Sabita Bhattacharya

In vitro multiplication of Coleus forskohlii Briq. : An approach towards shortening the protocol

SummaryA protocol has been developed that leads to the development of complete plantlets of Coleus forskohlii within 35–40 d by culturing stem tip explants in MS medium containing 0.57 μM indole-3-acetic acid and 0.46 μM kinetin through direct multiplication at the rate of 12.5 shoots per explant. About 100% shoots rooted and micropropagated plants were successfully established in soil after hardening with a high survival rate. The significance of the present micropropagation protocol of C. forskohlii is the formulation of growth regulators which effected very fast multiplication of the plant (time reduced to one-third of the hitherto known methods).

DEVELOPMENT OF A POTENT IN VITRO SOURCE OF PHYLLANTHUS AMARUS ROOTS WITH PRONOUNCED ACTIVITY AGAINST SURFACE ANTIGEN OF THE HEPATITIS B VIRUS

SummaryIn vitro culture of hairy roots of Phyllanthus amarus induced by Agrobacterium rhizogenes was established. Their growth and ability for in vitro inactivation of hepatitis B virus surface antigen was studied and compared with adventitious roots grown in vitro. The selected hairy root clone HR-1 was capable of growing at a very fast rate, and an approximately 900-fold increase in weight of root biomass was achieved after 4 wk of culture in hormone-free quarter-strength liquid Murashige and Skoog medium with continuous agitation. Non-transformed roots cultured in the presence of 1.0 mg l−1 (5.71 μM) indole-3-acetic acid increased by 330-fold. The immuno-inactive property of roots was maximal in the crude extract. The hairy roots were shown to possess 85% inhibition (in contrast to 15% in the control) in binding of hepatitis B surface antigen (HBsAg) to its antibody (anti-HBs) after 24 h of incubation with HbsAg-positive sera in vitro at 37°C. Out of three fractions selected on the basis of molecular weight components of the extract, the Fraction III containing comparatively lower molecular weight substances (≤3500) yielded the highest activity. The extract from non-transformed roots was found to possess similar efficiency (87% inhibition). The levels of activity in both types of in vitro-raised roots were higher than those of naturally occurring roots and leafy shoots. The ability of P. amarus hairy root cultures to yield high biomass with the anti-viral property at high levels may provide an alternative source of raw material for more detailed study in the field of pharmaceutical research.

In Vitro propagation of Jasminum officinale L.: a woody ornamental vine yielding aromatic oil from flowers.

The growing demand for flower extracts in perfume trade can primarily be met by increasing flower production and multiplying planting material. The major commercial aromatic flower yielding plants including Jasminum officinale L., a member of the Family Oleaceae have drawn the attention of a large section of the concerned sectors leading to a thrust upon developing advanced propagation technologies for these floral crops, in addition to conventional nature-dependent agro-techniques. This chapter describes concisely and critically, a protocol developed for in vitro propagation of Jasminum officinale by shoot regeneration from existing as well as newly developed adventitious axillary buds via proper phytohormonal stimulation. To start with nodal segments as explants, March-April is the most ideal time of the year when planting material suitable for in vitro multiplication is abundantly available. Prior to inoculation of explants in the culture medium, special care is needed to reduce microbial contamination by spraying on selected spots of the donor plant with anti-microbial agents 24 h prior to collection; treatment with antiseptic solution after final cleaning and surface sterilization by treating explants with mercuric chloride. Inoculated explants are free from brown leaching from cut ends by two consecutive subcultures within 48 h in MS basal medium. Multiplication of shoots, average 4-5 at each node, takes place in MS medium containing 4.0 mg/L BAP, 0.1 mg/L NAA, and 40 g/L sucrose over a period of 8 weeks. For elongation of regenerated shoots, cultures are transferred to MS medium, supplemented with a single growth hormone, kinetin at 2.0 mg/L. Emergence and elongation of roots from shoot base is facilitated by placing on the notch of a filter paper bridge. The hardened in vitro propagated plants are able to grow normally in soil like other conventionally propagated Jasminum officinale.

Clonal propagation of Zephyranthes grandiflora using bulbs as explants

Zephyr lily (Zephyranthes grandiflora), an important ornamental plant has been micropropagated in vitro after controlling microbial contamination by a pretreatment with 0.2 % Bavistin and 0.1 % Pantomycin for 4 h before final sterilization with 0.1 % mercuric chloride. In 67 % of the sterile cultures, 11 shoots on average were regenerated directly from basal half of bulb scales in Murashige and Skoog (MS) medium containing 3 % sucrose and 2 mg dm−3 benzylaminopurine (BAP). Shoots emerged in bunches on a basal achlorophyllous bulbous part. Combination of 2 mg dm−3 BAP with 1 mg dm−3 gibberellic acid (GA3) enhanced shoot growth. Stout roots (maximum of 5–6 per shoot) were developed in presence of 1 mg dm−3 indole-3-butyric acid (IBA). Micro-bulbs showed potential of regeneration and could be used as secondary explants. The morphologically identical plants derived by in vitro propagation were genetically identical as shown by PCR based ISSR marker analysis of genomic DNA.

Conservation and documentation of the medicinal plant resources of India

In India, activities in the field of medicinal plants, including conservation of germplasm, have been enhanced significantly during the past couple of decades and a huge volume of data is being generated out of these works. For maintaining the records in a consolidated form, documentation is required to store and manage all information on the related studies. In accordance with the implementation of various plans and programmes, some pioneer organisations started developing databases on medicinal plants. Based on the knowledge on contemporary works, as collected from published literature and websites, this article presents information on current activities in India in two important aspects of the field, namely, (1) conservation of medicinal plants; and (2) management of data generated from such studies. Another important aspect of the article is the announcement of a plant conservation related software, ‘PlantCon’. This digitised database contains data of 40 selected nationally prioritised medicinal plants (list enclosed). The notable difference of ‘PlantCon’ from other databases lies in its conservation-related information which is up-to-date and covers a wide area of Indian geographical sites. The database provides information in a user-friendly manner.

Rapid multiplication of Jasminum officinale L. by in vitro culture of nodal explants

Jasminum officinale L. (Fam. - Oleaceae) a shrub, noted specially for its aroma, has been micropropagated successfully by culturing nodal segments. MS basal media with 3% sucrose and supplemented with 6-benzyladenine, at 17.76 µM and 1-naphthaleneacetic acid at 0.53 µM concentration was used for shoot proliferation. For elongation, BA and kinetin, individually and also in combination with NAA were supplemented in the medium. 4 to 5 cm long shoots were transferred to liquid rooting medium. Rooted plants grew normally under natural conditions.