evaluated for their heterosis and combining ability effects. Parental genotypes AVTS-06-5, AVTS-06-10, IVTS-06-12, KYM-1, Tilak and TMV-6 were identified as high combiners based on general combining ability (gca) effects. Two combinations viz. AVTS-06-5 X KYM-1 and IVTS-06-12 X TMV-3 had significant values of per se performance, specific combining ability (sca) effects and standard heterosis for seed yield per plant. They can be evaluated for their hybrid vigour over locations and seasons. The crosses AVTS-06-5 X TMV-3, AVTS-06-5 X TMV-6 and TCR 3279A X KYM-1 have been identified as potential cross combinations for isolation of promising segregants as the parents involved in these crosses had high significant gca effects for seed yield per plant but the hybrids recorded non significant sca effects. Interspecific hybridization between S.malabaricum and S.indicum was attempted to develop male sterile lines. Seed set was noticed in three interspecific hybrids which failed to germinate due to embryo abortion. Hence these embryos were rescued and raised in vitro to obtain the hybrids.ThesisItem Open Access Gene pyramiding for bacterial blight resistance in rice variety Uma (Mo 16)(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2016) Tintumol, Joseph; KAU; Rose Mary, FranciesExploiting host-plant resistance through pyramiding of resistance genes have been recommended as the best approach to impart durable resistance to rice varieties in order to combat the bacterial blight (BB) disease caused by Xanthomonas oryzae pv.oryzae (Xoo). In lieu of this, F1s were produced by hybridizing the susceptible elite rice variety Uma with resistant donor parent Improved Samba Mahsuri (ISM) harbouring three R-genes xa5, xa13 and Xa21. BC1F1 individuals were generated by backcrossing the F1s using variety Uma as the recurrent parent. The present study aimed to identify the R-genes introgressed individuals in the BC1F1 population as well as to produce BC2F1s and BC1F2s of the identified R-genes introgressed BC1F1s. Foreground selection of the BC1F1 individuals was done using the R gene linked molecular markers. Restriction digestion of the PCR product of STS marker RG 556, linked to R gene xa5, with Dra1 restriction enzyme, resulted in production of alleles of size 128 bp, 514 bp, 587 bp, 624 bp, 650 bp and 836 bp in all the BC1F1 individuals as well as the parents indicating the presence of R gene xa5 in all the individuals studied. Amplification of DNA of the individuals with the functional marker xa5 SR further confirmed the presence of R gene xa5 in both the parents as well as in all the BC1F1s. Restriction digestion of the PCR product of STS marker RG 136, linked to R gene xa13, with Hinf1, produced alleles similar to that of the donor parent ISM in three BC1F1s namely, plant no. 8.3.2, plant no. 8.3.3 and plant no. 8.3.9, indicating the presence of R gene xa13 in these plants. The presence of gene xa13 in the identified BC1F1s was further affirmed by using the functional marker xa13 promoter. The analysis had resulted in the production of 560bp allele associated with the resistant allele of gene xa13 in homozygous state from donor parent ISM in the three BC1F1s mentioned above. Out of the 95 BC1F1 individuals scored with the STS marker pTA 248 linked to R gene Xa21, only BC1F1s plant no. 8.3.2, plant no. 8.3.3 and plant no. 8.3.9 were found to possess Xa 21. Results thus obtained revealed thatBC1F1plant no. 8.3.2, plant no. 8.3.3 and plant no. 8.3.9 were R gene pyramids (xa 5+xa 13+ Xa 21). Background profiling of the three R-genes introgressed BC1F1s using 22 rice microsatellite markers, revealed presence of the donor parent allele in the homozygous state. PCR analysis with the marker RM 307, however, revealed the presence of alleles from both the parents, ISM and Uma in the BC1F1 plant no. 8.3.2. This indicated that the plant was heterozygous at the marker locus and can be expected to segregate for the alleles at this locus in subsequent generations. Considering the segregation of the 22 markers the per cent recurrent parent genome recovery in the R-genes introgressed BC1F1s was estimated to be higher in BC1F1 plant no.8.3.2 but lower than the expected estimate of 75 per cent. This was also confirmed by graphical genotyping. The dendrogram thus generated out of the marker data, grouped the R-genes introgressed BC1F1s with ISM indicating that the three R-genes introgressed BC1F1s exhibited greater similarity with donor parent parent ISM at the genome level. Evaluation of BC1F1 individuals for morphological traits revealed presence of wide variability. The three R-genes introgressed