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
收稿日期: 2017-11-09
修回日期: 2018-01-30
网络出版日期: 2018-09-10
基金资助
国家重点研发计划资助项目(2017YFD0100400);镇江市科技支撑计划资助项目(NY2015018);江苏省重点研发计划资助项目(BE2015363);江苏省农业科技自主创新资金资助项目[CX(16)1029]
Development of Specific Markers for Pigm in Marker-Assisted Breeding of Panicle Blast Resistant japonica Rice
Received date: 2017-11-09
Revised date: 2018-01-30
Online published: 2018-09-10
【目的】Pigm是一个广谱的稻瘟病抗性基因,源自持久抗性品种谷梅4号,与Piz、Piz-t、Pi2、Pi9和Pi40等互为复等位基因但抗谱存在差异。为了更好地在分子辅助选择育种中利用Pigm基因,开发与Pigm特异性标记具有重要意义。【方法】在已有文献报道定位结果的基础上,通过随机测序获得了一段谷梅4号基因组的特异序列,并据此开发了一组用于筛选Pigm基因的分子标记,进一步选取江淮稻区3个不同生态型代表性粳稻品种作为受体,利用分子标记辅助选择结合对抗性基因的背景检测将Pigm基因导入受体品种。【结果】Pigm-4标记位于Pigm基因簇内部,与抗病功能元件PigmR紧密连锁,对不同类型的品种检测发现该标记特异性强,且利用该标记可将Pigm与Piz、Piz-t、Pi2、Pi9以及Pi40区分开来。对受体亲本稻瘟病抗性基因的检测和接种结果分析发现江淮稻区粳稻品种虽然携带了Pib、Pi54、Pita、Pb1中的2~3个基因,但是对强毒力的稻瘟病小种抗性普遍不强,而3种代表性粳稻背景下导入Pigm基因均可显著提高其对穗颈瘟的抗性水平。【结论】Pigm可以作为抗稻瘟病粳稻育种的有利基因资源加以利用,而Pigm-4是分子标记辅助筛选Pigm的优异标记。
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[J]. 中国水稻科学, 2018 , 32(5) : 453 -461 . DOI: 10.16819/j.1001-7216.2018.7135
【Objective】 Pigm is a broad-spectrum resistance gene to rice blast, which is isolated from Gumei 4, a variety with durable resistance. Pigm is allelic to Piz, Piz-t, Pi2, Pi9, Pi40 and so on, but shows different resistance spectrum. To breed resistance variety carrying Pigm with molecular marker-assisted selection, discovering Pigm-tightly-linked markers is important.【Method】We identified a unique DNA sequence of Gumei 4 by genomic sequencing, and developed a set of specific markers tagged to Pigm base on the sequence divergence. Then, by means of molecular-assisted selection and background gene screening, Pigm was pyramided to three representative japonica varieties belonging to different ecotypes from Yangtze-Huai River Area. 【Result】 An InDel marker Pigm-4 which is closely linked to the functional unit of Pigm was developed. Pigm-4 showed distinct specificity by genotype screening of various rice varieties, and can distinguish the genotype of Pigm from its alleles Piz, Piz-t, Pi2, Pi9 and Pi40. Although receptors carry with resistant genes such as Pib+Pi54 (
Key words： Pigm; specific-marker; japonica rice; panicle blast; resistance
| [1] | Liu Q, Yang J Y, Zhang S H, Zhao J L, Feng A Q, Yang T F, Wang X F, Mao X X, Dong J F, Zhu X Y, Leung H, Leach J E, Liu B.OsGF14b positively regulates panicle blast resistance but negatively regulates leaf blast resistance in rice. Mol Plant Microbe In, 2016, 29(1): 46-56. |
| [2] | Zhuang J Y, Ma W B, Wu J L, Chai R Y, Lu J, Fan Y Y, Jin M Z, Leung H, Zheng K L.Mapping of leaf and neck blast resistance genes with resistance gene analog, RAPD and RFLP in rice.Euphytica, 2002, 128(3): 363-370. |
| [3] | 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. |
| [3] | Zhu F, Tian Z H, Tan D L, Liu Y F.How to improve the countermeasures-Based on the rice blast outburst in 2014.Jiangsu Agric Sci, 2016, 44(8): 155-158. (in Chinese) |
