Browsing by Author "Reghunath, B R"

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Agrobacterium rhizogenes mediated genetic ransformation in Koduveli (Plumbago spp. Linn.)
(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2006) Roshna Bhaskar; Reghunath, B R
Investigations on “Agrobacterium rhizogenes mediated genetic transformation in Koduveli (Plumbago spp. L.)” were carried out at the Department of Plant Biotechnology, College of Agriculture, Vellayani during 2003-2006. Two plant species viz., Plumbago zeylanica and Plumbago rosea having great medicinal value by virtue of plumbagin, a naphthoquinone, present in them were selected for the study. Standardisation of rapid in vitro propagation of these medicinal plants was attempted in the present study. Enhanced release of axillary buds from nodal explants, with the highest shoot proliferation of 5.5 was obtained in MS medium supplemented with BA 1.0 mg l-1 + IAA 0.05 mg l-1 for P. zeylanica . In P. rosea the highest shoot proliferation of 5.0 was recorded in MS medium supplemented with BA 1.50 mg l-1 + IAA 0.05 mg l-1. Among the different basal media tested full strength MS medium was found to be the best for shoot proliferation. In indirect organogenesis, the highest callus index (300) was recorded in MS medium with BA 2.00 mgl-1 and 2,4-D 0.50 mg l-1 for P. zeylanica. In P. rosea also maximum callus index (300) was obtained in the same medium. In P. zeylanica BA 1.75 mg l-1, IAA 0.05 mgl-1 and adenine sulphate 20 mg l-1 in MS medium was identified as the best medium for shoot regeneration from leaf derived callus. Whereas in P. rosea BA 2.00 mg l-1, IAA 0.05 mg l-1 and adenine sulphate 20 mg l-1 in MS medium obtained the highest rate of shoot regeneration from callus. Coconut water (100 ml l-1) was found to increase the rate of shoot regeneration in P. rosea. Rooting of in vitro raised shoots was achieved by sub culturing them on MS medium containing IAA 0.50 mg l-1. In vitro root culture was carried out successfully in semi solid MS medium containing 1.50 mg l-1 NAA. Genetic transformation was carried out by co-culture method. Agrobacterium rhizogenes strains A4, MTCC 532 and MTCC 2364 were utilized for transformation. YEP medium was used for culturing the bacteria. The strain A4 was the best for transforming Plumbago spp. There was no significant difference between the two species P. zeylanica and P. rosea in their response to transformation. Nodal explants recorded the highest transformation percentage followed by, leaf segments. Calli and root explants did not respond to transformation. Bacterial density of one (O.D600) during transformation resulted in high transformation percentage. Co-culturing for two days resulted in high transformation percentage, whereas co culturing for more than two days resulted in bacterial over growth on the tissues and low transformation per cent. After two days of co-culture, the bacteria were killed by transferring the tissues to MS medium containing cefotaxime or ampicillin 500 mg l-1. The transformed tissues induced hairy roots in a period of seven to ten days. The hairy roots produced by transformation were negatively geotropic and possessed numerous root hairs. Transformation was confirmed by opine analysis, which was carried out by high voltage paper electrophoresis. Opines were detected only in transformed samples and not in normal roots. The hairy roots induced in P. zeylanica and P. rosea is a potential source for production of plumbagin in vitro.
Collection, description and performance evaluation of herbaceous medicinal leguminous plants of Kerala
(Department of Horticulture, College of Agriculture, Vellayani, 1996) Sunitha, C; Reghunath, B R
The present study titled 'Collection, description and performance evaluation of herbaceous medicinal leguminous plants of Kerala' was carried out at the Department of Horticulture, College of Agriculture, Vellayani from June 1995 to January 1996. Seeds of sixteen different species of herbaceous leguminous medicinal plants could be collected from authentic sources. Twelve different species with high rate of germination were raised and maintained till flowering and pod set in the medicinal plant garden of the Department of Horticulture. Detailed computerised descriptive blank supplemented with line drawings and authentic herbarium were prepared for each species. Cultural trial of eleven selected species of herbaceous leguminous medicinal plants, conducted as pure crop in open condition and as intercrop under shade in a twenty five year old coconut garden, indicated that under open condition the growth and yield are significantly superior in Abrus precatorius, Clitoria ternatea and Atylosia scarabaeoides than under shaded condition. Similarly, in Crotalaria verrucosa and Desmodium triangulare where leaves are the medicinally important plant part, the number of branches and number of leaves were significantly higher under open condition than under shade. The biomass yield of Crotalaria verrucosa was significantly superior under open condition than under shade. This revealed the limited chance of raising these five plant species as intercrops in coconut garden. Growth and yield were significantly superior under shaded condition or were comparable under both conditions in Mucuna prurita, Indigo/era tinctoria, Cassia mimosoides and Pseudarthria viscida. In Desmodium gangeticum, where root is the medicinally important plant pari, no significant difference was noticed in the root characteristics, namely, root length, tap root girth and number of lateral roots under open and shaded conditions. The content of active chemical ingredient in the medicinally important plant parts of the different plant species studied, showed no significant difference under open and shaded conditions. The presence of biochemical substances in these medicinal plants did not affect the association of rhizobia and mycorrhizae. Such positive association in native soil without artificial inoculation opens the scope for utilizing the micro symbiont technology for exploiting the full potential of growth and yield of medicinal plants. The study, thus indicated the feasibility of growing herbaceous, leguminous medicinal plant species as intercrops in coconut garden, with the dual benefit of contributing a part of the requirement of raw materials in the indigenous pharnaceutical industry and at the same time enriching soil fertility by way of nitrogen fixation.
