水稻广谱抗稻瘟病基因PigmR功能标记的开发及应用

doi:10.3864/j.issn.0578-1752.2019.06.001

中国农业科学 ›› 2019, Vol. 52 ›› Issue (6): 955-967.doi: 10.3864/j.issn.0578-1752.2019.06.001

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

水稻广谱抗稻瘟病基因PigmR功能标记的开发及应用

王芳权^1,²,陈智慧^1,²,许扬^1,²,王军^1,²,李文奇^1,²,范方军^1,²,陈丽琴¹,陶亚军^1,²,仲维功¹,杨杰^1,²()

1 江苏省农业科学院粮食作物研究所/国家水稻改良中心南京分中心/江苏省优质水稻工程技术研究中心,南京 210014
2 扬州大学/江苏省粮食作物现代产业技术协同创新中心,江苏扬州 225009

收稿日期:2018-11-18 接受日期:2019-01-18 出版日期:2019-03-16 发布日期:2019-03-22
通讯作者: 杨杰
作者简介:王芳权,Tel：025-84390320;E-mail： wfqjaas@163.com。
基金资助:
国家重点研发计划(2017YFD0100400-3);江苏省农业科技自主创新资金项目(CX182022);江苏省现代农业重点研发项目(BE2017368);江苏省现代农业重点研发项目(BE2018388);江苏省自然科学基金面上项目(BK20171326)

Development and Application of the Functional Marker for the Broad-Spectrum Blast Resistance Gene PigmR in Rice

WANG FangQuan^1,²,CHEN ZhiHui^1,²,XU Yang^1,²,WANG Jun^1,²,LI WenQi^1,²,FAN FangJun^1,²,CHEN LiQin¹,TAO YaJun^1,²,ZHONG WeiGong¹,YANG Jie^1,²()

1 Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R & D Center, Nanjing 210014;
2 Yangzhou University/Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou 225009, Jiangsu

Received:2018-11-18 Accepted:2019-01-18 Online:2019-03-16 Published:2019-03-22
Contact: Jie YANG

摘要/Abstract

摘要：

【目的】稻瘟病是世界上最严重的水稻病害之一。通过功能标记的开发,为加快广谱持久抗稻瘟病基因PigmR在水稻育种中的应用提供依据。【方法】利用Snapgene 2.3.2软件分析PigmR的碱基变异特征,用Oligo 7设计特异功能标记。为了避免因PCR扩增失败引起的假阴性,设计扩增内参基因Actin1的引物作为参照,对功能标记进行优化。用功能标记对水稻亲本材料、丽江新团黑谷的单基因系材料、育种中间材料和南粳53045/谷梅4号BC₁F₃群体株系进行鉴定。稻瘟病鉴定的供试菌株为江苏省稻瘟病代表菌株（2018-4、2018-65、2018-102、2018-222和2018-241）的混合菌。将供试菌株移植RCA培养基上,25℃培养7 d,用黑光灯照射72 h,待稻瘟病菌产生孢子后,再用无菌水洗下,配成10×10倍显微镜下每视野30—40个孢子的悬浮液。于水稻抽穗前3—4 d注射混合菌株,每穗注射1 mL菌液,做好标记。在水稻灌浆饱满后进行抗性调查。【结果】根据PigmR与PigmS、Pigm-R4序列比对及差异分析,设计了8对分子标记引物。通过分子检测,筛选获得的PigmR功能标记GMR-3,能特异扩增来源于谷梅4号的PigmR,获得98 bp产物,而不携带PigmR扩增不出产物。以不同浓度配比优化GMR-3与内参基因Actin1检测引物Actin1-1,发现以0.4 μmol·L ^-1 GMR-3与0.1 μmol·L ^-1 Actin1-1组合浓度扩增出PigmR和Actin1特征条带的效率相当,效果最优,将该标记命名为GMRA。用该标记扩增水稻样品,携带PigmR的样品能扩增出146和98 bp条带,不携带PigmR的样品仅能扩增出146 bp条带。利用GMRA标记检测229份水稻材料,只有谷梅4号能扩增出146和98 bp条带,其他籼稻和粳稻均只能扩增出146 bp条带。进一步对29份丽江新团黑谷的单基因系材料进行检测发现,GMRA标记能有效区分PigmR与Pi9、Piz、Piz-t等同源性较高的基因,有很好的特异性。利用GMRA标记,从240份育种中间材料中筛选到3份携带PigmR的材料,可作为该基因的供体材料。利用该标记进行分子标记辅助选择,将PigmR通过回交转育到优质食味粳稻南粳53045。对南粳53045/谷梅4号BC₁F₃群体单株进行分子标记检测及稻瘟病人工接种鉴定,发现携带PigmR的单株均表现为抗或中抗,不携带PigmR的单株均表现为高感,表明导入PigmR能显著改良南粳53045的穗颈瘟抗性。【结论】PigmR的功能标记能有效用于抗稻瘟病遗传改良和资源筛选。

