Genetic Analysis of Rice Neck Blast Resistance in Huang-Huai-Hai Region

doi:10.16819/j.1001-7216.2019.8104

Abstract

Abstract:

【Objective】 To identify the rice blast resistance and the genotype of rice varieties in Huang-Huai-Hai Region, 【Method】the rice blast resistance of the rice varieties collected from five districts of this region was identified for two consecutive years through artificial injection and natural induction in rice blast nursery. The genotypes were analyzed with the functional markers of Pita, Pib, Pi54 and Pikm. And the cluster analysis was carried out according to the rice blast resistance level. 【Result】 85.2% and 65.9% of the tested varieties were sensitive to rice blast through artificial identification and the blast nursery identification, respectively. 84% and 65% of the tested varieties carry Pi54 and Pib, respectively. But the cluster analysis result showed that Pi54 and Pib have low contribution rate of rice blast resistance in this region.【Conclusion】 The rice varieties in the region have low resistance to rice blast, as these genes confer weaker or lost resistance to rice blast. It is crucial to screen new resistant varieties and utilize new resistant genes for breeding.

Key words: rice, rice blast, resistant gene

摘要：

【目的】为明确黄淮海稻区育成水稻品种的稻瘟病抗性及其基因型,【方法】收集了来自本稻区5个不同地区的水稻品种,连续两年通过稻瘟病菌接种鉴定和田间病圃自然诱发对供试品种进行了稻瘟病抗性鉴定,并利用4个抗病基因Pita, Pib, Pi54和Pikm的分子标记,对水稻品种基因型进行了统计分析,并以抗性等级为表型值对品种资源进行聚类分析。【结果】黄淮海稻区水稻品种发病率较高,分别达85.2%和65.9%。Pi54和Pib检出率分别达84%、65%。同时抗性聚类分析表明Pi54和Pib在本稻区的抗性贡献率却很低。【结论】黄淮海稻区水稻品种稻瘟病抗性弱,携带的抗瘟基因的抗性在不断减弱或丧失,需加强稻瘟病主效抗性的发掘、开发与利用。

关键词: 水稻, 稻瘟病, 抗性基因

CLC Number:

S435.111.4⁺1

Yan LIU, Baoxiang WANG, Bo YANG, Tingmu CHEN, Yungao XING, Zhiguang SUN, Bo XU, Ming CHI, Baiguan LU, Jian LI, Jinbo LIU, Zhaowei FANG, Derong QIN, Dayong XU. Genetic Analysis of Rice Neck Blast Resistance in Huang-Huai-Hai Region[J]. Chinese Journal OF Rice Science, 2019, 33(4): 377-382.

刘艳, 王宝祥, 杨波, 陈庭木, 邢运高, 孙志广, 徐波, 迟铭, 卢百关, 李健, 刘金波, 方兆伟, 秦德荣, 徐大勇. 黄淮海稻区水稻品种的穗颈瘟抗性[J]. 中国水稻科学, 2019, 33(4): 377-382.

