In Vitro Direct Organogenesis Using Mature Embryo With Cotyledons In Chickpea

Abstract

The present study was conducted to find out an in vitro efficient method for multiple shoot regeneration of two local chickpea varieties. The mature embryos were excised of two chickpea varieties i.e. Bittle-98 and Dasht-2000 (with cotyledon and without cotyledon) used as explants. The explants were cultured on Murashige and Skoog (MS) medium supplemented with three concentrations of (2, 3, 4 mg/l) 6-benzylaminopurine (BAP) using explant with and without cotyledon. Further, 0.5 mg/l α-naphthaleneacetic acid (NAA) along with varied concentrations of BAP (2, 3, 4 mg/ l) was also tested using explant with cotyledon. 3 mg/l BAP alone and 3 mg/l BAP with 0.5 mg/l NAA were found the most effective cytokinin in multiple shoot induction in both tested varieties. Bittle-98 and Dasht-2000 showed 82% and 76% elongation in shoots induction with 0.2 mg/l Indole-3-acetic acid (IAA). Root formation was recorded 80% and 60% in Bittle-98 and Dasht-2000 with 1.0 mg/l Indole-3-butyric acid (IBA). Whereas, recorded root formation was 40 and 20% in Bittle-98 and Dasht-2000 with 1.0 mg/l NAA. The best response for rooting observed in Bittle-98 as its roots were thick, long and strong. Plantlets of Bittle-98 were acclimatized in solid medium for 7-14 days. The successful invitro regeneration of Bittle-98 was observed, when excised embryo with cotyledon were used as explant, made it valuable for genetic transformation.