关键词: 水稻, 稻瘟病, Pigm, 功能标记, 分子标记辅助选择

Abstract:

【Objective】Rice blast is one of the most serious rice diseases in the world. The objective of this study was to develop the functional marker for broad-spectrum blast resistance gene PigmR in rice. The marker improved the application of PigmR in blast resistance rice breeding. 【Method】The characteristics of PigmR were analyzed by Snapgene 2.3.2, and the specific function markers were designed with Oligo 7. To avoid mistake results of the PCR amplification failure, the functional marker was optimized by the specific primers of Actin1 as a reference. The parent materials, the monogenic lines of Lijiangxintuanheigu (LTH), the bridge materials, and the BC₁F₃ population lines of Nangeng 53045/Gumei 4 were identified by the functional markers. The blast isolates in this study were the mixed representative strains of rice blast in Jiangsu Province (2018-4, 2018-65, 2018-102, 2018-222, and 2018-241). The tested isolates were transplanted into RCA medium, cultured at 25 ℃ for 7 d, and irradiated for 72 h. After spores were produced, they were washed with sterile water and then formulated 30-40 spores per field in 10×10 microscope. The mixed spores were injected into each panicle with 1 mL of blast isolates solution at 3-4 days before heading. The resistance was investigated after the rice grains were matured.【Result】Eight pairs of molecular markers were designed, according to the sequence difference of PigmR and PigmS, Pigm-R4. By molecular detected, the function marker of PigmR, GMR-3, could specifically amplify PigmR from Gumei 4 with 98 bp fragment, and no fragment was amplified in the samples without PigmR. The functional marker was optimized by different concentrations of GMR-3 and Actin1-1 (a marker for the internal reference gene Actin1), results shown that the marker consist of 0.4 μmol·L ^-1 GMR-3 and 0.1 μmol·L ^-1 Actin1-1 had the best effect. The functional marker was named GMRA. Samples carrying PigmR were amplified the expected size of 146 and 98 bp by GMRA. By contrast, samples without PigmR were only amplified a 146 bp fragment. The 229 rice materials were detected with GMRA, only Gumei 4 amplified the size of 146 and 98 bp, others only amplified a 146 bp fragment. Furthermore, the results detected the monogenic lines of LTH by GMRA suggested that the marker had a strong specificity and could effectively distinguish PigmR from homology genes, such as Pi9, Piz, and Piz-t. Moreover, three donor materials carrying PigmR were obtained from 240 bridge materials by GMRA. By molecular marker-assisted selection (MAS) of GMRA, PigmR was transferred to the good eating quality rice cultivar Nangeng53045 by backcrossing. The BC₁F₃ plants with PigmR showed resistant/middle resistant to the panicle blast, while others showed high susceptible. It was suggested that PigmR observably improved the resistance of Nangeng53045-Pigm lines in the panicle blast.【Conclusion】In conclusion, the functional marker of PigmR can be effectively used for genetic improvement in breeding and germplasm screening.

Key words: rice (Oryza sativa L.), rice blast, Pigm, functional marker, molecular marker-assisted selection

王芳权,陈智慧,许扬,王军,李文奇,范方军,陈丽琴,陶亚军,仲维功,杨杰. 水稻广谱抗稻瘟病基因PigmR功能标记的开发及应用[J]. 中国农业科学, 2019, 52(6): 955-967.

WANG FangQuan,CHEN ZhiHui,XU Yang,WANG Jun,LI WenQi,FAN FangJun,CHEN LiQin,TAO YaJun,ZHONG WeiGong,YANG Jie. Development and Application of the Functional Marker for the Broad-Spectrum Blast Resistance Gene PigmR in Rice[J]. Scientia Agricultura Sinica, 2019, 52(6): 955-967.

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图/表 11

图1

表1

图2

图3

图4

图5

表2

GMRA标记鉴定部分水稻材料"

