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Open Access
Bioactivity of carotenoids from shrimp shell waste
(Department of Processing Technology,College of Fisheries,Panangad, 2010) Sindhu, S; KAU; Sherief, P M
Shrimp processing waste is the single largest industrial waste in the country causing diverse environmental problems. A study was carried out to assess the extractability of astaxanthin from shrimp waste in different organic solvents and vegetable oils. Extraction was tried using wet and dried waste, with and without deproteinisation. Waste was subjected to deproteinisation using alkali and enzyme (pancreatin). The different solvent systems tried were ether:acetone:water (15:75:10 v/v/v), acetone, hexane:isopropanol (3:2 v/v) and 90% acetone v/v. Astaxanthin in the extract was quantified by measuring the OD at 470 nm in hexane. Extraction was also done using vegetable oils viz. coconut oil, soybean oil and sunflower oil. Quantification of astaxanthin in pigmented oil was done by measuring the absorbance at 485 nm using 2155 as extinction coefficient. Astaxanthin yields from deproteinised samples were significantly lower than those from non deproteinised samples. The highest astaxanthin yield of 87.14 ± 4.55μg/g was obtained with non deproteinised wet waste extracted using acetone. The astaxanthin yield was significantly lower when oil was used as the extraction medium. Of the three oils coconut oil gave the highest yield. The results showed that acetone is the best solvent for extracting astaxanthin from shrimp shell waste in wet condition. The astaxanthin content in Aristeus alcocki shell waste is double that of Pandalus borealis shell waste, which is currently used as the commercial source of astaxanthin. The deep sea species Aristeus alcocki can thus be considered as a better source of astaxanthin for commercial exploitation than Pandalus borealis. TLC analysis of the shell waste extract showed that it contains free astaxanthin, astaxanthin monoester and astaxanthin diester in the ratio 1:1:2. GLC identification of the fatty acids esterified with astaxanthin revealed that saturated fatty acids, MUFA and PUFA are in the ratio 5:3:2 in monoester, whereas in diester they are in the ratio 4:3:3. The main fatty acids in monoester and diesters are palmitic acid, oleic acid, stearic acid and PUFAs: DHA and EPA. The in vitro antioxidant activity of the astaxanthin extract showed significant hydroxyl radical scavenging activity, superoxide anion scavenging activity and inhibition of lipid peroxidation. The IC50 values obtained were 56.43 ± 1.06 ng/ml, 27.91 ± 0.54 ng/ml and 26.54 ± 0.42 ng/ml, respectively. The antioxidant activity of astaxanthin from Aristeus alcocki was obtained at nanogram levels. This powerful antioxidant function may be due to the unique molecular structure of astaxanthin and synergistic effect of astaxanthin and PUFAs present in the astaxanthin monoester and diester fractions. The astaxanthin extract from shrimp shell waste significantly reduced carageenan induced paw edema in mice, percentage inhibition being 47.83 and 67.11 percent at astaxanthin concentrations of 0.5 mg/kg body weight and 1.0 mg/kg body weight, respectively. The inhibition of inflammation at 1.0mg/kg body weight was greater than that produced by the standard reference drug diclofenac. Cardioprotective effect of astaxanthin was examined in isoproterenol induced myocardial infarction in rats. Levels of diagnostic marker enzymes, LDH, CPK, GOT, GPT, CK, CK-MB in plasma, lipid peroxides, ascorbic acid, reduced glutathione and the activities of glutathione-dependent antioxidant enzymes GPx, GR, GST and antiperoxidate enzymes CAT, SOD and the membrane bound enzyme Na+ - K+ ATPase in the heart tissues of experimental groups of rats were determined. The prior administration of astaxanthin @ 10mg/kg feed for 45 days significantly prevented the isoproterenol-induced elevation in the levels of diagnostic marker enzymes in plasma, induction of lipid peroxidation and alterations in the level of reduced glutathione and in the activities of glutathione dependent antioxidant enzymes and antiperoxidative enzymes of experimental rats. Feeding astaxanthin caused a decrease in the inhibition of Na+ - K+ ATPase activity against isoproterenol induced myocardial infarction. The powerful cardioprotective effect of astaxanthin can be attributed to the multiple independent mechanisms viz. antioxidant effects, singlet oxygen quenching ability and inhibition of lipid peroxidation of membranes, increased functional gap junctional intercellular communication, anti-inflammatory effects etc. Immunostimulatory action of astaxanthin extract was evaluated in experimental mice. Astaxanthin administration was found to enhance the proliferation of spleen cells and bone marrow cells. Esterase activity was found to be enhanced in bone marrow cells indicating increased maturation of cells of lymophoid linkage. Astaxanthin also enhanced number of antibody forming cells and circulating antibody titre. Thus astaxanthin exhibits strong immunomodulating properties. A significant reduction in the viability of ascites tumour cells DLA in vitro was noted in the current study. The % viability was reduced to 4.34 % at a concentration of 15μg astaxanthin/ml. The cytotoxic action of astaxanthin against DLA may be through induction of apoptosis or through a different pathway. Antitumour activity of astaxanthin was studied by ascite and solid tumour models in mice. An increase in life span of about 67 % was noted in DLA bearing mice administered with astaxanthin at 5 mg/kg body weight. The tumour volume and tumour weight were significantly lower in mice injected with 5 mg/kg body weight astaxanthin. In vitro studies revealed that astaxanthin from shrimp shell waste of Aristeus alcocki inhibited the proliferation of cervical cancer cells HeLa in a dose dependent manner.
