Molecular cloning and characterization of the promoter for the multiple stress-inducible gene <Emphasis Type="Italic">BjCHI1</Emphasis> from <Emphasis Type="Italic">Brassica juncea</Emphasis> |
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Authors: | Xue-Feng Wu Chun-Lian Wang En-Bei Xie Ying Gao Ying-Lun Fan Pi-Qing Liu Kai-Jun Zhao |
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Institution: | (1) National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), 100081 Beijing, People’s Republic of China;(2) Graduate School of the Chinese Academy of Agricultural Sciences, 100081 Beijing, People’s Republic of China;(3) Agricultural College, Guangxi University, 530005 Nanning, Guangxi Province, People’s Republic of China;(4) Present address: Agricultural Vocation-Technical College, 530007 Nanning, Guangxi Province, People’s Republic of China;(5) Present address: School of Agriculture, Liaocheng University, 252059 Liaocheng, Shandong Province, People’s Republic of China; |
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Abstract: | We have previously isolated a Brassica juncea cDNA encoding a novel chitinase BjCHI1 with two chitin-binding domains (Zhao and Chye in Plant Mol Biol 40:1009–1018, 1999). The expression of BjCHI1 was highly inducible by methyl jasmonate (MeJA) treatment, wounding, caterpillar feeding, and pathogenic fungal infection.
These observations suggest that the promoter of BjCHI1 gene might contain specific cis-acting elements for stress responses. Here, we report the cloning and characterization of the BjCHI1 promoter. A 1,098 bp BjCHI1 genomic DNA fragment upstream of the ATG start codon was isolated by PCR walking and various constructs were made by fusing
the BjCHI1 promoter or its derivatives to β-glucuronidase reporter gene. The transgenic Arabidopsis plants showed that the BjCHI1 promoter responded to wounding and MeJA treatment, and to treatments with either NaCl or polyethyleneglycol (PEG 6000), indicating
that the BjCHI1 promoter responses to both biotic and abiotic stresses. A transient gene expression system of Nicotiana benthamiana leaves was adopted for promoter deletion analysis, and the results showed that a 76 bp region from −695 to −620 in the BjCHI1 promoter was necessary for MeJA-responsive expression. Furthermore, removal of a conserved T/G-box (AACGTG) at −353 to −348
of the promoter greatly reduced the induction by MeJA. This is the first T/G-box element identified in a chitinase gene promoter.
Gain-of-function analysis demonstrated that the cis-acting element present in the 76 bp region requires coupling with the T/G-box to confer full magnitude of BjCHI1 induction by MeJA. |
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Keywords: | Inducible promoter Methyl jasmonate Osmotic stress T/G-box Transgenic Arabidopsis W-box |
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