Figures/Tables 5

References 22

[1]	李俊周, 王书玉, 尹海庆, 姬生栋, 王生轩, 张栩, 陈楠, 赵全志. 黄海稻区优质抗病粳稻“六位一体”育种技术体系的构建与实践. 河南农业科学, 2017, 46(12): 42-47.
	Li J Z, Wang S Y, Yin H Q, Ji S D, Wang S X, Zhang X, Chen N, Zhao Q Z.Construction and practice of Six-in-one Breeding Technology System for japonica rice with high quality and disease resistance in Huang-Huai Region.J Henan Agric Sci, 2017, 46(12): 42-47. (in Chinese with English abstract)
[2]	国家水稻数据中心.稻瘟病主效抗性基因列表 [2019-04-24] .
	National Rice Data Center. List of major resistance genes in rice blast [2019-04-24]. .
[3]	田红刚, 陈红旗, 胡江, 雷财林, 朱旭东, 钱前. 抗稻瘟病基因Pigm导入对寒地粳稻抗病性和产量性状的影响. 沈阳农业大学学报, 2016, 47(5): 520-526.
	Tian H G, Cheng H Q, Hu J, Lei C L, Zhu X D, Qian Q.Effect of introgressed Pigm gene on rice blast resistance and yield traits of japonica rice in cold area. J Shenyang Agric Univ, 2016, 47(5): 520-526. (in Chinese with English abstract)
[4]	张礼霞, 王林友, 范宏环, 王建军. 利用Pigm基因改良粳稻保持系的稻瘟病抗性研究. 核农学报, 2017, 31(3): 424-431.
	Zhang L X, Wang L Y, Fan H H, Wang J J.Study on improving rice blast resistance of japonica maintainer line by introducing Pigm gene.Acta Agric Nucl Sin, 2017, 31(3): 424-431. (in Chinese with English abstract)
[5]	王飞, 王立广, 潘梅瑶, 钮中一, 周勇, 梁国华. 水稻抗稻瘟病Pigm (t)基因的分子标记辅助选择与利用. 华北农学报, 2016, 31(1): 51-56.
	Wang F, Wang L G, Pan M Y, Niu Z Y, Zhou Y, Liang G H.Marker-assisted selection and application of blast resistant gene Pigm(t) in rice.Acta Agric Boreali-Sin, 2016, 31(1): 51-56. (in Chinese with English abstract)
[6]	向小娇, 张建, 郑天清, 徐建龙. 应用分子标记技术改良京作1号的稻瘟病抗性. 植物遗传资源学报, 2017, 17(4): 773-780.
	Xiang X J, Zhang J, Zheng T Q, Xu J L.Improving blast resistance of Jingzuo 1 using molecular marker technique.Plant Genet Resour, 2017, 17(4): 773-780. (in Chinese with English abstract)
[7]	汪文娟, 周继勇, 汪聪颖, 苏菁, 封金奇, 陈炳, 冯爱卿, 杨健源, 陈深, 朱小源. 八个抗稻瘟病基因在华南籼型杂交水稻中的分布. 中国水稻科学, 2017, 31(3): 299-306.
	Wang W J, Zhou J Y, Wang C Y, Su J, Feng J Q, Chen B, Feng A Q, Yang J Y, Chen S, Zhu X Y.Distribution of eight rice blast resistance genes in indica hybrid rice in China.Chin J Rice Sci, 2017, 31(3): 299-306. (in Chinese with English abstract)
[8]	邹茜, 董丽英, 袁平荣, 刘慰华, 苏振喜, 金雁, 寇姝燕. 广谱持久抗病基因Pi40在云南高原粳稻的应用研究. 西南农业学报, 2016, 29(7): 1493-1498.
	Zou Q, Dong L Y, Yuan P R, Liu W H, Su Z X, Jin Y, Kou S Y.Application study on broad-spectrum blast resistance gene Pi40 in Yunnan Plateau rice breeding.Southwest China J Agric Sci, 2016, 29(7): 1493-1498. (in Chinese with English abstract)
[9]	邹德堂, 姜思达, 赵宏伟, 郭丽颖, 孙健, 刘化龙, 辛威. 广谱抗性基因Pi9在黑龙江省水稻品种中的分布. 东北农业大学学报, 2016, 47(7): 1-8.
	Zou D T, Jiang S D, Zhao H W, Guo L Y, Sun J, Liu H L, Xin W.Distribution of broad-spectrum resistance gene Pi9 in the rice cultivars of Heilongjiang Province.J Northeast Agric Univ, 2016, 47(7): 1-8. (in Chinese with English abstract)
[10]	王丽丽, 赵家铭, 马作斌, 郑文静, 马殿荣. 辽宁地区水稻资源抗稻瘟病基因的检测分析. 植物遗传资源学报, 2017, 18(2): 325-339.
	Wang L L, Zhao J M, Ma Z B, Zheng W J, Ma D R.Identificating and analyzing to rice blast resistant genes in rice germplasm resources of Liaoning province.Plant Genet Resour, 2017, 18(2): 325-339. (in Chinese with English abstract)
[11]	Wang Z, Jia Y, Rutger J N.Rapid survey for presence of a blast resistance gene Pita in rice cultivars using the dominant DNA markers derived from portions of the Pita gene.Plant Breed, 2007, 126: 36-42.
[12]	Fjellstrom R, Conaway B, Concetta A, McClung A M. Development of DNA markers suitable for marker assisted selection of three Pi genes conferring resistance to multiple Pyricularia grisea pathotypes.Crop Sci, 2004, 44(5): 1790-1798.
[13]	Ramkumar G, Srinivasarao K, Mohan M K. Development and validation of functional marker targeting an InDel in the major rice blast disease resistance genePi54(Pikh). Mol Breed, 2011(27): 129-135.
[14]	Ashikawa I, Hayashi N, Yamane H, Kanamori H, Wu J, Matsumoto T, Ono K, Yano M.Two adjacent nucleotide-binding site-leucine-rich repeat class genes are required to confer Pik-m-specific rice blast resistance.Genetics, 2008, 180(4): 2267-2276.
[15]	宋兆强, 刘艳, 王宝祥, 王芳权, 迟铭, 刘金波, 陈庭木, 方兆伟, 邢运高, 徐波, 杨波, 杨杰, 徐大勇. 稻瘟病抗性基因Pita、Pib、Pi54和Pik-m的育种利用价值评价. 江苏农业学报, 2017, 33(5): 968-974.
	Song Z Q, Liu Y, Wang B X, Wang F Q, Chi M, Liu J B, Chen T M, Fang Z W, Xing Y G, Xu B, Yang B, Yang J, Xu D Y.Application value of blast resistant genes Pita, Pib, Pi54 and Pik-m in rice breeding. Jiangsu J Agric Sci, 2017, 33(5): 968-974. (in Chinese with English abstract)
[16]	李刚, 袁彩勇, 曹奎荣, 孙祥良, 李军, 王健, 程保山, 罗伯祥, 徐卫军, 唐九友, 储成才. 544份水稻种质稻瘟病抗性鉴定及抗性基因的分布研究. 中国农业大学学报, 2018, 23(5): 22-28.
	Li G, Yuan C Y, Cao K R, Sun X L, Li J, Wang J, Cheng B S, Luo B X, Xu W J, Tang J Y, Chu C C.Evaluation and distribution of the blast resistance genes of 544 rice material.J China Agric Univ, 2018, 23(5): 22-28. (in Chinese with English abstract)
[17]	范方军, 王芳权, 刘永峰, 王军, 朱金燕, 李文奇, 仲维功, 杨杰. Pib、Pita、Pik-m 和Pi54 对水稻穗颈瘟的抗性评价. 华北农学报, 2014, 29(3): 221-226.
	Fan F J, Wang F Q, Liu Y F, Wang J, Zhu J Y, Li W Q, Zhong W G, Yang J.Evaluation of resistance to rice panicle blast with resistant genes Pib, Pita, Pik-m and Pi54.Acta Agric Boreali Sin, 2014, 29(3): 221-226. (in Chinese with English abstract)
[18]	王军, 宫丹妮, 杨杰, 朱金燕, 范方军, 李文奇, 王芳权, 仲维功. 江苏省粳稻品种抗稻瘟病基因型与穗茎瘟抗性分析. 江苏农业学报, 2016, 32(2): 250-256.
	Wang J, Gong D N, Yang J, Z Hu J Y, Fan F J, L I W Q, Wang F Q, Zhong W G. Relationship between rice blast resistance genotypes and neck blast resistance of Japonica rice in Jiangsu province.Jiangsu J Agric Sci, 2016, 32(2): 250-256. (in Chinese with English abstract)
[19]	李刚, 王健, 曹奎荣, 李军, 孙祥良, 程保山, 罗伯祥, 杨子博, 徐卫军,唐久友, 储成才, 袁彩勇. 江苏水稻主导品种的稻瘟病抗性及抗性基因遗传多样性. 湖南农业大学学报: 自然科学版, 2017, 43(3): 238-243.
	Li G, Wang J, Cao K R, Li J, Sun X L, Cheng B S, Luo B X, Yang Z B, Xu W J, Tang J Y, Chu C C, Yuan C Y.Blast resistance of the main rice cultivars in Jiangsu and genetic diversity of blast resistance genes.J Hunan Agric Univ: Nat Sci, 2017, 43(3): 238-243. (in Chinese with English abstract)
[20]	王生轩, 李俊周, 谢瑛, 李勇, 李梦琪, 曹炳武, 赵全志. 河南粳稻抗稻瘟病基因Pi9、Pita和Piz-t的分子检测. 分子植物育种, 2017, 15(3): 951-955.
	Wang S X, Li J Z, Xie Y, Li Y, Li M Q, Cao B W, Zhao Q Z.Molecular detection of rice blast resistance genes Pi9, Pita and Piz-t in Henan japonica rice.Mol Plant Breed, 2017, 15(3): 951-955. (in Chinese with English abstract)
[21]	王亚, 臧之光, 陈献功, 王生轩, 王越涛, 王付华, 付景, 杨文博, 白涛, 尹海庆. 6个抗稻瘟病基因在河南省主要稻种资源中的分布情况. 分子植物育种, 2017, 15(12): 4976-4984.
	Wang Y, Zang Z G, Chen X G, Wang S X, Wang Y T, Wang F H, Fu J, Yang W B, Bai T, Yin H Q.Distribution of six blast resistant genes in the main rice germplasms in Henan Province.Mol Plant Breed, 2017, 15(12): 4976-4984. (in Chinese with English abstract)
[22]	陈涛, 张亚东, 朱镇, 赵春芳, 赵庆勇, 赵凌, 周丽慧, 姚姝, 王才林. Pib和Pita基因在江苏省粳稻中的分布以及与穗茎瘟抗性的关系. 江苏农业学报, 2016, 32(1): 1-8.
	Chen T, Zhang Y D, Zhu Z, Zhao C F, Zhao Q Y, Zhao L, Zhou L H, Yao S, Wang C L.Distribution of Pib and Pita genes in japonica rice of Jiangsu Province and their relationship with neck blast resistance.Jiangsu J Agric Sci, 2016, 32(1): 1-8. (in Chinese with English abstract)

