1. KAUTIR (Kerala Agricultural University Theses Information and Retrieval)
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Item Cost- effective methods and devices for home scale adoption of plant tissue culture(Department of Horticulture, College of Agriculture, 1996) Deepa, V; Reghunath, B RAttempts 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.Item Improvement of propogation efficiency of anthurium species in Vitro(Department of Horticulture, College of Agriculture, Vellayani, 1992) Sreelatha, U; Ramachandran Nair, SAttempts were made, to improve the propagation efficiency of Anthurium species through enhanced release of axillary buds and callus-mediated somatic organogenesis/embryogenesis, in the plant tissue culture laboratory of the Department of Horticulture, College of Agriculture, Vellayani during 1990-92. Four species of Anthurium namely, A. andreanum, A. crystallinum, A. veitchii and A. grande were selected for the study. Shoot tips from in vitro grown seedling were used as explants for the enhanced release of axillary buds. Cent percent survival was observed in all the cytokinin treatments. The maximum number of shoots (4.50) was observed with kinetin 2.0 mg/1 as well as BA 1.0 mg/1. Treatments with kinetin was free of callus growth. In treatments with BA and 2ip, callus growth was observed at the base of the explant. Treatments with Ms inorganic salts as well as sucrose did not influence multiple shoot formation. One fourth strength of MS major rutrients with full strength of micro nutrients was ideal for multiple shoot induction. Glucose produced less number of shoots than sucrose. One percent sucrose did not influence multiple shoot induction. The longest shoot (0.95cm) was observed at 0.4 percent agar. Light was necessary for the enhancement of axillary buds. In darkness, callus growth was observed, from which many adventitious shoots were produced. Segments of leaf, petiole, spathe, spike and inflorescence stalk were used a explants for callus initiation. Combinations of 2, 4-D and BA were efficient in initiating callus. In A.andreanum, 2, 4-D 0.08 mg/1 and BA 1.0 mg/1 was ideal for callus initiation. Combination of 2, 4-D, 0.2 mg/1 and BA 1.0 mg/1 was the best for callus initiation in A. veitchii. In A. grande, the best callus initiation was observed with 2, 4-D 0.5 mg/1 and BA 1.0 mg/1. Modified MS medium with reduced salt concentrations was ideal for callus initiation in all the species. Inositol when reduced to half concentration (of the normal) influenced callus initiation. The leaf explant (with the smallest vascular bundles) among the other explants, had the highest number of cultures free of microbial contamination. Basal portions of leaf responded, better than the apical portions, to in vitro culture. Continuous darkness was necessary for callus initiation and growth. MS medium with ¼ strength major nutrients was ideal for callus multiplication. Attempts,made on callus-mediated somatic embryogenesis, were not successful. Shoot regeneration and growth of the shoots were the best in MS medium with BA 0.5 mg/1 and IAA 2.0 mg/1. No rooting treatments were required as the shoots rooted spontaneously. Plantlets survived, better than micro shoots, exvitro. The plantlets required less hardening treatments. Sand was the best potting medium for planting out. Nutrient solutions when used for the irrigation the plantlets, had a negative influence on the survival of plantlets. Treatments with VAM (Glomus constrictum and G. etunicatum) was beneficial for the survival as well as growth of the plantlets. Cytological examinations of the root tip squashes made on random number of plantlets, at planting out, showed a normal diploid chromosome count. Attempts, to correlate the biochemical properties with in vitro response, of different explants as well as species, were not successful. Based on the existing facilities of the plant tissue culture laboratory of the department of Horticulture, College of Agriculture, Vellayani, the cost of single anthurium plantlet was worked out to be Rs.3.00/=.Item Development of protocol for tissue culture of papaya (Carica papaya L) from mature explants(Department of pomology and