High frequency plant regeneration from embryogenic callus of a popular indica rice (Oryza sativa L.)

Abstract

The aim of the study is to establish a routine procedure for high frequency plant regeneration from in vitro raised embryogenic callus of abiotic salt sensitive indica rice (Oryza sativa L.) cultivar ADT 43. The effect of synthetic auxin 2,4-D on callus induction was optimized to achieve high frequency plant regeneration from fresh embryogenic callus without further subculture. Friable, nodular and creamish-white embryogenic callus cultures were raised from mature rice seeds on LS medium supplemented with 2.5 mg L -1 2,4-D and 1.0 mg L -1 thiamine-HCL. Plant regeneration was achieved by the 24 days old embryogenic callus on MS medium supplemented with 1.0 mg L -1 BAP and 1.5 mg L -1 NAA. In vitro regenerated plants with multiple tillers and roots were transferred to sterile soil and maintained in the growth chamber. The regenerated plants exhibited normal growth and were phenotypically similar to plants maintained in the garden. Using the present protocol, 25-30 plantlets were regenerated from 50 individual mature seed derived callus within two to three months. This protocol has the potential for large-scale production of elite plants after genetic transformation.

Optimization of regeneration using differential growth regulators in indica rice cultivars

“…Similarly, in Sambha mahsuri cultivar, maximum multiple shoots regeneration was achieved in supplementation of MS + BAP (5.0 mg L −1 ). Simultaneously, stimulatory effects of BAP in combination with IAA and NAA have previously been reported to facilitate regeneration in indica rice cell cultures (Mandal et al 2003; Karthekeyan et al 2009). The observed results revealed that kinetin was found to be effective for the induction and proliferation of shoots and roots.…”

Transgenic indica rice cv. ADT 43 expressing a Δ1-pyrroline-5-carboxylate synthetase (P5CS) gene from Vigna aconitifolia demonstrates salt tolerance

“…In the present study, we succeeded in obtaining efficient and short-term protocol for high frequency plant regeneration from mature seed derived embryogenic calli in order to use them for genetic transformation experiments as done previously in ADT 43 (Karthikeyan et al 2009). 2,4-D at 2.5 mg/l was found to be satisfactory in inducing regenerable calli from mature seed explants.…”

“…ADT 43 is indica type rice (IET 14879), medium slender white, abiotic stress sensitive, and dwarf rice of IR 509 improved ponni parentage yielding 56 Q/ha in 110 days. High frequency plant regeneration from embryogenic callus tissues have been reported from our group in this abiotic sensitive popular indica rice variety (ADT 43) in a shorter duration using embryogenic callus cultures (Karthikeyan et al 2009). In Tamil Nadu, rice is cultivated in 1.52 million hectares of which ADT 43 alone constitutes nearly 21% (Selvaraj and Ramasamy 2006).…”

“…In our earlier report Karthikeyan et al 2009, the effect of various concentrations of 2,4-D were analyzed for high frequency plant regeneration through direct organogenesis from mature seed derived callus explants. Of various auxin concentrations tried, seed explants showed high frequency callus induction on LS medium supplemented with 2.5 mg/l of 2,4-D (Fig.…”

Efficient In Vitro Somatic Embryogenesis and Plant Regeneration from Mature and Immature Embryos of Wheat ( Triticum aestivum L.)

“…It cultivates on approximately 17% of the cultivatable lands and plays a key role in economic development, food security and supply, human nutrition and is an important source of calories and proteins for human [1]. While, genetic engineering of wheat provides an efficient strategy for improvement of agronomic traits such as quality and disease resistance, but the graminaceous species are recalcitrant to in vitro regeneration and manipulations [2,3]. An efficient regeneration system is the main pre-requisite for the application of genetic transformation and functional genomics studies in the primomovente of important plants [4].…”