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  • ThesisItemOpen Access
    Genetic variability, path analysis and stability parameters in sesame
    (Department of Plant Breeding, College of Agriculture, Vellayani, 1985) Sverup, John; KAU; Gopinathan Nair, V
    Biometric analysis in a varietal collection of sesame was undertaken to study the genetic variability, correlations, path analysis and stability parameters. One hundred sesame types were evaluated in replicated trials at Vellayani in uplands during rabi and at Kayamkulam in rice fallows during summer. Genetic variability and correlations were estimated and path analysis worked out independently as both the locations. Location trials for estimating stability parameters were conducted at three places viz. in uplands during rabi at Pattambi and Vellayani and in rice fallows during summer at Kayamkulam. Large values for genotypic coefficients of variation were obtained for characters such as number of capsules on branches, number of capsules perplant, number of capsules on main stem and number of branches during rabi as well as summer. The lowest genotypic coefficient of variation was obtained for number of days to maturity during both rabi and summer. High values of heritability were recorded by seed protein content , seed oil content, height upto first capsule and weight of 1000 seeds under both conditions.
  • ThesisItemOpen Access
    Genetic divergence, prepotency and inbreeding depression in para rubber (Hevea brasiliensis Muell. Arg.)
    (Department of Plant Breeding, College of Agriculture, Vellayani, 1992) Kavitha Mydin, K; KAU; Gopinathan Nair, V
    A study on genetic divergence, prepotency and inbreeding depression in rubber was undertaken in an effort to identify clones for use as components of polyclonal seed gardens. Forty clones of Indian, Indonesian, Malaysian and Sri Lankan origin were evaluated in a replicated trial at the Rubber Research Institute of India. Genetic variability, correlations and the cause and effect relationships of dry rubber yield and its components were worked out. Genetic divergence was estimated employing the Mahalanobis' D2 techinique. The factors of divergence were identified through principal factor analysis. Twenty promising clones from genetically divergent clusters were subjected to seedling progeny analyses for the estimation of propotency based on performance of their open pollinated seedling progenies and inbreeding depression in the first generation of selfing. Significant clonal variation was revealed in respect of all the physiological, morphological and structural attributes studied as mean values for the fourth year of tapping for the stress period and for the peak yield period. High genetic variability for volume of latex under stress, plugging index under stress, annual mean dry rubber yield and dry rubber yield during the stress and peak periods was indicated by the high estimates of genotypic coefficient of variation. Additive gene effects offering scope for improvement through selection was indicated for dry rubber yield, latex flow rate and volume of latex during the three periods, girth increment rate, annual mean plugging index and plugging index under stress, by the moderate to high heritability estimates along with high genetic advance for these traits. Non- additive gene action was indicated by the high heritability and low genetic advance for dry rubber content during the three periods, girth and bark thickness. At both genotypic and phenotypic levels, annual mean dry rubber yield showed moderate to high positive correlations with dry rubber content and latex flow rate during the three periods, girth, girth increment rate, length of the tapping panel and bark thickness and negative correlations with yield depression under stress and plugging index during the three periods. Dry rubber yield under stress emerged as a more important component than peak dry rubber yield by its higher magnitude of positive direct effect on annual mean dry rubber yield. Latex flow rate during the stress and peak periods and annual mean volume of latex exhibited high positive direct effects on annual mean dry rubber yield while plugging index during the peak yield period, volume of latex under stress and girth increment rate had negative direct effects on annual mean dry rubber yield. The magnitude and direction of the effects of the components on dry rubber yield during the three periods varied indicating these relationships to be under different genetic control. Selection for a high dry rubber yield under stress, annual mean volume of latex and latex flow rate during the stress and peak periods and against a high plugging index during the peak period, volume of latex under stress and girth increment rate would help achieve improvement in annual mean dry rubber yield. Considerable genetic diversity was revealed by the wide range of D2 values and intra and inter cluster distances. The forty clones were grouped into eight genetically divergent clusters irrespective of their country of origin indicating the absence of any relationship between geographical diversity and genetic divergence. Volume of latex, plugging index, latex flow rate, dry rubber content and dry rubber yield contributed more towards divergence than the morphological and structural attributes. Supporting evidence was obtained from principal factor analysis which revealed the yield factor to be the main factor of divergence with respect to the clusters studied. Junveile rubber yield on test tapping, number of latex vessel rows and number of leaf flushes in seedling progenies exhibited high heritability and genetic advance indicating scope for their use as early selection parameters, while girth exhibited high heritability and low genetic advance. These three traits showed significant positive correlations with juvenile rubber yield, of which girth exhibited the strongest association. Juvenile rubber yield, number of latex vessel rows, girth and number of leaf flushes were identified as important traits for being accorded simultaneous emphasis in the computation of performance index and index scores for the determination of recovery of superior seedlings as estimates of prepotency. Nine clones were identified as likely preopotents on the basis of seedling progeny analysis at the age of two years. Selfing resulted in a lower fruit set than open pollination in the clones in general. No significant inbreeding depression was recorded for juvenile vegetative traits and rubber yield in seedlings. Clones PB 28/83, PB 215, RRII 105, AVT 73, PB 217, PB 252, Ch 26, PB 242 and PB 5/51 were identified as likely prepotents from three genetically divergent clusters. They recorded superiority for yield and various yield components. These clones exhibited synchrony in flowering and are suggested as components of a nine parent polyclonal seed garden. For a seven parent seed garden the clones suggested to be excluded are PB 5/51 and PB 242. A polyclonal seed garden comprising these nine or seven clones as components could generate good quality polycross seed material. Appropriate seed garden layouts have been suggested.