Cost- effective methods and devices for home scale adoption of plant tissue culture
(Department of Horticulture, College of Agriculture, 1996) Deepa, V; Reghunath, B R
Attempts were made to develop cost-effective methods and devices for home scale adoption of plant tissue culture in the plant tissue culture laboratory of the Department of Horticulture, College of Agriculture, Vellayani, during 1993-95. The test plant selected for the study was Anthurium andreanum Lind. (Pink). Segments of leaf were used as explant for cullus initiation. The callus so obtained was used for further regeneration studies. Various low cost equipment were fabricated and tested for their efficiency in comparison to the conventional expensive method. One-fourth strength of the major nutrients of MS medium along with full strength of micro nutrients was found good for the induction of multiple shoots. All the growth parameters were found adversely affected by the use of LR grade chemicals, when compared to AR grade chemicals. Confectionary grade sugar was found to be equally good to AR grade sucrose, while commercial grade crystal sugar was not. Rain water could be used as a substitute to double glass distilled water in the culture medium. Attempts to substitute agar-agar with less expensive playing marbles, as support matrix of the culture medium was also successful. Ordinary (gold-smith type) balance could be used to replace the expensive electronic balance in weighing chemicals for media preparation. The pH indicator paper could be effectively used instead of the pH meter, in adjusting pH of the medium. Ordinary colourless glass bottles and jam jars could be economically used, instead of expensive borosilicate glassware. The domestic pressure cooker was equally efficient as the electric autoclave in sterilising culture medium and containers. The expensive refrigerator could be effectively replaced with ice-packed thermocol boxes. Instead of the laminar airflow cabinet, the fabricated transfer hood could be effectively used. Attempts to substitute artificial flourescent light with natural light were successful. Rooted plantlets when planted out exhibited 30 per cent loss during various stages of hardening. The cost of producing a single anthurium plantlet was Rs. 5.16 in the conventional method, whereas in the cost-effective method it could be brought down to Rs. 1.82.
Effect of physiological pre-conditioning of explants and explant sources of Myristica fragrans Houtt. on Iinvitro culture establishment and growth
(Department of Horticulture, College of Agriculture, Vellayani, 1995) Geetha, S; Reghunath, B R
The present study was conducted to standardise pre – treatments to induce large number of orthotropic shoots and to enhance the in vitro response of explants taken from nutmeg threes by standardising suitable basal media and by incorporating suitable growth hormones. Among the various physical methods of mother tree pre – treatments tried, pruning and pruning followed by etiolation induced four orthotropic shoots each. Pre – treating explant sources (parent trees) with BAP and kinetin each at 500 mg/1 and 1000 mg/1 induced 12 orthotropic shoots per tree. However, the in vitro response of all these newly induced shoots were more or less similar, except those taken from 50 mg/1 and 100 mg/1 of Ga3 infused parent trees. Surface sterilisation of the explants with – 0.1 per cent mercuric chloride for 10 minutes and addition of 0.01 per cent of a Carbendazim based fungicide in the culture medium helped to increase the rate of contamination free cultures. Pre – treating the shoot tips with 2 per cent sugar and 0.7 per cent PVP gave significantly superior results in eliminating phenolic interference. Pre – treating the shoot tip explants with CH at 500 mg/1 for 10 minutes gave significantly superior results in the culture establishment of nutmeg explants. The results also revealed the significantly superior influence of seedling explants over mature tree explants. Among the various basal media tried, AM at half strength of major nutrients and full strength of minor nutrients was found to be the best with a hormonal combination of BAP, NAA and 2, 4 – Dat 2 mg/1, 1 mg/1 and 0.5 mg/1 respectively, giving 67 per cent culture establishment. Phloroglucinol (40 mg/1) in combination with IBA (2 mg/1)gave significantly superior results in the induction of roots in established shoot tip cultures.