编号 No.	品种 Variety	粳/籼稻 Geng/Xian	带型 Type	编号 No.	品种 Variety	粳/籼稻 Geng/Xian	带型 Type
1	谷梅4号 Gumei 4	X	2	40	盐粳10号 Yandao 10	G	1
2	日本晴 Niponbare	G	1	41	盐粳11号 Yangeng 11	G	1
3	黄华占 Huanghuazhan	X	1	42	镇稻99 Zhendao 99	G	1
4	IR36	X	1	43	扬粳113 Yanggeng 113	G	1
5	玉针香 Yuzhenxiang	X	1	44	WP16	G	1
6	南京16号 Nanjing 16	X	1	45	WP59	G	1
7	Kasalath	X	1	46	泗稻15号 Sidao 15	G	1
8	连粳7号 Liangeng 7	G	1	47	连糯1号 Liannuo 1	G	1
9	南粳9108 Nangeng 9108	G	1	48	皖粳糯1号 Liangengnuo 1	G	1
10	武运粳24号 Wuyungeng 24	G	1	49	圣稻19 Shengdao 19	G	1
11	华粳5号 Huageng 5	G	1	50	JX14-3	G	1
12	扬育粳2号 Yangyugeng 2	G	1	51	JX14-4	G	1
13	南粳44 Nangeng 44	G	1	52	K11	G	1
14	南粳45 Nangeng 45	G	1	53	N14-109	G	1
15	南粳46 Nangeng 46	G	1	54	宁粳7号 Ninggeng 7	G	1
16	南粳5055 Nangeng 5055	G	1	55	YJN1	G	1
17	金粳818 Jingeng 818	G	1	56	常粳144 Changgeng 144	G	1
18	苏秀867 Suxiu 867	G	1	57	常农粳4号 Changnonggeng 4	G	1
19	武运粳21号 Wuyungeng 21	G	1	58	常农粳6号 Changnonggeng 6	G	1
20	武运粳27号 Wuyungeng 27	G	1	59	常农粳7号 Changnonggeng 7	G	1
21	淮稻5号 Huaidao 5	G	1	60	Feng1326	G	1
22	南粳53045 Nangeng53045	G	1	61	华粳3号 Huageng 3	G	1
23	武运粳29号 Wuyungeng 29	G	1	62	华粳4号 Huageng 4	G	1
24	楚粳39 Chugeng 39	G	1	63	华粳6号 Huageng 6	G	1
25	丰粳3227 Fenggeng 3227	G	1	64	华瑞稻1号 Huaruidao 1	G	1
26	华粳7号 Huageng 7	G	1	65	淮1188 Huai 1188	G	1
27	淮稻7号 Huaidao 7	G	1	66	淮208 Huai 208	G	1
28	淮糯11号 Huainuo 11	G	1	67	淮稻10号 Huaidao 10	G	1
29	淮优粳2号 Huaiyougeng 2	G	1	68	淮稻11号 Huaidao 11	G	1
30	连粳11号 Liangeng 11	G	1	69	淮稻13号 Huaidao 13	G	1
31	连粳12号 Liangeng 12	G	1	70	淮稻8号 Huaidao 8	G	1
32	连粳4号 Liangeng 4	G	1	71	淮稻9号 Huaidao 9	G	1
33	连粳6号 Liangeng 6	G	1	72	淮粳096 Huaigeng 096	G	1
34	连粳147729 Liangeng 147729	G	1	73	淮香稻15号 Huaixiangdao 15	G	1
35	农香软米 Nongxiangruanmi	G	1	74	津稻263 Jindao 263	G	1
36	泗稻785 Sidao 785	G	1	75	连粳9号 Liangeng 9	G	1
37	苏秀326 Suxiu 326	G	1	76	连粳10号 Liangeng 10	G	1
38	徐稻3号 Xudao 3	G	1	77	南粳40 Nangeng 40	G	1
39	盐稻11号 Yandao 11	G	1	78	南粳41 Nangeng 41	G	1
79	南粳42 Nangeng 42	G	1	119	镇稻7号 Zhendao 7	G	1
80	南粳49 Nangeng 49	G	1	120	镇稻88 Zhendao 88	G	1
81	宁2600 Ning 2600	G	1	121	镇稻9424 Zhendao 9424	G	1
82	宁粳2号 Ninggeng 2	G	1	122	宁粳1号 Ninggeng 1	G	1
83	宁粳4号 Ninggeng 4	G	1	123	宁粳3号 Ninggeng 3	G	1
84	宁粳5号 Ninggeng 5	G	1	124	南粳47 Nangeng 47	G	1
85	迁稻11-72 Qiandao 11-72	G	1	125	通粳981 Tonggeng 981	G	1
86	泗稻12号 Sidao 12	G	1	126	武运粳7号 Wuyungeng 7	G	1
87	苏沪香粳 Suhuxianggeng	G	1	127	武运粳19号 Wuyungeng 19	G	1
88	苏粳5号 Sugeng 5	G	1	128	武运粳23号 Wuyungeng 23	G	1
89	苏秀10号 Suxiu 10	G	1	129	常农粳3号 Changnonggeng 3	G	1
90	武粳13号 Wugeng 13	G	1	130	常农粳5号 Changnonggeng 5	G	1
91	武粳15号 Wugeng 15	G	1	131	苏粳8号 Sugeng 8	G	1
92	武粳4号 Wugeng 4	G	1	132	苏香粳2号 Suxianggeng 2	G	1
