水稻抗稻瘟病基因Pi47的精细定位和候选基因分析

doi:10.3724/SP.J.1006.2018.00977

Abstract

Abstract:

Continuous mapping and cloning new blast resistance genes provided an important approach for revealing the molecular mechanism of resistance to rice disease and breeding new varieties resistant to rice blast. Previously, the blast resistance gene Pi47 was mapped between SSR markers RM224 and RM2956 on chromosome 11, from a broad-spectrum and durable resistance native cultivar Xiangzi 3150 in Hunan province. In this study, the fine mapping and candidate gene prediction were performed, showing that Pi47 was mapped in an interval of 0.24 cM between CAPS markers S32 and K33 with 171.2 kb on Nipponbare reference genome, and further narrowed in an interval of 67.8 kb between markers SC12 and K33, through background screening to Pi47 monogenic lines and susceptible parent CO39 with six STS markers. Eight genes were predicted in this region on the reference genome, among them two encoded NBS-LRR resistance-like proteins, which probably were Pi47 functional candidate genes. A blast resistance spectrum evaluation, using Pi47 monogenic lines and four near-isogenic lines with the Pik allelic gene Pik, Pikm, Pikh, and Pikp across Pi47 region, revealed that Pi47 shared different resistance spectra with these four alleles. These results shed light on further molecular cloning of Pi47, and the molecular marker will be useful for molecular maker assisted selection to breed new resistant cultivars.

Key words: rice, rice blast, resistance gene, Pi47, fine mapping

Xiang-Yi XIAO,Xue-Tao SHI,Hao-Wen SHENG,Jin-Ling LIU,Ying-Hui XIAO. Fine Mapping and Candidate Gene Analysis of Rice Blast Resistance Gene Pi47[J].Acta Agronomica Sinica, 2018, 44(7): 977-987.

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株系 Line	接种菌株193-1-1的抗性表型 Phenotype inoculated with isolate 193-1-1	Pi47连锁标记 Markers linked to Pi47			Pi48连锁标记 Markers linked to Pi48
株系 Line	接种菌株193-1-1的抗性表型 Phenotype inoculated with isolate 193-1-1	RM206	RM224	RM5926	RM3102	RM1337	RM7102
CX18	R	+	+	+	-	-	-
CX36	R	+	+	+	-	-	-
CX40	R	+	+	+	-	-	-
CX52	R	+	+	+	-	-	-
CX66	R	+	+	+	-	-	-
CX67	R	+	+	+	-	-	-
CX70	R	+	+	+	-	-	-
CX147	R	+	+	+	-	-	-
CX152	R	+	+	+	-	-	-
CX155	R	+	+	+	-	-	-
CX166	R	+	+	+	-	-	-
CX279	R	+	+	+	-	-	-

F₂群体 F₂ population	抗病单株 Resistant individuals	感病单株 Susceptible individuals	作图群体株数 Individual number of mapping population	卡方值 χ²for 3:1 ratio
CX18/CO39 F₂	1040	329	308	0.68
CX52/CO39 F₂	1116	340	247	2.21
CX155/CO39 F₂	1512	310	0	62.0
CX279/CO39 F₂	3820	1221	1132	1.66
总数 Total	/	/	1687	/

群体 Population	感病单株数目 No. of susceptible individuals	重组子分布 Distribution of recombinants
群体 Population	感病单株数目 No. of susceptible individuals	RM224	S11, S4, S32	S10	K33	K107	K134	RM5926
CX279/CO39F₂	1132	20	4	0	1	1	8	56
CX18/CO39F₂	308	5	2	0	0	0	1	8
CX52/CO39F₂	247	2	1	0	0	0	1	11
合计 Total	1687	27	7	0	1	1	10	75

标记 Marker	引物顺序 Primer sequence (5°-3°)	物理位置 Physical position	扩增大小 Expect size	酶 Enzyme	标记类型 Marker type
RM224	F: TCGATCGATCTTCACGAGG R: TGCTATAAAAGGCATTCGGG	27673251 -27673372	122	/	SSR
S11	F: TCTTTGAACAAGAGGACCAG R: AATTACCGAATTACTTAGCG	27824419 -27825439	1021	Mva I or Hinf I	CAPS
S4	F: CGAATGATGACGAGGACCCG R: ATCAGGCACGGCAGCAGAAG	27827786 -27828843	1058	Hinf I or Rsa I	CAPS
S32	F: GCCCAGTAACCGTAGGAGTC R: TTTGATGAGCGGCAAAGTAG	27835604 -27836295	692	Msp I	CAPS
S10	F: AGAGCAAGAAGGGCACGGTAT R: AGAATGGTCCTCCTGACATCG	27992866 -27993970	1105	Rsa I	CAPS
K33	F: TTTGGTGCTCCTCTTACGGG R: TCGGAGTTCACAGAGCCAAG	28005690 -28006842	1153	Hinf I	CAPS
K107	F: ACATCAATGGCTACAACT R: TGCTAACGGTGCTGGTAT	28012753 -28012940	184/188	/	STS
K134	F: GATGGCGAGATGGTTGTC R: GCCTTTGAGATAGGGATTGC	28163563 -28163742	180/168	/	STS
RM5926	F: TAGGTCCATCCAAATCTCGATCC R: TGGCAGAGGAGATTAGAGTAATACGG	28957564 -28957858	295	/	SSR
S6	F: TCAACCGAACGCAAGAATCAC R: GGCCCTCCAGCTCATCTACCT	27841529 -27842431	903	/	STS
SC1	F: CCCCACAATCCCACGAGAC R: CGGGCCGACAATCCAATAC	27848711 -27849635	925	/	STS
SC3	F: TTTCCCACCGATGACCCAG R: GCTCGCCATCCTTCCCTTT	27879005 -27879906	902	/	STS
SC6	F: TATTGGTAAGTGGTAACGGGTGA R: GCAGATTCGAGGAGGAAGG	27891111 -27892550	1440	/	STS
SC7	F: TGGCGTGGCGGTTTGGTAG R: GCTGGTTGGGCATGGGTTG	27909896 -27910602	707	/	STS
SC12	F: GGGCAACAAGCGAGCCATAA R: GTGGTGAGGCAGCGGAACAG	27939030 -27940473	1444	/	STS