Evaluation of genetic stock of mucuna pruriens baker non DC. for yield, L-DOPA content and nitrogen fixing potential in coconut garden
(Department of Horticulture, College of Agriculture, Vellayani, 2000) Ann Samuel; Reghunath, B R
The present study titled "Evaluation of genetic stock of Mucuna pruriens Baker non DC. for yield, L-DOPA content and nitrogen fixing potential in coconut garden" was carried out at the Department of Plantation Crops and Spices, College of Agriculture, Vellayani from May 1999 to January 2000. Seeds of twenty five different accessions were collected from inside and outside the state. They were subjected to seed characterization and seed germination studies. Based on the germination rate, ten accessrons with high germination percentage were selected. These were raised under coconut shade. Various biometric observations were taken at four different stages, viz., pre-flowering, flowering, seeding and seed maturation state. These observations included biomass yield, root yield and seed yield. Phytochemical analysis was carried after crop harvest to estimate the quantity of L-DOPA present in the seeds of each accession. Soil tests were carried out to determine the amount of nitrogen fixed in the soil by the accessions. 2 After analysis of the results it was evident that the accessions MP-62, MP-57, MP-44, MP-58, MP-46 and MP-45 had superior biomass yields compared to the others. The accessions showing superior seed yield were MP-60, MP-62, MP-58, MP-67, MP-57 and MP-66. The content of soil nitrogen fixed varied significantly among the plots of individual accessions. The accessions MP-62, MP-57, MP-67, MP-44, MP-58 and MP-60 were observed to have fixed higher amounts of nitrogen. After phytochemical tests, the accessions with significantly superior L-DOPA content (ie., greater than two per cent) were MP-60, MP-57, MP-46 and MP-66. From the study conducted, the most promising accessions, ie., those having superior biomass, root and seed yield, as well as superior L-DOPA content and nitrogen-fixation potential could be identified. They are MP-62, MP-60, MP-58, MP-57, MP-44 and MP-67. It can be reiterated that growing such accessions of this plant . species would be of much benefit to the cultivator enabling him to utilize existing areas under cultivation, enrich its soil fertility status as well as provide him with an additional source of income by way of selling seeds; the medicinally valuable plant part.
Evaluation of genetic stock of sanghupushpam (Clitoria ternatea L.) for yield, Alkaloid content and nitrogen fixing potential
(Department of Horticulture, College of Agriculture, Vellayani, 2000) Deepa S Nair; Reghunath, B R
The present study entitled “Evaluation of genetic stock of ‘Sanghupushpam’ (Clitoria ternatea L.) for yield, alkaloid content and nitrogen fixing potential” was carried out at the Department of Horticulture, College of Agriculture, Vellayani from June 1999 to January 2000. Seeds of twenty different accessions were collected from various locations from inside and outside the state. Thirteen accessions having high rate of seed germination were raised as intercrop in young coconut garden and maintained till seed maturation stage. The performance of the accessions in terms of growth, yield and physiological parameters were evaluated. Growth and yield parameters with respect to shoot, pod, root and root nodule characters were evaluated. Physiological parameters evaluated included leaf area index, leaf area duration, net assimilation rate, crop growth rate, relative growth rate, absolute growth rate and harvest index. Number of effective nodules was taken as an index for assessing nitrogen fixing potential of the plant. Leaf yield, shoot yield and root yield were significantly superior in the accession MP-90. MP-83 recorded significantly superior pod yield. The number of nodules was the highest in accessions MP-76 and MP-82. Crude alkaloid content was significantly superior in seeds of MP-74 and MP-76. Six accessions were selected based on yield , nodule characters and crude alkaloid content viz ., MP-90, MP-74, MP-83, MP-78, MP-76 and MP-82. Results of the present study indicated that Clitoria ternatea, is not a good proposition as an intercrop in young coconut garden. However, it may be worth studying the performance of the crop as a pure crop under open condition or as an intercrop in coconut gcirden with comparatively lesser shade (less than 50 per cent) than the situation of the present study.
