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ThesisItem Open Access Induction of heteroploidy in Dendrobium sp.(Department of Plant Breeding and Genetics, College of Agriculture ,Vellayani, 2021-12-27) Revathi, B S; Beena Thomas; KAUThe present study entitled “Induction of heteroploidy in Dendrobium sp.” was carried out at the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani during 2019-21. The study was undertaken to induce heteroploidy in Dendrobium crumenatum using antimitotic agent colchicine via in vitro and in vivo technique. In in vitro induction of heteroploidy in Dendrobium crumenatum, the explant used in the micropropagation was stem nodal segments containing one axillary bud and the basal medium chosen was VW medium. The best combination of surface sterilization agents identified were, 20 per cent sodium hypochlorite for 20 minutes followed by 0.1 per cent mercuric chloride for 5 minutes and 70 % alcohol for 2 minutes. The medium VW+1 mgL-1 BA + 0.5 mgL-1 2,4-D was chosen for callus induction, callus proliferation and PLB formation. The explants were maintained in dark for callus induction and proliferation and for subsequent stages a photoperiod of 16 hours light and eight hours dark was provided. For the induction of heteroploidy, PLBs were subjected to eight different colchicine treatments in basal VW suspension medium. Colchicine treatments were based on the concentration of colchicine (0.05%, 0.10%, 0.15%, 0.20%) and duration of exposure of PLBs to colchicine (24 and 48 hrs). For shoot induction, VW basal medium supplemented with the hormones BA (1mgL-1) and NAA (0.1 mgL-1) was used. The maximum survival percentage of cultures at shoot multiplication stage after different colchicine treatments was observed to be 83.33 per cent, and the minimum survival percentage noted was 25 per cent. In in vivo induction of heteroploidy in Dendrobium crumenatum, D. crumenatum plantlets were subjected to eight different colchicine treatments, based on colchicine concentration (0.05%, 0.1%) and the duration of exposure of plantlets to these doses of colchicine (24 hrs, 48 hrs, 72 hrs, 96 hrs). The highest percentage of plants showing response after colchicine treatment was found to be 83.33 per cent (observation recorded after 2 weeks of colchicine treatment) and the lowest percentage recorded was 16.67 per cent. The observations shoot length, diameter of pseudobulb, number of leaves and width of leaves showed mean values greater than their corresponding control means, whereas the observations, diameter of shoot and length of leaf noted lower mean values than corresponding control. The mean value for the character height of pseudobulb were found to be same for treated and control plantlets. The character, number of leaves recorded high PCV and GCV. Moderate PCV and GCV were recorded for the characters viz., diameter of pseudobulb, length of shoot, length of leaf and width of leaf. The characters, diameter of shoot and length of pseudobulb obtained low PCV and GCV. Significant positive genotypic correlation was observed between length of shoot and height of pseudobulb, diameter of shoot and width of leaf, length of pseudobulb and number of leaves, number of leaves and length of leaf and width of leaf and number of leaves. Flow cytometry analysis of the colchicine treated in vivo and in vitro D. crumenatum samples were performed at Rajiv Gandhi Institute for Biotechnology, Poojapura. The histogram peaks obtained after the analysis of control (diploid) and treated samples were compared for interpretation of the results. In the in vitro treated samples, the maximum tetraploids (66.67 %) were obtained from treatments 0.15% colchicine, 24 hrs and 0.10 % colchicine, 48 hrs. On analyzing the histogram peaks of in vivo treated samples, the highest tetraploid induction (50%) was noticed in treatment 0.05% colchicine, 96 hrs. It was concluded from the present investigation that, for Dendrobium crumenatum which was subjected to both in vitro and in vivo colchicine treatments, in vitro colchicine treatment was found effective than in vivo colchicine treatment, when the concentration and duration of exposure to colchicine used for the study were same.ThesisItem Open Access Evaluation of tomato genotypes for tomato leaf curl virus (tolcv) resistance(Department of Plant Breeding and Genetics, College of Agriculture, Vellanikkara, 2021) Anjitha, A R; KAU; Deepthy Antony, PTomato (Solanum lycopersicum, 2n=24) is a widely grown vegetable in India, exhibiting wide variability for plant traits. Worldwide, tomatoes are susceptible to many diseases like damping off, bacterial wilt, early blight, leaf curl virus disease, leaf spot etc. Among them, tomato leaf curl virus disease (ToLCD), caused by whitefly-transmitted begomoviruses is more severe and it can cause up to 100 per cent yield loss. Although, the cultivated tomatoes are susceptible to Tomato leaf curl virus (ToLCV), different levels of resistance were identified in wild relatives of tomato. Utilization of these natural resources will aid ToLCV management through crop improvement. The present study entitled ‘Evaluation of tomato genotypes for Tomato Leaf Curl Virus (ToLCV) resistance’ was carried out at Department of Plant Breeding and Genetics, College of Agriculture, Vellanikkara during 2019-21. The experimental material consisted of 27 tomato genotypes including eight NBPGR (National Bureau of Plant Genetic Resources) accessions, five breeding lines from the World Vegetable Centre, Taiwan, nine commercial hybrids, four KAU varieties and one local collection from Idukki. All 27 genotypes were screened for ToLCV resistance under natural conditions in randomized block design with two replications during January- May 2021. The disease response was assessed after 30 days of transplanting by adopting the score chart suggested by Banerjee and Kalloo (1987). Among the genotypes tested, Ansal was found