In vitro induction of multiple shoots and plant regeneration from shoot tips of mung bean (Vigna radiata (L.) Wilczek)

Abstract

Conditions for plant regeneration from excised shoot tips of Vigna radiata were studied. Complete plants were regenerated directly without an intervening callus phase from shoot tips on basal medium (MS salts + B 5 vitamins). Regeneration frequency varied with genotype, explant size and growth regulator combinations in the medium. Addition of cytokinins induced a variable amount of callus at the base of the shoot tip, followed by multiple shoot formation. Benzyladenine (BA), kinetin and zeatin at 5 x 10-6M each induced multiple shoots in 100% of the explants but the highest number of regenerants per explant (9) was produced with BA. The efficacy of BA for shoot multiplication was not improved when it was supplemented with naphthaleneacetic acid (NAA) or indoleacetic acid (IAA). NAA or adenine sulphate, when applied alone, induced complete plantlets. The growth regulator requirement of explants for the induction of multiple shoots varied with explant size. The shoot tip explants maintained proliferation ability on subculture. None of the treatments was effective in inducing shoot bud differentiation from callus. Regenerated shoots were rooted on MS basal medium and MS supplemented with either IAA or indolebutyric acid. The rooted plants were transferred to the field; 60% subsequently survived and grew.

In vitro plant regeneration from the petioles of primary leaves of mungbean Vigna radiata L.

“…The present regeneration protocol developed from preexisting meristem of petiole of primary leaf supports and confirms earlier reports (Gulati and Jaiwal 1992;Gulati and Jaiwal 1994;) that the BA could induce highly efficient multiple shoot development in mungbean followed by successful rooting, transplantation and survival. The concentration of BA at 1.0 mg 1 Ϫ1 was found to be the most responsive in the production of multiple shoot-buds (Table 1).…”

“…Earlier reports on direct multiple shoot regeneration from various explants (Gulati and Jaiwal 1992;Gulati and Jaiwal 1994;) have claimed a moderate regeneration frequency and a lesser number of shoots per explant compared to the present report. Previous studies on plant regeneration from various explants of mungbean obtained through callus also reported very low frequency of regeneration with low survival (Mathews 1987;Patel et al 1991;Mendoza et al 1992;Gulati and Jaiwal 1994).…”

“…Techniques making use of multiple shoot formation around pre-existing meristems may provide an ideal system for in vitro culture and transformation of grain legumes (Ramsay 1993;Olhoft and Somers 2001). There are a few reports on plant regeneration in mungbean from cotyledon explants (Gulati and Jaiwal 1990), shoot tip explants (Mathew 1987;Gulati and Jaiwal 1992) and cotyledonary node explants (Gulati and Jaiwal 1994;. Leaves obtained from in vitro grown seedlings eliminate the risk of contamination.…”

“…Hence, direct plant regeneration from meristems is the most ideal for genetic transformation. Previous reports on multiple shoot regeneration of mungbean have utilized shoot apices (Gulati and Jaiwal 1992) or cotyledonary node explants (Gulati and Jaiwal 1994;. The regenerated plantlets produced presently from leaf explants were healthy and produced normal flowers and pods and set viable seeds.…”

In Vitro Selection of Salt Tolerant Calli Lines and Regeneration of Salt Tolerant Plantlets in Mung Bean (Vigna radiata L. Wilczek)

“…Regeneration from various seedling explants viz shoot tip, cotyledons, cotyledonary node of mung on BAP supplemented medium has been reported earlier (Mathews, 1987;Gulati & Jaiwal, 1992;Kaviraj & Rao, 2009;Mendoza, et al, 1999).there is only one report of regeneration from callus cultures in mungbean , similarly plantlets have been regenerated from NaCl tolerant callus in many species like Citrus sinensis (Ben-Hyyim & Goffer,1989) Brassica juncea (Jain, et al,1991,Kerthi, et al, 1989Sashimadan, et el., 1995) Triticum durum (Zair, et al, 2003) Ipomea batats (He, et al, 2009) In the present study however, it was noticed that the formation of shoots from NaCl tolerant callai was lower (14.00% than that from NaCl-non selected calli (40.13%) and the number of multiple shoots per callai piece was reduced to 3.33 from 14.80, a phenomenon reported earlier by few workers (Basu, et al, 1997;Chen et al, 2001;Ochatt, et al,1999;Queiros, et al, 2007;Zair et al, 2003).…”