目的基因 Target gene	引物名称 Primer name	引物序列 Sequence(5′-3′)	预期片段长 Production size/bp
Pita	Pita-F	AGCAGGTTATAAGCTAGGCC	1042
Pita	Pita-R	CTACCAACAAGTTCATCAAA	1042
pita	NPita-F	AGCAGGTTATAAGCTAGCTAT	1042
pita	NPita-R	CTACCAACAAGTTCATCAAA	1042
Pib	Pib-F	GAACAATGCCCAAACTTGAGA	365
Pib	Pib-R	GGGTCCACATGTCAGTGAGC	365
pib	NPib-F	TCGGTGCCTCGGTAGTCAGT	803
pib	NPib-R	GGGAAGCGGATCCTAGGTCT	803
Pi54	Pi54-F	CAATCTCCAAAGTTTTCAGG	216/315
Pi54	Pi54-R	GCTTCAATCACTGCTAGACC	216/315
Pikm	Pikm1-F	TGAGCTCAAGGCAAGAGTTGAGGA	174/213
Pikm	Pikm1-R	TGTTCCAGCAACTCGATGAG	174/213
pikm	Pikm2-F	CAGTAGCTGTGTCTCAGAACTATG	290/332
pikm	Pikm2-R	AAGGTACCTCTTTTCGGCCAG	290/332