to be highly resistant, Kaustubh, EC 519806 (S. pimpinellifolium), Arka Rakshak and local collection (Idukki) were identified as resistant genotypes. The genotypes found to be highly resistant and resistant were subjected to whitefly mediated artificial inoculation. The resistant and highly resistant genotypes remained asymptomatic, whereas the susceptible genotypes (Anagha and Manuprabha) exhibited ToLCV symptoms after artificial inoculation. The morphological growth characters such as plant height, number of primary branches per plant, days to flowering and trichome density differed significantly among the genotypes evaluated in the present study. Density of both glandular and non-glandular trichomes on both abaxial as well as adaxial leaf surfaces were recorded. Correlation analysis between trichome density, and disease severity index and per cent disease incidence showed that glandular trichome density on abaxial and adaxial surfaces of leaf was significantly and negatively correlated with per cent disease incidence and disease severity index. However, the negative correlation was stronger between the abaxial glandular trichome density and ToLCV incidence. The glandular trichome density of the resistant genotypes identified in the field screening was found to be high. Non-glandular trichome density on abaxial and adaxial side of the leaf was significantly and positively correlated with per cent disease incidence and disease severity index. Screening for the reported Ty genes using the primers TG178 (SCAR), SCAR-2, P6-25, TY-1/3_K (SCAR), 18IY13 (Indel), SLM 4-34 (SSR) and SLM 10-46(SSR)linked to the ToLCV resistance genes Ty-1, Ty-2, Ty-3, Ty-1/3, Ty-4, Ty-5 and Ty-6 respectively was done in the present study. Seventeen genotypes produced amplicon using TG178 and ten genotypes produced amplicons using SLM4-34. However, there was no pattern in the amplification of the markers TG178 and SLM 4-34. Seven genotypes recorded the presence of Ty-1/3 in homozygous condition (AVTO 1727, AVTO 1726, AVTO 1706, AVTO 1707, EC 538153, EC 620428 and EC 620486) and seven genotypes in heterozygous condition (Ansal, Virang, Aryaman, Durg, Raymond, Abhiraj, Pranay). Four genotypes each carried the resistant allele for Ty-2 in the homozygous state (AVTO 1726, AVTO 1706, AVTO 1707 and AVTO 0922) and in the heterozygous state (AVTO 1727, Ansal, Arka Rakshak and Aryaman). Resistant allele for Ty-3 was present in the genotype AVTO 1726 and AVTO 1707 in homozygous state. Whereas, it was in heterozygous condition in AVTO 1727. None of the genotypes evaluated in the present study revealed the presence of amplicons corresponding to resistant allele of Ty-4 and Ty-6. The resistant genotypes identified in the present study viz., Ansal, Kaustubh, Arka Rakshak, EC 519806 and local collection (Idukki) can be used for resistance breeding. The present study revealed that trichome density can be used as a reliable morphological marker for ToLCV resistance. The donors for Ty-2, Ty-3 and Ty-1/3 identified in this study can be used for gene pyramiding in future breeding programmes.ThesisItem Open Access Genotype x environment interaction in advanced breeding lines of Cowpea (Vigna unguiculata (L.) Walp)(Department of Plant Breeding and Genetics, College of Agriculture, Vellanikkara, 2021) Swathi, S; KAU; Jiji JosephCowpea (Vigna unguiculata (L.) Walp) is an important tropical and subtropical annual legume crop grown for its green pods, grains and is also being used as a forage crop. It is one among the widely cultivated and consumed grain legumes, globally. As the grains contain high amount of protein (23.4 %) possessing better biological value on dry weight basis, cowpea is often considered as “vegetable meat”. Its ability to fix atmospheric nitrogen and drought tolerance makes it a suitable component in all major cropping systems. The present study entitled ‘Genotype x environment interaction in advanced breeding lines of Cowpea (Vigna unguiculata (L.) Walp)’ was an attempt to identify suitable and stable lines for general cultivation as a dual purpose type (both as grain purpose and vegetable purpose). Materials used for the study comprised of five cowpea cultures in stabilized F 7 generation developed as a result of pedigree selection from two crosses at the Department of Plant Breeding and Genetics, College of Agriculture, Vellanikkara, along with two check varieties, Anaswara and Kanakamony. The crop was raised during February 2021 to May 2021 over three environments viz., RARS Pattambi, CoA Vellanikkara and RRS Vyttila. Field experiments were laid out in plots of size 65 m 2 adopting randomized block design (RBD) with three replications. Observations were recorded on twelve characters: plant height (cm), number of branches, days to first flowering, days to first harvest, days to last harvest, number of pods per plant, pod length (cm), pod weight (g), number of seeds per pod, test weight (g), grain yield per plant (g) and protein content (%). All the observations were recorded at the time of harvest except for days to first flowering. The recorded observations were subjected to individual, location wise analysis of variance (ANOVA) followed by pooled analysis of variance (pooled ANOVA) over three locations. From the pooled ANOVA, the characters that exhibited significant genotype x environment (G x E) interaction were further assessed for stability using three models of stability. The Eberhart and Russell model, the Additive Main effects and Multiplicative Interaction effects (AMMI) model and the Genotype main effects plus Genotype-by- Environment interaction effect (GGE) biplot were the three models of stability used for the study.ANOVA revealed significant difference between lines for all the twelve characters considered. However, the Bartlett’s test for examining the homogeneity of error variance was found to be significant only for six characters. These six characters