Open Access
Genetic characterization, controlled breeding and development of transgenic varieties of puntius denisonii (day, 1865).
(Department of Aquaculture, College of Fisheries, Panangad, 2010) Manoj, C K; KAU; Mohanakumaran Nair, C
Puntius denisonii, a beautiful ornamental fish indigenous to the Western Ghats, which has been indiscriminately exploited from the different rivers of Kerala has been recently declared to be vulnerable by the IUCN. The population structure and genetic diversity of P. denisonii has not yet been studied and documented. Many previous attempts to breed this fish in captivity have yielded negative results. The increasing demand for this fish to decorate aquariums worldwide could be satisfied only by developing controlled breeding techniques and larval rearing of its fry. In the present study, the present population structure of P. denisonii has been studied combining both phenotypic and genotypic techniques. Fishes were collected from Irrity, Chaliyar and Periyar rivers of Kerala. Truss network analysis was conducted and the size adjusted morphometric variables were subjected to Principal Component Analysis and Canonical Variance Analysis. Scatter diagram and Dendrogram was plotted using PCA and CVA loadings. The Irrity and the Chaliyar populations were grouped on the positive sector of the PC and CV component showing morphological similarities between the two populations while the Periyar population was placed in the negative sector of the component separated far from the other two. The PC scores were used to find out the variables showing maximum variation between fishes collected from different rivers. RAPD PCR was conducted after isolating DNA from the fins of different populations of P. denisonii. Universal random primers were screened and the primers that produced reproducible bands were selected. Popgene analysis of the binary data yielded the genetic structure of different populations of P. denisonii. Number and percentage of polymorphic loci, Nei's (1973) gene diversity, Shannon's Information index Lewontin (1972), Nei's Unbiased Measures of Genetic Identity and Genetic distance and Dendrogram Based Nei's (1978) Genetic distance using UPGMA --Modified from NEIGHBOR procedure of PHYLIP Version 3.5 were studied. The results obtained supports the truss analysis in that the Irrity and Chaliyar populations in Northern Kerala are genetically more similar while that of Periyar population in Central Kerala are distinct. P. denisonii was successfully induced bred under controlled conditions with synthetic hormone preparations Ovaprim and WOVA-FH. Stress during transport and handling was minimized and live feed was supplemented to enhance maturation of the broodstock. The whole developmental sequence starting from fertilized eggs to hatching was photographed and documented. It took 29-30 hours for the eggs to hatch at 280C. Rearing of fry was successfully accomplished under laboratory conditions. In an attempt to develop transgenic varieties of P. denisonii, pCMV-GFP was electroporated into newly fertilized eggs, maintained in hypoosmolar electroporation buffer. The electroporation parameters that yielded best results were 20V, 3 bursts at 1 second interval. Fin clips were taken from the transgenic individuals reared for a period of 6 weeks. Dot blot test was positive showing integration of the GFP gene in P. denisonii, eventhough expression was not detected under blue or UV light. The genetic and phenotypic data of P. denisonii populations in the present study will aid as a base line for formulating conservation procedures to protect the genetic diversity of wild ones. Stock identification studies are recommended for more concise information on each population. Moreover, the larval rearing and controlled breeding techniques along with the genetic diversity studies will help to design captive breeding programs and enhance the production of hatchery bred ones to meet increasing demand. Further research is recommended for generating transgenic lines with uniform GFP expression.
Open Access
Heterosis breeding in sesame (Sesamum indicum L.).
