Establishment of regeneration and transformation system in Egyptian sesame ( Sesamum indicum L.) cv Sohag 1

Abstract

Sesame (Sesamum indicum L.) is an important oil crop in many tropical and sub-tropical regions of the world, yet has received little attention in applying modern biotechnology in its improvement due to regeneration and transformation difficulties. Here within, we report the successful production of transgenic fertile plants of sesame (cv Sohag 1), after screening several cultivars. Agrobacterium tumefaciens- carrying the pBI121 plasmid {neomycin phosphotransferase gene (NPTII) and a β-glucuronidase gene (GUS)} was used in all experiments. Recovery of transgenic sesame shoots was achieved using shoot induction medium (Murashige and Skoog MS basal salt mixture + Gamborg's B5 vitamins + 2.0 mg/l BA + 1.0 mg/l IAA + 5.0 mg/l AgNO3 + 30.0 g/l sucrose + 7.0 g/l agar + 200 mg/l cefotaxime and 25 mg/l kanamycin) and shoots were rooted on MS medium + B5 vitamins + 1.0 mg/l IAA + 10.0 g/l sucrose and 7.0 g/l agar. Rooted shoots were transplanted into soil and grown to maturity in greenhouse. Incorporation and expression of the GUS gene into T0 sesame plants was confirmed using polymerase chain reaction (PCR), reverse transcriptase-PCR (RT-PCR) and GUS histochemical assay. Several factors were found to be important for regeneration and transformation in sesame. The most effective were plant genotype and the addition of AgNO3 for successful recovery of sesame shoots. Co-cultivation time and optical density of the Agrobacterium were also critical for sesame transformation. This work is an attempt to open the door for further genetic improvement of sesame using important agronomic traits.

Lignans of Sesame (Sesamum indicum L.): A Comprehensive Review

“…The first attempt to transform S. indicum using Agrobacterium tumefaciens was undertaken in 1999 [ 438 ] but the difficulty of regenerating sesame plants from callus hampered progress in the field. Careful selection of accessions amenable to transformation and the optimization of regeneration conditions eventually allowed for the development of a reliable transformation protocols with an acceptable transformation efficiency [ 439 , 440 , 441 ]. A successful particle bombardment protocol for sesame was also developed [ 442 ].…”

The Emerging Oilseed Crop Sesamum indicum Enters the “Omics” Era

“…Genetic transformation would be an ideal opportunity to quickly transfer the functional genes into sesame elite cultivars. Actually, several successful attempts of sesame genetic transformation through Agrobacterium have led to up 42.66% of transformation efficiency (Yadav et al, 2010; Al-Shafeay et al, 2011; Chowdhury et al, 2014). However, improvements of the sesame genetic transformation protocol for reaching higher efficiency are imperative.…”

An efficient protocol for in vitro direct shoot organogenesis of Sesamum indicum L. using cotyledon as explant

“…Various efforts have been made over the past two decades for in vitro regeneration in sesame. Regeneration of S. indicum has been achieved from hypocotyl and cotyledon (Taskin and Turgut 1997;Were et al 2006;Seo et al 2007;Yadav et al 2010;Al-Shafeay et al 2011) by direct organogenesis and from hypocotyl (Kwon et al 1993;Wadeyar and Lokesha 2011) and leaf-derived callus (Raja and Jayabalan 2010) by indirect organogenesis. Somatic embryogenesis has also been reported from cotyledon, and hypocotyls-derived callus (Ram et al 1990;Xu et al 1997;Baskaran and Jayabalan 2006).…”