  • ThesisItemOpen Access
    Genetic analysis of biological nitrogen fixation traits and yield components in cowpea (Vigna unguiculata (Linn). walp)
    (Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 1995) Sreekumar, K; KAU; Manikantan Nair, P
    A study on the parameters of variability, correlations, path-coefficients, combining ability, gene action and heterosis in cowpea was undertaken at the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani during 1991 to 1993. Fifty three genotypes of cowpea collected from different sources were planted in a field experiment for the estimation of variability, correlation and path coefficient. Eleven biological nitrogen fixation characters, viz., number of days to flower, length of primary root, number of secondary roots, number of nodules in the primary root, number of nodules in the secondary roots, total number of nodules, weight of effective nodules in the primary root, weight of nodules in the secondary roots, total weight of nodules, nitrogen content in the plant at 50 per cent flowering and plant dry weight and six yield characters, viz., grain yield per plant, length of pods, number of pods per plant, number of seeds per pod, weight of 100 seeds and seed protein content were considered for this study. The ten selected varieties/types from the initial evaluation trial were crossed in a line x tester model, keeping the three high nitrogen fixing types and three high yielding types as lines (total six lines) and two low nitrogen fixing and two low yielding types as testers (total four testers). The F1’s along with their parents were compared in a field experiment and combining ability, gene action and heterosis were estimated. The study of combining ability and gene action were confined to six biological nitrogen fixation characters, viz., number of days to 50 per cent flowering, weight of nodules in the primary root, total weight of nodules, weight of effective nodules, dry weight of the plant and nitrogen content per plant and six yield characters, viz., length of pod, number of seeds per pod, number of pods per plant, hundred seed weight, seed protein content and grain yield per plant. The analysis of variance revealed that a considerable amount of variation among the varieties was present with respect to the characters under study. Characters like number of days to flower, total nodule weight, nitrogen content per plant, weight of 100 seeds and seed protein content had recorded narrow relative magnitude of difference of phenotypic and genotypic coefficient of variation along with high heritability estimate. Moderate magnitude of difference of PCV and GCV along with moderate heritability was recorded for the characters viz., number of nodules in the primary root, number of nodules in the secondary roots, total number of nodules, weight of effective nodules in the primary root, weight nodules in the secondary root, length of pods and number of pods per plant. Plant dry weight and grain yield registered a wider difference of PCV and GCV along with low heritability indication the greater influence of environment over these two characters. Genetic advance as percentage of mean was found to be high for the characters like number of nodules in the secondary roots, weight of effective nodules in the primary root, weight of nodules in the secondary roots, total nodule weight, number of pods per plant and 100 seed weight and moderate for number of nodules in the primary root, total number of nodules and grain yield. Low genetic advance was recorded by number of days to flower, plant dry weight, nitrogen content per plant, length of pod and seed protein content. Hence characters such as number and weight of nodules in the primary root, number and weight of nodules in the secondary roots, total number and weight of nodules, number of pods per plant and weight of 100 seeds may be controlled by additive genes whereas days to 50 per cent flowering, nitrogen content in plant, length of pod and seed protein content may be controlled by non-additive genes. Correlation coefficients were workedout at the genotypic and phenotypic levels. Based on the genetic correlation of characters studied, it was understood that high nitrogen fixing genotypes may not be high yielders because of the antagonistic relationship between grain yield and total nitrogen per plant. Weight of nodules in the primary root and total nodule weight were positively correlated with the nitrogen content in plant. Hence genotypes which was able to form effective large nodules on the primary root system seems to be a better nitrogen fixer. Number of days to 50 per cent flowering had negative genotypic correlation with grain yield. Hence an early flowering genotype may be better yielder than a late flowering type. Weight of hundred seeds and seed protein content exhibited very strong negative correlation indicating that small seeded genotypes may be better with respect to protein content. Grain yield recorded positive phenotypic and genotypic correlation with number of pods per plant. Path coefficient analysis at the genotypic level revealed that total number of nodules had the highest positive direct effect on nitrogen content per plant followed by weight of effective nodules in the primary root and weight of effective nodules in the secondary root. Highest positive direct effect was recorded for length of pod with grain yield. The combining ability analysis revealed that both additive and non-additive gene actions were important for all the characters under study. However GCA and SCA variance ratio which was less than unity for all the traits under study indicated the predominance of non-additive gene action in the inheritance of these traits. Considering the combining ability effects, VCP 4 was found to be the best general combiner for most of the biological nitrogen fixing characters and V 322 was the best general combiner for the grain yield. The cross combination of V 27 x C 152 and V 271 x Co Vu 85020 showed the best performance with respect to sea for the character number of days to 50 per cent flowering while VCP 4 x C 152 for weight of nodules in the primary root and nitrogen content in plant. DPLC 210 x PTB 2 recorded high sea for total weight of nodules, weight of effective nodules, dry weight of the plant and number of pods per plant, on the other hand Co Vu 358 x C 190 recorded high sca for length of pod, number of seeds per pod and seed protein content. The cross combination V 322 x C 190 exhibited high sca for hundred seed weight and V 27 X C 152 for grain yield per plant. Marked heterosis was observed in many cross combinations for most of the characters studied and pronounced heterotic expression was obtained for weight of nodules in the primary root, total weight of nodules, weight of effective nodules, dry weight of the plant at 50 per cent flowering, number of pods per plant and grain yield. It was already established that these characters are predominantly governed by the non – additive gene action. Hence the heterotic vigour expressed by the hybrid combination with respect to these characters are justified. Since the biological feasibility for the exploitation of heterosis is not economical as a plant improvement programme in this crop, genetic improvement of these trait can be brought about more effectively through combination breeding involving genetically diverse and high combining parants.
