[Transformation of embryogenic Calli of Siberian wildrye grass (Elymus sibiricus L. cv. Chuancao No.2) mediated by agrobacterium].
Formation of embryogenic calli of Siberian wildrye grass (Elymus sibiricus L. cv. Chuancao No.2) was induced from mature seeds as explants, and proliferated on MS medium containing 2,4-D 5.0 mg/L and KT 0.05 mg/L. An effective and stable callus regeneration system was established by optimizing the culture conditions (Tables 1, 2 and Fig.2). After the calli were subcultured 8 weeks, selected the whitish-yellow-coloured compact nodular calli that transformed with plasmid pCAMBIA1304 carrying hygromycin resistance gene (hptII) and Pseudomonas pseudoalcaligenes insecticidal protein gene (ppIP), which was mediated by an Agrobacterium strain EHA105. Resistant plants were obtained after hygromycin selection (Figs.3, 4). Some important factors that affect the transformation efficiency were studied, which included selection pressure, time of embryogenic calli proliferation, OD value of Agrobacterium suspension, temperature, medium and time of co-cultivation, and concentration of antibiotics used for suppressing the overgrowth of Agrobacterium in the course of transformation plant regeneration. This research is the first successful genetic transformation of Elymus sibiricus L. cv. Chuancao No.2 mediated by Agrobacterium.
Li
Da-Xu
DX
Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, China.
Zhang
Jie
J
Zhao
Jian
J
Zhang
Yi
Y
Li
Li
L
Liu
Su-Jun
SJ
Chen
Fei
F
Yang
Zhi-Rong
ZR
D003470:Culture Media; D031749:Elymus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D030821:Plants, Genetically Modified / Q000502:physiology; D010946:Plants, Medicinal; D006109:Poaceae; D012231:Rhizobium / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D014170:Transformation, Genetic / Q000502:physiology*
D003470:Culture Media