水稻Pi9位点6个稻瘟病抗性基因特异分子标记的开发及应用

doi:10.3864/j.issn.0578-1752.2023.21.007

中国农业科学 ›› 2023, Vol. 56 ›› Issue (21): 4219-4233.doi: 10.3864/j.issn.0578-1752.2023.21.007

水稻Pi9位点6个稻瘟病抗性基因特异分子标记的开发及应用

杨好¹(), 黄衍焱¹(), 易春霖¹, 石军², 谭楮湉¹, 任文芮¹, 王文明¹()

¹ 四川农业大学西南作物基因资源发掘与利用国家重点实验室，成都 611130
² 绵阳市农业科学研究院水稻研究所/厅市共建作物特色资源创制及应用四川省重点实验室，四川绵阳 621023

收稿日期:2023-07-23 接受日期:2023-09-06 出版日期:2023-11-01 发布日期:2023-11-06
通信作者:
王文明，E-mail：j316wenmingwang@163.com
联系方式: 杨好，E-mail：2365657091@qq.com。黄衍焱，E-mail：h1985yy@163.com。杨好和黄衍焱为同等贡献作者。
基金资助:
国家自然科学基金(U19A2033); 国家自然科学基金(31901839); 四川省青年科技创新研究团队(2022JDTD0023)

Development and Application of Specific Molecular Markers for Six Homologous Rice Blast Resistance Genes in Pi9 Locus of Rice

YANG Hao¹(), HUANG YanYan¹(), YI ChunLin¹, SHI Jun², TAN ChuTian¹, REN WenRui¹, WANG WenMing¹()

¹ State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130
² Institute of Rice Research, Mianyang Academy of Agricultural Sciences/Crop Characteristic Resources Creation and Utilization Key Laboratory of Sichuan Province, Mianyang 621023, Sichuan

Received:2023-07-23 Accepted:2023-09-06 Published:2023-11-01 Online:2023-11-06

摘要/Abstract

摘要：

【目的】水稻Pi9位点由多个串联的同源NLR基因组成，从中已克隆了超过10个优良的稻瘟病抗性基因。论文旨在鉴别水稻亲本Pi9位点抗性基因的组成，促进该位点基因快速、精准地应用于水稻抗性育种。【方法】对比Pi9位点已克隆抗性基因的序列，从中发掘各基因特异的核苷酸位点；然后将各目标基因分别与数据库（Rice Resource Center）中的155个水稻基因组进行比对分析，进一步从中筛选出特异最强的核苷酸位点，用于Pi2、Piz-t、Pi9、Pi9-type5、PigmR和Pid4 6个抗性基因特异分子标记的开发；以24个抗稻瘟病单基因系、阳性对照和110个四川盆地水稻亲本为鉴定对象，通过优化PCR扩增条件、测序或基因组数据分析检验分子标记鉴定结果的准确性。由于该位点基因的同源性较高、基因间常常拥有一些相同的特异位点，导致许多抗性基因很难用一对分子标记进行精准鉴定，因此采用多对分子标记共同鉴定的方式；另外许多特异位点为单碱基差异，因此需要对差异位点旁的碱基进行特异突变，使引物的3′端具有两个碱基的错配，以提高PCR扩增的特异性。【结果】最终为6个Pi9位点的抗性基因开发了有效的分子标记，发现32.09%的参试水稻材料含有Pi9位点抗性基因。Pi9、Pid4、PigmR、Piz-t、Pi2和Pi9-type5分别存在于1、7、8、14、23和33个水稻材料中，其中Pi9仅存在于单基因系中。这些抗性基因通常两个或多个组合在一起，同时存在于一个水稻材料中。其中Pi9-type5往往与Pi2或Piz-t成对出现，仅在成恢993、HR2168和绵恢365 3个亲本中单独存在。雨恢38含有的Pi9位点抗性基因最多，分别有Pi2、Pi9-type5、PigmR和Pid4。川谷B、川农4B、内香6B和双1B中均同时含有Piz-t、PigmR和Pid4 3个抗性基因。千乡654B中含有Piz-t和Pid4两个抗性基因。【结论】为Pi9位点的6个同源稻瘟病抗性基因开发了特异的分子标记，明确了四川盆地110个水稻亲本Pi9位点的抗性基因组成，揭示了Pi9位点抗性基因丰富的组合形式，为水稻育种的抗原选择提供了明确参考。

关键词: 水稻, 稻瘟病, Pi9位点, 同源基因, 分子标记

Abstract:

