Cloning and Expression of the Gene OC-XII Conferring Resistance to Aphelenchoides besseyi in Rice

doi:10.3969/j.issn.1001G7216.2015.06.013

Chinese Journal OF Rice Science ›› 2015, Vol. 29 ›› Issue (6): 658-664.DOI: 10.3969/j.issn.1001G7216.2015.06.013

Cloning and Expression of the Gene OC-XII Conferring Resistance to Aphelenchoides besseyi in Rice

Feng WANG, Qiao-li CHEN, Dan-lei LI^*(), Ya-ming LING, Bo-wen WANG, Rui-zhi ZHANG, Yan-yan JIA, Yong-hang LI, Yu-ying PEI, Xin-ming LU

Department of Forestry,Northeast Forestry University, Harbin 150040, China

Received:2014-11-11 Revised:2015-05-18 Online:2015-10-25 Published:2015-11-10
Contact: Dan-lei LI
About author:
^*Corresponding author：E-mail:danleili@126.com

水稻抗干尖线虫基因OC-XII的克隆及表达研究

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

东北林业大学林学系, 哈尔滨 150040

通讯作者: 李丹蕾
作者简介:
^*通讯录作者：E-mail:danleili@126.com
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)

Abstract

Abstract:

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

摘要：

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。

关键词: 半胱氨酸蛋白酶, 抑制剂, 水稻干尖线虫, 组织蛋白酶B

CLC Number:

Feng WANG, Qiao-li CHEN, Dan-lei LI, Ya-ming LING, Bo-wen WANG, Rui-zhi ZHANG, Yan-yan JIA, Yong-hang LI, Yu-ying PEI, Xin-ming LU. Cloning and Expression of the Gene OC-XII Conferring Resistance to Aphelenchoides besseyi in Rice[J]. Chinese Journal OF Rice Science, 2015, 29(6): 658-664.

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.

Figures/Tables 5

References 24

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物种 Species	GenBank号 NCBI No.	蛋白 Protein	E值 E-Value
大麦 Hordeum vulgare	CAG38124	Cystatin Hv-CPI3	4×10^-60
玉米 Zea mays	XP_008647964	Cystatin 12	9×10^-55
谷子 Setaria italica	XP_004962105	Cysteine protease inhibitor 3	7×10^-38
海枣 Phoenix dactylifera	XP_008795132	Cysteine protease inhibitor 12	2×10^-30
杨树 Populus balsamifera	ACO39758	Cysteine inhibitor 1	9×10^-26
中粒咖啡 Coffea canephora	CDP04913	Cystatin	2×10^-25
大豆 Glycine max	NP_001237734	Uncharacterized protein	4×10^-23

物种 Species	GenBank号 NCBI No.	蛋白 Protein	E值 E-Value
大麦 Hordeum vulgare	CAG38124	Cystatin Hv-CPI3	4×10^-60
玉米 Zea mays	XP_008647964	Cystatin 12	9×10^-55
谷子 Setaria italica	XP_004962105	Cysteine protease inhibitor 3	7×10^-38
海枣 Phoenix dactylifera	XP_008795132	Cysteine protease inhibitor 12	2×10^-30
杨树 Populus balsamifera	ACO39758	Cysteine inhibitor 1	9×10^-26
中粒咖啡 Coffea canephora	CDP04913	Cystatin	2×10^-25
大豆 Glycine max	NP_001237734	Uncharacterized protein	4×10^-23

Cloning and Expression of the Gene OC-XII Conferring Resistance to Aphelenchoides besseyi in Rice

水稻抗干尖线虫基因OC-XII的克隆及表达研究

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References 24

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