Medicinal plant biotechnology.

Biotechnology is described as methodology to enhance the formation and accumulation of desirable natural products and possible product modification in medicinal plants. Micropropagation, cell and hairy root culture as well as gene technology are being highlighted.

Enhancing the sensitivity of DNA detection and recovery from agarose gels.

By inserting nitrocellulose strips into agarose gels alongside the electrophoresed lanes and passing an electric current perpendicularly in the direction of the strips, highly efficient transfer of DNA bands onto the membrane in the form of concentrated dots is achieved. DNA detection limits by this technique are enhanced, at least three times as visualized by ethidium bromide fluorescence and at least twice more by radiolabeling.

Alkaloid formation by habituated and tumorous cell suspension cultures of Catharanthus roseus.

Habituated and tumorous Catharanthus roseus cells grown in the absence of hormones accumulated indole alkaloids. Total alkaloids and alkaloid pattern were the same when cells were cultured in medium without hormones or in alkaloid production medium with and without indole acetic acid. Treatment of cells with Pythium homogenate as elicitor did not increase total alkaloids or change the pattern of alkaloids produced.

Elicitor-mediated induction of tryptophan decarboxylase and strictosidine synthase activities in cell suspension cultures of Catharanthus roseus.

Treatment of one cell line (No. 615) of Catharanthus roseus c.v.

Plant Regeneration with Callus and Protoplasts of Solanum uporo Dun.

Vigorous plants exceding 1 m in height and producing fruits with seeds have been regenerated from callus derived from hypocotyl segments in less than one year, using B5-media containing 0.5 mg · l(-1) 2,4-D, 0.25 mg · l(-1) kinetin, and 0.

Liposomes as vehicles for transferring low molecular weight compounds into periwinkle protoplasts.

The uptake of liposomal contents into Catharanthus roseus protoplasts has been quantitated. Hi.phest uptake was obtained from positively charged vesicles (1.

Cryopreservation of Alkaloid-Producing Cell Cultures of Periwinkle (Catharanthus roseus).

A procedure for cryogenic storage of alkaloid producing cell lines of periwinkle, Catharanthus roseus (L.) G. Don.

Freezing Characteristics of Cultured Catharanthus roseus (L). G. Don Cells Treated with Dimethylsulfoxide and Sorbitol in Relation to Cryopreservation.

The freezing behavior of dimethylsulfoxide (DMSO) and sorbitol solutions and periwinkle (Catharanthus roseus) cells treated with DMSO and sorbitol alone and in combination was examined by nuclear magnetic resonance and differential thermal analysis. Incorporation of DMSO or sorbitol into the liquid growth medium had a significant effect in the temperature range for initiation to completion of ice crystallization. Compared to the control, less water crystallized at temperatures below -30 degrees C in DMSO-treated cells.

Quantitation of the delivery of liposome contents into plant protoplasts.

A procedure for the quantitation of the delivery of liposome contents into Catharanthus roseus protoplasts has been developed. The method is based on the uptake of liposome encapsulated methylumbelliferyl beta-D-glucoside and its enzymatic hydrolysis to yield fluorescent methylumbelliferone. Since the free glucoside is not taken up by the protoplasts to a significant extent, the delivery of material in the nanomole range can be measured with ease.

Organelles associated with the plasma membrane of tobacco leaf protoplasts.

Leaf protoplasts of tobacco (Nicotiana tabacum) heterozygous (Su/su) and normal (su/su) for the sulfur mutation exhibit a characteristic lag period before initiating cell wall formation. The early wall is composed of a network of Calcofluor positive fibrils. Large fragments of plasma membrane from freshly isolated protoplasts were examined by electron microscopy to determine the distribution of associated organelles.

Alkaloid production in Catharanthus roseus cell cultures : XII. Biosynthetic capacity of callus from original explants and regenerated shoots.

Callus derived from hypocotyls of periwinkle, Catharanthus roseus, responded to culture on nutrient media supplementedwith IAA, BA, and zeatin with shoot formation at low frequencies. However, shoot regenerating callus could be very successfully propagated and subcultured. Alkaloid profiles of callus derived from the original explants (hypocotyls) as well as callus derived from regenerated shoots were almost identical.

Cryopreservation of periwinkle, Catharanthus roseus cells cultured in vitro.

Unlabelled: A procedure for prolonged cryogenic storage of periwinkle cell cultures is described. Cells derived from periwinkle, Catharanthus roseus (L.) G.

Efficient incorporation of 10-hydroxygeraniol and 10-hydroxynerol into indole alkaloids by a cell suspension culture of Catharanthus roseus.

A cell suspension culture of Catharanthus roseus was evaluated with regard to its potential usage in studies of early stage indole alkaloid biosynthesis. Employing this material, 1,1,7-trideutero-10-hydroxygeraniol and 1,1,7-trideutero-10-hydroxynerol were incorporated into the alkaloids ajmalicine and strictosidine lactam in approximate proportions of 50% and 80% respectively.

Alkaloids of Vinca major cv. Variegata.

Analysis of the leaves of Vinca major L. cv. Variegata led to the isolation of seven alkaloids: reserpinine, majdine, akuammicine, strictosidine lactam, pseudoakuammigine, akuammine, and a new alkaloid tentatively assigned as 10-hydroxycathofoline.

Alkaloid production in Catharanthus roseus (L.) G. Don. : VI. Variation in alkaloid spectra of cell lines derived from one single leaf.

Analysis of 76 cell clones derived from one leaf of a periwinkle plant (Catharanthus roseus (L.) G. Don) showed the occurrence of Corynanthe-, Strychnos-, and Aspidosperma-type alkaloids.

Alkaloid Production in Catharanthus roseus cell cultures VIII.

A cell line of Catharanthus roseus (L.) G. Don coded PRL # 200, was characterized with respect to its biosynthetic capabilities for indolealkaloids, in particular catharanthine, in suspension cultures.

Fine structure of fusion products from soybean cell culture and pea leaf protoplasts.

Protoplasts from pea (Pisum sativum L.) leaves and cultured soybean (Glycine max L.) cells were fused by means of polyethylene glycol and subsequently cultured for one week.

Somatic hybridization in higher plants.

Somatic hybridization in higher plants has come into focus since methods have been established for protoplast fusion and uptake of foreign DNA and organelles by protoplasts. Polyethylene glycol (PEG) was an effective agent for inducing fusion. Treatment of protoplasts with PEG resulted in 5 to 30% heterospecific fusion products.

Ultrastructure of fusion products from soybean cell culture and sweet clover leaf protoplasts.

Protoplasts from cultured cells of soybean (Glycine max L.) and from sweet clover (Melilotus officinalis L.) mesophyll cells were fused with polyethylene glycol and subsequently cultured for six days.

Electron-microscope observations of mitosis and cytokinesis in multinucleate protoplasts of soybean.

Multinucleate soybean protoplasts produced by spontaneous fusion during enzyme digestion of the cell wall initiated cell division after approximately 40 h in culture. The structure of these protoplasts during mitosis and cytokinesis was studied with both light and electron microscopes. Most nuclei did not fuse but divided synchronously.