(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2011) Gayathri, G; KAU; Dijee, Bastian
The study entitled ‘Heterosis breeding in sesame (Sesamum indicum L.)’ was undertaken at the Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara. The objectives of the study were to collect and evaluate different genotypes of sesame for morphological traits and yield attributes, to identify useful parents producing heterotic crosses and developing hybrids in sesame. The study also intended to develop male sterile lines in sesame through interspecific hybridization with Sesamum malabaricum. Sesamum indicum and Sesamum malabaricum accessions were collected from Kerala and Tamil Nadu and evaluated for their morphological traits. Wide range of variation was noticed for characters like plant height, number of days to flowering and seed yield per plant which contributed maximum to genetic divergence. The genotypes studied were grouped into six clusters. High genotypic coefficient of variation (GCV) was recorded for number of capsules per plant, plant height, seed yield per plant and number of branches per plant. High heritability with high genetic advance as per cent of mean was recorded for number of days to flowering, plant height, number of branches per plant, number of capsules per plant and seed yield per plant. This indicates that the characters are governed by additive gene effects and selection for these traits will be effective. Association analysis revealed that seed yield per plant was correlated to plant height, number of capsules per plant and number of days to flowering. Path coefficient analysis indicated maximum positive direct effect by number of capsules per plant, capsule length, plant height and 1000 seed weight on seed yield per plant. In order to develop hybrids, fourteen parents were selected based on the per se performance of the genotypes. They were crossed in line X tester mating design. Forty eight hybrid combinations obtained were raised in the field along with the parents and evaluated for their heterosis and combining ability effects. Parental genotypes AVTS-06-5, AVTS-06-10, IVTS-06-12, KYM-1, Tilak and TMV-6 were identified as high combiners based on general combining ability (gca) effects. Two combinations viz. AVTS-06-5 X KYM-1 and IVTS-06-12 X TMV-3 had significant values of per se performance, specific combining ability (sca) effects and standard heterosis for seed yield per plant. They can be evaluated for their hybrid vigour over locations and seasons. The crosses AVTS-06-5 X TMV-3, AVTS-06-5 X TMV-6 and TCR 3279A X KYM-1 have been identified as potential cross combinations for isolation of promising segregants as the parents involved in these crosses had high significant gca effects for seed yield per plant but the hybrids recorded non significant sca effects. Interspecific hybridization between S.malabaricum and S.indicum was attempted to develop male sterile lines. Seed set was noticed in three interspecific hybrids which failed to germinate due to embryo abortion. Hence these embryos were rescued and raised in vitro to obtain the hybrids.
Open Access
Detergent potential of enzymes of dairy microflora and their effect on the shelf life of milk products
(Department of Dairy Science, College of Veterinary and Animal Sciences, Mannuthy, 2010) Beena, A K; KAU; Geevarghese, P I
A study was conducted to assess the detergent potential of a spoilage protease enzyme obtained from the microflora of dairy plant environment. An attempt was also made to study the impact of selected enzyme producers on the shelf life of curd (dahi) and sterilised skim milk. A total of 71 bacterial isolates obtained from dairy environment were screened for their ability to produce spoilage enzymes like proteases lipases and lecithinases. Based on the spoilage potential, Pseudomonas aeruginosa (P12) isolated from pasteurised milk and Bacillus cereus (S4) isolated from sterilized skim milk were selected for further work. The influence of spoilage enzymes on selected physico-chemical characteristics of curd (dahi) and sterilized skim milk was evaluated by preparing the products from milk precultured with isolate P12 and S4. In general, proteolysis of milk was found to have an adverse effect on the quality of products. The stimulatory effect of proteolytic products of P12 and S4 on curd starters was evident from the higher values of acidity, firmness and syneresis in treated curd. The spoilage enzymes adversely affected the overall quality and shelf life of curd. In treated sterilised milk, tyrosine and NPN values were highly elevated. A linear correlation was found to exist between off-flavour and proteolysis. Curd and sterilised skim milk prepared from milk precultured with proteolytic organism were significantly different from that of control. The possibility of exploiting an alkaline protease from spoilage organism in dairy plant sanitation was also looked into. Environmental conditions for the production of alkaline protease by a psychrotrophic strain of Bacillus cereus (S4) was optimised in whey based medium. The protease used in this trial preferred an alkaline medium to remain stable. The enzyme was found to be stable over a wide temperature range of -10°C to 80°C and a pH range of 7.0 to 12.0. The metal ions Ca++, Mg++, Zn++ and Hg++ enhanced the enzyme activity. Lack of inhibition by Hg++ suggested lack of disulphide bonds in the active site of enzyme. Significant inhibition of activity by serine inhibitors indicated an essential serine residue in the active site of enzyme. The deleterious effect of EDTA on enzyme activity showed the supportive role of divalent cations. Marked residual activity on treatment with β-mercaptoethanol indicated the absence of cysteine residue for the enzyme. Enhancement of protease activity in the presence of surfactants and stability in the presence of H2O2 signified its potential to be used as detergent additive. Qualitative assessment of cleaning efficiency of inbuilt formulation substantiated the superiority of enzyme based formulations. Ammonium sulphate fractionation, dialysis and gel filtation using seralose 4B and Seralose 6B were effective in purifying the protease preparation by 141.31 fold. The purified protease was found to be a homogenous preparation of molecular weight of 50.5 kDa as determined by SDS PAGE.
