Plant regeneration from stem-derived callus of the seed legume Lathyrus sativus L.

Abstract

Callus cultures from stem explants of six Lathyrus sativus L. cultivars were tested for their morphogenic capacity. Shoot-buds were formed in calli of only one cultivar. Maximum response was observed in the medium containing 10-SM picloram and 10-aM benzylaminopurin. Supplementation with adenine sulphate was required for shoot-bud formation. The greatest frequency of shoot-bud formation was detected at the second passage and complete lack of regeneration capacity was observed after the 8th passage. Cell regenerates were diploid.lntrodution Lathyrus sativus is a tropical seed legume crop which perform well in adverse agricultural conditions. The crop is widely grown in India despite the fact that the seeds contain neurotoxic substances known as 'Lathyrogens'. It might be possible to select toxin-free strains from mutagenized tissue cultures. Before such a programme can be initiated, totipotent tissue culture system must be available. However, legume tissue cultures were generally recalcitrant in regard to morphogenic capacity [7]. We therefore, initiated research to define the conditions required for high frequency plant regeneration from cultured tissues and protoplasts of L. sativus. In the present paper we report the regeneration of plants via organogenesis in stem-derived callus of L. sativus. It is also shown that regenerating ability is genotype dependent. /3-diaminopropionic acid) at levels (mg/100g seeds) of 245,426, 328,254, 423 and 295, respectively [12], were used in this study. All the cultivars contain 2n = 14 chromosomes in somatic cell.Seeds were surface sterilized in 0.02% mercuric chloride for 5 min and 67 Plant Cell Tissue Organ Culture 2: 67-76 (1983).

In vitro plant regeneration from callus derived from root explants of Lathyrus sativus

“…Earlier several workers attempted regeneration in Lathyrus sativus but in most cases shoots were obtained with no or negligible rooting from cultures of shoot apices, meristems or stem explants (Mukhopadhyay & Bhojwani 1978;Gharyal & Maheshwari 1983;Sinha et al 1983). Recently in vitro regeneration and rooting from leaf explants has been obtained (Roy et al 1991).…”

Callus Induction and Rhizogenesis in Lathyrus sativus L.

“…In contrast this cytokinin can inhibit root formation in higher concentration (Orinos and Mitrakos, 1991). The frequent and spontaneous rhizogenesis on media with diff erent combinations of growth regulators and in diff erent grass pea genotypes was also mentioned by Sinha et al (1983). These researchers observed also that the tissue producing roots diff erentiated shoots very rarely.…”

“…Tissue formed on the light was usually green colour, which obviously resulted from the chlorophyll synthesized under such conditions. Induction of callus from various grass pea explants and further tissue growth was carried out on the light (Barik and Kar, 2005;Roy et al, 1991Roy et al, , 1992Roy et al, , 1993Sinha et al, 1983) or in the dark for 2 to 7 days, and then on the light Maheshwari, 1980, 1983;Zambre et al, ). Just as the callus induction, its proliferation proceeded without major problems.…”

“…However, morphogenic capacity of grain legumes in tissue culture are generally poor (Sinha et al, 1983). Earlier studies showed that apart from the diffi culties in the regeneration of shoots from callus tissue of grass pea (Gharyal and Maheshwari, 1983;Sinha et al, 1983) it is very diffi cult to get the whole plant by indirect organogenesis and it succeeded only in a few cases in this species (Barik and Kar, 2005;Roy et al, 1991;Roy et al, 1992;Zambre et al, 2002).…”

Early stages of somatic embryogenesis in root callus of grasspea (Lathyrus sativus L.)

“…Sinha et al (1983) also mention a frequent formation of roots on media with various combinations of growth regulators and in various genotypes. These researchers observed no differentiation of other organs in the tissue that produced roots, in turn in our experiment rhizogenesis did not stop formation of ELSs.…”