viz., days to first flowering, days to last harvest, number of pods per plant, number of seeds per pod, grain yield per plant and protein content were subjected to pooled ANOVA across three environments. Significant G x E interaction was observed in the six characters considered and were hence forwarded for analysing the stability. The Eberhart and Russell model recognizes a stable genotype as the one with high mean performance, non-significant regression (bi) as well as deviation from regression (S 2 di) values. The genotypes were ranked according to their mean values and stability parameters and it was observed that genotype L 2 with the lowest score was the most stable one followed by L 1 and L 3 respectively. The AMMI model with its additive as well as multiplicative formulations could interpret the complex G x E patterns effectively through the AMMI biplots. Genotypes were scored and then ranked , according to their stability index (SI) value computed on the basis of rank of AMMI stability value (rASV) and the rank of performance for each character (rY). It was observed that L 2 with its lowest score ranked as the best and stable genotype followed by L 1 and L 5 . The GGE biplot model enabled effective interpretation of genotype x environment interaction by providing visual understanding of genotype and test- environment evaluation through mean versus stability graph, ranking of genotypes, ranking of environments and which won where analysis. On an overall basis it was - - identified from GGE biplot method that L 1 , L 2 and L 3 were the most stable and superior performing genotypes. Comparison of results from the three models of stability confirmed L 1 and L 2 as the most stable and promising genotypes. Hence these genotypes can be evaluated in large fields so as to confirm with the results and for checking the suitability of these genotypes to be released as a variety.ThesisItem Open Access Gene action and gene expression analysis in yardlong bean(Vigna unguiculata ssp. sesquipedalis(L.) Verdcourt) for drought tolerance(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2022) Rahana, S N; KAU; Beena ThomasYard long bean (Vigna unguiculata ssp. sesquipedalis (L.) Verdcourt) is a highly remunerative legume vegetable of Kerala. Due to climate change and erratic rainfall, in summer season the crop growth and pod production is heavily affected by moisture stress. Development of high yielding varieties of yard long bean with drought tolerance is essential for its sustainable production. In this context, the present study entitled "Gene action and gene expression analysis in yard long bean (Vigna unguiculata ssp. sesquipedalis (L.) Verdcourt) for drought tolerance" was carried out in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, with an objective to identify drought tolerant genotype from the available germplasm and to understand the nature and magnitude of gene action and gene expression involved in the inheritance of drought tolerance in yard long bean. The study comprised four experiments. First experiment dealt with the seedling stage evaluation of 100-yard-long bean genotypes for drought tolerance in field. The moisture stress was imposed by withholding irrigation and later irrigation was restored in order to ensure the survival of the tolerant lines. The results of the analysis showed significant variations among genotypes. Out of the 100 genotypes screened, 15 drought tolerant genotypes were identified based on their better performance in terms of high RLW, low PWP, more number of days for reaching critical stress level and high recovery percentage. The genotypes identified were G1 (Acc 5), G5 (Acc 1339), G6 (Adoor local), G14 (Anchal local II) G15 (Aranmula local), G24 (Elamadu local), G36 (Kattampally local), G42 (Kollam local), G45 (Kottarakara local), G46 (Kottayam local), G50 (Kulashegarapuram local), 051 (Kulathupuzha local), G60 (Muttathukonam local), G74 (Nilamcl local) and G89 (Pongamoodu local). The fifteen selected genotypes from the first experiment were evaluated for drought tolerance in the second experiment by imposing moisture stress at the reproductive stage. Based on the biometric and physiological evaluations, the top seven genotypes with high yield and drought tolerance A4 (Anchal local II), A5 (Aranmula local), A7 (Kattampally local). All (Kulashegarapuram local), A13 (Muttathukonam local), A14 (Nilamel local), and A15 (Pongamoodu local) were selected as parents for further hybridization in experiment in. In the third experiment, LxT crosses were performed by using seven selected tolerant genotypes as lines with three popular yard long bean varieties as testers to generate twenty one hybrids. The genetic analysis of hybrids and parents were evaluated based on eight morphological and eight physiological parameters. Mean performance, combming ability, gene action and heterosis were estimated. Based on the mean performance and gca effects, L4 (Kulashegarapuram local) and L3 (Kattampally local) are identified as outstanding general combiners and can be exploited as parents for drought tolerance breeding in yard long bean. Three superior crosses, Kattampally local x Vellayani Jyothika (L3 x T3), Nilamel local X Lola (L6 X T2) and Kulashegarapuram local x Lola (L4 x T2) were identified as promising combinations for drought tolerance and yield xmder water stress. In the final experiment quantitative real time PCR was conducted to analyze the gene expression of drought responsive genes in tolerant hybrids and parents. The elevated expression of DREBs and NCEDl genes in tolerant hybrids and genotypes in gene expression analysis reflects the increased drought tolerance ability of those genotypes. The gene expression analysis was in conformity with the field studies. All the hybrids mamfested significant amount of dominance variance for commercial exploitation. Existence of significant amoimt of dominance variance and