93	武陵粳1号 Wulinggeng 1	G	1	133	嘉33 Jia 33	G	1
94	武香粳14号 Wuxianggeng 14	G	1	134	嘉991 Jia 991	G	1
95	武香粳9号 Wuxianggeng 9	G	1	135	镇稻10号 Zhendao 10	G	1
96	武育粳18号 Wuyugeng 18	G	1	136	镇稻16号 Zhendao 16	G	1
97	武育粳3号 Wuyugeng 3	G	1	137	镇稻17号 Zhendao 17	G	1
98	武运粳11号 Wuyungeng 11	G	1	138	镇稻18号 Zhendao 18	G	1
99	兴化紫稻 Xinghuazidao	G	1	139	苏粳9号 Sugeng 9	G	1
100	徐稻8号 Xudao 8	G	1	140	宁2602 Ning 2602	G	1
101	徐稻9号 Xudao 9	G	1	141	宁2604 Ning 2604	G	1
102	盐稻8号 Yandao 8	G	1	142	嘉58 Jia 58	G	1
103	盐稻9号 Yandao 9	G	1	143	苏05-1176 Su 05-1176	G	1
104	盐粳16号 Yangeng 16	G	1	144	苏12-130 Su 12-130	G	1
105	盐粳9号 Yangeng 9	G	1	145	沪香软268 Huxiangruan 268	G	1
106	扬9709 Yang9700	G	1	146	沪香软386 Huxiangruan 386	G	1
107	扬辐粳1号 Yangfugeng 1	G	1	147	软大穗 Ruandaoli	G	1
108	扬辐粳8号 Yangfugeng 8	G	1	148	大粒糯 Dalinuo	G	1
109	扬粳4227 Yanggeng 4227	G	1	149	太湖香粳稻 Taihuxianggengdao	G	1
110	扬粳4308 Yanggeng 4308	G	1	150	JX14-5	G	1
111	扬粳805 Yanggeng 805	G	1	151	南粳43 Nangeng 43	G	1
112	扬农210 Yangnong210	G	1	152	农垦57 Nongken 57	G	1
113	扬中稻1号 Yangzhongdao 1	G	1	153	农垦58 Nongken 58	G	1
114	镇糯19号 Zhennuo 19	G	1	154	秀水63 Xiushui 63	G	1
115	镇稻12号 Zhendao 12	G	1	155	徐81698 Xu 81698	G	1
116	镇稻13号 Zhendao 13	G	1	156	徐稻4号 Xudao 4	G	1
117	镇稻15号 Zhendao 15	G	1	157	盐稻980 Yandao 980	G	1
118	镇稻1号 Zhendao 1	G	1	158	盐丰2号 Yanfeng 2	G	1
159	扬粳9224 Yanggeng 9224	G	1	195	14CZY43	G	1
160	扬育粳116 Yangyugeng 116	G	1	196	14CZY47	G	1
161	银玉2239 Yinyu 2239	G	1	197	14CZY49	G	1
162	镇稻615 Zhendao 615	G	1	198	14CZY53	G	1
163	隆粳968 Longgeng 968	G	1	199	14CZY60	G	1
164	大华香糯 Dahuaxiangnuo	G	1	200	14CZY65	G	1
165	华粳8号 Huageng 8	G	1	201	14CZY71	G	1
166	盐粳13号 Yangeng 13	G	1	202	14CZY73	G	1
167	盐粳15号 Yangeng 15	G	1	203	14CZY83	G	1
168	南粳505 Nangeng 505	G	1	204	14CZY84	G	1
169	盐丰稻2号 Yanfengdao 2	G	1	205	14CZY85	G	1
170	软香玉 Ruanxiangyu	G	1	206	14CZY86	G	1
171	苏香粳100 Suxianggeng 100	G	1	207	14CZY87	G	1
172	苏2100 Su 2100	G	1	208	15ZZY2	G	1
173	沪粳137 Hugeng 137	G	1	209	15ZZY7	G	1
174	沪香粳165 Huxianggeng 165	G	1	210	15ZZY8	G	1
175	松粳9号 Songgeng 9	G	1	211	15ZZY79	G	1
176	14CZY1	G	1	212	15ZZY89	G	1
177	14CZY8	G	1	213	15ZZY16	G	1
178	14CZY11	G	1	214	15ZZY48	G	1
179	14CZY13	G	1	215	15ZZY53	G	1
180	14CZY15	G	1	216	15ZZY58	G	1
181	14CZY16	G	1	217	15ZZY73	G	1
182	14CZY17	G	1	218	15ZZY90	G	1
183	14CZY21	G	1	219	15ZZQ6	G	1
184	14CZY22	G	1	220	15ZZQ12	G	1
185	14CZY23	G	1	221	15CZY3	G	1
186	14CZY28	G	1	222	15CZY31	G	1
187	14CZY30	G	1	223	15CZY65	G	1
188	14CZY31	G	1	224	15CZY70	G	1
189	14CZY36	G	1	225	15CZY71	G	1
190	14CZY37	G	1	226	15CZY77	G	1
191	14CZY38	G	1	227	15CZY84	G	1
192	14CZY39	G	1	228	15CZY86	G	1
193	14CZY40	G	1	229	15CZY91	G	1
194	14CZY41	G	1