BC1F1s were late in flowering compared to the recurrent parent Uma. Two of these genotypes i.e.,plant no. 8.3.2 (234 days) and plant no. 8.3.3 (228 days) flowered later than the donor parent. However, the three R-genes introgressed BC1F1s resembled the recurrent parent Uma with respect to grain and kernel characteristics. Backcrossing the three R-genes introgressed BC1F1s i.e., plant no. 8.3.2, plant no. 8.3.3 and plant no. 8.3.9 to the recurrent parent Uma resulted in 28 BC2F1s. Simultaneously, selfing of these individuals produced 850 BC1F2 seeds. Foreground and background profiling of these generations can ensure precise identification of genotypes that resembles the recurrent parent Uma possessing the resistance genes of interest with maximum recovery of recurrent parent genome.ThesisItem Open Access Evaluation of superior cultures for yield and yellow vein Mosaic resistance in okra(Department of Plant Breeding and Genetics College of Agriculture, Vellayani, 2016) Nikitha, J; KAU; Arya, KThe present study entitled “Evaluation of superior cultures for yield and yellow vein mosaic resistance in okra (Abelmoschus esculentus (L.) Moench)” was carried out in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani during 2014-2016, with the objective to identify high yielding and yellow vein mosaic resistant cultures of okra from those evolved through inter-varietal hybridization programme. Fifteen superior cultures of okra obtained from the previous project in the Department viz., VLYA 1, VLYA 2, VLYA 3, VLYA 4, VLYA 5, VLYA 6, VLYA 7, VLYA 8, VLYA 9, VLYA 10, VLYA 11, VLYA 12, VLYA 13, VLYA 14 and VLYA 15 along with two check varieties Varsha Uphar and Kiran were evaluated in a Randomized Block Design (RBD) with three replications during summer season of 2015. The analysis of variance was calculated for the traits under study viz., days to 50 per cent flowering, number of fruits plant-1, fruit weight (g), fruit length (cm), fruit girth (g), yield plant-1 (g), plant height (cm) and duration (days) and these were found to be highly significant for all the genotypes evaluated. The maximum yield was recorded by the genotype VLYA 2 which was on par with genotypes VLYA 5, VLYA 10, VLYA 11, VLYA 13 and VLYA 15 and the minimum yield was observed by the check variety Kiran. The yield plant-1 exhibited moderate GCV (18.42%) and PCV (19.05%), high heritability (94.00%) coupled with high genetic advance (36.69%). The yield plant-1 was found to be significantly and positively correlated with number of fruits plant-1, fruit length, fruit girth, fruit weight and plant height both at genotypic and phenotypic levels. Days to 50 per cent flowering and yellow vein mosaic disease incidence was found to be negatively correlated with yield plant-1. Very high positive and significant inter-correlation was noticed between height of plant and number of fruits plant-1. The path analysis showed that number of fruits plant-1 and fruit weight showed the maximum positive direct effect towards yield. The number of fruits plant-1 had high indirect effect through fruit length. The scoring for yellow vein mosaic disease and the vulnerability index revealed that the genotypes VLYA 5, VLYA 11 and VLYA 13 were resistant to the disease during all stages of crop growth. Number of white flies was found to be highest in VLYA 10 and lowest in VLYA 2. The incidence of fruit and shoot borer was also scored and five genotypes viz., VLYA 2, VLYA 4, VLYA 11, VLYA 13 and VLYA 14 were found to be immune and VLYA 3, VLYA 5, VLYA 15 and Varsha Uphar were found to be resistant. The glass house experiment of vector transmission and graft transmission was conducted for the confirmation of disease resistance and the vulnerability index was calculated to check the severity of the disease. The genotypes VLYA 5, VLYA 11 and VLYA 13 received a score ‘0’ which indicated that these genotypes were highly resistant. Hence the genotypes which obtained a vulnerability index of ‘0’ during both field evaluation and glass house experiment were confirmed to be resistant to yellow vein mosaic disease. Hence the present study revealed that the cultures VLYA 5, VLYA 11 and VLYA 13 were having high yield and yellow vein mosaic disease resistance. So these genotypes can be used for further trials before releasing for field cultivation.ThesisItem Open Access Combination breeding for high protein cowpea (Vigna unguiculata L. Walp)(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2015) Sarath, P S; KAU; Jiji, JosephCowpea (Vigna unguiculata L. Walp) is one of the most important legume crops grown in India. It is a versatile pulse crop