Expression of calcium-dependent protein kinase (CDPK) genes under abiotic stress conditions in wild-growing grapevine Vitis amurensis
Expression of calcium-dependent protein kinase (CDPK) genes under abiotic stress conditions in wild-growing grapevine Vitis amurensis
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DOI：
10.1016/j.jplph.2013.06.014
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发表时间：
2013-11-15
影响因子：
4.3
通讯作者：
Khristenko, Valeriya S.
中科院分区：
文献类型：
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作者：
Dubrovina, Alexandra S.;Kiselev, Konstantin V.;Khristenko, Valeriya S.
钙依赖性蛋白激酶（CDPKs）是植物钙离子通量的重要传感器，在植物发育和适应非生物胁迫中起着重要作用。本研究研究了野生葡萄（Vitis amurensis Rupr）在渗透胁迫和温度胁迫下CDPK基因的表达。这是已知具有高适应潜力和对不利环境条件的高水平抵抗。在本研究中，我们利用带简并引物的RT-PCR、DNA测序和RT-PCR产物的频率分析，鉴定出13个CDPK基因在高盐、高甘露醇、干燥和温度胁迫条件下在健康的紫茎甘蓝插枝中活跃表达。12个CDPKs分别为VaCPK1、VaCPK2、VaCPK3、VaCPK9、VaCPK13、VaCPK16、VaCPK20、VaCPK21、VaCPK25、VaCPK26、VaCPK29和VaCPK30，为维科新发现的CDPKs，并获得了它们的完整cdna序列。实时荧光定量RT-PCR结果显示，渗透和温度胁迫下，10个VaCPK基因的mRNA水平有差异上调，而VaCPK3a、VaCPK25和VaCPK30 3个转录本变体的丰度没有显著变化。VaCPK基因在白杨叶片、叶柄、茎、花序、果实和种子中的表达谱分析表明，这些基因具有不同的器官特异性表达模式。非生物胁迫对VaCPK1、2、3、9、13、16、20、21、26和VaCPK29基因表达的刺激作用暗示了它们与葡萄对渗透和温度胁迫的反应有关，而它们器官特异性表达模式的可变性表明这些酶具有不同的生物学功能。(C) 2013爱思唯尔有限公司版权所有。
Calcium-dependent protein kinases (CDPKs), which are important sensors of Ca2+ flux in plants, are known to play essential roles in plant development and adaptation to abiotic stresses. In the present work, we studied expression of CDPK genes under osmotic and temperature stress treatments in wild-growing grapevine Vitis amurensis Rupr., which is known to possess high adaptive potential and a high level of resistance against adverse environmental conditions. In this study, using RT-PCR with degenerate primers, DNA sequencing and frequency analysis of RT-PCR products, we identified 13 CDPK genes that are actively expressed in healthy V. amurensis cuttings under high salt, high mannitol, desiccation, and temperature stress conditions. 12 CDPKs, namely VaCPK1, VaCPK2, VaCPK3, VaCPK9, VaCPK13, VaCPK16, VaCPK20, VaCPK21, VaCPK25, VaCPK26, VaCPK29 and VaCPK30, were novel for Vitaceae, and their full cDNAs were obtained and described. Quantitative real-time RT-PCR demonstrated that mRNA levels of 10 VaCPK genes were differentially up-regulated under the osmotic and temperature stress treatments, while the abundance of 3 VaCPK transcript variants, VaCPK3a, VaCPK25, and VaCPK30, was not markedly changed. Expression profiling of the VaCPK genes in leaves, leaf petioles, stems, inflorescences, berries, and seeds of V. amurensis revealed that the genes exhibit different organ-specific expression patterns. The stimulatory effect of abiotic stress on the expression of the VaCPK1, 2, 3, 9, 13, 16, 20, 21, 26, and VaCPK29 genes is suggestive of their implication in the grapevine response to osmotic and temperature stresses, while the variability in their organ-specific expression patterns indicates that the enzymes perform distinct biological functions. (C) 2013 Elsevier GmbH. All rights reserved.