Open Access
Development and evaluation of different vaccines against duck pasteurellosis
(Department of Dairy Science, College of Veterinary and Animal Sciences, Mannuthy, 2010) Jesto, George; KAU; Krishnan Nair
This study was undertaken to develop biofilm vaccines against duck pasteurellosis using oil, saponin and aluminium hydroxide as adjuvants and to experimentally evaluate their immunogenicity in ducks. Identity of P. multocida serotype A: 1 (DP1) used for study was confirmed by biochemical tests and by PM-PCR and pathogenicity was established in Swiss albino mice before vaccine production. The LD50 (11 w) of the DP1 isolate determined was 10 CFU / bird and MDT was 23.75 h in 11 week old ducklings when a high dose of 3 X 10 6 CFU of P. multocida per bird was given. At 11 Weeks age MDT gradually increased as the dose of inoculum decreased. In 21 week old ducks, the LD50 (21w) of the isolate DP1 was estimated to be 3 ×108 CFU of P. multocida and it showed that the Kuttanad duck had decreased susceptibility to pasteurellosis with age. On light microscopic studies planktonic cells appeared to be Gram negative coccobacillary, while biofilm cells were Gram negative and pleomorphic. Electron microscopic studies revealed that P. multocida could form well differentiated classic biofilm and 0.32 per cent TSB media supplemented 0.5 per cent chitin seemed to be excellent medium for biofilm formation by P. multocida. Four different vaccines viz. OV, OBV, SV and SAV were prepared and all of them were found to be sterile and safe. The oil adjuvanted vaccines (OV and OBV) offered better protection compared to saponin and aluminium hydroxide adjuvanted vaccine groups (SV and SAV) following primary vaccination, up to seven weeks. The modified oil adjuvanted vaccine prepared was not only found to be homogenous, but also more efficient in stimulating a humoral immune response and hence may be recommended. The oil adjuvanation gave better protection than saponin and aluminium hydroxide adjuvanation. The SAV gave better protection than SV which might be due to the presence of aluminium hydroxide which potentiated the immunostimulating ability of saponin. The combined vaccine (SAV) although was found to be better than single vaccine (SV) they cannot be used as a substitute to oil adjuvanted vaccines. The booster vaccination was found to have added advantageous effect on protection and is a must, to prevent losses. Pasteurella biofilms although found to be weak in inducing a primary immune response had the potency to evoke a more powerful secondary response compared to planktonic cells. Vaccination done at six weeks age followed by booster vaccination at 16 weeks age seemed to be a better modification of existing schedule and may be recommended. In histopathological studies, lymphoid hyperplasia was observed in spleen in survived control birds and in SV and SAV vaccine groups that did not survive challenge test, which indicated the persistence of Pasteurella organisms through mild infection in them following experimental challenge. Lymphoid depletion was observed in caecal tonsil in experimental pasteurellosis as in spleen. As the survived vaccinated birds following challenge test showed normal intact caecal tonsil, the course of disease and lesions might be less prominent in vaccinated birds during infection process. The well developed bursa observed in OV and OBV birds that survived challenge test indicated that the humoral immune response was well induced in them compared to other groups. The designed primers E1 and E2 amplified the gene E and hence, this pair of primers could be used for the production of amplified Gene E sequences for further studies on recombinant ghost system. In conclusion, 0.32 per cent TSB media supplemented 0.5 per cent chitin seemed to be an excellent medium that support classical biofilm formation by P. multocida. Booster vaccination definitely had added advantageous effect on protection. Immunization at 6 weeks of age with OV followed by booster vaccination at 16 weeks age with OBV seemed to be a better modification of existing schedule and may be recommended. In histopathological studies, the lesions were less prominent in vaccinated birds than control birds which indicated that the vaccines were effective.