owing to its nutritional value, weed smothering nature, drought tolerant characters, soil restoring properties and multi-purpose uses. Protein deficiency has been reported to be one of the main nutritional problems in the developing world. About one billion people are reported to be suffering from protein deficiency and malnutrition worldwide. Studies have revealed that protein content in cowpea grain ranges between 18 to 40 per cent depending on the genotypes. Identifying high yielding varieties of cowpea with high protein content will not only contribute to food security and alleviate poverty but could also contribute to the alleviation of protein deficiencies. Review on the earlier research pointed to poor emphasis on the genetic improvement of cowpea for improved protein content. Hence, the present study was envisaged to combine the high protein trait with high yielding genotypes. The study was grouped under three experiments, i) Evaluation of cowpea genotypes, ii) Hybridisation of the selected genotypes in line x tester design, iii) Evaluation of F1 hybrids. The experiments were conducted at College of Horticulture, Kerala Agricultural University, Vellanikkara during 2014-2015. All the crop management practices were followed as per KAU (2011). Evaluation of cowpea genotypes revealed that there was wide variability for all the traits studied except branches per plant among bushy and semi trailing types of cowpea. In trailing type, variability was observed for plant height, number of pods per plant, pod weight grain yield per plant and seed protein content. Among the twenty two genotypes evaluated, ten genotypes that exhibited a protein content of above 25 per cent were selected for hybridization programme. The selected genotypes (Vellayani Jyothika, Bhagyalakshmi, Anaswara, Kanakamony, Lola, Vyjayanthi, AV-5, PKB-3, PKB-4 and Sharika) were hybridized in line x tester (6 x 4) design. High magnitude of phenotypic coefficient of variation (PCV), genotypic coefficient of variation (GCV), heritability and genetic advance was observed for plant height, grain yield per plant and length of pods were observed in parents and hybrids suggesting scope for genetic improvement of these traits through selection. Seeds per pod and protein content exhibited low PCV and GCV but high heritability and low genetic gain, indicating that these traits were governed by non-additive gene action and therefore breeding method other than simple selection is required. Combining ability analysis revealed that higher magnitude of specific combining ability (SCA) variances than the general combining ability (GCA) variances for the traits like pods per plant, length of pod, test weight, grain yield per plant and seed protein content indicating preponderance of non-additive gene action in the inheritance of these traits. Higher estimates of GCA variances over SCA variances for plant height, days to flowering, days to first harvest, days to last harvest, pod-weight and seeds per pod implies the preponderance of additive gene action in controlling these traits. The hybrids were ranked based on mean value, sca effects and estimates of heterosis for the traits like days to flowering, pods per plant, seeds per pod, test weight, grain yield per plant and protein content. Four hybrids with the lowest scores were selected. Hybrids H2 (Vellayani Jyothika x PKB-3), H10 (Anaswara x PKB-3), H11(Anaswara x PKB-4) and H12 (Anaswara xSharika) were found to be the most promising. The transgressive segregants with high yield and protein content can be identified on further evaluation of the segregating population from the above hybridsThesisItem Open Access In vitro mutagenesis and evaluation of somatic embryo derived plantlets in cassava (Manihot esculenta Crantz.)(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2016) Riya, Antony; KAU; Jiji, JosephThe present study ‘In vitro mutagenesis and evaluation of somatic embryo derived plantlets in cassava (Manihot esculenta Crantz.)’ was conducted in the Department of Plant Breeding and Genetics, College of Horticulture during 2014-15. They study attempted to assess the variability existing among the in vitro derived plantlets of cassava genotypes Sree Jaya and CC1 as well as to generate further variability through in vitro mutagenesis. With the aim to induce variability, in vitro mutagenesis of callus derived from cassava genotypes Sree Jaya and CC1 was attempted earlier in the department. This had resulted in 10 somatic embryo derived plantlets in primary hardening stage and 58 plantlets yet to be transferred for hardening. The above plantlets formed the basic material for field evaluation undertaken in the present study. Sree Jaya and CC1 genotypes were planted as control. Out of the 58 plantlets transferred from the in vitro cultures to hardening, only 34 plantlets survived i.e., the survival was found to be 58.62 per cent. All the ten plants that were already in the primary hardening stage survived. Observations were recorded during field evaluation of the in vitro mutagen treated plants at three, six and nine months after planting as per the descriptor of cassava (Fukuda et al., 2015). The plants varied with respect to qualitative characters like colour of apical leaves, leaf retention, shape of leaf lobe, petiole colour, leaf orientation, colour of stem exterior, extend of root peduncle, shape of tuber, root colour, colour of root pulp and colour of root cortex. Variability was also observed for quantitative characters like length and width of leaf lobe, length to width ratio of leaf lobe, petiole length, distance between leaf scars, height of plant, tuber weight per plant, tuber girth, stem girth, extend of root peduncle, starch content and dry matter content. Among the mutagen treated plants of Sree Jaya none of in vitro mutagen treated plants were found to be superior with respect to tuber yield while in CC1 genotype, six plants yielded better than the control. Sensory evaluation of the tubers produced by in vitro mutagen treated plants as well as control plants was done by twelve panelists to assess consumer perception. The tubers from plant 32 (Sree Jaya; 1.2 % EMS), followed by plant 31 (Sree Jaya; 1.2 % EMS) and plant 42 (CC1; 0.9 % EMS) were most preferred for various sensory attributes evaluated. In vitro mutation being a potential method to induce variability, mutation of callus derived from genotypes was as attempted to create more variants. The callus cultures of Sree Jaya and CC1 genotypes were established as per the protocol standardised by Magaia, (2015). Friable embryogenic calli production was higher in the media MS + 8 mg L-1 2,4-D + 1 mg L-1 NAA + 0.5 mg L-1 BAP using leaf explants. Calli were subjected to physical (γ irradiation at 30- 60 Gy at an interval of 10Gy) and chemical (EMS 0.1 - 0.9 % at an interval of 0.1%) mutagens as advocated by Magaia, (2015). However, regeneration of mutagen treated friable calli was not obtained in both genotypes. Quantum of variability expressed in the in vitro mutated plants of cassava with respect quantitative traits shows the efficiency of in vitro mutagenesis in creating variability in cassava. In vitro mutagenesis is a potential tool in the hands of plant breeder to create variability especially in vegetative propagated crops. The evaluation of in vitro mutagen treated plants in the field showed wide variation with respect to most morphological as well as biometrical traits. All the plants evaluated can hence, be advanced to next generation of evaluation (M1V1) with replication to identify the mutants. From the results obtained on induction of in vitro mutation to create more variants in cassava, it is concluded that the friable callus in both the genotypes need to be cultured in alternate regeneration medium for successful regeneration of somatic embryos.ThesisItem Open Access Evaluation of cowpea [Vigna unguiculata (L.) Walp.] genotypes for yield and resistance to pulse beetle [Callosobruchus spp.](Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2017) Thouseem, N; KAU; Beena, ThomasThe present study entitled “Evaluation of cowpea [Vigna unguiculata (L.) Walp.] genotypes for yield and resistance to pulse beetle [Callosobruchus spp.]” was carried out at farmer’s field, Kayamkulam during 2015-2017, with an objective to evaluate cowpea genotypes for yield and resistance to pulse beetle. The study was conducted in two experiments. In experiment-I, yield and resistance to pulse beetle was evaluated in a Randomised Block Design (RBD) with three replications using 30 genotypes collected from different places. Experiment-II was to study the seed morphological and biochemical factors associated with resistance to pulse beetle in five relatively resistant genotypes and five relatively susceptible ones which were identified through experiment-I. The field experiment revealed that the genotype T1 (Ambalappuzha local) showed the highest mean values for days to 50 per cent flowering, length of main stem, pod weight, pod length, number of seeds pod-1, 100 seed weight and crop duration. The maximum yield plant-1 (32.91g) was obtained for the genotype T29 (Sreya) followed byT14 (Alathur local) and T7 (Dhavengarae local), whereas the minimum yield plant-1 (11.07g) was