  • ThesisItemOpen Access
    Intra and inter generic hybridization and molecular charatrization in monopodial orchids
    (Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2008) Beena, Thomas; KAU; Lekha Rani, C
    A research programme entitled “Intra and inter generic hybridization and molecular characterization in monopodial orchids” was carried out in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani during 2005-2008 with the objective of undertaking intra and intergeneric hybridization, in vitro embryo culture and molecular characterization in monopodial orchids, as a preliminary step to develop novel hybrids. Fifteen monopodial orchid genotypes comprising of six genera viz., Aranda, Aranthera, Kagawara, Mokara, Renanthera and Vanda, with good cut flower qualities and high demand in the market were selected as parents after initial evaluation. They were evaluated adopting completely randomized design with five replications. Analysis of variance revealed significant differences for almost all the characters studied. Genotypic and phenotypic coefficients of variation were high for thickness of leaf, leaf area and number of aerial roots. High heritability (>70 %) combined with high genetic advance (>70 %) was observed for number of aerial roots, width of leaf, thickness of leaf, leaf area, number of spikes per shoot and number of flowers per inflorescence. Significant positive inter-correlation at genotypic and phenotypic levels was observed for length of flower and width of flower with number of spikes per shoot. The character number of spikes per shoot recorded significant positive correlation with leaf area. Number of flowers per inflorescence was positively correlated with number of leaves per shoot and length of inflorescence. Out of the six genera studied, four viz., Aranda, Aranthera, Kagawara and Mokara, exhibited free-flowering nature. Seasonal flowering was observed mainly from June to December in Vanda, whereas it was confined to two seasons, from February to March and August to October in Renanthera. Inflorescence axis was found to be arching in Aranthera and Renanthera while the rest of the genotypes produced erect inflorescence axis. The 15 parental genotypes were crossed in all possible combinations after preliminary studies on floral biology. A total of 225 cross combinations were attempted including 105 crosses, 105 reciprocals and 15 selfs. Incompatibility reactions were noticed at different stages ranging from flower abscission before the onset of any visible post pollination change to instances where seeds germinated but aborted in culture. Mature green capsules were harvested from 70 combinations at 70 to 90 per cent maturity. Among them 15 combinations did not yield any seeds in the capsule while the remaining 55 combinations were cultured axenically. Among the 55 combinations inoculated in vitro, no germination was obtained from seeds of 12 combinations. Out of the 43 combinations that germinated successfully, seven combinations showed arrested development. Thus out of the total 55 combinations inoculated in vitro 36 combinations developed successfully. These were subcultured three to four times. Seedlings having 2-3 leaves and 2-3 roots were deflasked and planted out. MS half strength was selected as the best basal medium. For improving the in vitro growth of hybrid monopodial orchid seedlings refinement of this medium by supplementing with IAA (8 mg l-1) and NAA (2 mg l-1) was beneficial. Significant differences among the combinations were observed with respect to number of days taken for germination initiation, number of days taken for development of protocorms, chlorophyll, first leaf, first shoot and first root primordia and for deflasking. Significant differences in seedling morphology were observed among the 36 hybrid combinations at deflasking. These were kept in humidity chamber for acclimatization for one month, transferred to net house for hardening and maintained there for further growth. In the present study, RAPD was employed for studying the genetic diversity and for the fingerprinting of 20 monopodial orchid hybrids, making use of arbitrary primers to amplify random DNA sequences in the genome. To identify the promising primers for RAPD analysis, 70 decamer primers of kit A, B, C and D were screened using the DNA of hybrid H-2. Based on the performance in DNA amplification, eight decamer primers were identified for RAPD analysis. Primers that produced highest number of polymorphic bands which were intense and reproducible were selected. They were OPB-07, OPB-15, OPB-l7, OPC-04, OPC-05, OPC-08, OPC-15 and OPD-02. a total of 57 scorable bands (average of 7.125 bands per primer) were generated by the selected eight primers of which six were monomorphic and the remaining 51 were polymorphic (89.47%). The estimation of Jaccard’s coefficients and construction of dendrogram by using UPGMA revealed the presence and extent of genetic similarities among the 20 monopodial orchid hybrids. The overall similarity coefficients ranged from 0.40 to 0.84. Cluster analysis revealed that at 0.69 similarity coefficient, the 20 monopodial orchid hybrids got divided into six groups. Among the 20 hybrids, H-18, H-7, H-13, H-17, H-19 and H-20 stood separately in clusters II, III A, III B, IV, V and cluster VI respectively. This substantiates the moderately broad distribution of genetic variability, which can be attributed to the broad genetic base in their ancestry. Cluster I A contained four hybrids viz., H-1, H-10, H-3 and H-9. This grouping is justified by the presence of a common parent i.e., Arachnis Maggie Oei Red Ribbon in their parentage. Moreover, two common species viz., Arachnis hookeriana and Arachnis flos-aeris are involved in their ancestry. All these support their belonging to the same cluster. The three hybrids such as H-5, H-6 and H-8 fell in cluster I B. RAPD technique is relatively simpler, quicker, less expensive and non-radioactive than other molecular characterization techniques. The results of present