Open Access
Sulphur and boron nutrition and their foliar diagnosis in sesame
(Department of Soil Science and Agricultural Chemistry,College of Agriculture, Vellayani, 2010) Jeena, Mathew; KAU; Sumam, George
A laboratory cum field experiment was conducted to study the effect of S and B on the growth, yield and quality of sesame var. Thilarani and to standardize the foliar diagnosis of these elements in Onattukara sandy loam soil. The study included an incubation study and two field experiments. The treatments comprising the different levels of S and B laid out in 42factorial RBD. The treatments were T1(S0B0), T2(S0B1), T3(S0B2), T4(S0B3), T5(S1B0), T6(S1B1), T7(S1B2), T8(S1B3), T9(S2B0), T10(S2B1), T11(S2B2), T12(S2B3), T13(S3B0), T14(S3B1), T15(S3B2), T16(S3B3). The different levels of S were S0 (0 kg ha-1), S1 (7.5 kg ha-1), S2, (15 kg ha-1) and S3 (30 kg ha-1) and B0 (0 kg ha-1), B1(2.5 kg ha- 1), B2 (5 kg ha-1) and B3 (7.5 kg ha-1). The incubation study was conducted at College of Agriculture, Vellayani to understand the dissolution and release pattern of S and B from their sources gypsum and borax respectively in Onattukara sandy soil. The results revealed that the release of S and B was maximum at the 30th DOI. Increasing levels of S and B has a positive influence on the S content of the soil. T16 (S3B3) recorded the highest value at all the sampling stages for S whereas in the case of B, the treatment combinations which received B at the highest levels in combination with S3 or S2 showed the highest value. The field experiments were laid out at ORARS, Kayamkulam in 42 factorial RBD having two replications using Thilarani as the test crop. It was observed that application of S and B favourably influenced the yield and yield attributes of sesame. T16 was found to be the treatment which gave the highest grain yield and oil yield in both the years and was found to be on par with T14 (S3B1). S3 was the superior S level. As for the different levels of B, B1 can be inferred as the best level. The content of saturated fatty acids such as palmatic and stearic acid showed a decreasing trend with increasing levels of S and B whereas the content of the unsaturated fatty acids showed an increasing trend. The quality attributes of oil such as acid value, iodine value and saponification value was also studied and it was found that there is a decreasing trend with regard to acid and saponification value and an increasing trend for iodine number. The grain protein content also showed an increasing trend with the increase in rate of application of S and B. Regarding the content and uptake of N, P, K, S, B, Fe, Mn, Cu and Zn, a favourable influence for the different levels of S and B was recorded. Results regarding the S and B use efficiency and their apparent recovery showed that with increase in levels of S, an increasing trend was observed for S. In the case of B, increase was noticed up to B1 (2.5 kg ha-1) and there after showed a decreasing trend. This positive influence was also reflected on the available nutrient status of the soil such as organic carbon content, available N, P, K, S, B and DTPA extractable micronutrients. Correlation studies conducted to standardize the part and stage of sampling for the foliar diagnosis of sesame showed petiole at 30 DAS and 20 DAS in the case of S and B respectively. The same stages were found for the soil sampling also for both the nutrients. The critical nutrient level in the part standardised for these two nutrients were standardized using the graphical method proposed by Cate and Nelson (1965). In the case of S, it had been standardized as 0.088 per cent and for B, it had been found to be 28 mg kg-1. The critical nutrient level in soil was also estimated using the scatter diagram technique and was found to be 23 kg ha-1 at 30 DAS for S and 1.4 ppm at 20 DAS for B. Hence the application of S @ 30 kg ha-1 and B @ 2.5 kg ha-1 could faourably enhance growth of sesame with regard to the growth characters, yield and yield attributes and the quality aspects. Moreover, analysis of the plant and soil samples at the critical stages fixed for the respective nutrients will provide the necessary data for the sustainable management of the crop in Onattukara sandy loam soil.