基因名称 Gene name	推测功能 Putative functions
LOC_Os11g46150	Expressed protein
LOC_Os11g46180	Transposon protein
LOC_Os11g46190	Transposon protein
LOC_Os11g46200	Leucine rich repeat family protein, expressed
LOC_Os11g46210	NB-ARC domain containing protein, expressed
LOC_Os11g46220	Hypothetical protein
LOC_Os11g46230	Tetratricopeptide repeat domain containing protein
LOC_Os11g46240	Retrotransposon protein

Fine Mapping and Candidate Gene Analysis of Rice Blast Resistance Gene Pi47

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基因 Gene	Pik1-KA	Pi7-1	Piks-1	Pik-2	Pikp-1	Pikm1-TS	Pi1-5
Pi47-1 DNA	0.997	0.975	0.998	0.997	0.975	0.998	0.997
Pi47-1蛋白 Pi47-1 protein	0.987	0.947	0.993	0.989	0.946	0.994	0.995

基因 Gene	Pik2-KA	Pi7-2	Piks-2	Pik-1	Pikp-2	Pikm2-TS	Pi1-6
Pi47-2 DNA	0.999	0.998	0.999	0.999	0.998	0.999	1.000
Pi47-2蛋白 Pi47-2 protein	0.999	0.996	0.999	0.999	0.996	0.999	1.000

稻瘟菌小种 Isolate	来源 Origin	水稻品种 Rice cultivar
稻瘟菌小种 Isolate	来源 Origin	CO39	CX18 (Pi47)	IRBLk-Ka (Pik)	IRBLkm-Ts (Pikm)	IRBLkh-K3 (Pikh)	IRBLkp-K60 (Pikp)
318-2	中国湖南 Hunan, China	S	R	R	R	R	R
CHL473	中国湖南 Hunan, China	S	R	R	R	R	R
X2007A-3	中国湖南 Hunan, China	S	R	R	R	R	S
CHL440	中国湖南 Hunan, China	S	R	R	R	R	S
CHL438	中国湖南 Hunan, China	S	R	R	R	R	R
X2007A-7	中国湖南 Hunan, China	S	R	R	R	R	R
195-2-2	中国湖南 Hunan, China	S	S	R	R	R	S
87-4	中国湖南 Hunan, China	S	R	R	R	R	R
193-1-1	中国湖南 Hunan, China	S	R	R	R	R	R
110-2	中国湖南 Hunan, China	S	R	R	R	R	R
236-1	中国湖南 Hunan, China	S	R	R	R	R	R
RB4	中国广东 Guangdong, China	S	R	R	R	R	S
RB6	中国广东 Guangdong, China	S	R	R	R	R	S
RB18	中国福建 Fujian, China	S	R	S	R	R	S
RB19	中国福建 Fujian, China	S	R	R	R	R	R
RB12	中国 China	MR	MR	MR	MR	MR	MR
KOH	日本 Japan	S	S	S	S	S	S