目的基因 Target gene	引物名称 Primer name	引物序列 Sequence(5′-3′)	预期片段长 Production size/bp
Pita	Pita-F	AGCAGGTTATAAGCTAGGCC	1042
Pita	Pita-R	CTACCAACAAGTTCATCAAA	1042
pita	NPita-F	AGCAGGTTATAAGCTAGCTAT	1042
pita	NPita-R	CTACCAACAAGTTCATCAAA	1042
Pib	Pib-F	GAACAATGCCCAAACTTGAGA	365
Pib	Pib-R	GGGTCCACATGTCAGTGAGC	365
pib	NPib-F	TCGGTGCCTCGGTAGTCAGT	803
pib	NPib-R	GGGAAGCGGATCCTAGGTCT	803
Pi54	Pi54-F	CAATCTCCAAAGTTTTCAGG	216/315
Pi54	Pi54-R	GCTTCAATCACTGCTAGACC	216/315
Pikm	Pikm1-F	TGAGCTCAAGGCAAGAGTTGAGGA	174/213
Pikm	Pikm1-R	TGTTCCAGCAACTCGATGAG	174/213
pikm	Pikm2-F	CAGTAGCTGTGTCTCAGAACTATG	290/332
pikm	Pikm2-R	AAGGTACCTCTTTTCGGCCAG	290/332

分类 Group	抗性等级均值 Resistance grade	所占比例 Percentage / %
分类 Group	抗性等级均值 Resistance grade	Pita	Pib	Pikm	Pi54
Ⅰ	3.2	17	65	12	88
Ⅱ	3.0	0	0	0	100
Ⅲ	2.0	58	67	8	58

分类 Group	抗性等级均值 Resistance grade	所占比例 Percentage / %
分类 Group	抗性等级均值 Resistance grade	Pita	Pib	Pikm	Pi54
Ⅰ	3.2	17	65	12	88
Ⅱ	3.0	0	0	0	100
Ⅲ	2.0	58	67	8	58

抗病基因 Genotype	江苏 Jiangsu		安徽 Anhui		河南 Henan		山东 Shandong		京津地区 Beijing and Tianjin
抗病基因 Genotype	材料数 n	比例Percent /%	材料数 n	比例 Percent /%	材料数 n	比例Percent /%	材料数 n	比例Percent /%	材料数 n	比例Percent /%
无目标基因None	0	0.0	0	0.0	4	20.0	0	0.0	0	0.0
Pita	1	2.9	0	0.0	0	0.0	0	0.0	0	0.0
Pib	2	5.9	0	0.0	0	0.0	1	5.9	0	0.0
Pi54	4	11.8	1	33.3	6	30.0	2	11.8	5	35.7
Pita+Pib	2	5.9	1	33.3	0	0.0	0	0.0	3	21.4
Pita+Pi54	0	0.0	0	0.0	5	25.0	1	5.9	0	0.0
Pib+Pi54	18	52.9	0	0.0	4	20.0	9	52.9	2	14.3
Pib+Pikm	1	2.9	1	33.3	0	0.0	0	0.0	0	0.0
Pi54+Pikm	0	0.0	0	0.0	0	0.0	3	17.6	0	0.0
Pita+Pi-b+Pi54	4	11.8	0	0.0	0	0.0	1	5.9	2	14.3
Pib+Pi54+Pikm	1	2.9	0	0.0	0	0.0	0	0.0	2	14.3
Pita+Pib+Pi54+Pikm	1	2.9	0	0.0	1	5.0	0	0.0	0	0.0