Molecular cloning and characterization of the mitogen-activated protein kinase kinase gene (MKK4) and its promoter sequence from oilseed rape (Brassica campestris L.)
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- 作者：Teng Guo Zhang (1)
Qiong Qiong Chen (1)
Ning Wang (1)
Xiao Hui Xia (2)
Juan Wang (1)
Yan Chang (1)
Ying Li Yang (1)
Ning Yang (1)
Wan Cang Sun (3) - 关键词：Brassica campestris L. ; MKK4 ; Promoter ; Abiotic stress ; Quantitative real ; time polymerase chain reaction (qRT ; PCR)
- 刊名：Plant Cell, Tissue and Organ Culture
- 出版年：2013
- 出版时间：December 2013
- 年：2013
- 卷：115
- 期：3
- 页码：341-353
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- 作者单位：Teng Guo Zhang (1)
Qiong Qiong Chen (1)
Ning Wang (1)
Xiao Hui Xia (2)
Juan Wang (1)
Yan Chang (1)
Ying Li Yang (1)
Ning Yang (1)
Wan Cang Sun (3)
1. School of Life Sciences, Northwest Normal University, Lanzhou, 730070, China
2. Lanzhou City University, Lanzhou, 730070, China
3. College of Agronomy, Gansu Agricultural University, Lanzhou, 730070, China - ISSN：1573-5044
文摘
Mitogen-activated protein kinase (MAPK) cascades are involved in various processes, including plant growth and development as well as biotic and abiotic stress responses. MAPK kinases (MKKs), which link MPKs and MAPKK kinases (MKKKs), are crucial in MAPK cascades because these kinases mediate various stress responses in plants. However, only few MKKs in Brassica campestris (rape) have been functionally characterized. In this study, a novel gene, MKK4 that belongs to a C MKK group, was isolated and characterized from rape. Bioinformatics analysis revealed that the length of cDNA was 1,317?bp with an open reading frame of 993?bp, which encodes a polypeptide containing 330 amino acids, including a putative signal peptide with 27 amino acid residues and a mature protein with 303 amino acids. The obtained MKK4 exhibited a predicted molecular mass of 36.5?kDa and an isoelectric point of 9.01. Quantitative real-time polymerase chain reaction analysis revealed that MKK4 expression could be induced by cold and salt. We also found that the MKK4 protein is localized in the nucleus. In addition, a 999?bp promoter fragment of MKK4 was cloned. Sequence analysis revealed that several putative regulatory elements were found in the MKK4 promoter. Transient expression assay showed that the MKK4 promoter fragments exhibited promoter activity and stimulated GFP expression. The effects of GFP gene expression at different temperatures and in different onion epidermis culture patterns were compared. Results showed that the MKK4 promoter could respond to low temperature and salt stress. These results suggested that MKK4 is possibly important for the regulation of cold- and salt-stress responses in plants.