Spinach is one of the dioecious plant which is consid- ered as a model plant in genetic and molecular studies of sex determination because of its special character- istics such as low chromosome number and short life cycle. An efficient protocol for Spinacia oleracea Agro- bacterium-mediated gene transformation was devel-oped. The leaf disks, roots, hypocotyls and cotyledons of this plant were inoculated with LBA4404. LBA4404 carrying pCAMBIA3301 binary vector with 35SCaMV gus-int and 35SCaMV bar cassettes. Effects of two preparation condition (induction of vir genes and noninduction) were considered. Also effects of differ-ent number days of co-cultivation and pre-culture of explants were examined. After co-cultivation, the ex-plants were transferred to regeneration medium con-taining 250 mg·L–1 Carbeniciline. Transient expres-sion efficiency was calculated based on the number of blue spots per explants one week after inoculation. Based on the results of transient expression, stable transformation was carried out. After formation of callus the histochemical GUS assay was carried out on some parts of them and other parts were leaved for being regenerated.
کلید واژگان :Spinacia Oleracea; Transformation; Gus Gene; Transient Expression; Stable Expression
ارزش ریالی : 300000 ریال
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