Genetic transformation of Pinellia ternata with Agrobacterium tumefaciens-mediated sHSP genes.
- Author:
Zhao-Yang GUO
1
;
Ting-Ting CUI
;
Jian-Ping XUE
;
Yan-Fang ZHU
;
Ai-Min ZHANG
;
Wei SHENG
;
Jing-Tong TENG
Author Information
- Publication Type:Journal Article
- MeSH: Agrobacterium tumefaciens; genetics; DNA, Plant; genetics; Genetic Engineering; methods; Glucuronidase; genetics; metabolism; Heat-Shock Proteins, Small; genetics; Pinellia; genetics; growth & development; metabolism; Plant Leaves; genetics; growth & development; metabolism; Plants, Genetically Modified; Polymerase Chain Reaction; Reproducibility of Results; Tissue Culture Techniques; methods; Transformation, Genetic
- From: China Journal of Chinese Materia Medica 2012;37(24):3758-3762
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish an efficient genetic transformation system of Pinellia ternata.
METHODWith petioles from test-tube seedlings of P. ternata as explants, Agrobacterium tumefaciens mediation method was adopted to explore the effect of phenolic substances, A. tumefaciens's concentration, infection time, pre-incubation time and co-cultivation time on genetic transformation efficiency of P. ternata.
RESULT AND CONCLUSIONThe genetic transformation efficiency could be effectively enhanced by infecting in A. tumefaciens culture containing AS 40 mg x L(-1) for 15 min for three days. The petioles were put into the differentiation medium containing 150 mg x L(-1) Kan and 350 mg x L(-1) Carb to screening and cultivation. After around 30 days, microtubers could be observed at both sides of the petioles. Gus staining and PCR verification on the regenerated plants showed that the exogenous gene sHSP had been integrated into genome of P. ternata.