【Objective】The Pi9 resistance gene locus, conferring a broad-spectrum resistance against Magnaporthe oryzae, is consist by several tandem homologous genes. Over 10 resistance genes have been cloned from this gene locus. This study aims to clarify the R gene composition at Pi9 locus in rice resource materials and promote the application of those genes in rice resistance breeding.【Method】Comparing the DNA sequence of cloned R genes at Pi9 locus, the specific nucleotide polymorphism sites were screened as the candidate sites. Subsequently, each R gene was blasted with 155 rice genomes in the database of Rice Resource Center. The most specific nucleotide polymorphism sites were picked out from the candidate site in each gene to develop primer pair of molecular markers. The PCR product of primer pairs was used to mark indicated R gene in tested rice materials via parameter optimization. To verify the results, the R genes were cloned from indicated rice variety randomly and examined by Sanger sequencing, or analyzed the R genes from the genome database if the genome sequence of indicated rice variety exists in Rice Resource Center. The R genes in Pi9 locus have high homology, which cause same specific nucleotide polymorphism sites existing in different R genes. Therefore, some R genes are hardly identified by one molecular marker. For this case, several molecular markers were employed to identify the indicated R gene simultaneously. Moreover, some specific nucleotide polymorphism sites are single nucleotide polymorphism (SNP), in where the primers of molecular markers have a mismatched base. In order to improve the specificity of PCR amplification, the adjacent base of SNP was mutated to generate two mismatched bases at 3′ site of primer.【Result】Finally, the valid molecular markers were developed for each R gene and identified 32.09% tested materials containing R genes at Pi9 locus. Pi9, Pid4, PigmR, Piz-t, Pi2 and Pi9-type5 are present in 1, 7, 8, 14, 23 and 33 tested materials, respectively. The Pi9 only presents in monogenic line but not in rice parent lines. The other genes are usually present in two or more gene combinations in rice parent lines. The Pi9-type5 often presents in pair with Pi2 and Piz-t, and presents alone in three rice parents, Chenghui 993, HR2168 and Mianhui 365. Yuhui 38 contains the most R genes at Pi9 locus, including Pi2, Pi9-type5, PigmR and Pid4. Chuangu B, Chuannong 4B, Neixiang 6B and Shuang 1B contain Piz-t, PigmR and Pid4. Qianxiang 654B contains Piz-t and Pid4.【Conclusion】This study successfully developed specific molecular markers for six homologous rice blast resistance genes in Pi9 locus and identified the R gene composition in Pi9 locus for 110 rice parent lines that used in rice breeding in Sichuan basin. It also discovered different types of R genes combination at Pi9 locus and provided a clear reference for choosing the resistance source in rice breeding.

Key words: rice, rice blast, Pi9 locus, homologous gene, molecular marker

杨好, 黄衍焱, 易春霖, 石军, 谭楮湉, 任文芮, 王文明. 水稻Pi9位点6个稻瘟病抗性基因特异分子标记的开发及应用[J]. 中国农业科学, 2023, 56(21): 4219-4233.

YANG Hao, HUANG YanYan, YI ChunLin, SHI Jun, TAN ChuTian, REN WenRui, WANG WenMing. Development and Application of Specific Molecular Markers for Six Homologous Rice Blast Resistance Genes in Pi9 Locus of Rice[J]. Scientia Agricultura Sinica, 2023, 56(21): 4219-4233.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2023.21.007

https://www.chinaagrisci.com/CN/Y2023/V56/I21/4219

图/表 12

表1

供试水稻品种"

