Isolation of salt stress-related genes from Aspergillus glaucus CCHA by random overexpression in Escherichia coli.
The halotolerant fungus Aspergillus glaucus CCHA was isolated from the surface of wild vegetation around a saltern with the salinity range being 0-31%. Here, a full-length cDNA library of A. glaucus under salt stress was constructed to identify genes related to salt tolerance, and one hundred clones were randomly selected for sequencing and bioinformatics analysis. Among these, 82 putative sequences were functionally annotated as being involved in signal transduction, osmolyte synthesis and transport, or regulation of transcription. Subsequently, the cDNA library was transformed into E. coli cells to screen for putative salt stress-related clones. Five putative positive clones were obtained from E. coli cells grown on LB agar containing 1 M NaCl, on which they showed rapid growth compared to the empty vector control line. Analysis of transgenic Arabidopsis thaliana lines overexpressing CCHA-2142 demonstrated that the gene conferred increased salt tolerance to plants as well by protecting the cellular membranes, suppressing the inhibition of chlorophyll biosynthesis. These results highlight the utility of this A. glaucus cDNA library as a tool for isolating and characterizing genes related to salt tolerance. Furthermore, the identified genes can be used for the study of the underlying biology of halotolerance.
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China ; Key Lab of Tree Genomics, The Research Institute of Subtropical of Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400, China.
Jie
J
Fang
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China ; Key Lab of Tree Genomics, The Research Institute of Subtropical of Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400, China.
Xiaojiao
X
Han
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China ; Key Lab of Tree Genomics, The Research Institute of Subtropical of Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400, China.
Lihua
L
Xie
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China ; Key Lab of Tree Genomics, The Research Institute of Subtropical of Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400, China.
Mingying
M
Liu
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China ; Key Lab of Tree Genomics, The Research Institute of Subtropical of Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400, China.
Guirong
G
Qiao
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China ; Key Lab of Tree Genomics, The Research Institute of Subtropical of Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400, China.
Jing
J
Jiang
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China ; Key Lab of Tree Genomics, The Research Institute of Subtropical of Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400, China.
Renying
R
Zhuo
D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development; D001230:Aspergillus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D002241:Carbohydrates / Q000096:biosynthesis / Q000235:genetics; D002364:Caseins / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D008055:Lipids / Q000096:biosynthesis / Q000235:genetics; D014674:Plant Proteins, Dietary / Q000096:biosynthesis / Q000235:genetics; D030821:Plants, Genetically Modified; D054712:Salinity; D055049:Salt Tolerance / Q000235:genetics*; D015398:Signal Transduction; D012965:Sodium Chloride / Q000633:toxicity; D013312:Stress, Physiological / Q000235:genetics*
D002241:Carbohydrates; D002364:Caseins; D008055:Lipids; D014674:Plant Proteins, Dietary; C080652:Osmolite; D012965:Sodium Chloride