Characterization of VvPAL-like promoter from grapevine using transgenic tobacco plants

被引:0
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作者
SongTao Jiu
Chen Wang
Ting Zheng
Zhongjie Liu
XiangPeng Leng
Tariq Pervaiz
Abolfazl Lotfi
JingGui Fang
XiaoMin Wang
机构
[1] Nanjing Agricultural University,Key Laboratory of Genetics and Fruit development, College of Horticulture
[2] National Institute of Genetic Engineering and Biotechnology (NIGEB),Department of Plant Molecular Biotechnology, Institute of Agricultural Biotechnology
[3] Institute of Botany,undefined
[4] Jiangsu Province and Chinese Academy of Sciences,undefined
来源
关键词
Phenylalanine ammonia-lyase-like; Promoter; Transgenic tobacco; Spatial and temporal expression; Abiotic stress;
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中图分类号
学科分类号
摘要
A 2000-bp 5′-flanking region of VvPAL-like was isolated from ‘Summer Black’ grapevine by PCR amplification, named pVvPAL-like. To gain a better understanding of the expression and regulatory mechanism of VvPAL-like, a chimeric expression unit consisting of the β-glucuronidase (GUS) reporter gene under the control of a 2000-bp fragment of the VvPAL-like promoter was transformed into tobacco via Agrobacterium tumefaciens. Histochemical staining showed that the full-length promoter directs efficient expression of the reporter gene in cotyledons and hypocotyls, stigma, style, anthers, pollen, ovary, trichomes, and vascular bundles of transgenic plants. A series of 5′ progressive deletions of the promoter revealed the presence of a negative regulatory region (−424 to −292) in the VvPAL-like promoter. Exposure of the transgenic tobacco plants to various abiotic stresses demonstrated that the full-length construct could be induced by light, copper (Cu), abscisic acid (ABA), indole-3-acetic (IAA), methyl jasmonate (MeJA) (N-1-naphthylphthalamic acid), ethylene, and drought. Furthermore, the ethylene-responsive region was found to be located in the −1461/−930 fragment, while the element(s) for the MeJA-responsive expression may be present in the −424/−292 region in the VvPAL-like promoter. These findings will help us to better understand the molecular mechanisms by which VvPAL-like participates in biosynthesis of flavonoids and stress responses.
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页码:595 / 617
页数:22
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