Gene manipulation of a heavy metal hyperaccumulator species Thlaspi caerulescens L. via Agrobacterium-mediated transformation.
Thlaspi caerulescens L. is well known as a Zn/Cd hyperaccumulator. The genetic manipulation of T. caerulescens through transgenic technology can modify plant features for use in phytoremediation. Here, we describe the efficient transformation of T. caerulescens using Agrobacterium tumefaciens strain EHA105 harboring a binary vector pBI121 with the nptII gene as a selectable marker, the gus gene as a reporter and a foreign catalase gene. Based on the optimal concentration of growth regulators, the shoot cluster regeneration system via callus phase provided the basis of the genetic transformation in T. caerulescens. The key variables in transformation were examined, such as co-cultivation period and bacterial suspension density. Optimizing factors for T-DNA delivery resulted in kanamycin-resistant transgenic shoots with transformation efficiency more than 20%, proven by histochemical GUS assay and PCR analysis. Southern analysis of nptII and RT-PCR of catalase gene demonstrated that the foreign genes were integrated in the genome of transformed plantlets. Moreover, the activity of catalase enzyme in transgenic plants was obviously higher than in wild-type plants. This method offers new prospects for the genetic engineering of this important hyperaccumulator species.
Guan
Zi Qiu
ZQ
Department of Biology, Graduate University of Chinese Academy of Sciences, Beijing, 100049, China.
Chai
Tuan Yao
TY
Zhang
Yu Xiu
YX
Xu
Jin
J
Wei
Wei
W
Han
Lu
L
Cong
Lin
L
D001483:Base Sequence; D015139:Blotting, Southern; D002374:Catalase / Q000235:genetics; D003470:Culture Media; D017931:DNA Primers; D017930:Genes, Reporter; D019216:Metals, Heavy / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012231:Rhizobium / Q000235:genetics*; D031227:Thlaspi / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D003470:Culture Media; D017931:DNA Primers; D019216:Metals, Heavy; D002374:Catalase