云南疣粒野生稻稻瘟病抗性

doi:10.11983/CBB17112

植物学报 ›› 2018, Vol. 53 ›› Issue (4): 477-486.DOI: 10.11983/CBB17112 cstr: 32102.14.CBB17112

云南疣粒野生稻稻瘟病抗性

王韵茜, 苏延红, 杨睿, 李鑫, 李晶, 曾千春^*(), 罗琼^*()

云南农业大学, 农业生物多样性与病虫害控制教育部重点实验室, 云南生物资源保护与利用国家重点实验室, 昆明 650201

收稿日期:2017-06-05 接受日期:2017-08-07 出版日期:2018-07-01 发布日期:2018-09-11
通讯作者: 曾千春,罗琼
作者简介:† 共同第一作者。
基金资助:
国家重点研发计划(No.2016YFD0100600)和国家自然科学基金-云南联合基金(No.U1302261)

Rice Blast Resistance of Wild Rice in Yunnan

Wang Yunqian, Su Yanhong, Yang Rui, Li Xin, Li Jing, Zeng Qianchun^*(), Luo Qiong^*()

State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Ministry of Education Key Laboratory of Agriculture Biodiversity for Plant Disease Management, Yunnan Agricultural University, Kunming 650201, China

Received:2017-06-05 Accepted:2017-08-07 Online:2018-07-01 Published:2018-09-11
Contact: Zeng Qianchun,Luo Qiong
About author:† These authors contributed equally to this paper

摘要/Abstract

摘要： 野生稻(Oryza rufipogo)保存有许多栽培稻(O. sativa)不具备或已经消失的优异基因资源, 是扩大栽培稻遗传背景、改良产量与品质、提高抗病虫害及抗逆境能力的重要基因库。疣粒野生稻(O. meyeriana)是中国3种野生稻资源之一, 主要分布在云南。为进一步了解其稻瘟病抗性, 首先利用来自不同稻作区的稻瘟病菌株, 通过注射接种法对疣粒野生稻进行系统的稻瘟病抗性鉴定, 发现疣粒野生稻对接种的所有稻瘟病菌株都感病。进一步采用3'/5' RACE方法, 从疣粒野生稻中克隆了水稻同源基因Pid2和Pid3, 并构建过表达转基因株系对基因功能进行了研究。结果表明, Pid2和Pid3与疣粒野生稻中同源基因间在DNA和氨基酸水平上有较大的序列差异, 过表达转基因的日本晴植株对稻瘟病菌的敏感性与对照相似。推测疣粒野生稻在自然接种条件下, 表现出的抗稻瘟病表型很可能是其旱生叶片结构特征形成了对稻瘟病菌侵染的天然屏障。对控制疣粒野生稻这一类性状基因资源的挖掘和利用, 有利于优良抗性水稻品种的培育。研究结果为疣粒野生稻的研究利用提供了新信息和新思路。

关键词: 疣粒野生稻, 稻瘟病抗性, 抗病基因, 基因克隆, 稻瘟病菌

Abstract: Wild species of Oryza contain numerous genes of economic importance and are being used as an important cultivated rice gene pool to expand the genetic background, improve yield and quality, and increase the ability for resistance or tolerance to pests as well as bioic and abiotic stresses. O. meyeriana is one of three native wild rice species in China, mainly distributed in Yunnan province. To further understand the disease resistance to rice blast, we performed a systematic investigation of rice blast resistance with O. meyeriana. O. meyeriana plants were inoculated by injection with multiple blast strains from different rice planting areas. Then, we cloned and functionally identified Pid2 and Pid3 orthologous genes in O. meyeriana. O. meyeriana was sensitive to all blast isolates used in this experiment. The DNA and amino acid sequences significantly differed between Pid2 and its orthologs in O. meyeriana, as did Pid3 and its orthologs. Nipponbare plants overexpressing the orthologous genes of Pid2 and Pid3 showed mildly enhanced susceptibility to Magnaporthe oryzae. The orthologs of blast-resistant genes in O. meyeriana may be susceptible alleles. O. meyeriana exhibited resistance to rice blast under natural inoculation likely due to a xeromorphic leaf structure that reduces susceptibility to M. oryzae. Characterization of these genes will help breed elite rice. Our study provides new information and ideas for research and utilization of O. meyeriana.