编号 No.	品种 Variety	编号 No.	品种 Variety	编号 No.	品种 Variety	编号 No.	品种 Variety
1	IRBLa-A (Pi-a)	35	绵恢146 Mianhui 146	69	泰恢808 Taihui 808	103	德香074B Dexiang 074B
2	IRBLi-F5 (Pi-i)	36	绵恢725 Mianhui 725	70	五山丝苗Wushansimiao	104	赣香B Ganxiang B
3	IRBLks-F5 (Pi-ks)	37	南恢558 Nanhui 558	71	西科恢768 Xikehui 768	105	广8B Guang 8B
4	IRBLk-Ka (Pi-k)	38	南恢968 Nanhui 968	72	雅禾Yahe	106	泸香618B Luxiang 618B
5	IRBLkp-K60 (Pi-kp)	39	蜀恢257 Shuhui 257	73	雅恢2115 Yahui 2115	107	绵5B Mian 5B
6	IRBLkh-K3 (Pi-kh)	40	蜀恢308 Shuhui 308	74	雅恢2116 Yahui 2116	108	忠香B Zhongxiang B
7	IRBLz-Fu (Pi-z)	41	蜀恢451 Shuhui 451	75	雅恢2117 Yahui 2117	109	内香6B Neixiang 6B
8	IRBLz5-CA (Pi-z5)	42	蜀恢498 Shuhui 498	76	雅恢2118 Yahui 2118	110	千乡654B Qianxiang 654B
9	IRBLzt-T (Pi-zt)	43	蜀恢505 Shuhui 505	77	雅恢2119 Yahui 2119	111	荃9311B Quan 9311B
10	IRBLta-K1 (Pi-ta)	44	蜀恢548 Shuhui 548	78	雅恢2199 Yahui 2199	112	蓉18B Rong 18B
11	IRBLb-B (Pi-b)	45	蜀恢573 Shuhui 573	79	雅恢2275 Yahui 2275	113	蓉7B Rong 7B
12	IRBLt-K59 (Pi-t)	46	蜀恢586 Shuhui 586	80	雅恢2348 Yahui 2348	114	双1B Shuang 1B
13	IRBLsh-B (Pi-sh)	47	蜀恢589 Shuhui 589	81	雅恢2816 Yahui 2816	115	泰丰B Taifeng B
14	IRBL1-CL (Pi-1)	48	猪毛糯Zhumaonuo	82	雅恢2918 Yahui 2918	116	武金4B Wujin 4B
15	IRBL3-CP4 (Pi-3)	49	HR2168	83	雅恢5049 Yahui 5049	117	武香B Wuxiang B
16	IRBL5-M (Pi-5)	50	R900	84	雅恢5217 Yahui 5217	118	雅1B Ya 1B
17	IRBL7-M (Pi-7)	51	R9311	85	雨恢38 Yuhui 38	119	雅3B Ya 3B
18	IRBL9-W (Pi-9)	52	成恢19 Chenghui 19	86	岳恢9113 Yuehui 9113	120	雅7B Ya 7B
19	IRBL12-M (Pi-12)	53	乐恢188 Lehui 188	87	粤农丝苗Yuenongsimiao	121	宜香1B Yixiang 1B
20	IRBL19-M (Pi-19)	54	乐恢584 Lehui 584	88	D297B	122	泰国香稻Thai fragrant rice
21	IRBLkm-Ts (Pi-km)	55	泸恢17 Luhui 17	89	D35B	123	楚粳28 Chugeng 28
22	IRBL20-IR24 (Pi-20)	56	泸恢8258 Luhui 8258	90	D62B	124	野香B Yexiang B
23	IRBLta2-Pi (Pi-ta2)	57	绵恢523 Mianhui 523	91	D79B	125	绵恢6139 Mianhui 6139
24	IRBL11-Zh (Pi-11)	58	绵恢528 Mianhui 528	92	F-32B	126	绵恢313 Mianhui 313
25	E33	59	绵恢662 Mianhui 662	93	F59B	127	绵恢357 Mianhui 357
26	成恢3203 Chenghui 3203	60	明恢63 Minghui 63	94	G46B	128	绵恢365 Mianhui 365
27	成恢727 Chenghui 727	61	明恢86 Minghui 86	95	川106B Chuan 106B	129	绵恢821 Mianhui 821
28	成恢993 Chenghui 993	62	内恢99-14 Neihui 99-14	96	川608B Chuan 608B	130	绵恢823 Mianhui 823
29	德恢6099 Dehui 6099	63	蜀恢137 Shuhui 137	97	川谷B Chuangu B	131	绵恢837 Mianhui 837
30	多恢1号 Duohui 1	64	蜀恢208 Shuhui 208	98	川华B Chuanhua B	132	绵恢857 Mianhui 857
31	航恢1179 Hanghui 1179	65	蜀恢527 Shuhui 527	99	川农1B Chuannong 1B	133	绵恢815 Mianhui 815
32	航恢570 Hanghui 570	66	蜀恢538 Shuhui 538	100	川农4B Chuannong 4B	134	地谷B Digu B
33	华占Huazhan	67	蜀恢569 Shuhui 569	101	川种3B Chuanzhong 3B
34	乐恢433 Lehui 433	68	蜀恢882 Shuhui 882	102	德66B De 66B