investigation proved that it can detect sufficient polymorphisms in genetic distance studies in monopodial orchids. A research programme entitled “Intra and inter generic hybridization and molecular characterization in monopodial orchids” was carried out in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani during 2005-2008 with the objective of undertaking intra and intergeneric hybridization, in vitro embryo culture and molecular characterization in monopodial orchids, as a preliminary step to develop novel hybrids. Fifteen monopodial orchid genotypes comprising of six genera viz., Aranda, Aranthera, Kagawara, Mokara, Renanthera and Vanda, with good cut flower qualities and high demand in the market were selected as parents after initial evaluation. They were evaluated adopting completely randomized design with five replications. Analysis of variance revealed significant differences for almost all the characters studied. Genotypic and phenotypic coefficients of variation were high for thickness of leaf, leaf area and number of aerial roots. High heritability (>70 %) combined with high genetic advance (>70 %) was observed for number of aerial roots, width of leaf, thickness of leaf, leaf area, number of spikes per shoot and number of flowers per inflorescence. Significant positive inter-correlation at genotypic and phenotypic levels was observed for length of flower and width of flower with number of spikes per shoot. The character number of spikes per shoot recorded significant positive correlation with leaf area. Number of flowers per inflorescence was positively correlated with number of leaves per shoot and length of inflorescence. Out of the six genera studied, four viz., Aranda, Aranthera, Kagawara and Mokara, exhibited free-flowering nature. Seasonal flowering was observed mainly from June to December in Vanda, whereas it was confined to two seasons, from February to March and August to October in Renanthera. Inflorescence axis was found to be arching in Aranthera and Renanthera while the rest of the genotypes produced erect inflorescence axis. The 15 parental genotypes were crossed in all possible combinations after preliminary studies on floral biology. A total of 225 cross combinations were attempted including 105 crosses, 105 reciprocals and 15 selfs. Incompatibility reactions were noticed at different stages ranging from flower abscission before the onset of any visible post pollination change to instances where seeds germinated but aborted in culture. Mature green capsules were harvested from 70 combinations at 70 to 90 per cent maturity. Among them 15 combinations did not yield any seeds in the capsule while the remaining 55 combinations were cultured axenically. Among the 55 combinations inoculated in vitro, no germination was obtained from seeds of 12 combinations. Out of the 43 combinations that germinated successfully, seven combinations showed arrested development. Thus out of the total 55 combinations inoculated in vitro 36 combinations developed successfully. These were subcultured three to four times. Seedlings having 2-3 leaves and 2-3 roots were deflasked and planted out. MS half strength was selected as the best basal medium. For improving the in vitro growth of hybrid monopodial orchid seedlings refinement of this medium by supplementing with IAA (8 mg l-1) and NAA (2 mg l-1) was beneficial. Significant differences among the combinations were observed with respect to number of days taken for germination initiation, number of days taken for development of protocorms, chlorophyll, first leaf, first shoot and first root primordia and for deflasking. Significant differences in seedling morphology were observed among the 36 hybrid combinations at deflasking. These were kept in humidity chamber for acclimatization for one month, transferred to net house for hardening and maintained there for further growth. In the present study, RAPD was employed for studying the genetic diversity and for the fingerprinting of 20 monopodial orchid hybrids, making use of arbitrary primers to amplify random DNA sequences in the genome. To identify the promising primers for RAPD analysis, 70 decamer primers of kit A, B, C and D were screened using the DNA of hybrid H-2. Based on the performance in DNA amplification, eight decamer primers were identified for RAPD analysis. Primers that produced highest number of polymorphic bands which were intense and reproducible were selected. They were OPB-07, OPB-15, OPB-l7, OPC-04, OPC-05, OPC-08, OPC-15 and OPD-02. a total of 57 scorable bands (average of 7.125 bands per primer) were generated by the selected eight primers of which six were monomorphic and the remaining 51 were polymorphic (89.47%). The estimation of Jaccard’s coefficients and construction of dendrogram by using UPGMA revealed the presence and extent of genetic similarities among the 20 monopodial orchid hybrids. The overall similarity coefficients ranged from 0.40 to 0.84. Cluster analysis revealed that at 0.69 similarity coefficient, the 20 monopodial orchid hybrids got divided into six groups. Among the 20 hybrids, H-18, H-7, H-13, H-17, H-19 and H-20 stood separately in clusters II, III A, III B, IV, V and cluster VI respectively. This substantiates the moderately broad distribution of genetic variability, which can be attributed to the broad genetic base in their ancestry. Cluster I A contained four hybrids viz., H-1, H-10, H-3 and H-9. This grouping is justified by the presence of a common parent i.e., Arachnis Maggie Oei Red Ribbon in their parentage. Moreover, two common species viz., Arachnis hookeriana and Arachnis flos-aeris are involved in their ancestry. All these support their belonging to the same cluster. The three hybrids such as H-5, H-6 and H-8 fell in cluster I B. RAPD technique is relatively simpler, quicker, less expensive and non-radioactive than other molecular characterization techniques. The results of present investigation proved that it can detect sufficient polymorphisms in genetic distance studies in monopodial orchids.