[1]	TIAN Tian, CHEN Li-Juan, HE Hua-Qin. Identification of rice blast resistance candidate genes based on integrating Meta-QTL and RNA-seq analysis [J]. Acta Agronomica Sinica, 2022, 48(6): 1372-1388.
[2]	ZHENG Chong-Ke, ZHOU Guan-Hua, NIU Shu-Lin, HE Ya-Nan, SUN wei, XIE Xian-Zhi. Phenotypic characterization and gene mapping of an early senescence leaf H5(esl-H5) mutant in rice (Oryza sativa L.) [J]. Acta Agronomica Sinica, 2022, 48(6): 1389-1400.
[3]	ZHOU Wen-Qi, QIANG Xiao-Xia, WANG Sen, JIANG Jing-Wen, WEI Wan-Rong. Mechanism of drought and salt tolerance of OsLPL2/PIR gene in rice [J]. Acta Agronomica Sinica, 2022, 48(6): 1401-1415.
[4]	ZHENG Xiao-Long, ZHOU Jing-Qing, BAI Yang, SHAO Ya-Fang, ZHANG Lin-Ping, HU Pei-Song, WEI Xiang-Jin. Difference and molecular mechanism of soluble sugar metabolism and quality of different rice panicle in japonica rice [J]. Acta Agronomica Sinica, 2022, 48(6): 1425-1436.
[5]	YAN Jia-Qian, GU Yi-Biao, XUE Zhang-Yi, ZHOU Tian-Yang, GE Qian-Qian, ZHANG Hao, LIU Li-Jun, WANG Zhi-Qin, GU Jun-Fei, YANG Jian-Chang, ZHOU Zhen-Ling, XU Da-Yong. Different responses of rice cultivars to salt stress and the underlying mechanisms [J]. Acta Agronomica Sinica, 2022, 48(6): 1463-1475.
[6]	YANG Jian-Chang, LI Chao-Qing, JIANG Yi. Contents and compositions of amino acids in rice grains and their regulation: a review [J]. Acta Agronomica Sinica, 2022, 48(5): 1037-1050.
[7]	DENG Zhao, JIANG Nan, FU Chen-Jian, YAN Tian-Zhe, FU Xing-Xue, HU Xiao-Chun, QIN Peng, LIU Shan-Shan, WANG Kai, YANG Yuan-Zhu. Analysis of blast resistance genes in Longliangyou and Jingliangyou hybrid rice varieties [J]. Acta Agronomica Sinica, 2022, 48(5): 1071-1080.
[8]	YANG De-Wei, WANG Xun, ZHENG Xing-Xing, XIANG Xin-Quan, CUI Hai-Tao, LI Sheng-Ping, TANG Ding-Zhong. Functional studies of rice blast resistance related gene OsSAMS1 [J]. Acta Agronomica Sinica, 2022, 48(5): 1119-1128.
[9]	ZHU Zheng, WANG Tian-Xing-Zi, CHEN Yue, LIU Yu-Qing, YAN Gao-Wei, XU Shan, MA Jin-Jiao, DOU Shi-Juan, LI Li-Yun, LIU Guo-Zhen. Rice transcription factor WRKY68 plays a positive role in Xa21-mediated resistance to Xanthomonas oryzae pv. oryzae [J]. Acta Agronomica Sinica, 2022, 48(5): 1129-1140.
[10]	WANG Xiao-Lei, LI Wei-Xing, OU-YANG Lin-Juan, XU Jie, CHEN Xiao-Rong, BIAN Jian-Min, HU Li-Fang, PENG Xiao-Song, HE Xiao-Peng, FU Jun-Ru, ZHOU Da-Hu, HE Hao-Hua, SUN Xiao-Tang, ZHU Chang-Lan. QTL mapping for plant architecture in rice based on chromosome segment substitution lines [J]. Acta Agronomica Sinica, 2022, 48(5): 1141-1151.
[11]	WANG Ze, ZHOU Qin-Yang, LIU Cong, MU Yue, GUO Wei, DING Yan-Feng, NINOMIYA Seishi. Estimation and evaluation of paddy rice canopy characteristics based on images from UAV and ground camera [J]. Acta Agronomica Sinica, 2022, 48(5): 1248-1261.
[12]	KE Jian, CHEN Ting-Ting, WU Zhou, ZHU Tie-Zhong, SUN Jie, HE Hai-Bing, YOU Cui-Cui, ZHU De-Quan, WU Li-Quan. Suitable varieties and high-yielding population characteristics of late season rice in the northern margin area of double-cropping rice along the Yangtze River [J]. Acta Agronomica Sinica, 2022, 48(4): 1005-1016.
[13]	CHEN Yue, SUN Ming-Zhe, JIA Bo-Wei, LENG Yue, SUN Xiao-Li. Research progress regarding the function and mechanism of rice AP2/ERF transcription factor in stress response [J]. Acta Agronomica Sinica, 2022, 48(4): 781-790.
[14]	WANG Hao-Rang, ZHANG Yong, YU Chun-Miao, DONG Quan-Zhong, LI Wei-Wei, HU Kai-Feng, ZHANG Ming-Ming, XUE Hong, YANG Meng-Ping, SONG Ji-Ling, WANG Lei, YANG Xing-Yong, QIU Li-Juan. Fine mapping of yellow-green leaf gene (ygl2) in soybean (Glycine max L.) [J]. Acta Agronomica Sinica, 2022, 48(4): 791-800.
[15]	WANG Lyu, CUI Yue-Zhen, WU Yu-Hong, HAO Xing-Shun, ZHANG Chun-Hui, WANG Jun-Yi, LIU Yi-Xin, LI Xiao-Gang, QIN Yu-Hang. Effects of rice stalks mulching combined with green manure (Astragalus smicus L.) incorporated into soil and reducing nitrogen fertilizer rate on rice yield and soil fertility [J]. Acta Agronomica Sinica, 2022, 48(4): 952-961.