Browsing by Author "Asha Sankar, M"

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Evaluation of seedlings and clonal progenies of kattupatavalam (trichosanthes cucumerina l) for yiels and quality
(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2007) Sreerekha, M V; Asha Sankar, M
A study entitled “Evaluation of seedlings and clonal progenies of Kattupatavalam (Trichosanthes cucumerina L.) for yield and quality” was carried out at the Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, during 2005-2006, to evaluate the seedlings and clonal progenies of Kattupatavalam for growth and yield in open and under shade. The study also aimed at assessing the quality attributes of seedlings and clonal progenies, grown under shade and in the open by comparison with the drug source in the user industry. Results of the study revealed that shade and open conditions had a significant influence on various parameters of growth, yield and quality attributes of seedlings and clonal progenies. Seedlings and clonal progenies also showed significant difference in some growth parameters as well as in yield and quality attributes. Considering main vine length and primary branches per plant, seedlings were superior as compared to clonal progenies. Growing in the open, conferred earliness in flowering, wherein the species registered 66.53 days for flowering. Between subplot treatments, earlier flowering was recorded for clonal progenies as compared to seedlings. Plants grown in the open attained fruit maturity earlier as compared to plants grown under shade. But seedlings and clonal progenies recorded values on par with each other. Highest mean fruit number was recorded for plants grown under shade as compared to plants in the open. Clonal progenies recorded 45.47 mean number of fruits, which was superior to fruit number per plant in seedlings. No significant difference was observed in fruit yield per plant, in plants grown under shade as well as in the open. Similar trend was exhibited by seedlings and clonal progenies as well. Total fresh yield and total dry yield of whole plant, did not differ significantly under shade and in the open. It was also revealed that, seedlings and clonal progenies registered values on par with each other, on the above two yield parameters. Shorter crop duration was recorded in plants grown under shade (111.07 days) as compared to plants in the open. Clonal progenies recorded shorter crop duration as compared to seedlings. Fresh weight of soxhlet extractables was highest for plants grown under shade (0.312g). Clonal progenies recorded 0.358 g of soxhlet extractables, which was superior to that of seedlings. It was found that, shade and open conditions had no influence on the fresh weight of hot water extractables. Considering subplot treatments, clonal progenies recorded higher mean weight of hot water extractables (0.929). Market samples yielded 0.718g of hot water extractables and 0.213 g of soxhlet extractables. Tissue culture derived plants grown under shade, registered cucurbitacin of low molecular weight, while high molecular weight cucurbitacin was recorded in seedlings grown in the open. Intermediate types of cucurbitacin were expressed by clonal progenies in the open, seedlings under shade and market samples. Cucurbitacin content was more for clonal progenies grown under shade, while least quantity was observed in seedlings grown in the open. Market sample yielded five units of cucurbitacin, which was superior to cucurbitacin content of seedlings grown in open. On further biochemical analyses, it was revealed that, other secondary metabolites like alkaloids, phenols, tannins and saponins were present in domesticated samples as well as in market samples. Quantification of terpene revealed that tissue culture derived plants grown under shade registered higher amount of terpenes (13 units) than others. With regard to anatomical studies of experimental samples, no difference in anatomy of stem, root and leaf sections of domesticated samples and market samples was noted. The study conclusively proved that, Trichosanthes cucumerina can be ideally raised as a profitable intercrop in the coconut gardens of Kerala and that performance of tissue culture derived plantlets was comparable to seedlings. Also, market sample did not reveal presence of adulterants, as indicated by qualitative as well as anatomical investigations.
