Abstract
BackgroundRough lemon (Citrus jambhiri Lush.) is the most commonly used Citrus rootstock in south Asia. It is extremely sensitive to salt stress that decreases the growth and yield of Citrus crops in many areas worldwide. Over expression of the yeast halotolerant gene (HAL2) results in increasing the level of salt tolerance in transgenic plants.ResultsTransformation of rough lemon was carried out by using Agrobacterium tumefaciens strains LBA4404 harboring plasmid pJRM17. Transgenic shoots were selected on kanamycin 100 mg L-1 along with 250 mg L-1 each of cefotaxime and vancomycin for effective inhibition of Agrobacterium growth. The Murashige and Skoog (MS) medium containing 200 μM acetoseryngone (AS) proved to be the best inoculation and co-cultivation medium for transformation. MS medium supplemented with 3 mg L-1 of 6-benzylaminopurine (BA) showed maximum regeneration efficiency of the transformed explants. The final selection of the transformed plants was made on the basis of PCR and Southern blot analysis.ConclusionRough lemon has been successfully transformed via Agrobacterium tumefaciens with β-glucuronidase (GUS) and HAL2. Various factors affecting gene transformation and regeneration efficiency were also investigated.
Highlights
Rough lemon (Citrus jambhiri Lush.) is the most commonly used Citrus rootstock in south Asia
Rough lemon (C. jambhiri Lush.) originated from Himalayan foothills in India and Pakistan, is the commonly used Citrus rootstock in riverland of South Australia, lemon growing area of Arizona [14], south Asia, and in Pakistan it is used as rootstock for
We have previously reported micropropagation of rough lemon [15] which is a prerequisite for Agrobacterium mediated transformation
Summary
Rough lemon (Citrus jambhiri Lush.) is the most commonly used Citrus rootstock in south Asia. In Citrus, gene transformation is carried out by three different techniques i.e., particle bombardment [1] protoplast transformation [2] and Agrobacterium [3,4,5,6,7]. Agrobacterium mediated transformation is the most commonly used method for gene transfer to Citrus as it does not need embryogenic calli and is protoplast independent [2]. Agrobacterium mediated gene transformation have been reported for a number of Citrus species by different groups [8,9,10]. Rough lemon (C. jambhiri Lush.) originated from Himalayan foothills in India and Pakistan, is the commonly used Citrus rootstock in riverland of South Australia, lemon growing area of Arizona [14], south Asia, and in Pakistan it is used as rootstock for
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