non-additive gene action suggests that hybridization as the best strategy for improving the drought tolerance character in yard long bean. The identified genotypes can be used for isolation of purelines with enhanced drought tolerance and the transgressive segregants from the identified crosses can be used for the development of drought tolerant high yielding cultivars in the fixture. The work can be continued with the identified genotypes and crosses for the development of climate smart drought tolerant varieties of yard long bean.ThesisItem Open Access Gene action and gene expression analysis in tomato (Solanum iycopersicum L.) under moisture stress(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2021) Chippy, A K; KAU; Beena, ThomasThe present investigation entitled ‘‘Gene action and gene expression analysis in tomato (Solanum lycopersicum L.) under moisture stress” was conducted during the period 2017-2021, in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani. Thirty-two genotypes including local genotypes of tomato were collected from different sources and studied under three different experiments. In the first experiment, twenty-one F1 hybrids derived from the line x tester mating and their ten parents were transplanted in the field and the treatment mean sum of square due to genotypes was found to be highly significant for all the characters studied which would ultimately indicate diverse nature of selected genotypes. The mean performance showed wide range of variation for most of the characters studied. Among lines, L7 (Pusa Ruby) showed higher mean performance for all-out of nine characters viz., plant height, fruit length, fruit girth, fruit volume, fruit weight, yield per plant, yield per plot, root length, root volume and relative water content. Among three testers, T2 (Kuttichal Local) showed favourable mean performance for seventeen traits viz., plant height, primary branches per plant, number of leaves per plant, number of flowering clusters per plant, number of fruits per cluster, fruit length, fruit girth, fruit volume, fruit weight, yield per plant, yield per plot, vitamin C, total acidity, stomatal frequency, root length, root volume and proline content. Under moisture stress (Second experiment), among the lines, L7 (Pusa Ruby) showed higher mean performance for eleven characters viz., plant height, fruit length, fruit girth, fruit volume, fruit weight, yield per plant, yield per plot, total soluble solids, root length, root volume and pollen viability. Among three testers, T2 (Kuttichal Local) showed favourable mean performance for sixteen traits viz., plant height, primary branches per plant, number of leaves per plant, number of flowering clusters per plant, number of fruits per cluster, number of fruits per plant, fruit length, fruit girth, fruit volume, total soluble solids, total acidity, stomatal frequency, root length, root volume and canopy temperature. Based on general combining ability analysis, the line L6 (Arka Alok) ranked as top by exhibiting significant gca effects for six traits viz., fruit length, fruit girth, fruit volume, fruit weight, specific leaf area and root length. Among the testers, T3 (Kottayam Local) had superior gca effects for Days to 50% flowering, number of flowering clusters per plant, number of fruits per cluster, fruit length, fruit volume, yield per plot, total soluble solids, lycopene, titrable acidity canopy temperature and proline content. Under moisture stress, the line L3 (Akshaya) ranked as top by exhibiting significant gca effects for ten traits viz., primary branches per plant, number of fruits per cluster, number of fruits per plant, fruit weight, yield per plant, yield per plot, stomatal frequency, specific leaf area, canopy temperature and pollen viability. Among the testers, T1 (Palakkad Local) had superior gca effects for plant height, number of fruits per cluster, number of fruits per plant, fruit girth, fruit volume, yield per plant, yield per plot, total soluble solids, relative water content, canopy temperature and proline content. In the case of hybrids, Based on mean performance, the crosses L1 x T1 (Vellayani Vijay x Palakkad Local), L2 x T3 (Anagha x Kottayam Local), L3 x T1 (Akshaya x Palakkad Local), L3 x T3 (Akshaya x Kottayam Local), L4 x T1 (PKM 1 x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T1 (Arka Meghali x Palakkad Local), L5 x T2 (Arka Meghali x Kuttichal Local), L5 x T3 (Arka Meghali x Kottayam Local), L6 x T2 (Arka Alok x Kuttichal Local) and L6 x T3 (Arka Alok x Kottayam Local) were superior for different yield characters. Under moisture stress, Hybrids L3 x T1 (Akshaya x Palakkad Local), L3 x T3 (Akshaya x Kottayam Local), L4 x T1 (PKM 1 x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T2 (Arka Meghali x Kuttichal Local), L7 x T2 (Pusa Ruby x Kuttichal Local) and L7 x T3 (Pusa Ruby x Kottayam Local) were superior for yield based on mean value. Based on combining ability studies under normal field condition, identified superior crosses such as L1 x T1 (Vellayani Vijay x Palakkad Local), L2 x T1 (Anagha x Palakkad Local), L2 x T3 (Anagha x Kottayam Local), L3 x T1 (Akshaya x Palakkad Local), L3 x T3 (Akshaya x Kottayam Local), L4 x T1 (PKM 1 x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L6 x T2 (Arka Alok x Kuttichal Local), L6 x T3 (Arka Alok x Kottayam Local), L7 x T1 (Pusa Ruby x Palakkad Local) and L7 x T3 (Pusa Ruby x Kottayam Local) exhibited higher sca effects for yield per plant. Under moisture stress, identified superior crosses such as L1 x T2 (Vellayani Vijay x Kuttichal Local), L2 x T3 (Anagha x Kottayam Local), L3 x T1 (Akshaya x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T2 (Arka Meghali x Kuttichal Local), L6 x T2 (Arka Alok x Kuttichal Local) and L7 x T3 (Pusa Ruby x Kottayam Local) exhibited higher sca effects for yield per plant. Based on heterosis, Overall, nineteen hybrids (L1 x T1, L1 x T2, L1 x T3, L2 