Exploitation of invitro cultures of Indian madder (Rubia cordifolia.Linn) for anticancerous compounds
(Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2009) Labade Dinesh Sitaram; Asha Sankar, M
The present investigation on “Exploitation of in vitro cultures of Indian Madder (Rubia cordifolia L.) for anticancerous compounds” was carried out at the Plant Tissue Culture Laboratory of the Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara and Amala Cancer Research Centre, Thrissur during the period 2006-2008. The study was undertaken with the objective to standardize the in vitro techniques for initiation and proliferation of static and suspension cultures of Rubia cordifolia and to screen the in vitro cultures for synthesis of naphthoquinone and quantify it. It was also envisaged to enhance the level of product synthesis in in vitro cultures and to assess the anticancerous activity of in vitro and in vivo extracts in terms of cytotoxicity, antioxidant and prooxidant activities in vitro. Leaf, nodal and root derived callus cultures of Rubia cordifolia were established in vitro. Explants were pre treated with the fungicide, Bavistin 2.5 per cent for 15 minutes. Surface sterilization with mercuric chloride (HgCl2) at 0.1 per cent for 1 min and 30 sec was effective for yielding healthy, contamination free cultures from nodal segments and leaves, respectively. MS medium at full strength, supplemented with NAA at 2 mg l-1 along with BA at 0.5 mg l-1 was observed ideal for initiation and proliferation of calli. The auxin synergist phloroglucinol, when supplemented to the medium, did not not yield encouraging results, with respect to callusing in the experimental species. Root derived cultures were inferior with respect to callus initiation and proliferation, registering low values for all the parameters studied. Incubating in vitro cultures under illuminated condition at 26 ± 2 C was superior to dark incubation, with respect to callus initiation and proliferation. Chloroform – methanol at 8.5 :1.5 ratio was indentified as the appropriate solvent system for detection of naphthoquinone on thin layer chromatograms in the test extracts of the experimental species, with alcoholic KOH (10 per cent) as the spray reagent. Ms medium at full strength, fortified with NAA and BA at 2.0 mg l-1 and 0.5 mg l-1 respectively, which recorded maximum naphthoquinone synthesis, was standardized as the production medium. Enhancing concentration of sucrose to 5 per cent in the production medium, did not elicit a positive response on naphthoquinone production in vitro. Reducing nitrate concentration of the production medium, to half and one fourth the original concentration, resulted in enhanced in vitro synthesis of the target compound. Supplementing the production medium with yeast extract (1 per cent and 2 per cent) as well as precursor feeding with phenyl alanine and tyrosine each at levels of 50 mg l-1, 100 mg l-1 and 150 mg l-1 exerted a favourable influence on synthesis of naphthoquonines, in vitro. Incubation in dark resulted in marginal increase in in vitro production of naphthoquinones. Incorporation of autoclaved mycelia of Pythium aphanidermeatum at levels of 2.0 per cent and 5.0 per cent resulted in enhanced in vitro production of naphthoquinone. The abiotic elicitor, salicylic acid at concentration of 10 μM and 100 μM resulted in maximum synthesis of naphthoquinones in in vitro root cultures (8.76 units g -1 calli) of Rubia cordifolia. Immobilization of test calli with sodium alginate – calcium chloride complex as well as subjecting the in vitro cultures to stress conditions, as imposed by sorbitol failed to bring about an enhancement in the in vitro production of naphthoquinones. None of the explants employed in the study induced hairy roots, when co- cultured with the Agrobacterium rhizogenes strains, MTCC 2364 and MTCC 532. Based on cell count, subculturing intervals of leafs, nodal and root derived suspension were fixed as 24, 27 and 27 days respectively with the respective packed cell volume as 0.93 per cent, 0.83 per cent and 0.80 per cent. Naphthoquinone was detected, in ex vitro and in vitro test extracts at all levels of maturity tested. Both ex vitro and in vitro root extracts exihibited maximum cytotoxicity, as revealed by the percentage of cell death on DLA and EAC cell lines as well as their IC50 values. As compared to whole plant extract, in vitro systems of the experimental species exhibited least antioxidant action. Extent of pro-oxidant activity was higher in in vitro root extract of the experimental species.
Hydration-dehydration technique, a mid-storage correction to prolong viability of rice seeds
(Kerala Agricultural University, 1996) Girija, T; Sukumara Dev, V P; Asha Sankar, M; Rajappan Nair, N
In vivo and in vitro screening of sida spp. for ephedrine content
(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 1998) Asha Sankar, M; Sreekandan Nair, G
Standardization of good agricultural practices (GAP) in Kacholam (Kaempferia galanga L) for yield and quality
(Department of Plantation crops and spices, College of Horticulture, Vellanikkara, 2011) Chandana, R; Asha Sankar, M
Survey, collection and characterization of 'Kizharnelli' (Phyllanthus spp.) of Kerala
(Department of plantation crops and spices, College of horticulture, Vellanikkara, 2015) Shafna Kalarikkal; Asha Sankar, M
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, M
The 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.