表1

表2

图1

表3

表4

图2

表5

图3

表6

图4

表7

表8

参考文献 32

[1]	DENG Y, ZHAI K, XIE Z, YANG D, ZHU X, LIU J, WANG X, QIN P, YANG Y, ZHANG G, et al. 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
[2]	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 Pi2/9locus. Theoretical and Applied Genetics, 2015, 128(11): 2213-2225. doi: 10.1007/s00122-015-2579-9
[3]	XIE Z, YAN B, SHOU J, TANG J, WANG X, ZHAI K, LIU J, LI Q, LUO M, DENG Y, HE Z. A nucleotide-binding site-leucine-rich repeat receptor pair confers broad-spectrum disease resistance through physical association in rice. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences, 2019, 374(1767): 20180308.
[4]	黄衍焱, 李燕, 王贺, 王文明. 水稻小种特异性抗稻瘟病基因的等位性变异研究进展. 植物病理学报, 2023, doi: 10.13926/j.cnki.apps.000866.
	HUANG Y Y, LI Y, WANG H, WANG W M. Allelic variation in the race-specific blast resistance genes in rice. Acta Phytopathologica Sinica, 2023, doi: 10.13926/j.cnki.apps.000866. (in Chinese)
[5]	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
[6]	ZHOU B, DOLAN M, SAKAI H, WANG G L. The genomic dynamics and evolutionary mechanism of the Pi2/9locus in rice. Molecular Plant-Microbe Interactions, 2007, 20(1): 63-71. doi: 10.1094/MPMI-20-0063
[7]	ZHOU Y, LEI F, WANG Q, HE W, YUAN B, YUAN W. Identification of novel alleles of the rice blast-resistance gene Pi9 through sequence-based allele mining. Rice, 2020, 13(1): 80. doi: 10.1186/s12284-020-00442-z
[8]	ZHOU B, QU S, LIU G, DOLAN M, SAKAI H, LU G, 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
[9]	CHEN Z, ZHAO W, ZHU X, ZOU C, YIN J, CHERN M, ZHOU X, YING H, JIANG X, LI Y, et al. Identification and characterization of rice blast resistance gene Pid4by a combination of transcriptomic profiling and genome analysis. Journal of Genetics and Genomics, 2018, 45(12): 663-672. doi: 10.1016/j.jgg.2018.10.007
[10]	马军韬, 李文华, 张国民, 王永力, 张丽艳, 孟文凯. 水直播条件下黑龙江省不同稻区稻瘟病菌致病性分析. 植物保护学报, 2020, 47(5): 987-996.
	MA J T, LI W H, ZHANG G M, WANG Y L, ZHANG L Y, MENG W K. Pathogenicity of Magnaporthe oryzae from different rice growing regions of Heilongjiang Province under water direct seeding. Journal of Plant Protection, 2020, 47(5): 987-996. (in Chinese)
[11]	田珂, 杨武, 李泌, 徐鑫, 刘新琼, 刘学群, 王春台. 鄂西南地区2012-2014年稻瘟病菌致病性变化分析. 华中农业大学学报, 2017, 36(5): 10-14.
	TIAN K, YANG W, LI M, XU X, LIU X Q, LIU X Q, WANG C T. Pathogenicity changes of Magnaporthe oryzae in south-western Hubei Province during 2012-2014. Journal of Huazhong Agricultural University, 2017, 36(5): 10-14. (in Chinese)
[12]	马继琼, 孙一丁, 杨奕, 李进斌, 许明辉. 适用于云南粳稻种植区的稻瘟病抗性基因分析. 分子植物育种, 2021, 19(5): 1556-1568.
	MA J Q, SUN Y D, YANG Y, LI J B, XU M H. Analysis of rice blast resistance genes suitable for japonica rice planting areas in Yunnan. Molecular Plant Breeding, 2021, 19(5): 1556-1568. (in Chinese)
[13]	张姝, 钟雪莲, 乔贵艳, 沈丽, 周天云, 彭云良. 川渝黔稻瘟病菌毒性的区域分化研究. 西南农业学报, 2017, 30(2): 359-365.
	ZHANG S, ZHONG X L, QIAO G Y, SHEN L, ZHOU T Y, PENG Y L. Difference in virulence of Magnaporthe oryzae from Sichuan, Chongqing and Guizhou. Southwest China Journal of Agricultural Sciences, 2017, 30(2): 359-365. (in Chinese)
[14]	SHI J, LI D, LI Y, LI X, GUO X, LUO Y, LU Y, ZHANG Q, XU Y, FAN J, HUANG F, WANG W. Identification of rice blast resistance genes in the elite hybrid rice restorer line Yahui 2115. Genome, 2015, 58(3): 91-97. doi: 10.1139/gen-2015-0005
[15]	HASSAN B, PENG Y T, LI S, YIN X X, CHEN C, GULZAR F, ZHOU S X, PU M, JI Y P, WANG Y P, ZHAO W, HUANG F, PENG Y L, ZHAO Z X, WANG W M. Identification of the blast resistance genes in three elite restorer lines of hybrid rice. Phytopathology Research, 2022, 4(1): 15. doi: 10.1186/s42483-022-00120-6
[16]	TSUNEMATSU H, YANORIA M J T, EBRON L A, HAYASHI N, ANDO I, KATO H, IMBE T, KHUSH G S. Development of monogenic lines of rice for blast resistance. Breeding Science, 2000, 50(3): 229-234. doi: 10.1270/jsbbs.50.229
[17]	陈嘉林, 余敏祥, 何旎清, 黄凤凰, 杨德卫. 稻瘟病抗病基因分子标记的开发与育种利用. 分子植物育种, http://kns.cnki.net/kcms/detail/46.1068.S.20220112.1946.012.html.
	CHEN J L, YU M X, HE N Q, HUANG F H, YANG D W. Development and breeding application of molecular markers for rice blast resistance genes. Molecular Plant Breeding, http://kns.cnki.net/kcms/detail/46.1068.S.20220112.1946.012.html. in Chinese)
[18]	HAYASHI K, YOSHIDA H, ASHIKAWA I. Development of PCR- based allele-specific and InDel marker sets for nine rice blast resistance genes. Theoretical and Applied Genetics, 2006, 113(2): 251-260. doi: 10.1007/s00122-006-0290-6
[19]	华丽霞, 汪文娟, 陈深, 汪聪颖, 曾烈先, 杨健源, 朱小源, 苏菁. 抗稻瘟病Pi2/9/z-t基因特异性分子标记的开发. 中国水稻科学, 2015, 29(3): 305-310.