Evaluation of neelayamari(indigofera tinctoria L.) for yield and glysocide content under open and shade conditions
(Department of Plantation Crops and Spices, College of Agriculture , Vellayani, 2004) Sarada, S; Reghunath, B R
Evaluation of selected sankhupushpam (Clitoria ternatea L.) lines for yield, alkaloid content and nodulation
(Department of Plantation Crops and Spices, College of Agriculture, Vellayani, 2001) Reshmi, C R; Reghunath, B R
The present study entitled 'Evaluation of selected Sankhupushpam (Clitoria ternatea L.) lines for yield, alkaloid content and nodulation' was carried out at the Department of Plantation Crops and Spices, College of Agriculture, Vellayani from June to December 2000. Six superior lines evolved from a preliminary screening of genetic stock conducted by Nair (2000) were raised under shade, as an intercrop in coconut garden and in open as a pure crop. Various biometric observations were taken at four different stages of plant growth, viz., pre-flowering (45 DAS), flowering (90 DAS), seed formation (135 DAS) and seed maturation (180 DAS) stages. These included observations on shoot yield, root yield and pod yield. Nodule characteristics were also recorded. Phytochemical analysis was carried out after harvest of the crop to estimate the crude alkaloid content in the seeds of each accession. Analysis of the results revealed maximum shoot yield for MP-73 and MP-8I under shade and open conditions respectively. The highest pod yield was recorded for MP-76 under shade and for MP-73 under open condition. Root yield on fresh weight basis was found to be the highest for MP-76 under shade and MP-74 under open condition. On dry weight basis, root yield was maximum for MP-74 under shade and for MP-8I under open condition. Maximum nodulation at flowering stage was observed in MP-73 and MP-7& under shade and open conditions respectively. MP-73 and MP-76 recorded higher crude alkaloid content in seeds under shade condition. In open, maximum crude alkaloid was obtained in MP-73. Both yield and nodulation were found to be higher under open condition whereas the crude alkaloid content showed no significant variation between shade and open conditions. From the study, the accessions MP-73 (Thiruvattar White) and MP-76 (Vattiyoorkavu White) were found to show superior performance under both conditions.
Genetic variability analysis in Indian indigo(indigoferatinctoria L.)
(Department of Plantation Crops and Spices, College of Agriculture,Vellayani, 2004) Neema, M; Reghunath, B R
Hairy caterpillar euchromia polymena Linn.- A serious pest of Tylophora Indica (Burm.F.) Merr. and Ipomoea Mauritiana Jacq.
(Kerala Agricultural University, 1992) Lyla, K R; Reghunath, B R; Joy, P J
In vitro production of microrhizomes in Curcuma aromatica Salisb
(Department of plantation crops and spices,College of Agriculture,, 2012) Shameena, S; Reghunath, B R
In vitro studies on the propagation of cardamom (Elettaria cardamomum maton)
(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 1989) Reghunath, B R; Gopalakrishnan, P K
Lily caterpillar polytela gloriosae fab.(Noctuidae : Lepidoptera), a serious pest of gloriosa superba L.
(Kerala Agricultural University, 1992) Lyla, K R; Reghunath, B R; Viswanathan, T V
New leaf blight disease of Catharanthus Roseus(L.) G. Don (L)
(Kerala Agricultural University, 1981) Padmakumary, G; Suharban, M; Nair, M C; Reghunath, B R; Sreekantan Nair, G
Quantification of medicinally valuable weeds in oil palm plantations of Kerala
(Kerala Agricultural University, Vellanikara, 2002) Sarada, S; Sreekandan Nair, G; Reghunath, B R
The medicinally valuable weeds naturally growing in young, medium and mature plantations of oil palm were identified and quantified by stratified random sampling technique. Quantification of plants was done in comparison with that in the open condition. Chrysopogon aciculatus Trin. dominates in all the four strata. Naregamia alata Wight & Arn. is another dominant species in mature plantation. Hemidesmus indicus R. Br. occurs frequently in all the four strata. Another frequently occurring species in young oil palm plantation is Elephantopus scaber Linn, and in medium plantations is Chromolaena odorata King & Robinson. Phyllanthus amarus Schum.& Thonn. is very frequent in medium plantation and open condition. Holostemma adakodien Schult occurs frequently in mature plantation. Young oil palm plantation and open condition were found to be the most similar strata whereas mature plantation and open condition were the most dissimilar strata in vegetation pair-wise analysis. Medium oil palm plantation was found to have high concentration of dominance. Species diversity was the highest in mature oil palm plantation. Species evenness was maximum in open condition.
Rapid propogation and conservation of selected leguminous medicinal plants using in vitro techniques
(Department of Plantation Crops and Spices,College of Agriculture, Vellayani, 2004) Deepa Nair, S; Reghunath, B R
Synseed production, in vitro conservation and plant conversion in banana
(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2017) Nazrin Nizar; Reghunath, B R