Isolation and promoter analysis of a chalcone synthase gene PtrCHS4 from Populus trichocarpa
Isolation and promoter analysis of a chalcone synthase gene PtrCHS4 from Populus trichocarpa
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毛果杨查耳酮合酶基因PtrCHS4的分离和启动子分析
DOI：
10.1007/s00299-011-1075-1
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发表时间：
2011-05
影响因子：
6.2
通讯作者：
中科院分区：
文献类型：
--
作者：
杨树作为多年生植物，经常受到环境胁迫，如创伤和病原菌的侵袭，导致产生木质素、类黄酮和植物抗毒素等化合物。查尔酮合成酶（CHS）是黄酮类化合物生物合成途径中的关键酶。本研究利用RT-PCR技术从大叶杨中克隆了一个编码CHS基因的cDNA克隆。PtrCHS 4基因全长为1，314 bp，开放阅读框为1，173 bp，编码391个氨基酸。多重序列比对表明，PtrCHS 4与其他植物CHS蛋白具有较高的同源性。系统进化分析表明，PtrCHS 4与矮牵牛PhCHS和烟草NaCHS的亲缘关系最近。半定量RT-PCR分析表明，PtrCHS 4基因在叶和茎中大量表达，而在根中表达量显著降低。PtrCHS 4的转录丰度在创伤处理的24小时内被刺激2.5倍。启动子分析证实PtrCHS 4启动子能够指导GUS报告基因在转基因毛白色白杨（P. tomentosa卡尔.）的受伤和未受伤叶片中表达，表明PtrCHS 4启动子对创伤刺激有系统性应答。此外，启动子缺失分析表明，从转录起始位点的近端1,592 bp是茉莉酸激活启动子所必需的，并且证明-1,096至-148区域是建立创伤诱导表达模式所必需的。
As perennial plants, Populus species are constantly exposed to environmental stresses, such as wounding and pathogen attack, which lead to production of compounds including lignin, flavonoids and phytoalexins. Chalcone synthase (CHS) is a key enzyme in the flavonoid biosynthesis pathway. In this study, a cDNA clone encoding CHS was isolated from Populus trichocarpa by reverse transcription-polymerase chain reaction (RT-PCR). The full-length cDNA, named PtrCHS4, was 1,314 bp with a 1,173 bp open reading frame that corresponded to a deduced protein of 391 amino acid residues. Multiple sequence alignments showed that PtrCHS4 shared high homology with CHS proteins from other plants. Phylogenetic analysis revealed that PtrCHS4 was most closely related to PhCHS from Petunia hybrida and NaCHS from Nicotiana attenuata. Semi-quantitative RT-PCR analysis identified that the PtrCHS4 gene was abundantly expressed in the leaves and stems, while its expression was drastically reduced in the roots. Transcript abundance of PtrCHS4 was stimulated by 2.5-fold within 24 h of wounding treatment. Promoter analysis confirmed that the PtrCHS4 promoter was capable of directing expression of the GUS reporter in both wounded and unwounded leaves of transgenic Chinese white poplar (P. tomentosa Carr.), indicating that the PtrCHS4 promoter is systemically responsive to wounding stimuli. Furthermore, promoter deletion analysis showed that the proximal 1,592 bp from the transcription start site were required for promoter activation by jasmonic acid and the -1,096 to -148 region was proved to be necessary for establishing wound-induced pattern of expression.
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影响因子：
7.4
作者：
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通讯作者：
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作者：
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通讯作者：
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