Open Access
Variability in asoka (Saraca asoca(Roxb.) de wilde)
(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2010) Vidhu Francis, Palathingal; KAU; Radhakrishnan, V V
Asoka (Saraca asoca) is a sacred tree among the Buddhists and Hindus. It is called sorrow-less tree as it removes the grief. The tree has immense medicinal properties. Its bark is mainly used for correcting uterine problems. The well-known Ayurvedic preparations of asoka bark are Asokarishtam and Asokaghrutham. Due to over exploitation of this tree for its bark, this has now become almost extinct. The International Union for Conservation of Nature and Natural Resources (IUCN) has listed this species under ‘globally vulnerable’ category. It is also enlisted among the 36 threatened and endangered medicinal plants of India. Due to its acute short supply compared to its demand, various development and research activities are being prioritized to conserve, utilize and improve this species. Therefore the present study was undertaken to assess morphological variations in existing germplasm and to study the reproductive biology of S. asoca. Collection of seeds and evaluation of seed and seedling traits, evaluation of therapeutical components and molecular characterisation of asoka were the other objectives of the study. Variability studies for morphological traits of asoka indicated that height of plant and stem girth have high correlation with bark yield as well as higher direct effect. These traits can be used for identifying better genotypes for higher bark yield. In discriminant function analysis, the selection index involving height of plant along with bark yield constituted for selection criteria among asoka genotypes. Using this selection index, accessions IC566463, IC566489, IC566488, IC566482 were selected as better accessions for higher bark yield. The 43 accessions maintained in germplasm of asoka at AICRP on M&AP were grouped into two major clusters based on morphological traits. These clusters further formed 6 clusters. Accession IC566488 grouped alone in one cluster indicated that this accession was different from all other accessions. In all the other 5 clusters, accessions from Thrissur was included. This indicated that accessions did not follow geographical distributions. Some of the accessions from Thrissur may have relationship with accessions from other districts. Reproductive biology of asoka was studied. It indicated that the reproductive traits like floral biology, anthesis, pollen morphology, its viability have variability among eight trees studied in KAU campus. Among the eight trees studied, ‘KAU8’ have better reproductive traits compared to the rest. Preliminary studies were carried out in pollination system and agents for pollination in asoka. The studies indicated that ants may be one of the pollinating agents. Seed and seedling behaviour of asoka were studied on seeds collected from 80 trees located in different districts of Kerala. Seed breadth, seed volume, height of plant and stem girth were identified as selection traits for better seedlings. Hence at seed stage, bigger sized seeds with higher seed volume will result into better seedlings. Vigorous seedlings is produced from tall seedlings with higher stem girth and in mature plants for higher bark yield, height of plant and stem girth can be selection traits. Thus the selection parameters were worked out in three stages in asoka. Based on seed and seedling selection parameters, OKL4, OKL2 from Odakkali, KKL2 from Kottakkal and TVM2 from Thiruvananthapuram were selected as better mother plants for higher bark yielding accessions. The biochemical constituents (phenol and tannin content) imparting medicinal properties were estimated among the different age groups of asoka. Both phenol and tannin contents were higher in bark compared to flower and leaves. About 50 per cent of therapeutical constituents are available in flowers and hence at non destructive level, flowers of asoka can be substituted for bark. The trees KAU8, KAU7, KAU6 and among the accessions IC566474, IC566467 and among the seedlings VKA6, VKA7, VKA8, KMK3 indicated higher phenol and tannin content compared to rest. Molecular characterisation among the selected asoka accessions representing the different districts of Kerala were studied. RAPD analysis was attempted among ten accessions selected. Dendrogram was constituted based on pooled RAPD data. The ten selected accessions were grouped into two major clusters as done in morphological grouping. In grouping at molecular level, the accession IC566488 kept apart as a single group indicating the same trend at its morphological level.The clustering pattern based on molecular characterisation did not follow geographical distribution of accessions.
Open Access
Cross compatibility analysis for production of hybrids in anthurium andreanum linden
(Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, 2010) MadhuKumar, K; KAU; Mayadevi, P