  • ThesisItemOpen Access
    In vitro multiplication and DNA fingerprinting of selected hybrids and their parents in anthurium andreanum linden
    (Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2007) Yasin Jeshima, K; KAU; Mayadevi, P
    Anthurium is the largest genus in the family Araceae, encompassing more than 800 species, native to tropical America, from Mexico, Costa Rica, Cuba to Brazil and Argentina. The spadix is composed of a multitude of flowers, which are perfect having two-carpelled ovary and four anthers. A few commercially grown plants are classified as "rat tail" anthuriums as their inflorescences have a long spadix and a small non-descript spathe. Anthuriums with colourful inflorescences have been grown for cut flowers. With the introduction of compact interspecific hybrids through breeding and the selection of somaclonal variants, the new commercially available types were developed. Propagation is not easy for anthuriums and is considered a long-term crop which will take long time for the propagator to multiply. In light of afore said views, an attempt was made to standardize in vitro multiplication and DNA fingerprinting of selected hybrids and their parents in Anthurium andreanum Linden. The explants after standardizing for the surface sterilization and survival were cultured on selected media with different hormone concentrations to get maximum callus induction. For callus induction the culture flasks were kept in dark at 25˚C and subcultured every third week. Calli were transferred to regeneration medium and embryogenic calli induction medium. Regenerants were selected and placed in rooting medium; further hardened and transferred to the field condition. Preconditioned embryos were suspended in calcium free half strength NN medium supplemented with 1.5 per cent sodium alginate and 0.5 M sucrose. This mixture was dispensed with a micropipette into 0.1 M calcium chloride. Twenty minutes after encapsulation beads were pre cultured on modified half strength NN liquid medium supplemented with 0.75 M sucrose and three per cent DMSO into 100 ml Erlenmeyer flasks for one day without agitation. Beads were then transferred to fresh medium of same composition and incubated in darkness at 4ºC for three days. Beads were desiccated in a sterile laminar air flow chamber. Dehydrated beads were transferred to 4 ml cryo vials and stored at – 80˚C. On rewarming over a water bath at 25˚C, the beads were transferred to culture medium for germination. Surface disinfection treatments were standardized for the different explants, irrespective of the explants and varieties and double sterilization was found to be effective. Among the explants, the highest number of sterile cultures was observed in double sterilization, followed by the treatment with 70 per cent Ethyl alcohol for 20 minutes. Majority of the contamination found in the cultures was due to the presence of systemic infection of Xanthomonas compestris pv dieffenbachiae. This directly influences the percentage of contamination occurred in the culturing condition and the size of explants which also play a major role in creating the bacterial contamination. Candle explants were found to exhibit more systemic infections than other explants and seed explants were found to be free from systemic infections. Leaf explants are highly vulnerable to exhibit systemic infections and are more sensitive; unable to recover even after treatments with antibiotics. The callus cultures exhibiting systemic infections can be recovered by kanamycin 50 mg per litre containing multiplication medium. In most studies of in vitro culture of anthurium, MS medium has been used. In the present study also, it was observed that Nitsch and Nitsch medium was better than MS medium for multiple shoot induction. Nitsch and Nitsch medium is especially suitable for morphogenesis, meristem culture and regeneration. As the genotype showed different nutrient requirements for their survival and growth, the present investigation was planned to standardize the media by screening with modified MS, half strength MS, modified NN and half strength NN medium. Modified NN with activated charcoal and coconut water showed better response. Half strength NN with coconut water and activated charcoal, modified MS with activated charcoal and coconut water and half strength MS activated charcoal and coconut water were also found to support the explants without hindering the survival. Addition of inositol and glycine along with folic acid was found to be essential but the presence of small amount was inefficient. In the present investigation no callus initiation was observed when inositol was reduced to half of the reported quantity. Various treatments were tried for callus multiplication. The maximum fresh weight of callus 4.2458 g was observed in PR X DT inoculated in NN medium with major nutrients at normal strength followed by 4.1325 g in OG X DT for the same composition in NN medium. From the economic point of view NN medium can be recommended for callus multiplication. Among the treatments, combination of 2, 4-D and zeatin was found to be the best. It stimulates callus formation and strongly antagonizes