x T1, L2 x T2, L2 x T3, L3 x T1, L3 x T3, L4 x T1, L4 x T2, L4 x T3, L5 x T1, L5 x T2, L5 x T3, L6 x T1, L6 x T2, L6 x T3, L7 x T1 and L7 x T3) from twenty one crosses revealed significant and positive heterobeltiosis for yield per plant under normal field condition. In the case of gene action, dominance genetic variance (σ2D) is higher than additive genetic variance (σ2A) was observed for all the traits under normal field condition and water stress condition, indicating that traits are controlled by non-additive gene action. Correlation study revealed yield per plant was positively and significantly correlated with the traits viz., primary branches per plant, number of flowering clusters per plant, number of fruits per cluster, number of fruits per plant, fruit girth, fruit volume, fruit weight and yield per plot. Based on qRT-PCR, the expression of both genes (moisture stress related genesSlDREB 1 and SlWRKY 4) was upregulated under water stress in all selected genotypes and the expression was more than moisture stress tolerant check variety. The result is in accordance with the analysis of moisture stress related physiological characters in these genotypes in the field under stress. From the study, identified tolerant sources for moisture stress viz., Akshaya, Pusa Ruby and Kuttichal Local, these lines and tester can be used in breeding programmes for moisture stress tolerance. Based on mean performance and gca, the genotypes Vellayani Vijay, Akshaya, Arka Meghali, Arka Alok and Kottayam local which showed superiority in yield and fruit quality traits can be used for breeding for improvement of yield and quality traits. The hybrids L3xT1 (Akshaya x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T2 (Arka Meghali x Kuttichal Local) and L7 x T3 (Pusa Ruby x Kottayam Local) which showed superiority for yield and quality characters with tolerance to moisture stress can be recommended for release after yield trials.ThesisItem Open Access Gene action and gene expression analysis in tomato (Solanum iycopersicum L.) under moisture stress(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2022) Chippy, A K; KAU; Beena, ThomasThe present investigation entitled ‘‘Gene action and gene expression analysis in tomato (Solanum lycopersicum L.) under moisture stress” was conducted during the period 2017-2021, in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani. Thirty-two genotypes including local genotypes of tomato were collected from different sources and studied under three different experiments. In the first experiment, twenty-one F1 hybrids derived from the line x tester mating and their ten parents were transplanted in the field and the treatment mean sum of square due to genotypes was found to be highly significant for all the characters studied which would ultimately indicate diverse nature of selected genotypes. The mean performance showed wide range of variation for most of the characters studied. Among lines, L7 (Pusa Ruby) showed higher mean performance for all-out of nine characters viz., plant height, fruit length, fruit girth, fruit volume, fruit weight, yield per plant, yield per plot, root length, root volume and relative water content. Among three testers, T2 (Kuttichal Local) showed favourable mean performance for seventeen traits viz., plant height, primary branches per plant, number of leaves per plant, number of flowering clusters per plant, number of fruits per cluster, fruit length, fruit girth, fruit volume, fruit weight, yield per plant, yield per plot, vitamin C, total acidity, stomatal frequency, root length, root volume and proline content. Under moisture stress (Second experiment), among the lines, L7 (Pusa Ruby) showed higher mean performance for eleven characters viz., plant height, fruit length, fruit girth, fruit volume, fruit weight, yield per plant, yield per plot, total soluble solids, root length, root volume and pollen viability. Among three testers, T2 (Kuttichal Local) showed favourable mean performance for sixteen traits viz., plant height, primary branches per plant, number of leaves per plant, number of flowering clusters per plant, number of fruits per cluster, number of fruits per plant, fruit length, fruit girth, fruit volume, total soluble solids, total acidity, stomatal frequency, root length, root volume and canopy temperature. Based on general combining ability analysis, the line L6 (Arka Alok) ranked as top by exhibiting significant gca effects for six traits viz., fruit length, fruit girth, fruit volume, fruit weight, specific leaf area and root length. Among the testers, T3 (Kottayam Local) had superior gca effects for Days to 50% flowering, number of flowering clusters per plant, number of fruits per cluster, fruit length, fruit volume, yield per plot, total soluble solids, lycopene, titrable acidity canopy temperature and proline content. Under moisture stress, the line L3 (Akshaya) ranked as top by exhibiting significant gca effects for ten traits viz., primary branches per plant, number of fruits per cluster, number of fruits per plant, fruit weight, yield per plant, yield per plot, stomatal frequency, specific leaf area, canopy temperature and pollen viability. Among the testers, T1 (Palakkad Local) had superior gca effects for plant height, number of fruits per cluster, number of fruits per plant, fruit girth, fruit volume, yield per plant, yield per plot, total soluble solids, relative water content, canopy temperature and proline content. In the case of hybrids, Based on mean performance, the crosses L1 x T1 (Vellayani Vijay x Palakkad Local), L2 x T3 (Anagha x Kottayam Local), L3 x T1 (Akshaya x Palakkad Local), L3 x T3 (Akshaya x Kottayam Local), L4 x T1 (PKM 1 x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T1 (Arka Meghali x Palakkad Local), L5 x T2 (Arka Meghali x Kuttichal Local), L5 x T3 (Arka Meghali x Kottayam Local), L6 x T2 (Arka Alok x Kuttichal Local) and L6 x T3 (Arka Alok x Kottayam Local) were superior for different yield characters. Under moisture stress, Hybrids L3 x T1 (Akshaya x Palakkad Local), L3 x T3 (Akshaya x Kottayam Local), L4 x T1 (PKM 1 x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T2 (Arka Meghali x Kuttichal Local), L7 x T2 (Pusa Ruby x Kuttichal Local) and L7 x T3 (Pusa Ruby x Kottayam Local) were superior for yield based on mean value. Based on combining ability studies under normal field condition, identified superior crosses such as L1 x T1 (Vellayani Vijay x Palakkad Local), L2 x T1 (Anagha x Palakkad Local), L2 x T3 (Anagha x Kottayam Local), L3 x T1 (Akshaya x Palakkad Local), L3 x T3 (Akshaya x Kottayam Local), L4 x T1 (PKM 1 x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L6 x T2 (Arka Alok x Kuttichal Local), L6 x T3 (Arka Alok x Kottayam Local), L7 x T1 (Pusa Ruby x Palakkad Local) and L7 x T3 (Pusa Ruby x Kottayam Local) exhibited higher sca effects for yield per plant. Under moisture stress, identified superior crosses such as L1 x T2 (Vellayani Vijay x Kuttichal Local), L2 x T3 (Anagha x Kottayam Local), L3 x T1 (Akshaya x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T2 (Arka Meghali x Kuttichal Local), L6 x T2 (Arka Alok x Kuttichal Local) and L7 x T3 (Pusa Ruby x Kottayam Local) exhibited higher sca effects for yield per plant. Based on heterosis, Overall, nineteen hybrids (L1 x T1, L1 x T2, L1 x T3, L2 x T1, L2 x T2, L2 x T3, L3 x T1, L3 x T3, L4 x T1, L4 x T2, L4 x T3, L5 x T1, L5 x T2, L5 x T3, L6 x T1, L6 x T2, L6 x T3, L7 x T1 and L7 x T3) from twenty one crosses revealed significant and positive heterobeltiosis for yield per plant under normal field condition. In the case of gene action, dominance genetic variance (σ2D) is higher than additive genetic variance (σ2A) was observed for all the traits under normal field condition and water stress condition, indicating that traits are controlled by non-additive gene action. Correlation study revealed yield per plant was positively and significantly correlated with the traits viz., primary branches per plant, number of flowering clusters per plant, number of fruits per cluster, number of fruits per plant, fruit girth, fruit volume, fruit weight and yield per plot. Based on qRT-PCR, the expression of both genes (moisture stress related genesSlDREB 1 and SlWRKY 4) was upregulated under water stress in all selected genotypes and the expression was more than moisture stress tolerant check variety. The result is in accordance with the analysis of moisture stress related physiological characters in these genotypes in the field under stress. From the study, identified tolerant sources for moisture stress viz., Akshaya, Pusa Ruby and Kuttichal Local, these lines and tester can be used in breeding programmes for moisture stress tolerance. Based on mean performance and gca, the genotypes Vellayani Vijay, Akshaya, Arka Meghali, Arka Alok and Kottayam local which showed superiority in yield and fruit quality traits can be used for breeding for improvement of yield and quality traits. The hybrids L3xT1 (Akshaya x Palakkad Local), L4 x T2 (PKM 1 x Kuttichal Local), L5 x T2 (Arka Meghali x Kuttichal Local) and L7 x T3 (Pusa Ruby x Kottayam Local) which showed superiority for yield and quality characters with tolerance to moisture stress can be recommended for release after yield trials.ThesisItem Open Access Breeding brinjal (Solanum melongena L.) for resistance to brinjal shoot and fruit borer(Department of Plant Breeding and Genetics, College of Agriculture, Vellanikkara, 2021) Neeraja, Puthiamadom; KAU; Jiji, JosephBrinjal shoot and fruit borer is one of the most devastating pests of brinjal. It is a major bottle neck in the cultivation of egg plant. Identification of resistant sources and transfer of these genes into the high yielding varieties is an important step in mitigating the damage caused by this pest. Keeping the above facts in view, the present investigation entitled “Breeding brinjal (Solanum melongena L.) for resistance to brinjal shoot and fruit borer” was taken up in the Department of Plant Breeding and Genetics from Sept 2017- April 2021. Thirty one accessions belonging to eight different species of Solanum were evaluated in the field for resistance to brinjal shoot and fruit borer during Aug- Nov 2018. The treatments showed significant difference for all the characters studied. The highest number of primary branches was observed in IC618034 belonging to S. virginianum and the lowest number was observed in IC618035 belonging to Solanum melongena. Infestation of shoots by borer started nearly 7th week after transplanting. The number of damaged shoots was highest in Surya. The per cent shoot infestation ranged from 0 to 75 per cent. The wild accessions were resistant to brinjal shoot and fruit borer and only the melongena lines exhibited considerable damage. The extent of per cent fruit damage ranged from 0 per cent in the wild accessions to 64.67 per cent in Arka Neelkanth. Yield per plant and marketable yield per plant was highest in Vengeri. Based on the observation from the screening experiment conducted during August –November 2018, eleven accessions were evaluated further for a confirmatory screening during March- July 2020. As there was considerable infestation by borer, artificial inoculation was not carried out. There was significant difference among the treatments for the characters studied. Highest yield per plant and marketable yield per plant was observed in Vengeri. Lowest yield was observed in IC624241 and IC624239. Yield loss was highest in Ponni and lowest in IC624239. The resistance index developed by Mishra et al. (1988) was followed to classify the accessions. The accessions IC618035, IC618044, IC624241, Arka Kusumkar and Vengeri were graded as resistant (1-10 %) with respect to per cent shoot damage. Based on the extent the of fruit damage, the accessions IC618035, IC618044 and