<it>Agrobacterium</it>-mediated transformation of rough lemon (<it>Citrus jambhiri</it> Lush) with yeast <it>HAL2</it> gene

• Main Authors: Ali Shawkat, Mannan Abdul, El Oirdi Mohamed, Waheed Abdul, Mirza Bushra
• Format: Article
• Language: English
• Published: BMC 2012-06-01
• Series: BMC Research Notes
• Subjects: Genetic transformation; Yeast halotolerant gene (<it>HAL2</it>); <it>Citrus jambhiri</it> Lush and <it>Agrobacterium tumefaciens</it>
• Online Access: http://www.biomedcentral.com/1756-0500/5/1/285

<p>Abstract</p> <p>Background</p> <p>Rough lemon (<it>Citrus jambhiri</it> Lush.) is the most commonly used <it>Citrus</it> rootstock in south Asia. It is extremely sensitive to salt stress that decreases the growth and yield of <it>Citrus</it> crops in many areas worldwide. Over expression of the yeast halotolerant gene (<it>HAL2)</it> results in increasing the level of salt tolerance in transgenic plants.</p> <p>Results</p> <p>Transformation of rough lemon was carried out by using <it>Agrobacterium tumefaciens</it> 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 <it>Agrobacterium</it> 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.</p> <p>Conclusion</p> <p>Rough lemon has been successfully transformed via <it>Agrobacterium tumefaciens</it> with β-glucuronidase (<it>GUS)</it> and <it>HAL2.</it> Various factors affecting gene transformation and regeneration efficiency were also investigated.</p>