for the genotype T13 (Hridya). Thirteen genotypes were found to be infested under field condition and only very low seed damage via carry over population was noticed. The characters studied were found to be significant for all the genotypes evaluated. The pod weight exhibited the highest GCV (47.73%) and PCV (52.12%). Heritability was high for all the characters except number of primary branches plant-1, number of pods plant-1 and seed yield plant-1 which possessed moderate heritability. GA (% mean) was high for all the characters except crop duration. The association analysis revealed highly significant positive correlation for 100 seed weight, days to 50 per cent flowering, crop duration, number of seeds pod-1, pod girth, and pod length with seed yield both at genotypic and phenotypic levels. The path analysis revealed that 100 seed weight, number of pods plant-1 and number of seeds pod-1 had the maximum positive direct effect on seed yield plant-1. In no choice confinement test under experiment-I, there were significant differences among the genotypes in terms of oviposition, percentage seed damage and percentage seed weight loss. T2 (Kayamkulam local-1) had the least egg load while more eggs were laid on seeds of T29 (Sreya) and T1 (Ambalappuzha local). The highest percentage seed damage and the highest percentage weight loss were recorded for T13 (Hridya) and T27 (Bijapur local) respectively. The lowest percentage seed damage and percentage seed weight loss were observed for the genotype T7 (Dhavengarae local). In the study of seed morphological and biochemical factors associated with resistance under experiment-II, it was found that seed coat texture, seed coat colour and protein content of seeds were found to have no influence on resistance to pulse beetle. However, total phenol content of seed was having significant negative correlation with percentage seed weight loss. An index score was worked out to find out genotypes with good yield and relative resistance to pulse beetle. It was observed that the genotype T7 (Dhavengarae local) had the highest index score followed by T14 (Alathur local). The present study revealed that the genotype T7 (Dhavengarae local) was found to be a good yielder with relative resistance to pulse beetle followed by the genotype T14 (Alathur local). Hence these genotypes can be recommended for future breeding programmes.ThesisItem Open Access Genetic divergence in kiriyat (andrographis paniculata nees)(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2017) Prathibha, S S; KAU; Arya, KThe study entitled “Genetic divergence in kiriyat (Andrographis paniculata Nees)." was under taken at the College of Agriculture, Vellayani during 2015-17 with the objective to assess the genetic variability present in the natural ecotypes of kiriyat from different regions and identifying the superior ecotypes in terms of herbage yield and quality. Thirty accessions of kiriyat were collected from different parts of India and were evaluated for genetic variability with respect to herbage yield (fresh weight) and quality in terms of total extractives (%). Accession A10 from Aruvipuram showed highest mean values for seedling height (15 DAT), number of primary branches, herbage yield (137.25 g), yield plant-1 (dry herbage yield plant-1) (37.79 g) and total extractives (13.6 %) followed by the A14 (Kottakkal) and A7( Kottakkunnu). The accessions A14 (Kottakkal) and A15 (Nilambur) showed highest mean values for number of secondary branches and number of leaves seedlings-1 (15 DAT) respectively. Mean value for days to 50 percent flowering was least for accession A10. The lowest yield was exhibited by A3 (Coimbatore) accession. Average duration of the plants exhibited ranged between 182.67 and 213 days. The earliest accession was A10 (Aruvipuram) with an average duration of 182.67 days. Seedling height, number of leaves seedling-1, number of secondary branches, leaf length and width, stem girth, leaf/stem ratio, plant height, herbage yield, yield plant-1 and total extractives exhibited high coefficient of variations. Heritability was high for all the characters except number of leaves seedling-1 (15 DAT), herbage yield and yield plant-1 which possessed moderate heritability. GA (% mean) was high for all the characters except plant duration. The association analysis revealed a significant correlation among almost all characters and also with yield. Path coefficient analysis revealed that plant height, herbage yield, number of primary and secondary branches had high positive direct effect on yield. The genetic divergence was studied using Mahanalobis D2 statistics and accessions were grouped into seven clusters. Cluster VII accommodated maximum number of accessions (13) followed by cluster VI (5), cluster V (4), cluster IV (3), clusters III and II (2) and cluster I (1). Highest inter cluster distance was between clusters VI and VII while intra cluster distance was highest for cluster IV. The study revealed that variability existed among the different ecotypes of kiriyat and the ecotype collected from Aruvipuram (A10) was found to be superior in terms of herbage yield and quality followed by ecotypes from Kottakkal (A14) and Kottakkunnu (AThesisItem Open Access Invitro propagation in ashoka : saraca asoca (Roxb.) de wilde.