	HUA L X, WANG W J, CHEN S, WANG C Y, ZENG L X, YANG J Y, ZHU X Y, SU J. Development of specific DNA markers for detecting the rice blast resistance gene alleles Pi2/9/z-t. Chinese Journal of Rice Science, 2015, 29(3): 305-310. (in Chinese)
[20]	杨立明, 纪剑辉, 周颖君, 方继朝, 刘永锋, 罗玉明. 水稻稻瘟病抗性基因Pi2-InDel标记的开发与评价. 分子植物育种, 2017, 15(2): 594-598.
	YANG L M, JI J H, ZHOU Y J, FANG J C, LIU Y F, LUO Y M. The development of the rice blast resistance genes Pi2-InDel markers and its evaluation in application. Molecular Plant Breeding, 2017, 15(2): 594-598. (in Chinese)
[21]	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. 中国农业科学, 2019, 52(6): 955-967. doi: 10.3864/j.issn.0571-1752.2019.06.001.
	WANG F Q, CHEN Z H, XU Y, WANG J, LI W Q, FAN F J, CHEN L Q, TAO Y J, ZHONG W G, YANG J. Development and application of the functional marker for the broad-spectrum blast resistance gene PigmR in rice. Scientia Agricultura Sinica, 2019, 52(6): 955-967. doi: 10.3864/j.issn.0571-1752.2019.06.001. (in Chinese)
[22]	谢旺有, 陈锦文, 谢少和, 陈惠清, 王天生, 余文昌, 侯凯强. 抗稻瘟病基因Pi9导入籼稻恢复系泉恢039的应用研究. 福建农业学报, 2022, 37(10): 1266-1274.
	XIE W Y, CHEN J W, XIE S H, CHEN H Q, WANG T S, YU W C, HOU K Q. Introduction of blast resistant Pi9 in indica rice restorer line Quanhui 039. Fujian Journal of Agricultural Sciences, 2022, 37(10): 1266-1274. (in Chinese)
[23]	柳絮, 张华, 宣宁, 李平波, 张梦琦, 姚方印. 水稻抗虫(Cry1C)和抗稻瘟病(Pi1,Pi2)材料的创制. 山东农业科学, 2021, 53(2): 89-93.
	LIU X, ZHANG H, XUAN N, LI P B, ZHANG M Q, YAO F Y. Breeding of new rice varieties with rice blast and insect resistances. Shandong Agricultural Sciences, 2021, 53(2): 89-93. (in Chinese)
[24]	王哉, 蒋英健, 谢留杰, 王敏天, 黄丛林, 陈嘉乐, 阮文晓. 分子标记辅助选育含Pigm抗稻瘟病基因的水稻新品系. 浙江农业科学, 2023, 64(8): 1945-1948.
	WANG Z, JIANG Y J, XIE L J, WANG M T, HUANG C L, CHEN J L, RUAN W X. The new rice lines containing rice blast resistance gene Pigm were selected by molecular marker assisted breeding. Journal of Zhejiang Agricultural Sciences, 2023, 64(8): 1945-1948. (in Chinese)
[25]	于连鹏. 黑龙江省主栽水稻品种Pita、Pia和Piz-t抗瘟基因检测和抗性评价[D]. 大庆: 黑龙江八一农垦大学, 2017.
	YU L P. Pita, Pia and Piz-t genes detection and blast resistance evaluation of main rice varieties in Heilongjiang Province[D]. Daqing: Heilongjiang Bayi Agricultural University, 2017. (in Chinese)
[26]	张亚玲, 高清, 赵羽涵, 刘瑞, 付忠举, 李雪, 孙宇佳, 靳学慧. 黑龙江省水稻种质稻瘟病抗性评价及抗瘟基因结构分析. 中国农业科学, 2022, 55(4): 625-645. doi: 10.3864/j.issn.0578-1752.2022.04.001.
	ZHANG Y L, GAO Q, ZHAO Y H, LIU R, FU Z J, LI X, SUN Y J, JIN X H. Evaluation of rice blast resistance and genetic structure analysis of rice germplasm in Heilongjiang Province. Scientia Agricultura Sinica, 2022, 55(4): 625-645. doi: 10.3864/j.issn.0578-1752.2022.04.001. (in Chinese)
[27]	陈晴晴, 杨雪, 张爱芳. 长江中下游区试水稻品种稻瘟病抗性评价及抗性基因检测. 南方农业学报, 2022, 53(1): 21-28.
	CHEN Q Q, YANG X, ZHANG A F. Evaluation of rice blast resistance and detection of resistance genes of rice varieties in regional trials in the middle and lower reaches of Yangtze River. Journal of Southern Agriculture, 2022, 53(1): 21-28. (in Chinese)
[28]	董丽英, 刘树芳, 田维逵, 周伍民, 张先闻, 李迅东, 杨勤忠. 云南省稻瘟病菌群体的致病性分析及交配型分布. 植物保护学报, 2023, 50(2): 316-324.
	DONG L Y, LIU S F, TIAN W K, ZHOU W M, ZHANG X W, LI X D, YANG Q Z. Pathogenicity and mating type of rice blast fungus Magnaporthe oryzae isolates in Yunnan Province. Journal of Plant Protection, 2023, 50(2): 316-324. (in Chinese)
[29]	桑世飞, 王亚男, 王君怡, 曹梦雨, 孙晓涵, 姬生栋. 9个抗稻瘟病基因在291份水稻种质资源中的分布及组合特征. 河南农业科学, 2022, 51(12): 19-27.
	SANG S F, WANG Y N, WANG J Y, CAO M Y, SUN X H, JI S D. Distribution and combination features of nine blast resistant genes in 291 rice germplasm resources. Journal of Henan Agricultural Sciences, 2022, 51(12): 19-27. (in Chinese) doi: 10.15933/j.cnki.1004-3268.2022.12.003
[30]	何弯弯, 王健康, 丁成伟, 郭荣良, 吴玉玲, 王友霜, 赵轶鹏, 胡婷婷. Pib、Pi9、Pi2、Pi54和Pish在粳稻品种(系)中的分布及对穗颈瘟的抗性. 西南农业学报, 2022, 35(3): 497-502.
	HE W W, WANG J K, DING C W, GUO R L, WU Y L, WANG Y S, ZHAO Y P, HU T T. Distribution of Pib, Pi9, Pi2, Pi54 and Pish genes in japonica rice varieties (lines) and resistance to panicle blast. Southwest China Journal of Agricultural Sciences, 2022, 35(3): 497-502. (in Chinese)
[31]	沈浙南, 时焕斌, 邱结华, 王聪聪, 孟帅, 寇艳君. 江苏省212份水稻材料中抗稻瘟病基因Pi2/Pi9/Piz-t/Pigm的情况分析. 分子植物育种, 2023, 21(13): 4344-4351.
	SHEN Z N, SHI H B, QIU J H, WANG C C, MENG S, KOU Y J. Analysis of rice blast resistance gene Pi2/Pi9/Piz-t/Pigm in 212 rice materials in Jiangsu Province. Molecular Plant Breeding, 2023, 21(13): 4344-4351. (in Chinese)
[32]	刘军化, 黄成志, 蒋静玥, 吕直文, 刘忠贤, 蔡钟亚, 雷树凡. 87份水稻材料中抗稻瘟病基因的分子检测. 西南农业学报, 2022, 35(9): 2030-2037.
	LIU J H, HUANG C Z, JIANG J Y, LÜ Z W, LIU Z X, CAI Z Y, LEI S F. Molecular detection of blast resistance genes in 87 rice materials. Southwest China Journal of Agricultural Sciences, 2022, 35(9): 2030-2037. (in Chinese)