表2

表3

图6

图7

图8

参考文献 32

[1]	KHAN M A I, BHUIYAN M R, HOSSAIN M S, SEN P P, ARA A, SIDDIQUE M A, ALI M A . Neck blast disease influences grain yield and quality traits of aromatic rice. Comptes Rendus Biologies, 2014,337(11):635-641. doi: 10.1016/j.crvi.2014.08.007 pmid: 25444707
[2]	朱凤, 田子华, 邰德良, 刘永锋 . 从2014年稻瘟病重发谈今后防控对策的改进. 江苏农业科学, 2016,44(8):155-158. doi: 10.15889/j.issn.1002-1302.2016.08.042
	ZHU F, TIAN Z H, TAI D L, LIU Y F . Improvement of prevention and control measures from the outburst of rice blast in 2014. Jiangsu Agricultural Sciences, 2016,44(8):155-158. (in Chinese) doi: 10.15889/j.issn.1002-1302.2016.08.042
[3]	SHARMA T R, RAI A K, GUPTA S K, VIJAYAN J, DEVANNA B N, RAY S . Rice blast management through host-plant resistance: Retrospect and prospects. Agricultural Research, 2012,1(1):37-52. doi: 10.1007/s40003-011-0003-5
[4]	ROYCHOWDHURY M, JIA Y, JIA M H, FJELLSTROM R, CARTWRIGHT R D . Identification of the rice blast resistance gene Pib in the national small grains collection. Phytopathology, 2012,155(2):700-706. doi: 10.1080/00222935808697054 pmid: 22667447
[5]	BRYAN G T, WU K S, FARRALL L, JIA Y, HERSHEY H P, MCADAMS S A, FAULK K N, DONALDSON G K, TARCHINI R, VALENT B . A single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pi-ta. The Plant Cell, 2000,12(11):2033-2046. doi: 10.2307/3871103 pmid: 11090207
[6]	SHARMA T R, RAI A K, GUPTA S K, SINGH N K . Broad-spectrum blast resistance gene Pi-k(h) cloned from rice line Tetep designated as Pi54. Journal of Plant Biochemistry and Biotechnology, 2010,19(1):87-89. doi: 10.1007/BF03323441
[7]	FUKUOKA S, SAKA N, KOGA H, ONO K, SHIMIZU T, EBANA K, HAYASHI N, TAKAHASHI A, HIROCHIKA H, OKUNO K, YANO M . Loss of function of a proline-containing protein confers durable disease resistance in rice. Science, 2009,325(5943):998-1001. doi: 10.1126/science.1175550 pmid: 19696351
[8]	LI W, ZHU Z, CHERN M, YIN J, YANG C, RAN L, CHENG M, HE M, WANG K, WANG J, ZHOU X, ZHU X, CHEN Z, WANG J, ZHAO W, MA B, QIN P, CHEN W, WANG Y, LIU J, WANG W, WU X, LI P, WANG J, ZHU L, LI S, CHEN X . A natural allele of a transcription factor in rice confers broad-spectrum blast resistance. Cell, 2017,170(1):114-126. doi: 10.1016/j.cell.2017.06.008 pmid: 28666113
[9]	ZHOU B, QU S H, LIU G F, DOLAN M, SAKAI H, LU G D, BELLIZZI M, WANG G L . The eight amino-acid differences within three leucine-rich repeats between Pi2 and Piz-t resistance proteins determine the resistance specificity to Magnaporthe grisea. Molecular Plant-Microbe Interactions, 2006,19(11):1216-1228. doi: 10.1094/MPMI-19-1216 pmid: 17073304
[10]	QU S, LIU G, ZHOU B, BELLIZZI M, ZENG L, DAI L, HAN B, WANG G L . The broad-spectrum blast resistance gene Pi9 encodes a nucleotide-binding site-leucine-rich repeat protein and is a member of a multigene family in rice. Genetics, 2006,172(3):1901-1914. doi: 10.1534/genetics.105.044891 pmid: 16387888
[11]	ZHU X, CHEN S, YANG J, ZHOU S, ZENG L, HAN J, SU J, WANG L , PAN, Q . The identification of Pi50(t), a new member of the rice blast resistance Pi2/Pi9 multigene family. Theoretical and Applied Genetics, 2012,124(7):1295-1304. doi: 10.1007/s00122-012-1787-9 pmid: 22270148
[12]	SU J, WANG W, HAN J, CHEN S, WANG C, ZENG L, FENG A, YANG J, ZHOU B, ZHU X . Functional divergence of duplicated genes results in a novel blast resistance gene Pi50 at the Pi2/9 locus. Theoretical and Applied Genetics, 2015,128(11):2213-2225. doi: 10.1007/s00122-015-2579-9 pmid: 26183036
[13]	DENG Y, ZHU X, SHEN Y, HE Z . Genetic characterization and fine mapping of the blast resistance locus Pigm(t) tightly linked to Pi2 and Pi9 in a broad-spectrum resistant Chinese variety. Theoretical and Applied Genetics, 2006,113(4):705-713. doi: 10.1007/s00122-006-0338-7 pmid: 16832648
[14]	DENG Y, ZHAI K, XIE Z, YANG D, ZHU X, LIU J, WANG X, QIN P, YANG Y, ZHANG G, LI Q, ZHANG J, WU S, MILAZZO J, MAO B, WANG E, XIE H, THARREAU D, HE Z . Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance. Science, 2017,355(6328):962-965. doi: 10.1126/science.aai8898 pmid: 28154240
[15]	ZHAO H, WANG X, JIA Y, MINKENBERG B, WHEATLEY M, FAN J, JIA M H, FAMOSO A, EDWARDS J D, WAMISHE Y, VALENT B, WANG G L, YANG Y . The rice blast resistance gene Ptr encodes an atypical protein required for broad-spectrum disease resistance. Nature Communications, 2018,9(1):2039. doi: 10.1038/s41467-018-04369-4
[16]	WU Y, XIAO N, YU L, PAN C, LI Y, ZHANG X, LIU G, DAI Z, PAN X, LI A . Combination patterns of major R genes determine the level of resistance to the M. oryzae in rice( Oryza sativa L.). PLoS ONE, 2015,10(6):e0126130. doi: 10.1371/journal.pone.0126130 pmid: 26030358
[17]	杨杰, 杨金欢, 王军, 范方军, 朱金燕, 曹卿, 田胜尼, 仲维功 . 稻瘟病抗病基因Pita和Pib在中国水稻地方品种中的分布. 华北农学报, 2011,26(3):1-6. doi: 10.7668/hbnxb.2011.03.001
	YANG J, YANG J H, WANG J, FAN F J, ZHU J Y, CAO Q, TIAN S N, ZHONG W G . Distribution of two blast resistant genes Pita and Pib in landrace rice in China. Acta Agriculture Boreali-Sinica, 2011,26(3):1-6. (in Chinese) doi: 10.7668/hbnxb.2011.03.001