IC624239 and Arka Kusumkar were graded as moderately resistant (11-20%). IC624241 and Vengeri were graded as resistant (0-10%). Haritha, Surya, Arka Keshav, Ponni and Arka Neelkanth were graded as highly susceptible (>40 %). The biophysical, anatomical and biochemical basis of resistance was elucidated in the selected accessions. Trichomes in Solanum sps. were stellate in shape with six trichome strands forming one asterisk. Trichome density on calyx was negatively and significantly correlated with shoot damage while trichome density on leaf was positively and significantly correlated with shoot damage. Pericarp thickness, fruit breadth, total sugar content and total nitrogen content were positively and significantly correlated with per cent fruit damage. Total phenol content showed a significant negative correlation with the per cent fruit infestation. The resistant lines IC618035, IC618044, IC624241, Arka Kusumkar and Vengeri were characterized by thick epidermis and cuticle, dense and compact cortex and cambium, well developed vascular bundles and small pith. IC618044, IC624241, IC618035, Arka Kusumkar and Vengeri falling under the resistant and moderately resistant category were selected as testers to cross with highly susceptible lines Haritha, Ponni and Arka Neelkanth. The twelve hybrids and parents were screened in the field along with the hybrid ‘Neelima’ as check during Dec 2020- April 2021 to check the per cent infestation of brinjal shoot and fruit borer. On the basis of per cent fruit infestation the hybrids Haritha x JRPH IC624241 and Ponni x IC624241 were graded as immune. Haritha x IC618044, Haritha x IC618035, Ponni x IC618035 and Ponni x Vengeri were graded as resistant. The hybrids Haritha x Vengeri , Haritha x Arka Kusumkar, Ponni x IC618044 and Neelima were graded as moderately resistant. Ponni x Arka Kusumkar was graded as tolerant. The hybrid Arka Neelkanth x IC618035 Arka Neelkanth x IC618044 were graded as highly susceptible. L x T analysis revealed that Vengeri can be considered as a good combiner with respect to per cent shoot damage, yield per plant and marketable yield per plant. While Arka Kusumkar is a good combiner with respect to number of fruits per plant, number of damaged fruits and per cent fruit damage. Among the lines, Ponni can be considered as a good combiner in terms of yield and marketable yield. Ponni x Vengeri can be considered to be superior as it exhibited high positive heterosis over the mid parent, better parent as well as the commercial check for yield as well as marketable yield and high negative heterosis for per cent shoot and fruit damage. Almost all the hybrids exhibited negative heterosis for per cent shoot and fruit damage. Ponni x Vengeri was ranked as ‘one’ on the basis of organoleptic evaluation conducted with a panel of ten judges. DNA fingerprinting of the superior F1 hybrids and the respective parents done using nineteen ISSR markers revealed that Arka Neelkanth was unique in a way that it produced 10 bands for UBC 836. Comparing other hybrids highest number of bands for UBC 868, UBC 816, UBC 836 and ISSR 08 were produced by Arka Neelkanth. UBC 826 was used to identify bands common in hybrids and parents in Haritha x IC618044, Haritha x Arka Kusumkar, Ponni x Arka Kusumkar and Arka Neelkanth x IC618044. The bands similar in parents and hybrids were identified in all the hybrids except Ponni x IC624241. Superior hybrids viz., Ponni x Vengeri, Ponni x Arka Kusumkar and Haritha x Vengeri can be evaluated further. Mapping populations can be developed from the segregating generations to develop functional markers for resistance to brinjal fruit and shoot borer.ThesisItem Open Access Genetic diversity analysis of horse gram [Macrotyloma uniflorum (Lam.) Verdc.] for moisture stress tolerance in south central laterites of Kerala(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2022) Visakh, R L; KAU; Bindu, M RThe study entitled “Genetic diversity analysis of horse gram [Macrotyloma uniflorum (Lam.) Verdc.] for moisture stress tolerance in south central laterites of Kerala” was carried out at Farming Systems Research station, Sadanandapuram during October, 2020 – March, 2021 with the objective to identify superior genotypes of horse gram with moisture stress tolerance having high yield and quality, suitable for south central laterites of Kerala. The 30 horse gram genotypes collected from RARS Pattambi under KAU and other SAUs were evaluated for moisture stress tolerance at seedling stage in laboratory using two different concentrations of PEG 6000 (10% and 20%), in factorial completely randomized design replicated thrice. These 30 genotypes were further evaluated for water stress tolerance in field by withholding irrigation for 15 days at reproductive stage, in randomized block design replicated thrice. The seeds were sown on raised beds of 2.25 m2 size at a spacing of 30 cm х 30 cm and cultural operations were adopted as per the “Package of Practices Recommendations Crops 2016” of Kerala Agricultural University. The soil moisture was also measured during water stress-imposed period at weekly interval by following gravimetric method. In laboratory experiment, genotypes were evaluated for six different morphophysiological characters and in field experiment, genotypes were evaluated for 31 characters which included biometric, physiological, biochemical and seed quality characters. The genotypes showed significant differences for all the characters studied. Genetic parameter analysis was performed for nineteen characters and for all the characters PCV values were higher than GCV values indicating the influence of environment. High heritability coupled with high GAM was observed in days to 50% flowering, number of primary branches per plant, plant height, number of pods per plant, haulm yield per plant, harvest index, days to maturity, leaf area index, total phenol content of the seed, root dry weight, proline content, total chlorophyll content and seed yield per plant. 