编号 No.	材料 Material	引物Primer
编号 No.	材料 Material	Pi2/z-t-F Piz-t-R2	Pi2/z-t-F Piz-t-R3	Piz-t-F4 Piz-t-R4
9	IRBLzt-T	√	√	√
42	蜀恢498 Shuhui 498	√	√	√
52	成恢19 Chenghui 19	√	√	√
56	泸恢8258 Luhui 8258	√	√	√
61	明恢86 Minghui 86	√	√	√
63	蜀恢137 Shuhui 137	√	√	√
64	蜀恢208 Shuhui 208	√	√	√
78	雅恢2199 Yahui 2199	√	√	√
97	川谷B Chuangu B	√	√	√
100	川农4B Chuannong 4B	√	√	√
109	内香6B Neixiang 6B	√	√	√
110	千乡654B Qianxiang 654B	√	√	√
113	蓉7B Rong 7B	√	√	√
114	双1B Shuang 1B	√	√	√
阳性对照Positive control	中花11 ZH11	√	√	√

编号 No.	材料 Material	引物Primer
编号 No.	材料 Material	Pi9-type5-F1 Pi9-type5-R1	Pi9-type5-F2 Pi9-type5-R2	Pi9-type5-F3 Pi9-type5-mR
8	IRBLz5-CA	√	√	√
9	IRBLzt-T	√	√	√
25	E33	√	√	√
28	成恢993 Chenghui 993	√	√	√
29	德恢6099 Dehui 6099	√	√	√
31	航恢1179 Hanghui 1179	√	√	√
33	华占Huazhan	√	√	√
40	蜀恢308 Shuhui 308	√	√	√
42	蜀恢498 Shuhui 498	√	√	√
43	蜀恢505 Shuhui 505	√	√	√
44	蜀恢548 Shuhui 548	√	√	√
46	蜀恢586 Shuhui 586	√	√	√
50	R900	√	√	√
52	成恢19 Chenghui 19	√	√	√
56	泸恢8258 Luhui 8258	√	√	√
61	明恢86 Minghui 86	√	√	√
63	蜀恢137 Shuhui 137	√	√	√
64	蜀恢208 Shuhui 208	√	√	√
70	五山丝苗Wushansimiao	√	√	√
72	雅禾Yahe	√	√	√
73	雅恢2115 Yahui 2115	√	√	√
74	雅恢2116 Yahui 2116	√	√	√
77	雅恢2119 Yahui 2119	√	√	√
78	雅恢2199 Yahui 2199	√	√	√
79	雅恢2275 Yahui 2275	√	√	√
82	雅恢2918 Yahui 2918	√	√	√
83	雅恢5049 Yahui 5049	√	√	√
84	雅恢5217 Yahui 5217	√	√	√
85	雨恢38 Yuhui 38	√	√	√
87	粤农丝苗Yuenongsimiao	√	√	√
118	雅1B Ya 1B	√	√	√
128	绵恢365 Mianhui 365	√	√	√
133	绵恢815 Mianhui 815	√	√	√

编号 No.	材料 Material	引物Primer
编号 No.	材料 Material	Pigm-F2+ Pigm-newmR1	Pigm-newmF1+ Pigm-newmR1	Pigm-mF4+ Pigm-mR5	Pigm-mF6+ Pigm-mR7
85	雨恢38 Yuhui 38	√	√	√	√
97	川谷B Chuangu B	√	√	√	√
100	川农4B Chuannong 4B	√	√	√	√
109	内香6B Neixiang 6B	√	√	√	√
112	蓉18B Rong 18 B	√	√	√	√
113	蓉7B Rong 7B	√	√	√	√
114	双1B Shuang 1B	√	√	√	√
134	地谷B Digu B	√	√	√	√
阳性对照Positive control	谷梅4号Gumei 4	√	√	√	√