[18]	王军, 赵婕宇, 许扬, 范方军, 朱金燕, 李文奇, 王芳权, 费云燕, 仲维功, 杨杰 . 水稻稻瘟病抗性基因Bsr-d1功能标记的开发和利用. 作物学报, 2018,44(11):1612-1620.
	WANG J, ZHAO J Y, XU Y, FAN F J, ZHU J Y, LI W Q, WANG F Q, FEI Y Y, ZHONG W G, YANG J . Development and application of functional markers for rice blast resistance gene Bsr-d1 in rice. Acta Agronomica Sinica, 2018,44(11):1612-1620. (in Chinese)
[19]	曾生元, 李闯, 杜灿灿, 孙立亭, 景德道, 林添资, 余波, 钱华飞, 姚维成, 周义文, 龚红兵 . Pigm特异性选择标记的开发及其在粳稻穗颈瘟抗性育种中的利用. 中国水稻科学, 2018,32(5):453-461. doi: 10.16819/j.1001-7216.2018.7135
	ZENG S Y, LI C, DU C C, SUN L T, JING D D, LIN T Z, YU B, QIAN H F, YAO W C, ZHOU Y W, GONG H B . Development of specific markers for Pigm in marker-assisted breeding of panicle blast resistant japonica rice. Chinese Journal of Rice Science, 2018,32(5):453-461. (in Chinese) doi: 10.16819/j.1001-7216.2018.7135
[20]	潘存红, 李爱宏, 戴正元, 朱俊凯, 余玲, 肖宁, 李育红, 张小祥, 刘广青, 赵步洪, 王宝和, 黄年生, 周长海, 谭长乐, 季红娟, 刘晓静 . 一种用于检测谷梅4号抗稻瘟病基因Pigm(t)的分子标记InDel587. 中国, ZL201310428162.0, 2015 -11-25.
	PAN C H, LI A H, DAI Z Y, ZHU J K, YU L, XIAO N, LI Y H, ZHANG X Y, LIU G Q, ZHAO B H, WANG B H, HUANG N S, ZHOU C H, TAN C L, JI H J , LIU X J. A molecular marker InDel587 for detecting the rice blast resistance gene Pigm(t) of Gumei 4. China, ZL201310428162.0, 2015 -11-25. (in Chinese)
[21]	戴小军, 杨远柱, 陈良碧, 胡小淳, 秦鹏, 符辰建 . 水稻抗稻瘟病基因Pigm的分子标记方法 . 中国, ZL201210325874.5, 2014 -11-05.
	DAI X J, YANG Y Z, CHEN L B, HU X C, QIN P ,FU C J . Molecular marker for rice blast resistance gene Pigm. China, ZL201210325874.5, 2014 -11-05. (in Chinese)
[22]	田大刚, 王锋, 陈松彪, 陈子强, 林艳, 陈在杰, 杨立明, 胡昌泉 . 一种稻瘟病抗性基因Pigm功能特异性分子标记及其应用 . 中国, ZL201810054317.1, 2018 -05-04.
	TIAN D G, WANG F, CHEN S B, CHEN Z Q, LIN Y, CHEN Z J, YANG L M, HU C Q . A specific molecular marker for rice blast resistance gene Pigmand its application.China, ZL201810054317.1, 2018 -05-04. (in Chinese)
[23]	International Rice Research Institute . Standard Evaluation System for Rice (SES). Manila: International Rice Research Institute, 2013.
[24]	刘海涛, 徐倩, 何炜, 魏林艳, 张建福, 谢华安 . 水稻稻瘟病抗性变化及抗性基因克隆的研究进展. 福建农业学报, 2016,31(5):545-552. doi: 10.3969/j.issn.1008-0384.2016.05.019
	LIU H T, XU Q, HE W, WEI L Y, ZHANG J F, XIE H A . Recent progress on the variation of blast resistance and cloning of the resistance genes in rice. Fujian Journal of Agricultural Sciences, 2016,31(5):545-552. (in Chinese) doi: 10.3969/j.issn.1008-0384.2016.05.019
[25]	任世龙, 白辉, 王永芳, 全建章, 董志平, 李志勇, 邢继红 . 谷瘟病菌无毒基因型鉴定及分析. 中国农业科学, 2018,51(6):1079-1088.
	REN S L, BAI H, WANG Y F, QUAN J Z, DONG Z P, LI Z Y, XING J H . Identification and analysis of Magnaporthe oryzae of foxtail millet avirulence genes. Scientia Agricultura Sinica, 2018,51(6):1079-1088. (in Chinese)
[26]	袁熹, 李大勇, 宋凤鸣 . 水稻对稻瘟病的广谱抗性: 分子机制及其育种应用. 植物生理学报, 2017,53(8):1348-1358.
	YUAN X, LI D Y, SONG F M . Broad-spectrum blast resistance in rice: Molecular mechanism and its breeding application. Plant Physiology Journal, 2017,53(8):1348-1358. (in Chinese)
[27]	于苗苗, 戴正元, 潘存红, 陈夕军, 余玲, 张晓祥, 李育红, 肖宁, 龚红兵, 盛生兰, 潘学彪, 张洪熙, 李爱宏 . 广谱稻瘟病抗性基因Pigm和Pi2的抗谱差异及与Pi1的互作效应. 作物学报, 2013,39(11):1927-1934. doi: 10.3724/SP.J.1006.2013.01927
	YU M M, DAI Z Y, PAN C H, CHEN X J, YU L, ZHANG X X, LI Y H, XIAO N, GONG H B, SHENG S L, PAN X B, ZHANG H X, LI A H . Resistance spectrum difference between two broad-spectrum blast resistance genes, Pigm and Pi2, and their interaction effect on Pi1. Acta Agronomica Sinica, 2013,39(11):1927-1934. (in Chinese) doi: 10.3724/SP.J.1006.2013.01927
[28]	梁毅, 杨婷婷, 谭令辞, 文婷, 吴俊, 江南, 李智强, 戴良英, 王国梁, 刘雄伦 . 水稻广谱抗瘟基因Pigm紧密连锁分子标记开发及其育种应用. 杂交水稻, 2013,28(4):63-68.
	LIANG Y, YANG T T, TAN L C, WEN T, WU J, JIANG N, LI Z Q, DAI L Y, WANG G L, LIU X L . Development of the linked molecular marker for the broad-spectrum blast resistance gene Pigm in rice. Hybrid Rice, 2013,28(4):63-68. (in Chinese)
[29]	ZHANG J, ZHOU X, YAN W, ZHANG Z, LU L, HAN Z, ZHAO H, LIU H, SONG P, HU Y, SHEN G, HE Q, GUO S, GAO G, WANG G, XING Y . Combinations of the Ghd7, Ghd8 and Hd1 genes largely define the ecogeographical adaptation and yield potential of cultivated rice. New Phytologist, 2015,208(4):1056-1066. doi: 10.1111/nph.13538 pmid: 26147403
[30]	LEE Y S, AN G . Regulation of flowering time in rice. Journal of Plant Biology, 2015,58(6):353-360. doi: 10.1007/s12374-015-0425-x
[31]	YE J, NIU X, YANG Y, WANG S, XU Q, YUAN X, YU H, WANG Y, WANG S, FENG Y, WEI X . Divergent Hd1, Ghd7, and DTH7 alleles control heading date and yield potential of Japonica rice in northeast china. Frontiers in Plant Science, 2018,9:35. doi: 10.3389/fpls.2018.00035
[32]	YANG J, WANG J, FAN F J, ZHU J Y, CHEN T, WANG C L, ZHENG T Q, ZHANG J, ZHONG W G, XU J L . Development of AS-PCR marker based on a key mutation confirmed by resequencing of Wx-mp in Milky Princess and its application in japonica soft rice( Oryza sativa L.) breeding. Plant Breeding, 2013,132(6):595-603. doi: 10.1111/pbr.12088

