Enhanced cadmium resistance and accumulation in Pseudomonas putida KT2440 expressing the phytochelatin synthase gene of Schizosaccharomyces pombe.
Phytochelatins (PCs) are cysteine-rich peptides with high binding affinity for toxic metals. Expressing the PC synthase gene (PCS) in plant growth-promoting bacteria may enhance its metal resistance and accumulation, consequently increasing phytoremediation efficiency in heavy metal pollution. In this study, PCS from Schizosaccharomyces pombe was cloned and expressed in Pseudomonas putida KT2440, which was confirmed by real-time RT-PCR through an increase in SpPCS mRNA expression level when induced by 20 μmol of CdCl2 in the transformed Ps. putida cells. The recombined strain KT2440-SpPCS exhibited enhanced Cd, Ag and Hg resistance. Compared with the original strain, KT2440-SpPCS also displayed a threefold to fivefold increase in Cd accumulation (14·32 μmol g(-1) to 17·38 μmol g(-1) ; dry weight) when grown in 30 and 50 μmol CdCl2 , along with an increase in nonprotein thiols. Further experiments showed significantly enhanced germination rates and growth of wheat seeds in 0·1 mmol to 1·0 mmol Cd when inoculated with KT2440-SpPCS. This study shows potential use of Ps. putida KT2440-SpPCS in plants to construct a symbiotic system for an enhanced phytoremediation of heavy metal-contaminated environments.


CONCLUSIONS
The symbiotic system of using plant growth-promoting bacteria Pseudomonas putida to express phytochelatin synthase gene of Schizosaccharomyces pombe together in plants resulted in high heavy metal resistance and high accumulation capacity, suggesting potential enhancement in phytoremediation of heavy metal-contaminated environments.
College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, China; Bioenergy Research Institute, Nanjing University of Technology, Nanjing, China.
X
X
Yong
Y
Y
Chen
W
W
Liu
L
L
Xu
J
J
Zhou
S
S
Wang
P
P
Chen
P
P
Ouyang
T
T
Zheng
D019881:Aminoacyltransferases / Q000235:genetics* / Q000378:metabolism; D002104:Cadmium / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D019216:Metals, Heavy / Q000378:metabolism; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000235:genetics*; D012639:Seeds / Q000254:growth & development / Q000382:microbiology; D014908:Triticum / Q000254:growth & development / Q000382:microbiology
D005656:Fungal Proteins; D019216:Metals, Heavy; D002104:Cadmium; D019881:Aminoacyltransferases; C093784:glutathione gamma-glutamylcysteinyltransferase
Pseudomonas putida; accumulation; cadmium; phytochelatin synthase; resistance