编号 No.	材料 Material	抗性基因Resistance gene
编号 No.	材料 Material	Pi2	Piz-t	Pi9	Pi9-type5	PigmR	Pid4
18	IRBL9-W			√
8	IRBLz5-CA	√			√
25	E33	√			√
29	德恢6099 Dehui 6099	√			√
31	航恢1179 Hanghui 1179	√			√
33	华占Huazhan	√			√
40	蜀恢308 Shuhui 308	√			√
43	蜀恢505 Shuhui 505	√			√
44	蜀恢548 Shuhui 548	√			√
46	蜀恢586 Shuhui 586	√			√
50	R900	√			√
70	五山丝苗Wushansimiao	√			√
72	雅禾Yahe	√			√
73	雅恢2115 Yahui 2115	√			√
74	雅恢2116 Yahui 2116	√			√
77	雅恢2119 Yahui 2119	√			√
79	雅恢2275 Yahui 2275	√			√
82	雅恢2918 Yahui 2918	√			√
83	雅恢5049 Yahui 5049	√			√
84	雅恢5217 Yahui 5217	√			√
85	雨恢38 Yuhui 38	√			√	√	√
87	粤农丝苗Yuenongsimiao	√			√
118	雅1B Ya 1B	√			√
133	绵恢815 Mianhui 815	√			√
9	IRBLzt-T		√		√
42	蜀恢498 Shuhui 498		√		√
52	成恢19 Chenghui 19		√		√
56	泸恢8258 Luhui 8258		√		√
61	明恢86 Minghui 86		√		√
63	蜀恢137 Shuhui 137		√		√
64	蜀恢208 Shuhui 208		√		√
78	雅恢2199 Yahui 2199		√		√
97	川谷B Chuangu B		√			√	√
100	川农4B Chuannong 4B		√			√	√
109	内香6B Neixiang 6B		√			√	√
113	蓉7B Rong 7B		√			√	√
114	双1B Shuang 1B		√			√	√
110	千乡654B Qianxiang 654B		√				√
28	成恢993 Chenghui 993				√
49	HR2168				√
128	绵恢365 Mianhui 365				√
134	地谷B Digu B					√	√
112	蓉18B Rong 18B					√

水稻Pi9位点6个稻瘟病抗性基因特异分子标记的开发及应用

Development and Application of Specific Molecular Markers for Six Homologous Rice Blast Resistance Genes in Pi9 Locus of Rice

RichHTML

PDF

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 12

参考文献 32

相关文章 15

Metrics

本文评价

推荐阅读 0

R基因 R gene	引物名称 Primer name	引物序列 Primer sequence	产物长度 Product length (bp)	退火温度 Annealing temperature (℃)	延伸时间 Extension time (s)
Pi2	Pi2/z-t-F	CTCTTCGTTGTATAGGACAGTTTCA	424	57	30
Pi2	Pi2-R	GTTCCACCATCTGAGATTCCTG	424	57	30
Pi9	Pi9-F	CTCTTCGTTGTAGTAGAAAGTTTGT	788	45	50
Pi9	Pi9-R	TTCCAACCTGCAGCAAGAG	788	45	50
Pi9-type5	Pi9-type5F1	CTAAGAACAATAAGAAGGGCAG	454	62	30
	Pi9-type5R1	CCTACTAGACGATTCCTTTGT	454	62	30
	Pi9-type5F2	CGTTGTATAAGAGAGCTTGAATT	709	60	50
	Pi9-type5R2	GCTGATCCAAATTGAAAATCAC	709	60	50
	Pi9-type5F3	ATCTTTCCTGAGGATTTTGAAATAC	185	55	25
	Pi9-type5mR	CAAGTCTTAATTTTTCCTGCAG	185	55	25
Piz-t	Pi2/z-t-F	CTCTTCGTTGTATAGGACAGTTTCA	548	57	35
	Piz-t-R3	GTAGATCTTCTTCAGGTGCA	548	57	35
	Pi2/z-t-F	CTCTTCGTTGTATAGGACAGTTTCA	502	59	30
	Piz-t-R2	TCCTCAAGAATTCCATCCAACAC	502	59	30
	Piz-t-F4	CAGAGAACGCCACCGAATT	169	59	20
	Piz-t-R4	CACATTGCGAGCTGACTGATTA	169	59	20
Pi9-type9	Pi9-type9F2	AGAATGGGAGAAATTCTATGAAAAG	230	59	25
	Pi9-type9R2	TAGTCATCATCCCAACCTGC	230	59	25
	Pi9-type9F	ACCTGAAGAAGATCTACTTATTGAA	305	59	25
	Pi9-type9R	AATCTCCTTGAGCTTTGGAT	305	59	25
	Pi9-type9mF	ACCTGAAGAAGATCTACTTATTGTA	305	59	25
	Pi9-type9mR	AATCTCCTTGAGCTTTGGTT	305	59	25
PigmR	Pigm-F2	CACCTGAAGAAGTTCTACTTACG	111	59	20
	Pigm-newmR1	ATAAGAATTATGATAAAGAGAAAGGTA	111	59	20
	Pigm-newmF1	TCCTCCTCCCCTACTTAGT	180	59	20
	Pigm-newmR1	ATAAGAATTATGATAAAGAGAAAGGTA	180	59	20
	Pigm-mF4	GGAGAAGCTAGTATTCAAAAGG	297	59	20
	Pigm-mR5	TGATACCTTCGGTCACTGAC	297	59	20
	Pigm-mF6	GTATATGGATAGCAGAAGGGTAG	98	59	20
	Pigm-mR7	CATACTTCGGTTGATTAGCTCAAA	98	59	20
Pid4	Pid4-mF	GAATCTGTTTTTCCTGACTCTACA	797	59	50
Pid4	Pid4-mR	GGATTGTAAAAGAAACTTAATGAGT	797	59	50

