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Cloning and expression analyses of LEA2 gene from Ipomoea batatas(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2014 02
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Cloning and expression analyses of LEA2 gene from Ipomoea batatas
CHEN Jiao JIANG Yusong ZHANG Yizheng TAN Xuemei
1College of Life Sciences, Sichuan University, Key Laboratory of Resource Biology and Eco-environment, Ministry of Education, Chengdu 610064, China 2Chongqing University of Art and Sciences, Yongchuan 402160, China
sweet potato LEA2 gene cloning digital gene expression profiling salt stress
Q78 : S531.034

Late embryogenesis abundant (LEA) proteins, which are produced in plant under stress, have high hydrophilic activity and thermal stability. They are closely related to plant defense reactions. Researches on LEA proteins are important for molecular breeding of sweet potato under stress. This research analyzed the expression pattern of Ib-LEA2 in vivo, and investigated the function of LEA2 in defense of salt stress. Transcripts encoding LEA2 proteins were found when searching the transcriptome database of sweet potato. Subsequently, Ib-LEA2 was cloned and sequenced. Digital gene expression profiling (DGE) and semi-quantitative RT-PCR were chosen for study expression of Ib-LEA2 in different tissues and under different developmental stages. In order to further study the function in response to salt stress, Ib-LEA2 coding sequence was cloned into pET-32a expression vector and then expressed in E. coli BL21(DE3). Result shows that Ib-LEA2 had an open reading frame (ORF) of 942 bp in length, which encoded 313 amino acids. Further analysis revealed three highly homologous genotypes for Ib-LEA2 (nucleotide sequences similarity > 99%), named Ib-LEA2a, Ib-LEA2b and Ib-LEA2c. Digital gene expression profiling (DGE) and semi-quantitative assay showed that Ib-LEA2 proteins were differentially expressed in different tissues and under different developmental stages, and that the expression in stems was much higher than in leaves and roots. The growth curves of E. coli BL21(DE3) showed that the strain expressing Ib-LEA2 can better tolerate high concentration of salt than the control. Our work revealed that Ib-LEA2 could response to stress, protecting cells from high concentration of salt. The results laid a theoretical foundation to the molecular breeding of sweet potato, promoting the researches in salt-tolerance in plants.


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Last Update: 2014-05-04