organized development. The low auxin requirement may be due to the high potency of the auxin which was used for callus initiation. The young developing leaf may be a rich source of endogenous auxins due to which lower exogenous application is required. Irrespective of the source of explant all the callus cultures were able to be converted into plantlets by redifferentiation. The number of days taken for regeneration ranges from 55.5 to 82. This variation is due to the varietal difference and difference in hormonal effect. Modified NN with activated charcoal and coconut water along with 22.2 μM BA, 11.42 μM IAA and 4.09 μM biotin was found to produce regenerants. Each genotype is varying with the response to change in media composition in producing somatic embryogenesis. The treatments with MS and modified MS media were found to be insignificant when compared with NN and modified NN media. Among these modified NN was found to be the best one. Within two weeks on embryo development medium, the globular embryos developed a bipolar shape. Embryos at this stage were comprised of cells larger than those at the globular stage. Bipolar embryos had an extended upper region that formed the cotyledon and the epicotyl, and a lower region that formed the radicle.The main difference between the mature embryos of monocotyledons in vitro and in vivo is the absence or presence of suspensor. The presence of single cotyledon which is the terminal structure and the shoot initials present at the sides or hidden creating a heart shape. When the cotyledon starts growing the embryo will have a single cotyledon at the terminal end which is some what cylindrical in shape. In anthurium tissue culture, no special rooting treatments were needed and the shoots developed in vitro were found to develop roots spontaneously even in the absence of additional growth hormones in the supporting medium. The spontaneous root formation was not due to the carry over effect of the hormones supplied in the previous cultures for shoot formation. Irrespective of the supporting medium the shoots were able to form roots even in sterile sand supplied with sterilized compost materials. For in vitro studies Completely Randomized Block Design (CRD) was followed for statistical analysis wherever necessary. Molecular characterization of twelve hybrids and their parents were carried out with RAPD using AP-PCR. Young leaf samples from each genotype were collected for DNA isolation.Young copper coloured leaf tissues were used immediately after collection for DNA extraction. Leaf samples were pre-chilled at -80ºC for half an hour with pestle and mortar and then pulverized in liquid nitrogen by rapid grinding to a fine powder.The frozen powder was used to extract the total genomic DNA using CTAB extraction buffer. The purity of the DNA was analysed by running in 0.8 per cent agarose gel with 1 X TAE buffer. The optimized PCR mixture with 50ng of template DNA for a final volume of 20μl was used in thermal cycling in a PCR machine. The amplified products were run in 1.6 per cent agarose gel with 1X TAE (Tris buffer, Glacial acetic acid and EDTA pH 8.0) buffer. A total of 114 AP-PCR bands were generated by the 25 primers, of which 74.56 per cent were polymorphic (88 bands) and 26 were monomorphic. Ten primers showed high level of polymorphism out of which seven were selected.Seven promising primers were identified for AP-PCR analysis based on performance in DNA amplification, production of highest number of polymorphic bands as well as intense bands and reproducibility viz. OPA 10, OPB15, OPA13, OPB20, OPB6, OPB8 and OPB18 primers were found to produce polymorphism in Anthurium andreanum Linden.A total of 50 scorable bands (average of 7.143 bands per primer were generated by the selected seven primers of which only 8 were monomorphic and the rest were polymorphic. The number of bands ranged form 4 to 11 with an average of 7.143 per primer. The reproducible bands were scored for their presence (1) or absence (0) for all the hybrids and parents. A genetic similarity matrix was constructed using Jaccards’s similarity co-efficient methods. From the cluster analysis based on the dendrogram, TR X MW was found to be extreamly different from the other accessions and its own parents showing the significance of hybridization. The hybrids like OO X KR, PR X DT, OG X DT, FK X LR and PR X MW are not closely related to either of their parents and hybrids were distinguished from others. Some hybrids like LJ X W and PR X MW; PR X LR and FK X DT shows 30 to 39 per cent similarity. This shows there is considerable variability among the genotypes selected and can be further utilized for crop improvement. Confirming that, they were quite different from the other hybrids and varieties. Pair wise genetic distances based on RAPD [(AP-PCR) (Nei and Li Genetic Distance GDNL)] genetic distance co-efficient values for twelve varieties and twelve hybrids ranged from 0.1935 to 0.7037 indicating the wider diversity. The AP-PCR profiles show the relatedness and diversity of the hybrids and varieties. The bands were found within 1.5kb from 100bp. Most of the bands were concentrated between 300bp and 1200bp.
