In vitro regeneration of chickpea (Cicer arietinum L.): Stimulation of direct organogenesis and somatic embryogenesis by thidiazuron

Abstract

In vitro regeneration in chickpea (Cicer arietinum L.) was achieved by direct culture of mature seeds on Murashige and Skoog (MS) medium supplemented with either N-phenyl-N' (-1,2,3-thidiazol-5-yl) urea (thidiazuron, TDZ) or N6-benzylaminopurine (BAP). Multiple shoots formed de novo without an intermediary callus phase at the cotyledonary notch region of the seedlings within 2 to 3 weeks of culture initiation. TDZ was found to be more effective compared to BAP as an inductive signal of regeneration. The former induced multiple shoot formation at all the concentrations tested (1 PM to 100 PM), although, maximum morphogenic response was observed at 10 PM concentration. Addition of naphthaleneacetic acid (NAA) alone or in combination with BAP to the MS medium failed to invoke a similar response. When the TDZ supplemented medium was amended with L-proline, the resultant regenerants were mostly somatic embryos. Histological investigations confirmed the switch in the regeneration pathway from directly formed adventitious shoots to embryogenesis. For obtaining plantlets, adventitious shoots were rooted on MS medium supplemented with 2.5 ,uM NAA; somatic embryos were germinated and established on MS medium. Normal plants were regenerated from both adventitious shoots and somatic embryos and transferred to soil.Abbreviations: BAP = 6-benzylaminopurine; MS = Murashige and Skoog [ 141 basal medium; NAA = naphthaleneacetic acid; TDZ = thidiazuron [N-phenyl-N'(-1,2,3,-thidiazol-5yl)-urea]

Bauhinia forficata link shoot regeneration: histological analysis of organogenesis pathway

“…It has been observed for various legumes that high concentration of cytokinin stimulated callus and subsequently shoot bud formation (Malik & Saxena, 1992;Dorneles et al, 1994), increased their survival (Dorneles et al, 1994) but also inhibited shoot growth (Thomé et al 1995). Other studies with legumes in vitro (Dewan et al, 1992;Mohamed et al, 1992;Murthy et al, 1996) have determined the need of cytokinin for shoot differentiation and bud multiplication. The percentage of buds elongating into shoot regenerants was 68.2% in this work, compared to values of 81 and 94% obtained by Murthy et al (1996) with Cicer arietinum.…”

“…Other studies with legumes in vitro (Dewan et al, 1992;Mohamed et al, 1992;Murthy et al, 1996) have determined the need of cytokinin for shoot differentiation and bud multiplication. The percentage of buds elongating into shoot regenerants was 68.2% in this work, compared to values of 81 and 94% obtained by Murthy et al (1996) with Cicer arietinum. The choice of MS culture medium to establish a protocol for B. forficata in vitro plant regeneration was due to its frequent use and good success for shoot induction in tissue culture studies of legumes trees like Swartzia madagascariensis (Berger & Schaffner, 1995) and other species of Bauhinia genus (Upreti & Dhar, 1996).…”

Thidiazuron: A multi-dimensional plant growth regulator

“…The modulation of endogenous ABA and GA by TDZ was also reported (Murthy et al, 1996;Murch et al, 1999). But unfortunately, no progress was made in this field later on.…”

“…Tapan et al (1998) observed the callus of tea overgrowth and later necrosis when the responsive explants were continuously grown on the medium containing TDZ. The deleterious effect of the continued presence of TDZ on the growth and multiplication have been reported for chickpea (Murthy et al, 1996), Pisum sativum (Bohmer et al, 1995), Anoectochilus formosanus (Ket et al, 2004), Rauvolfia tetraphylla (Faisal et al, 2005) and Psoralea corylifolia (Faisal and Anis, 2006). Chitra and Padmaja (2005) reported that only few buds derived leaves of mulberry developed into shoots of 0.5 to 1.5 cm at the end of 30 days, and growth of the rest of the adventitious shoot buds remained arrested on medium containing 18.17 and 22.7 mM TDZ.…”

In vitro regeneration through organogenesis and somatic embryogenesis in pigeon pea [Cajanus cajan (L.) Millsp.] cv. JKR105

“…TDZ has been used to elicit multiple shoot formation in a broad range of plant species (Malik and Saxena 1992a,b), and somatic embryogenesis in a few crop species Lu 1993). While other studies indicated successful utilization of cytokinins for somatic embryogenesis not only in pigeon pea (Patel et al 1994;Sreenivasu et al 1998;Mohan and Krishnamurthy 2002;Chandra et al 2003;Singh et al 2003) but also other leguminous crops (Maheswaran and Williams 1984;Gill and Saxena 1992;Malik and Saxena 1992b;Murthy et al 1996). The MS medium comprising TDZ and BAP was found less effective for globular embryo formation with glucose than sucrose.…”