Key words: wild rice (Oryza meyeriana), rice blast resistance, R gene, gene cloning, Magnaporthe oryzae

王韵茜, 苏延红, 杨睿, 李鑫, 李晶, 曾千春, 罗琼. 云南疣粒野生稻稻瘟病抗性. 植物学报, 2018, 53(4): 477-486.

Wang Yunqian, Su Yanhong, Yang Rui, Li Xin, Li Jing, Zeng Qianchun, Luo Qiong. Rice Blast Resistance of Wild Rice in Yunnan. Chinese Bulletin of Botany, 2018, 53(4): 477-486.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB17112

https://www.chinbullbotany.com/CN/Y2018/V53/I4/477

图/表 4

图1 疣粒野生稻稻瘟病抗性鉴定(A) 注射接种; (B) 喷雾接种; (C) 分别注射接种ZB15和Zhong-10-8-14

Figure 1 Identification of blast resistance in Oryza meyeriana(A) Injection inoculation; (B) Spray inoculation; (C) Inoculated by blast strains ZB15 and Zhong-10-8-14, respectively

图2 OmPid2与水稻同源蛋白的氨基酸序列比对OmPid2: 疣粒野生稻; R-Pid2: 地谷; S-pid2: 日本晴。信号肽结构域(21-52氨基酸、B凝集素结构域(68-184氨基酸)、PAN结构域(355-436氨基酸)和丝氨酸-苏氨酸激酶(STK)结构域(519-789氨基酸)在图中已用下划线标注。箭头示区分稻瘟病抗病基因Pid2抗感等位的关键氨基酸。

Figure 2 Amino acid sequence alignment of OmPid2 and orthologs in riceOmPid2: Oryza meyeriana; R-Pid2: Digu; S-pid2: Nipponbare. Signal peptide domain (21-52 amino acids), B-lectin domain (68-184 amino acids), PAN domain (355-436 amino acids) and Serine/threonine kinase (STK) domain (519-789 amino acids) are underlined. Arrow indicates the amino acid that distinguishes resistant and susceptible alleles of rice blast resistance gene Pid2.

图3 OmPid3与水稻同源蛋白的氨基酸序列比对OmPid3: 疣粒野生稻; R-Pid3: 地谷; S-pid3: 日本晴。核酸结合位点(NBS)结构域(158-445氨基酸)和亮氨酸重复(LRR)结构域(563-923氨基酸)在图中已用下划线标注。

Figure 3 Amino acid sequence alignment of OmPid3 and orthologs in riceOmPid3: Oryza meyeriana; R-Pid3: Digu; S-pid3: Nipponbare. Nucleotide-binding site (NBS) domain (158-445 amino acids) and Leucine-rich repeat (LRR) domain (563-923 amino acids) are underlined.

图4 转基因水稻植株的鉴定OmPid2 (A)和OmPid3 (B)过表达转基因株系潮霉素和基因引物分子检测; OmPid2 (C)和OmPid3 (D)过表达转基因株系喷雾接种抗性鉴定; OmPid2和OmPid3 (E)过表达转基因株系注射接种ZB15和Zhong-10-8-14抗性鉴定。HYG: 潮霉素; Di: 水稻地谷; N: 野生型对照日本晴

Figure 4 Characterisation of transgenic rice linesMolecular detection of transgenic rice lines with Hygromycin primer, OmPid2 gene-specific primer (A) and OmPid3 gene-specific primer (B); Blast resistance idetification of OmPid2 (C) and OmPid3 (D) overexpressing transgenic rice lines inoculated by blast strains; Overexpressing transgenic rice lines OmPid2 and OmPid3 (E) were inoculated by blast strains ZB15 and Zhong-10-8-14, respectively. HYG: Hygromycin; Di: The resistant rice variety Digu; N: Wild-type susceptible rice variety Nipponbare

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云南疣粒野生稻稻瘟病抗性

Rice Blast Resistance of Wild Rice in Yunnan

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