Mechanism Analysis of Rice Resistance Protein OsRRK1 Againstthe Brown Planthopper

doi:10.16819/j.1001-7216.2020.0406

Abstract

Abstract:

【Objective】OsRRK1 (Rop-interacting receptor-like kinase 1) protein in rice is the first RLCK VI family protein in rice that has been studied.It plays an important role inregulating the curling of rice leaves and the defense against brown planthoppers. However, the mechanism of its regulation of resistance to the brown planthopperremains unclear. Weattempt to further explore the mechanism of OsRRK1gene resistance to the brown planthoppers.【Method】The interaction between OsRRK1 protein and OsLecRK (lectin-like receptor kinase), OsLecRK1, OsLecRK2 and OsLecRK3 was analyzed by yeast two-hybrid method.OsLecRK is a candidate gene forBPH15 range of anti-brown planthopper, and is an important member of rice innate immune system. OsLecRK is involved innot only rice innate immune response, including resistance to brown planthoppers, white leaf blight and rice blast, but alsothe development of rice. The gene clusters of OsLecRK1, OsLecRK2 and OsLecRK3 constitute the anti-brown planthopper gene BPH3. Rice containing the BPH3 gene has a broad spectrum and lasting insect resistance. Meanwhile, DNAMAN software was used to analyze the homology of OsLecRK with OsLecRK1, OsLecRK2 and OsLecRK3.【Result】The analysis results of DNAMAN showed that OsLecRK has high homology with OsLecRK1, OsLecRK2 and OsLecRK3, and the proteinsshared more than 50% identity.The results of yeast two-hybrid analysis showed that OsRRK1 interacted with OsLecRK, OsLecRK2 and OsLecRK3.【Conclusion】OsRRK1 participates in the process of resistance to the brownplanthoppersthroughinteracting with BPH15 and BPH3 candidate genes.

Key words: rice, OsRRK1, brown planthopper, yeast two-hybrid system

摘要： 目的 OsRRK1（Rop-interacting receptor-like kinase 1）蛋白是水稻中第一个被研究的类胞质受体激酶RLCKⅥ家族蛋白。在调控水稻叶片的卷曲和对褐飞虱的防御中都起着重要作用,但是其调控机理尚不清楚。本研究试图进一步探究OsRRK1基因抗褐飞虱的机理。方法利用酵母双杂交的方法分析OsRRK1蛋白与OsLecRK（lectin-like receptor kinase）、OsLecRK1、OsLecRK2、OsLecRK3的互作关系,OsLecRK是抗褐飞虱基因BPH15区间的候选基因,是水稻先天免疫系统中一个重要成员,既参与水稻先天免疫反应,包括对褐飞虱、白叶枯病和稻瘟病的抗性,又参与水稻的发育过程。OsLecRK1、OsLecRK2、OsLecRK3的基因簇组成抗褐飞虱基因BPH3,含BPH3基因的水稻具有广谱、持久的抗虫性。同时利用DNAMAN软件分析OsLecRK蛋白与OsLecRK1、OsLecRK2、OsLecRK3的同源性。结果 DNAMAN的分析结果显示OsLecRK与OsLecRK1、OsLecRK2、OsLecRK3的同源性都很高,蛋白一致性在50%以上;酵母双杂交结果显示OsRRK1与OsLecRK、OsLecRK2和OsLecRK3互作。结论 OsRRK1通过与抗褐飞虱基因BPH15和BPH3候选基因的相互作用参与水稻抗褐飞虱过程。

关键词: 水稻, OsRRK1, 褐飞虱, 酵母双杂交系统

CLC Number:

Q786

Yinhua MA, Pingfang LI, Wenjing DONG, Songwang YI, Fang YANG, Bo DU, Chenzhong JIN. Mechanism Analysis of Rice Resistance Protein OsRRK1 Againstthe Brown Planthopper[J]. Chinese Journal OF Rice Science, 2020, 34(6): 512-519.

马银花, 李萍芳, 董文静, 易松望, 杨芳, 杜波, 金晨钟. 水稻抗性蛋白OsRRK1抗褐飞虱机理分析[J]. 中国水稻科学, 2020, 34(6): 512-519.

Figures/Tables 6

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引物 Primer	引物序列（5'-3'） Primer sequence（5'-3'）	限制性内切酶酶切位点 Restriction endonuclease site
AD-LecRKF	CGGAATTCATGGTTGCTCTGCTACTCTTT	EcoR I
AD-LecRKR	CGGAATTCTTAATGAGAAAACAGAGGATTACATTCC	EcoR I
AD-LecRK1F	TCCCCCGGGATGGTTGCTCTGCTACTCTTTCCC	Sma I
AD-LecRK1R	TCCCCCGGGCTATGGAAGTGAGCTGATGAAGG	Sma I
AD-LecRK2F	TCCCCCGGGTATGGCACCTATCCTGTTCTTGCCC	Sma I
AD-LecRK2R	TCCCCCGGGTCATGCGAGTGAACTGATATAGG	Sma I
AD-LecRK3F	TCCCCCGGGTATGGCTCATCTCCTGTTCTTGCCC	Sma I
AD-LecRK3R	TCCCCCGGGTCATGCAAGTGAACTGATATAGG	SmaI
BK-OsRRK1F	CGGAATTCATGAGGCCTCTGTACCTGCGCAGC	EcoR I
BK-OsRRK1R	CGGAATTCCTAATTGCTCAAAGATGATGAGCA	EcoR I
T7	TAATACGACTCACTATAGGGCGA
3′AD	AGATGGTGCACGATGCACAG
3′BD	TTTTCGTTTTCCTAAGAGTC

引物 Primer	引物序列（5'-3'） Primer sequence（5'-3'）	限制性内切酶酶切位点 Restriction endonuclease site
AD-LecRKF	CGGAATTCATGGTTGCTCTGCTACTCTTT	EcoR I
AD-LecRKR	CGGAATTCTTAATGAGAAAACAGAGGATTACATTCC	EcoR I
AD-LecRK1F	TCCCCCGGGATGGTTGCTCTGCTACTCTTTCCC	Sma I
AD-LecRK1R	TCCCCCGGGCTATGGAAGTGAGCTGATGAAGG	Sma I
AD-LecRK2F	TCCCCCGGGTATGGCACCTATCCTGTTCTTGCCC	Sma I
AD-LecRK2R	TCCCCCGGGTCATGCGAGTGAACTGATATAGG	Sma I
AD-LecRK3F	TCCCCCGGGTATGGCTCATCTCCTGTTCTTGCCC	Sma I
AD-LecRK3R	TCCCCCGGGTCATGCAAGTGAACTGATATAGG	SmaI
BK-OsRRK1F	CGGAATTCATGAGGCCTCTGTACCTGCGCAGC	EcoR I
BK-OsRRK1R	CGGAATTCCTAATTGCTCAAAGATGATGAGCA	EcoR I
T7	TAATACGACTCACTATAGGGCGA
3′AD	AGATGGTGCACGATGCACAG
3′BD	TTTTCGTTTTCCTAAGAGTC