Plant regeneration and propagation of chickpea (Cicer arietinum L.) through tissue-culture techniques

Culture conditions have been developed to facilitate callus induction in chickpea seeds and various plant parts, such as apices, stem portions, leaves and roots. Callus was induced by keeping seeds, or any plant part, in B5 or Murashige and Skoog (MS) basal medium containing 5x10-7M kinetin or benzyladenine (BA), 10-5M naphthaleneacetic acid (NAA) and 10-6M 2,4-dichlorophenoxyacetic acid (2,4-D). Regeneration of plantlets was possible only till the second subculture. Shoots regenerated when the calli were transferred to medium containing BA (10-6-10-7M) alone. After growth, roots were induced by putting the shoots in a medium containing BA (5x10-7M) and NAA (10-5-10-6M). A method was established to obtain plantlets from nodal or apical explants without callus formation. Shoots were developed by keeping explants in B5 or MS basal medium containing BA (10-6M). After a ten-day period, the growing shoots were transferred for four days to a medium containing BA (5x10-7M) and NAA (10-6M) for root induction. Then they were transferred to basal medium lacking any hormones, where a root system developed. The root system was well developed within seven days and plantlets were transferred to soil in earthen or plastic pots. For anther cultures, buds at the tetrad stage were collected from field-grown crops. Best results were achieved when the buds were kept at 4 deg. C for 3-7 d. The buds were then centrifuged and sterilized with ethanol, the anthers being dissected and put into callus inducing media. The most suitable medium was B5 or MS basal salt containing BA (10-6M) and 2,4-D(10-5M), where compact calli of plant-green colour were produced. Roots were occasionally formed upon gradual removal of exogenous auxins