The present study, ‘Cross compatibility analysis for production of hybrids in Anthurium andreanum Linden’ was undertaken to identify suitable parents with commercial qualities and to determine the cross compatibility among the selected parents in anthurium. The present investigation was carried out in the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani during the period 2006-2009. The analysis of variance revealed significant variation among the 40 genotypes for the eighteen characters studied. This reveals the high genetic potential for the improvement in this crop. Variability studies indicated high phenotypic and genotypic coefficients of variation for the characters for anthocyanin content, pollen fertility, leaf size/leaf area, spathe size and spadix length. The high heritability coupled with high genetic advance values were found for characters plant height, leaf size/leaf area, internodal length, spathe size, total anthocyanin content, spadix length, inclination of candle with spathe, number of flowers per spadix, life of spadix, days to initiation of female phase, days to inter phase, duration of male phase, pollen fertility and pollen size. These characters are therefore controlled by additive gene action and amenable to genetic improvement through selection. Pollen fertility estimated using acetocarmine method indicated that most of the genotypes had low fertility values. Liver Red had the highest pollen fertility of 43.01 per cent followed by Lady Jane (36.14 per cent). Pollen emergence was low in the months from March to June, during which the average maximum and minimum temperatures were relatively high. Pollen emergence was highest during October to December months. A study of six qualitative characters such as colour of young leaf and petiole, spathe colour, spathe texture, candle colour and type of inflorescence axis also showed considerable variation among the genotypes studied. Plant height was found to have highly significant positive phenotypic and genotypic correlation with leaf size/leaf area, internodal length, total Anthocyanin content, spadix length, number of flowers per spadix and life of spadix. Number of flowers per spadix had significant positive genotypic correlation with plant height, leaf size/leaf area, internodal length, spathe size, spadix length, total anthocyanin content, life of spadix, pollen fertility and pollen size. Spadix length had significant positive genotypic correlation with plant height, leaf size/leaf area, internodal length, spathe size, number of flowers per spadix, life of spadix and pollen size. Days to initiation of female phase exhibited significant negative correlation with plant height, leaf size/leaf area, spadix length and pollen fertility. The environmental correlations were absent for almost all pairs of characters except for number of leaves/spadices per plant with spathe size. Path analysis revealed that spadix length, plant height, leaf size and life of spadix had high positive direct effect on number of flowers per spadix. Mahalanobis D2 analysis clustered the 40 genotypes into seven clusters. The maximum number of genotypes (17) were included in Cluster I, followed by cluster II (7), cluster III (5), cluster IV (5) and cluster V (4). Clusters VI and VII had one genotype each. Maximum divergence was shown between the Clusters II and VI, while the minimum divergence between clusters I and II. The intracluster distance was highest for the Cluster II. Among the 13 characters considered, life of spadix contributed maximum towards divergence followed by days to initiation of female phase. Grouping of genotypes into different clusters did not reflect the geographical origin of the varieties. Selection index analysis revealed that genotype Liver Red attained the maximum selection index value followed by PR x LR and PR x DT (1) and the minimum estimates were recorded for Rembolina, W x LJ and Corolix. The grouping of genotypes by selection indices followed almost the same pattern as their clustering pattern in the D2 analysis. Intervarietal hybridization was done to analyse the cross compatibility between 15 genotypes based on the percentage of candles bearing fruits, fruit set and seed germination. A total of 127 crosses were attempted based on the availability of receptive spadices and fresh pollen, out of which 80 were found to be successful. In almost all cross combinations the percentage of candles bearing fruits ranged from 50 to 100 per cent. Among the 15 genotypes, the maximum percentage of candles bearing berries was obtained for PR x OG (90.00 per cent) followed by AW (66.67 per cent) and PR x LR (65.00 per cent). The lowest value 5 per cent was obtained for Esmeralda and 11.11 per cent for Dragon’s Tongue. The number of fruits per candle ranged from 5 in Esmeralda x (PR x OG) to 85 in (PR x OG) x (OG x DT). The crosses LR x DT, (OG x DT) x (PR x OG), LR x FR and (PR x OG) x DT also recorded higher number of fruits per candle. The average number of fruits per candles was highest for Liver Red and lowest for Esmeralda. The percentage of fruit set was below 50 per cent for all the crosses. The lowest and highest percentage of fruit set was observed for Ceasor Violet and ‘Liver Red respectively. The berries obtained from different cross combinations took 4.5 to 7 months to mature. Most of the crosses had a high percentage of single seeded berries compared to double seeded berries except for the cross FR x LR. Among single seeded berries maximum seed size was for AW x (PR x OG). In a berry largest seed among the two seeded berries were observed for the crosses AW x (PR x OG) followed by [(PR x LR) x (PR x DT (2)]. The number of days taken for germination varied from four to nine days. The seeds obtained from most of the crosses showed germination except three crosses i.e., DT x LR, (KR x LR) x LJ and Esmeralda x (PR x OG) did not germinate at all. Percentage of germination was lowest in [PR x DT (2)] x LJ and highest in (PR x LR) x C. Seedlings of 57 out of 73 crosses that germinated survived for more than four months. The seedling survival percentage ranged from 38.46 per cent in LJ x (PR