Agrobacterium-mediated transformation of rough lemon (Citrus jambhiri Lush) with yeast HAL2 gene.
BACKGROUND
Rough 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.


RESULTS
Transformation 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.


CONCLUSIONS
Rough 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.
Department of Biochemistry, Quaid-i-Azam University, Islamabad, Pakistan. shawkatali77@hotmail.com
Shawkat
S
Ali
Abdul
A
Mannan
Mohamed
M
El Oirdi
Abdul
A
Waheed
Bushra
B
Mirza
D060054:Agrobacterium / Q000187:drug effects / Q000502:physiology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002957:Citrus / Q000235:genetics*; D003470:Culture Media; D005800:Genes, Fungal; D009708:Nucleotidases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial
D000900:Anti-Bacterial Agents; D003470:Culture Media; D009708:Nucleotidases; C053354:bisphosphoadenylate 3'-nucleotidase