  • ThesisItemOpen Access
    Triallel analysis of yield and resistance to anthracnose in chilli ( Capsicum annuum L)
    (Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2007) Haridass, A; KAU; Manju, P
    . Chilli (Capsicum annuum L.) is an important spice cum vegetable crop grown commercially in India. It is an important constituent of many foods since it adds flavour, colour, vitamin C and pungency. Productivity of the crop remains low mostly due to destructive diseases, of which the most dreaded disease affecting chilli is anthracnose also known as dieback and fruit rot. The best method to tackle the disease is to grow resistant varieties and hence it is essential to breed high yielding anthracnose resistance varieties of chilli. Therefore an investigation was carried out at the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani to estimate the combining ability and heterosis through diallel pattern and combining ability analysis through general line effect of the first kind ((hi), general line effect of the second kind (gi), two line specific effect of the first kind (dij), two line specific effect of the second kind (sij/sji), three line specific effect (tijk), estimates of genetic components and heterosis by triallel analysis to assess the inheritance pattern of anthracnose resistance, yield and yield component traits, identify the order in which the parents should be combined to obtain maximum effect for a particular character and also to formulate an appropriate breeding programme for improving each trait. Six parents viz., Jwalamukhi (P1), Jwalasakhi (P2), Samkranthi local (P3), Vellayani Athulya (P4), Kidangoor local (P5) and Ujwala (P6) along with their 15 F1 hybrids, and their possible three way cross hybrids was evaluated during 2005-2007. Diallel analysis revealed that the magnitude of gca variance was higher than sca variance suggesting the predominance of additive gene action except for number of seeds per fruit, harvest index and vitamin C. On the basis of per se performance of the parents for different traits, P4 was superior with regard to average fruit weight, fruit length, fruit girth and hundred seed weight while P1 for fruit yield per plant and vitamin C. P6 showed high resistance to anthracnose with high enzymatic activity for anthracnose, phenol, peroxidase, polyphenol oxidase, phenylalanine ammonia lyase along with high capsaicin and oleoresin. The hybrid P1xP6 had highest per se performance for fruit yield per plant and number of fruits per plant, P2xP4 for fruit length, average fruit weight, fruit girth, number of seeds per fruit and hundred seed weight and P5xP6 for duration, incidence of anthracnose, disease intensity, biochemical characters, capsaicin and oleoresin. High values of gca effects were noticed for fruit yield per plant, number of fruits per plant and incidence of anthracnose at 45 DAT and 60 DAT. Highest sca effect was recorded for fruit yield per plant, number of fruits per plant and vitamin C. With respect to mean and general combining ability, P2 was superior for days to first flowering, average fruit weight, fruit girth, fruit yield per plant, hundred seed weight and harvest index while P6 for incidence of anthracnose, disease intensity and all the biochemical characters including capsaicin and oleoresin. Among the fifteen hybrids evaluated with respect to per se, standard heterosis and sca effects, P1xP6 was superior with regard to number of fruits per plant, fruit yield per plant, incidence of anthracnose, disease intensity, phenol, peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, capsaicin and oleoresin. In triallel analysis, P4xP6xP1 had highest per se performance for fruit yield per plant, P1xP5xP4 for average fruit weight, fruit length, number of seeds per fruit while P5xP6xP4 had significant per se performance for incidence of anthracnose and high enzyme activity. P6 had the highest general line effect of the first kind (hi) for fruit yield per plant and number of fruits per plant while P1 for days to first flowering, plant height, average fruit weight, fruit length, fruit girth, harvest index and vitamin C. P4 had highest general line effect of the first kind for incidence of anthracnose, disease intensity and for all the enzyme activity. The general line effect of the second kind (gi) was highest in P1 for fruit yield per plant, number of fruits per plant, fruit girth, hundred seed weight and oleoresin while P5 for low incidence of anthracnose, disease intensity and high activity of the enzymes. P6 had highest effect for number of branches per plant, capsaicin, and vitamin C. The two line specific effect of the first kind (dij) was highest in P4xP5 for number of fruits per plant, fruit yield per plant and duration while P1xP6 for incidence of anthracnose, disease intensity and all the enzyme activities. P1 x P2 had highest two line specific effect of the second kind (sij) for fruit yield per plant and number of fruits per plant whereas P4xP6 for incidence of anthracnose, disease intensity, phenol, peroxidase, phenylalanine ammonia lyase and polyphenol oxidase. The two line specific effect of the second kind (sji) reciprocal effect was highest in P5xP4 for fruit yield per plant and average fruit weight while P2xP1 for incidence of anthracnose, disease intensity, phenol, polyphenol oxidase and phenylalanine ammonia lyase. P1xP2xP3 recorded high value of three line specific effect for fruit yield per plant and average fruit weight and P5xP6xP4 for incidence of anthracnose at 60 DAT, phenol, and polyphenol oxidase and phenylalanine ammonia lyase. The hybrid P4xP6xP1 had high relative heterosis and heterobeltiosis for fruit yield per plant. Among the biometrical traits, the relative heterosis was high for fruit yield per plant followed by harvest index and number of fruits per plant whereas heterobeltiosis had maximum per cent for harvest index followed by number of fruits per plant and fruit yield per plant. At 45 DAT the incidence of anthracnose had high relative heterosis and heterobeltiosis. The enzyme polyphenol oxidase had highest heterosis followed by capsaicin and oleoresin for both type of heterosis. From the present investigation several promising three way cross hybrids were obtained with regard to fruit yield, yield attributes and resistance to anthracnose. Some of the promising three way cross hybrids are Jwalamukhi x Kidangoor local x Jwalasakhi, Jwalamukhi x Ujwala x Vellayani Athulya, Jwalamukhi x Ujwala x Kidangoor local, Jwalasakhi x Vellayani Athulya x Kidangoor local, Samkranthi local x Vellayani Athulya x Kidangoor local, Vellayani Athulya x Kidangoor local x Samkranthi local, Vellayani Athulya x Ujwala x Jwalamukhi, and Kidangoor local x Ujwala x Jwalasakhi. These can be directly used as hybrids or can be subjected to selection of superior types in the segregating generation to obtain stable varieties. The estimation of genetic components revealed the predominance of dominance x dominance gene effect for fruit yield per plant, number of branches per plant, number of fruits per plant, average fruit weight, fruit length, fruit girth, harvest index and capsaicin while the remaining traits days to first flowering, plant height, number of seeds per fruit, hundred seed weight, incidence of anthracnose, enzyme activity, phenol, oleoresin and vitamin C had additive x dominance type of epistatic effect. Heterosis method of breeding could be followed for the improvement of the traits which had dominance x dominance type of epistatic effect whereas the traits with additive x dominance component could be improved thereby postponing selection to later segregating generations and following recurrent selection.