标记名称 Marker name	引物名称 Primer name	引物序列 Primer sequence (5′-3′)	理论产物长度 Expected size (bp)
GMR-1	GMR-1F	TTCCTTTCTCTTTATCATAATT	180
GMR-1	GMR-1R	AGATTCCAACCTGCACTTGCCT	180
GMS-1	GMS-1F	GTCCTTGATCTTTATCAGAAAG	180
GMS-1	GMS-1R	TGGTTCCAACCTGCAGAATCTG	180
GMR-2	GMR-2F	TTCCTTTCTCTTTATCATAcTT	180
GMR-2	GMR-2R	AGATTCCAACCTGCACTTGCCT	180
GMS-2	GMS-2F	GTCCTTGATCTTTATCAGAcAG	180
GMS-2	GMS-2R	TGGTTCCAACCTGCAGAATCTG	180
GMR-3	GMR-3F	AGTTCTACTTACGGAGGAGC	98
GMR-3	GMR-3R	AGAATTATGATAAAGAGAAAGGAA	98
GMS-3	GMS-3F	AGATCAACTTATTGAGCAGC	98
GMS-3	GMS-3R	AGCTTTCTGATAAAGATCAAGGAC	98
GMR-4	GMR-4F	AGTTCTACTTACGGAGGcGC	98
GMR-4	GMR-4R	AGAATTATGATAAAGAGAAAGcAA	98
GMS-4	GMS-4F	AGATCAACTTATTGAGCcGC	98
GMS-4	GMS-4R	AGCTTTCTGATAAAGATCAAGcAC	98
Actin1-1	Actin1-1F	ACAAAGTTTTCAACCGGCCTA	146
Actin1-1	Actin1-1R	CTGGTACCCTCATCAGGCATC	146
Actin1-2	Actin1-2F	AGTCTGGCCCATCCATTGTG	149
Actin1-2	Actin1-2R	CGGTTGAAAACTTTGTCCACGCTA	149

