玉米 ZmKNOLLE基因的克隆及其增强烟草耐盐性研究
研究表明 SNARE蛋白（可溶性N-己基顺丁烯二酰亚胺敏感因子结合蛋白）参与植物抗逆性；然而，在对KNOLLE耐盐的调节作用研究还比较少。在这项研究中，我们描述了1个玉米SNARE蛋白ZmKNOLLE，其中包含一个SNARE特定的结构域。它的cDNA为945 bp，编码一个314个氨基酸的蛋白，预测分子量为34.54 kD，等电点（pI）为7.02。在ZmKNOLLE蛋白C端有一个跨膜结构域。实时定量PCR结果显示，ZmKNOLLE能被盐和H2O2强烈诱导。通过农杆菌介导把ZmKNOLLE基因转化到烟草（W38）中，启动子为CaMV 35S。在盐胁迫下，与野生型相比，转基因烟草植株表现出较长的主根和较高的萌发率，相对含水量、POD、SOD活性较高；而MDA含量较少。这些结果表明转基因烟草比野生型烟草对盐胁迫有更强的耐受性。这些结果将有助于玉米SNARE蛋白在抗逆性响应方面的进一步研究，并为玉米抗性育种提供极好的候选基因。
英文摘要:
Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are involved in plant resistance; however, the role of KNOLLE in the regulation of salt resistant is not well known. In this study, we characterized a plant-specific SNARE in maize, ZmKNOLLE, which contains a SNARE domain. Its cDNA was 945 bp and encoded a putative protein of 314 amino acids with a predicted molecular mass of 34.54 kD and an isoelectric point （PI） of 7.02. A transmembrane domain was found at the C-terminal region of ZmKNOLLE protein. Real-time PCR showed that the transcript of ZmKNOLLE was strongly induced by salinity and H2O2 to some extent. The ZmKNOLLE gene was transformed into tobacco cultivar Wisconsin 38 （W38） by Agrobacterium mediated under the control of the CaMV 35S promoter. Under salt stresses, transgenic tobacco lines carrying ZmKNOLLE gene developed the strong primary root and more germination rate, higher RWC, POD and SOD activity, less MDA content, and performed higher tolerance to these stresses than the wide-type tobaccos, The primary function verification showed that overexpression of ZmKNOLLE gene in tobaccos enhanced their tolerance to salt stresses. This study will help further research of maize SNARE protein reactions in the role of stress resistance, and bring maize resistance breeding excellent candidate genes.
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