| [4] | Gouda P K, Saikumar S, Varma C M K, Nagesh K, Thippeswamy S, Shenoy V, Ramesha M S, Shashidhar H E. Marker-assisted breeding of Pi-1 and Piz-5 genes imparting resistance to rice blast in PRR78, restorer line of Pusa RH-10 Basmati rice hybrid. Plant Breeding, 2013, 132(1): 61-69. |
| [5] | Costanzo S, Jia Y L.Sequence variation at the rice blast resistance gene Pi-km locus: Implications for the development of allele specific markers. Plant Sci, 2010, 178(6): 523-530. |
| [6] | Deng Y W, Zhu X D, Shen Y, He Z H.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. Theor Appl Genet, 2006, 113(4): 705-713. |
| [7] | Jiang N, Li Z Q, Wu J, Wang Y, Wu L Q, Wang S H, Wang D J, Wen T J, Liang Y, Sun P Y.Molecular mapping of the Pi2/9 allelic gene Pi2-2 conferring broad-spectrum resistance to Magnaporthe oryzae in the rice cultivar Jefferson. Rice, 2012, 5(21): 1-7. |
| [8] | Turnbull L A, Levine J M, Fergus A J F, Petermann J S. The Pi40 gene for durable resistance to rice blast and molecular analysis of Pi40-advanced backcross breeding lines. Phytopathology, 2009, 99(3): 243-250. |
| [9] | Zhu X Y, Shen C, Yang J Y, Zhou S C, Zeng L X, Han J L, Jing S, Ling W, Pan Q H.The identification of Pi50(t), a new member of the rice blast resistance Pi2/Pi9 multigene family. Theor Appl Genet, 2012, 124(7): 1295-1304. |
| [10] | Su J, Wang W J, Han J H, Chen S, Wang C Y, Zeng L X, Feng A Q, Yang J Y, Zhou B, Zhu X Y.Functional divergence of duplicated genes results in a novel blast resistance gene Pi50 at the Pi2/9 locus. Theor Appl Genet, 2015, 128(11): 1-13. |
| [11] | 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. Mol Plant Microbe In, 2006, 19(11): 1216-1228. |
| [12] | Wu Y Y, Xiao N, Yu L, Pan C H, Li Y H, Zhang X X, Liu G Q, Dai Z Y, Pan X B, Li A H.Combination patterns of majorR genes determine the level of resistance to the M. oryzae in rice(Oryza sativa L.). PloS ONE, 2014, 10(6): e0126130. |
| [13] | 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. 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. |
| [13] | 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. Aata Agron Sin, 2013, 39(11): 1927-1934. (in Chinese with English abstract) |
| [14] | 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. |
| [14] | 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) |
| [15] | 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. |
| [15] | 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). |
| [16] | 杨平, 邹国兴, 陈春莲, 黄永萍, 兰波, 熊运华, 尹建华. 利用分子标记辅助选择改良春恢350稻瘟病抗性. 分子植物育种, 2015, 13(4): 741-747. |
| [16] | Yang P, Zou G X, Chen C L, Huang Y P, Lan B, Xiong Y H, Yin J H. Improvement of rice blast resistance of Chunhui350 by using molecular-marker assisted selection.Mol Plant Breeding, 2015, 13(4): 741-747. (in Chinese with English abstract) |
| [17] | 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. |
| [17] | 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. J Nucl Agric Sci, 2017, 31(3): 424-431. (in Chinese with English abstract). |