124 The correlation studies revealed significant positive correlation of the characters RWC, number of pods per plant, number of seeds per pod, total chlorophyll content, harvest index, proline content, root dry weight, root length, LAI and hundred seed weight with seed yield. However, days to 50% flowering, days to maturity, plant height and number of primary branches showed a significant negative correlation with yield. Path analysis of the thirteen characters showing significant correlation with seed yield revealed high positive direct effect of the number of pods per plant on seed yield. High indirect effect on yield was observed for number of seeds per pod through number of pods per plant. Genetic divergence analysis using Mahanalobis D2 statistic following Tocher’s method grouped the 30 genotypes into four clusters. Cluster II with 14 genotypes was the largest followed by cluster I (9 genotypes), cluster III (6 genotypes) and clusters IV was solitary cluster. Highest intra cluster distance was recorded among the genotypes of cluster II and lowest in cluster IV whereas highest inter cluster distance was observed between the genotypes of the clusters I and III, while lowest inter cluster distance was between the genotypes of the clusters III and IV. The present study revealed that the genotypes IC71841, IC139464 and IC22785 as water stress tolerant ones in laboratory experiment and in field experiment, genotypes IC22785, IC221105, IC22759 and IC139464 were identified as high yielding and moisture stress tolerant ones. The superior genotypes identified in the study can be recommended for cultivation and future breeding programs.ThesisItem Open Access Genetics of shattering resistance in rice (Oryza sativa L.)(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2020) Akhil, K P; KAU; Biju, SRice is the staple food crop of Kerala. Among the plethora of rice varieties available Jyothi is by far the most popular in the state owing to its consumer preference. Easy seed shattering leads to significant loss in Jyothi which can be overcome by transferring shattering resistance to it. Screening of genotypes for shattering resistance identified donors like Athira and Triveni with good combining ability. The present experiment was thus envisaged to transfer shattering resistance to Jyothi from the donors and to study the nature of inheritance of seed shattering trait using six parameter model of generation mean analysis. Observations were recorded in the two crosses viz., Jyothi x Aathira and Jyothi x Triveni for eleven quantitative characters viz. days to 50 per cent flowering, plant height, tillers per plant, panicles per plant, panicle length, spikelets per panicle, seeds per panicle, days to maturity, test weight, grain yield per plant and shattering per cent in six generation (P1, P2, F1, F2, B1 and B2) obtained from the two crosses. Panicle per plant, test weight, grain yield per plant and shattering per cent recorded significant heterosis in the desirable direction in both the crosses. Mean effect (m) was significant and positive for all the characters studied in both the crosses. The additive component (d) was significant and positive for days to 50 per cent flowering, tillers per plant, panicles per plant, days to maturity, grain yield per plant and shattering per cent in the cross Jyothi x Triveni while the other characters recorded significant and negative additive gene effect. The characters tillers per plant, test weight and shattering per cent showed significant positive additive gene effect in the cross Jyothi x Aathira and rest of the characters showed significant and negative additive component. Dominance gene effect was observed to be significant and negative for most of the characters studied in both the crosses. Non-allelic interactions were observed to be significant in most of the characters except days to 50 per cent flowering, tillers per plant, days to maturity and shattering per cent in the cross Jyothi x Aathira. The epistatic interaction model of generation mean analysis was found adequate for obtaining gene actions for all the characters. Among the three type of interactions dominance x dominance interaction was found to be more important for all the characters. Additive x additive and additive x dominance gene interactions were found to be equally important for most of the characters studied in both the crosses. Dominance gene effect was observed controlling the genetic variance in most of the traits studied. Presence of additive and non-additive along with epistatic interaction revealed the complex nature of inheritance of the characters. Predominant dominant gene effect along with duplicate epistasis limits the scope of direct selection. In the F2 populations characters showed high heritability in both the populations studied. Plant height, panicle length and days to maturity were found to have low genetic advance. Plant height recorded moderate genetic advance, whereas, other characters exhibited high genetic advance in F2 population derived from the cross Jyothi x Triveni. Days to maturity showed low genetic advance, days to 50 per cent flowering and panicle length recorded moderate genetic advance and other characters recorded high genetic advance in F2 population derived from the cross Jyothi x Aathira. As shattering resistance is governed predominantly by additive gene effects selection in the early segregating generations will be highly rewarding. The selected lines may be advanced to further generations to identify lines with high yield coupled with shattering resistance and can be forwarded to develop high yielding varieties with low shattering or bi-parental mating among the selected lines may be undertaken to recover superior recombinants with high yield and shattering resistance.
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