[1]	李浩, 陈金, 王洪亮, 柳开楼, 韩天富, 都江雪, 申哲, 刘立生, 黄晶, 张会民. 红壤性水稻土有机无机复合体中碳氮特征对长期施肥的响应[J]. 中国农业科学, 2023, 56(7): 1333-1343.
[2]	温一博, 陈淑婷, 徐正进, 孙健, 徐铨. DEP1、Gn1a和qSW5组合应用调控水稻穗部性状[J]. 中国农业科学, 2023, 56(7): 1218-1227.
[3]	李儒香, 周恺, 王大川, 李巧龙, 向奥妮, 李璐, 李苗苗, 向思茜, 凌英华, 何光华, 赵芳明. 水稻CSSL-Z481代换片段携带的穗部性状QTL分析及次级代换系培育[J]. 中国农业科学, 2023, 56(7): 1228-1247.
[4]	赵梓钧, 吴如会, 王硕, 张君, 游静, 段倩楠, 唐俊, 张新芳, 韦秘, 刘金艳, 李云峰, 何光华, 张婷. 水稻PDL2的突变导致小穗外稃退化[J]. 中国农业科学, 2023, 56(7): 1248-1259.
[5]	朱洪慧, 李映姿, 高远卓, 林泓, 王成洋, 晏紫仪, 彭瀚平, 李田野, 熊茂, 李云峰. 水稻短宽粒基因SWG1的图位克隆[J]. 中国农业科学, 2023, 56(7): 1260-1274.
[6]	张绩, 周上铃, 何发, 刘莉莎, 张玉娟, 何晋宇, 杜晓秋. 水稻α-淀粉酶基因的表达模式与颖花开放的关系[J]. 中国农业科学, 2023, 56(7): 1275-1282.
[7]	贺江, 丁颖, 娄向弟, 姬东玲, 张向向, 王永慧, 张伟杨, 王志琴, 王伟露, 杨建昌. 水稻分蘖期干物质积累对大气CO₂浓度升高和氮素营养的综合响应差异及其生理机制[J]. 中国农业科学, 2023, 56(6): 1045-1060.
[8]	谢军, 尹学伟, 魏灵, 王子芳, 李清虎, 张晓春, 鲁远源, 王秋月, 高明. 垄作直播控制灌溉对水稻产量和温室气体排放的影响[J]. 中国农业科学, 2023, 56(4): 697-710.
[9]	刘刚, 夏快飞, 吴艳, 张明永, 张再君, 杨金松, 邱东峰. 水稻耐热新种质R203的创制与应用[J]. 中国农业科学, 2023, 56(3): 405-415.
[10]	刘瑞, 赵羽涵, 顾欣怡, 王艳霞, 靳学慧, 吴伟怀, 张亚玲. 黑龙江省和海南省稻瘟病菌中AVR-Pita家族的分布及变异分析[J]. 中国农业科学, 2023, 56(3): 466-480.
[11]	徐海, 李秀坤, 芦佳浩, 姜恺, 马玥, 徐正进, 徐铨. 籼型血缘渗入对北方粳稻产量和品质的影响[J]. 中国农业科学, 2023, 56(22): 4359-4370.
[12]	李莉, 孙玲, 张金花, 邹晓威, 孙辉, 任金平, 姜兆远, 刘晓梅. 基于稻瘟病菌小种变化的吉林省主要粳稻品种抗性评价及利用价值分析[J]. 中国农业科学, 2023, 56(22): 4441-4452.
[13]	张泽源, 李玥, 赵文莎, 顾晶晶, 张傲琰, 张海龙, 宋鹏博, 吴建辉, 张传量, 宋全昊, 简俊涛, 孙道杰, 王兴荣. 小麦粒重相关性状的QTL定位及分子标记的开发[J]. 中国农业科学, 2023, 56(21): 4137-4149.
[14]	王伟康, 张嘉懿, 汪慧, 曹强, 田永超, 朱艳, 曹卫星, 刘小军. 基于固定翼无人机多光谱影像的水稻长势关键指标无损监测[J]. 中国农业科学, 2023, 56(21): 4175-4191.
[15]	樊自尧, 李奎, 李家洋, 黄三文. “稻薯猪”生态循环农业的设想[J]. 中国农业科学, 2023, 56(20): 4067-4071.