Читать книгу Biotechnology of Fruit and Nut Crops - Группа авторов - Страница 42

Transformation of cashew callus was attempted using binary plasmid PBI426, having nopaline synthase poly-A (NOS) terminator and the CaMV 35S promoter. The GUS-nptII was the selection marker with β-glucuronidase (GUS) reporter. The protocol of transformation involved micro-projectile bombardment. Gold nanoparticles were coated with the plasmid and used for gene delivery. Transient Gus expression in cashew was observed (Cardoza and D’Souza, 2002a) at 900 psi. Stable expression and callus proliferation did not occur. Nivas et al. (2007) report Agrobacterium-mediated transformation using plasmid PBIN m-gfp5-ER as a vector carrying the green fluorescent protein (GFP) marker. Agrobacterium tumefaciens strain EHA-105 was used. Transformed bacteria were cocultured with germinating cashew seedlings. The seedlings were subsequently cultured on kanamycin-containing selection medium, and surviving plants were screened after 6 months for GFP expression. Successful transformation was confirmed on the basis of GFP and by PCR using marker gene-specific primers. The plants showed delayed growth and did not survive after 1 year.