(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2011) Brindha Devi, I; KAU; Sreenivasan, EAsoka (Saraca asoca) is an important medicinal and ornamental tropical tree currently facing the threat of extinction due to overexploitation of trees occurring in forests and other natural habitats. Unscientific and destructive extraction of bark from trees has lead to acute shortage of raw bark by ayurvedic industries. Hence, the International Union for Conservation of Nature and Natural Resources (IUCN) has listed this species under ‘globally vulnerable’ category. It is also enlisted among the 36 threatened and endangered medicinal plants of India. It is considered as the sacred tree of buddhists and Hindus. Literally the term ‘asoca’ means ‘sorrow-less’ and the tree is believed to remove the grief and unhappiness. The tree has immense medicinal properties. Its bark is considered as the primary medicinal part. Due to its acute short supply compared to its demand, various development and research activities are being prioritized to conserve, utilize and improve this species. It is mainly propagated by seeds. Due to heterozygous and cross pollinated nature of the species, it never gives a true to type progeny. Therefore the present study was undertaken to standardize the technique of in vitro propagation of Saraca asoca. Standardization of suitable explants, surface sterilization procedures and culture establishment protocol, Induction of multiple shoots and Elongation of root, hardening and planting out are the major objectives of the study. Nodal segment, Internodal segment and shoot tip were the three explants tried. Various surface sterilization procedures were tried using Chloramphenicol, ethyl alcohol, 0.1% mercuric chloride and combination of ethyl alcohol and mercuric chloride in various concentration and duration, using nodal segments as explant. Surface sterilization using 70% ethyl alcohol for 3 minutes followed by 0.1% mercuric chloride for minutes proved to be the best, which gave the maximum survival percentage of 80. The next part of the study was standardisation of suitable explants for culture establishment. Among the three explants, Nodal segments gave maximum response of 60 per cent in ½ MS medium with BAP 0.5mg/l. This was followed by shoot tips in the same medium, which gave 10 per cent response. Internodal segments did not respond in any of the media used. Standardisation of basal media for culture establishment was done using nodal segments as the explants. Three media supplemented with BA 0.5 mg.l-1 were tried viz. MS, Half strenght MS, Woody plant media. Among the three, ½ MS media was identified as the best basal medium followed by MS medium. No response was seen in WPM medium. Culture establishment as well as Shoot bud initiation was attempted in ½ MS and MS media with various growth regulator combinations. Maximum response of 60 per cent was obtained in ½ MS medium containing BAP 0.5 mg/l followed by 30 percent in the same medium containing BAP 1.5 mg/l. There were no response with 2,4-D. The response obtained was callusing in all cases. Induction of multiple shooting was tried in ½ MS medium supplemented with BAP, and Kn alone as well as combinations of BAP, IAA at various concentration. Here highest response of 30 per cent of single shoots was recorded in ½ MS media containing BAP 0.5 mg/l. Response was in the form of single shoot. The single shoots with a mean length of about 1.5mm after one week of growth was obtained. With BAP 2.0 mg/l, single shoots were produced in about 5% of cultures within 54 days. Effect of Kn in various concentration ranging from 0.5 to 2.0 mg/l was found to be low in shoot induction. The maximum length of shoot of about 1.6 cm was recorded in combination of BAP 0.5 mg/l and IAA0.5 mg/l. Various combinations of IAA and IBA at different concentrations were tried for rooting of in vitro shoots. However there was no response in any of the combinations tried.