floriculture, College of agriculture, Vellayani, 2012) Hutke Vikram Rajendra; Jayachandran Nair, C SItem Utilisation of in vitro cultures of Tinospora cordifolia Miers (chittamrithu) for berberine(Department of Plantation Crops, College of Horticulture, Vellanikkara, 2002) Kalimuthu, M; Asha Sankar, MThe present investigation on "Utilisation of in vitro cultures of Tinospora cordifolia Miers. (Chittamrithu) for berberine" was carried out in the Plant Tissue Culture and Biochemistry Laboratories, College of Horticulture, Vellanikkara, Thrissur during the period 1999-2001. The study was undertaken with the objective to standardise the in vitro techniques for initiation and proliferation of static and suspension cultures of T cordifolia and to screen the in vitro cultures for synthesis of berberine and quantify it. It was also envisaged to enhance the level of product synthesis in in vitro cultures. Leaf, petiole and stem derived callus cultures of Vellanikkara and Madurai ecotypes were established in vitro. Surface sterilisation with mercuric chloride (HgCh) at 0.1 per cent for 8 min was most effective in all the explants. MS medium at full strength supplemented with NAA at 4 mg r ' was observed ideal for initiation and proliferation of calli. Kinetin at 3 mg r' and BA at 4 mg r' enhanced the callus inducing property of NAA. Both the ecotypes responded equally for most of the parameters observed, with respect to callusing. The auxin synergist, phloroglucinol at levels of 100.0 mg r' and 125 mg r' and casein hyrdolysate at 100, 200 and 300 mg r' registered favourable influence on callusing. Incubating leaf and stem cultures under illuminated conditions at 26±1 QC was significantly superior to incubation in dark. Successful regeneration of roots from leaf and stem calli of the experimental ecotypes was achieved on MS medium at half strength supplemented with NAA or lAA each at 2 mg r'. Calli derived from Madurai ecotype performed better with respect to root regeneration. None of the treatments tried resulted in a positive response with respect to shoot initiation from callus cultures of both the ecotypes. Substituting sucrose with lactose in proportions of 2: 1 in MS medium at full strength fortified with NAA at 1 mg r ' and Kin at 4 mg r' initiated embryoids in both the ecotypes. MS media at full strength supplemented with NAA and BA or NAA and Kin each at 2 mg r', was standardised as the production medium, which recorded maximum berberine synthesis. Butanol-glacial acetic acid-water at 7:1:2 was identified as the appropriate solvent system for detecting the alkaloid with Dragendorff's reagent as the localizing spray. Substituting sucrose with lactose maintaining a proportion of 2: 1 and reducing the phosphorus level in basal medium to half the original strength resulted in increased levels of berberine synthesis. The precursor phenyl alanine at 100, 150 and 200 mg r' elicited synthesis of berberine. Addition of osmoregulants, polyethylene glycol at 2.0 and 3.0 per cent and mannitol at 1.5 per cent exerted a favourable influence on synthesis of berberine in Tinospora. Incorporation of autoclaved mycelia of Pythium aphanidermatum at 0.5, 1.0 and 1.5 g r' and immobilisation of calli with sodium alginate-calcium chloride complex revealed a positive influence on synthesis of berberine. Liquid suspensions of Vellanikkara and Madurai ecotypes registered 0.92 and 0.87 per cent of packed cell volume. Based on critical cell density, the liquid suspension were subcultured at 16 days interval. As compared to static cultures, suspensions synthesized lesser quantity of berberine. Berberine was detected only in stem extracts of ex vitro plants. When compared to ex vitro samples, in vitro cultures yielded higher quantities of berberine. The highest berberine yield (23.176 ug/g of callus) was obtained from stem cultures maintained in solid MS media supplemented with NAA 2 mg r' + BA 2 mg r ' and autoclaved mycelia of P. aphanidermatum at 0.5 g r'. Madurai ecotype performed better with respect to berberine synthesis with a mean value of 17.565 ug of berberine/gram of callus whereas Vellanikkara ecotype synthesized 16.051 ug/g of callus under positively responding treatments.