x OG) to 81.25 per cent in (KR x LR) x DT. The highest average survival was recorded by Liver Red and lowest was recorded for the genotype Lady Jane. Compatibility parameters estimated based on the performance of the fifteen genotypes as pollen parents. Highest percentage of candles bearing fruits was shown by PR x OG followed by Fla Red, Dragon’s Tongue and PR x LR. Lowest percentage were recorded for Flirt and KR x LR. Number of fruits per candle was the highest for the genotype PR x DT (2) followed by PR x OG, Dragon’s Tongue and Acropolis white. In Lady Jane, Flirt and KR x LR the number of fruits were relatively lower. Higher percentage fruit set was observed for the genotypes Dragon’s Tongue while it was lowest in Lady Jane. Percentage of seed germination was the highest for Fla Red and lowest for Lady Jane. Scoring of the compatibility reactions based on the percentage of fruiting candles, fruit set and seed germination on a scale ranging from zero to nine. After scoring the different genotypes, the crosses with highest compatibility score was for the crosses (PR x OG) x (OG x DT), (PR x LR) x C, FR x DT, AW x (PR x OG), (OG x DT) x (PR x OG), (PR x LR) x (PR x OG),PR x DT (2) x (PR x OG), (OG x DT) x LR, LJ x LR, (PR x OG) x DT, (PR x OG) x LR, (PR x OG) x (PR x LR) and (PR x LR) x FR. So these were the most compatible crosses. The performance of the variety as female parent was found to be different from its performance as male parent. The female parents PR x OG, PR x LR, Lady Jane, OG x DT and PR x DT (2) got the high varietal scores. Among male parents PR x OG, DT, OG x DT, FR and PR x LR got the high varietal scores. So PR x OG, OG x DT and PR x LR could be judged as the best male as well as female parents. Among the seedlings of the successful crosses maximum number of days for maturity of leaves was taken by the cross LJ x (PR x LR) and the least number of days was taken by C x (PR x OG). Number of leaves ranged from 4.50 in DT x FR to 6.33 in [(PR x OG) x (OG x DT)]. The leaf area was maximum for the cross LR x FR and it was minimum for LJ x (PR x OG). The colour of young leaf showed a range from brown to reddish brown to greenish brown to green. The colour of petiole also varied from brown to reddish brown to greenish brown to green.
Open Access
Genetic analysis of yield and leaf curl virus resistance in chilli (Capsicum spp.)
(Department of Plant Breeding, College of Agriculture, Vellayani, 2010) Anandhi, K; KAU; Abdul Khader, K M
Pepper fruits (Capsicum spp.) are among the most consumed vegetables as fresh green or red and dried whole or ground forms in the world for its pungency. Leaf curl virus is an important biotic stress transmitted by the vector, Bemisia tabaci. Controlling the vector can be the only way to manage the disease and results in only partial control of disease. The incidence of disease is more in summer season and makes the cultivation uneconomical. There is an immediate need to develop leaf curl resistant varieties especially for summer cultivation. Hence the present investigation was undertaken at the Department of Plant Breeding and Genetics, College of Agriculture, Vellayani during 2007-2009 with the objective of estimating the combining ability, heterosis and gene action involved in the inheritance of yield and leaf curl virus resistance. Five resistant C. frutescens accessions Mangalapuram Local, Thavanur Local, Kayamkulam Local, Mavelikkara Local and Nenmara Local were crossed in L x T pattern with three high yielding susceptible varieties Jwalamukhi, Jwalasakhi and Vellayani Athulya and 15 F1s were produced and evaluated along with the parents in randomized block design. Analysis of variance revealed highly significant genotypic difference for all the characters studied. The observations recorded were plant height (cm), number of branches, number of days to first flowering, plant spread (cm), duration of flowering (fruiting span), number of fruits per plant, fruit length (cm), fruit width (cm), pedicel - fruit ratio, fruit colour at intermediate stage, green fruit yield per plant (g), average fruit weight (g), number of seeds per fruit, hundred seed weight (g), duration of crop, vector population and virus disease scoring. Higher estimates of PCV and GCV were recorded for vulnerability index and green fruit yield per plant. High heritability coupled with high genetic advance were exhibited for days to first flowering, duration of flowering, number of fruits per plant, green fruit yield per plant, number of seeds per fruit, duration of crop and vulnerability index. Combining ability analysis showed that the line Mavelikkara Local and Nenmara Local were alone good general combiners for fruit yield along with leaf curl resistance. Four hybrids viz., Mavelikkara Local x Jwalasakhi, Nenmara Local x Vellayani Athulya, Kayamkulam Local x Jwalamukhi and Thavanur Local x Jwalamukhi exhibited significant sca effect for fruit yield. Considering per se performance, standard heterosis and sca effect two hybrids Mavelikkara Local x Jwalasakhi and Nenmara Local x Vellayani Athulya were found to be superior. Generation mean analysis was carried out using six-parameter model. Six generations viz. P1, P2, F1, F2, B1, B2 were built up among the crosses Mavelikkara Local x Jwalasakhi and Nenmara Local x Vellayani Athulya. Presence of additive, dominance and epistatic interaction for all the characters indicated that hybridization or recombination-breeding programme can be followed for future breeding.

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