Agrobacterium-mediated Genetic Transformation in Cucumber (Cucumis sativus L.)

N Selvaraj,A Vasudevan,C.W Choi,A Ganapathi

doi:10.3844/ajbbsp.2007.24.32

Abstract

The present study was undertaken to develop efficient transformation protocol for cucumber cv. Poinsett 76 using Agrobacterium strain EHA 105. Five-day-old mature cotyledon explants was used for transformation study. The infected explants were co-cultivated for 2 days in MS medium containing BA (1.0 mg Lˉ1). The selection of transformed shoots was carried out in MS medium fortified with BA (1.0 mg Lˉ1), Cefotaxime (300 mg Lˉ1) and PPT (2.0 mg Lˉ1). The transformed shoots were elongated in MS medium containing BA (1.0 mg Lˉ1), Cefotaxime (300 mg Lˉ1), PPT (2.0 mg Lˉ1) along with GA3 (0.5 mg Lˉ1). The rooting of elongated shoots was achieved in MS medium with BA (1.0 mg Lˉ1), Cefotaxime (300 mg Lˉ1), PPT (2.0 mg Lˉ1) and IBA (0.6mg Lˉ1). The transient GUS expression assay and leaf disc assay were carried out in order to find transformed shoots. The molecular confirmation of transformed shoots revealed the foreign gene integration into cucumber genome.

Highlights

Genetic engineering has many potential applications in fields such as medicine, agriculture and industries
Beyond 5th day, cotyledon explants produced shoots but most of them were escapes due to precultivar Poinsett 76 was used as target tissue for Agrobacterium-mediated gene transfer through direct regeneration
Three to seven-day-old in vitro grown seedlings were used as donors of cotyledon explants[9,10,15,16,27,29]

Summary

Introduction

Genetic engineering has many potential applications in fields such as medicine, agriculture and industries. The new application of genetic engineering includes the development of transgenic plants. In order to establish a successful programme of practical plant genetic engineering, it is important to develop systems for the recovery of whole plants in large members from primary explants. This needs to be done while optimizing the procedure for introducing foreign genes into the species of interest. There is no universally applicable method of culture, regeneration and transformation systems for all species, as tissues from different genotypes will differ in their response to culture. Culture and regeneration protocols must be modified appropriately for cultures of each species[7]

Methods

Results

Conclusion