| [18] | Cho Y C, Kwon S W, Choi I S, Lee S K, Jeon J S, Oh M K, Roh J H, Hwang H G, Yang S J, Kim Y G.Identification of major blast resistance genes in Korean rice varieties (Oryza sativa L.) using molecular markers. J Crop Sci Biotechnol, 2007, 10(4): 265-276. |
| [19] | IRRI. Standard evaluation system for rice(SES). Los Banos: International Rice Research Institute, 2002. |
| [20] | 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. |
| [20] | 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 markes for detecting the rice blast resisitance gene alleles Pi2/9/z-t. Chin J Rice Sci, 2015, 29(3): 305-310. (in Chinese with English abstract) |
| [21] | 孙立亭, 林添资, 龚红兵, 景德道, 余波, 钱华飞, 曾生元, 李闯, 姚维成, 盛生兰. 稻瘟病抗性基因Pb1功能特异性分子标记及其专用引物、检测方法和应用: 201610305124X.2016-08-24. |
| [21] | Sun L T, Lin T Z, Gong H B, Jing D D, Yu B, Qian H F, Zeng S Y, Li C, Yao W, C, Sheng S L. Specific marker for Pb1 and its primer, detection method and application: 201610305124X.2016-08-24. (in Chinese) |
| [22] | Deng Y W, Zhai K R, Xie Z, Yang D Y, Zhu X D, Liu J Z, Wang X, Qin P, Yang Y Z, Zhang G M, Li Q, Zhang J F, Wu S H, Milazzo J, Mao B Z, Wang E T, Xie H A, Tharreau D, He Z H.Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance.Science, 2017, 355(6328): 962-965. |
| [23] | Wang Z Y, Wu Z L, Xing Y Y, Zheng F G, Guo X L, Zhang W G, Hong M M.Nucleotide sequence of rice waxy gene. Nucleic Acids Res, 1990, 18(19): 5898-5898. |
| [24] | Gao Z Y, Zeng D L, Cui X, Zhou Y H, Yan M X, Huang D N, Li J Y, Qian Q.Map-based cloning of the ALK gene, which controls the gelatinization temperature of rice. Sci China: Ser C, 2003, 46(6): 661-668. |
| [25] | Yano M, Katayose Y, Ashikari M, Yamanouchi U, Monna L, Fuse T, Baba T, Yamamoto K, Umehara Y, Nagamura Y.Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS. Plant Cell, 2000, 12(12): 2473-2483. |
| [26] | Tamaki S, Matsuo S, Wong H L, Yokoi S, Shimamoto K.Hd3a protein is a mobile flowering signal in rice. Science, 2007, 316(5827): 1033-1036. |
| [27] | Chen J, Ding J, Ouyang Y, Du H, Yang J, Cheng K, Zhao J, Qiu S, Zhang X, Yao J, Liu K, Wang L, Xu C, Li X, Xue Y, Xia M, Ji Q, Lu J, Xu M, Zhang Q.A triallelic system of S5 is a major regulator of the reproductive barrier and compatibility of indica-japonica hybrids in rice. Proc Natl Acad Sci USA, 2008, 105(32): 11436-11441. |
| [28] | Inoue H, Hayashi N, Matsushita A, Liu X Q, Nakayama A, Sugano S, Jiang C J, Takatsuji H.Blast resistance of CC-NB-LRR protein Pb1 is mediated by WRKY45 through protein-protein interaction. Proc Natl Acad Sci USA, 2013, 110(23): 9577-9582. |
| [29] | Lee S K, Song M Y, Seo Y S, Kim H K, Ko S, Cao P J, Suh J P, Yi G, Roh J H, Lee S, An G, Hahn T R, Wang G L, Ronald P, Jeon J S.Rice Pi5-mediated resistance to magnaporthe oryzae requires the presence of two coiled-coil-nucleotide-binding-leucine-rich repeat genes. Genetics, 2009, 181(4): 1627-1638. |
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