*通讯录作者：E-mail:danleili@126.com
收稿日期: 2014-11-11
修回日期: 2015-05-18
网络出版日期: 2015-11-10
基金资助
中央高校基本科研业务费专项资金资助项目(2572015EY02,DL13BA01,2572015CA09)黑龙江省博士后基金资助项目(LBH-Z11248)高等学校博士学科点新教师基金资助项目(20100062120002)大学生创新性实验计划资助项目(201410225163)
Cloning and Expression of the Gene OC-XII Conferring Resistance to Aphelenchoides besseyi in Rice
*Corresponding author：E-mail:danleili@126.com
Received date: 2014-11-11
Revised date: 2015-05-18
Online published: 2015-11-10
抗性验证试验表明籼稻大白谷抗水稻干尖线虫侵染。为进一步探究此抗性与病程相关蛋白——半胱氨酸蛋白酶抑制剂(Cystatin)的相关性,基于水稻基因组数据克隆籼稻大白谷Cystatin家族的OC-XII基因,分别进行OC-XII、Japonica-OC-XII和Indica-OC-XII蛋白质三维结构构建,并与水稻干尖线虫组织蛋白酶B(Cathepsin B)进行分子对接以验证OC-XII基因编码蛋白对Cathepsin B的抑制功能。通过荧光定量qPCR验证OC-XII基因在线虫侵染前后的表达特性,OC-XII蛋白可以与水稻干尖线虫Cathepsin B蛋白质特异结合,线虫侵染后OC-XII基因在浸染12h~2d期间表达量持续上调,侵染3d后下调。表明OC-XII基因在大白谷抗水稻干尖线虫侵染过程中发挥功能,具有对植物寄生线虫进行生物防治的潜力,有必要对其进行深入研究。
王峰, 陈俏丽, 李丹蕾, 零雅茗, 王博文, 张瑞芝, 贾岩岩, 李永航, 裴雨滢, 昕明 . 水稻抗干尖线虫基因OC-XII的克隆及表达研究[J]. 中国水稻科学, 2015 , 29(6) : 658 -664 . DOI: 10.3969/j.issn.1001G7216.2015.06.013
The previous resistance verification indicated that Dabaigu (Oryza sativa ssp. indica) resists the infection of Aphelenchoides bessey. To reveal the interaction between the resistance and the pathogenesis-related protein Cystatin, the gene OC-XII was cloned, the docking between OC-XII and A. besseyi Cathepsin B was done, and the expression of OC-XII was tested by quantitative real-time PCR. Results showed that OC-XII bound with Cathepsin B specifily. The expression level of OC-XII was upregulated from 12 hours after infection to 2 days after infection, and downregulated 3 days after infection. These results indicated that OC-XII was involved in the resistance to A. besseyi. The gene has the potential for biological control of plant-parasitic nematodes, and it is necessary to conduct in-depth research.
Key words： cystatin; inhibitor; Aphelenchoides besseyi; Cathepsin B
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