编号No.	名称Designation	目标基因Target gene	带型Type
1	IRBLa-A	Pia	1
2	IRBLa-C	Pia	1
3	IRBLi-F5	Pii	1
4	IRBLks-F5	Pik-s	1
5	IRBLks-S	Pik-s	1
6	IRBLk-ka	Pik	1
7	IRBLkp-K60	Pik-p	1
8	IRBLkh-K3	Pik-h	1
9	IRBLz-Fu	Piz	1
10	IRBLzt-T	Piz-t	1
11	IRBLta-K1	Pita	1
12	IRBLta-CT2	Pita	1
13	IRBLb-B	Pib	1
14	IRBLt-K59	Pit	1
15	IRBLsh-S	Pish	1
16	IRBLsh-B	Pish	1
17	IRBL1-CL	Pi1	1
18	IRBL3-CP4	Pi3	1
19	IRBL5-M	Pi5(t)	1
20	IRBL7-M	Pi7(t)	1
21	IRBL9-W	Pi9	1
22	IRBL12-M	Pi12(t)	1
23	IRBL19-A	Pi19	1
24	IRBLkm-Ts	Pik-m	1
25	IRBL20-IR24	Pi20	1
26	IRBLta2-Pi	Pita2	1
27	IRBLta2-Re	Pita2	1
28	IRBLta-CP1	Pita	1
29	IRBL11-Zh	Pi11(t)	1
30	丽江新团黑谷 Lijiangxintuanheigu	-	1
31	谷梅4号 Gumei 4	Pigm	2

水稻广谱抗稻瘟病基因PigmR功能标记的开发及应用

Development and Application of the Functional Marker for the Broad-Spectrum Blast Resistance Gene PigmR in Rice

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