  • ThesisItemOpen Access
    Genetic variability, path analysis and stability parameters in sugarcane
    (Department of Plant Breeding, College of Agriculture, Vellayani, 1986) Sreekumar, S G; KAU; Gopinathan Nair, V
    Investigations on genetic variability, path analysis and stability parameters wore conducted at the Sugarcane Research Station, Thiruvalla, Sugarcane Research Centre, Chittoor and Horticultural Research Station, Ambalavayal. The analysis of data collected from 48 clones evaluated at Thiruvalla revealed that the clones showed significant differences for all the 21 characters except Juiciness at 10th month in the first ratoon crop. The germination count on the 45th day, shoot count on the 180th day, number of late shoots at harvest, number of millable canes per plot, cane yield per plot and sugar yield per plot recorded moderate to high heritabiiity in the broad sense coupled with high genetic advance both in the plant and in the first ratoon crops indicating that selection based on these characters will be effective in improving cane and sugar yields. Correlation studies revealed that cane yield per plot had significant positive genotypic correlations with germination count on the 43th day, shoot count on the 180th day, number of millable canes per plot, cane weight and length of cane both in the plant and the first ratoon crops. Therefore, improvement in any one or more of these characters will result In a simultaneous improvement in the yiold of cane. Tho quality characters such as Juiciness, brix, pol and C.C.S. percentages at 12th month had positive but non-significant correlations with cane yield In ths plant crop and the first ratoon crop, suggesting difficulty in the simultaneous improvement of cane yield and quality characters through direct selection. The number of late shoots at harvest had highly significant positive genotypic and phenotypic correlations with the yield of cane in the first ratoon crop indicating that the number of late shoots at the time of harvest can be considered as a reliable criterion for the selection of clones with good yielding ability in the ratoon crop. Sugar yield recorded significant positive gonotypic correlations with cane yield and tho components of cane yield such as germination count on the 45th day, number of millable canes per plot, cane weight and length of cane. Sugar yiola also had significant positive genotypic correlations with brix, pol and C.C.S,percentages at 12th month, Tho significant positive gonotypic correlations of sugar yield with cane yield and its important components suggests that by increasing the cane yield, sugar yield per plot can be increased.
  • ThesisItemOpen Access
    Genetic variability, path analysis and stability parameters in sesame
    (Department of Plant Breeding, College of Agriculture, Vellayani, 1985) Sverup John; KAU; Gopinathan Nair, V
    Biometric analysis in a varietal collection of sesame was undertaken to study the genetic variability, correlations, path analysis and stability parameters. One hundred sesame types were evaluated in replicated trials at Vellayani in uplands during rabi and at Kayamkulam in rice fallows during summer. Genetic variability and correlations were estimated and path analysis worked out independently as both the locations. Location trials for estimating stability parameters were conducted at three places viz. in uplands during rabi at Pattambi and Vellayani and in rice fallows during summer at Kayamkulam
  • ThesisItemOpen Access
    Genetic resources utilisation and biometric analysis in groundnut (Arachis hypogaea L.)
    (Department of Plant Breeding, College of Agriculture, Vellayani, 1983) Pushkaran, K; KAU; Gopinathan Nair, V
    Maximising the production, of vegetable oils in the country is one of the badly felt national needs for which the improvement in the productivity of groundnut, the major oilseed crop of the country, deserves the best attention. As a crop, groundnut has wide adaptability and a number of other favourable attributes. Breeding improved varieties suited to the two major areas of groundnut cultivation viz, the traditional uplands during kharif land the non-traditional, but potential rice fallows during summer is an immediate necessity for popularising the crop in this State. Germplasm collection, evaluation and utilisation constitute the basic approaches in the genetic improvement of any crop. Hence this investigation. Eighty eight divergent varieties of groundnut were collected from different sources and subjected to preliminary evaluation in uplands during kharif. Ninety three varieties were evaluated in rice fallows during summer. These varieties showed wide variability in general performance, yield and other attributes within and between the situations. AH-6915 in uplands and TG-3 in rice fallows were the highest yielders. Several varieties were better in yield than the recommended varieties, viz., TMV-2 and TMV-7. A few varieties such as TG-14 end Spanish Improved were consistently superior at both' the situations.