小麦秆锈病新抗源及抗病基因所在染色体特异分子标记

doi:10.3864/j.issn.0578-1752.2018.07.001

中国农业科学 ›› 2018, Vol. 51 ›› Issue (7): 1223-1232.doi: 10.3864/j.issn.0578-1752.2018.07.001

• 作物遗传育种·种质资源·分子遗传学 • 下一篇

小麦秆锈病新抗源及抗病基因所在染色体特异分子标记

韩冉¹，李天亚²，宫文萍¹，李豪圣¹，宋健民¹，刘爱峰¹，曹新有¹，程敦公¹，赵振东¹，刘成¹，刘建军¹

¹山东省农业科学院作物研究所/农业部黄淮北部小麦生物学与遗传育种重点实验室/小麦玉米国家工程实验室，济南 250100；²沈阳农业大学植物保护学院，沈阳 110866

收稿日期:2017-10-12 出版日期:2018-04-01 发布日期:2018-04-01
通讯作者: 刘成，E-mail：lch6688407@163.com。刘建军，E-mail：ljjsaas@163.com
作者简介:韩冉，E-mail：hr022cn@aliyun.com；李天亚，E-mail：litianya11@163.com。韩冉与李天亚为同等贡献作者。
基金资助:
国家重点研发计划（2017YFD0100600）、山东省自然科学基金（ZR2017MC004）、山东农科院青年拔尖人才计划（1-18-024）、现代农业产业技术体系（CARS-03）

New Resistance Sources of Wheat Stem Rust and Molecular Markers Specific for Relative Chromosomes That the Resistance Genes are Located on

HAN Ran¹, LI TianYa², GONG WenPing¹, LI HaoSheng¹, SONG JianMin¹, LIU AiFeng¹, CAO XinYou¹, CHENG Dungong¹, ZHAO Zhendong¹, LIU Cheng¹, LIU Jianjun¹

¹Crop Research Institute, Shandong Academy of Agricultural Sciences/Key Laboratory of Wheat Biology and Genetic Improvement in the North Yellow & Huai River Valley, Ministry of Agriculture/National Engineering Laboratory for Wheat & Maize, Jinan 250100; ²College of Plant protection, Shenyang Agricultural University, Shenyang 110866

Received:2017-10-12 Online:2018-04-01 Published:2018-04-01

摘要/Abstract

摘要： 【目的】小麦秆锈病是具潜在毁灭性的小麦病害之一，秆锈菌小种Ug99严重威胁全球小麦生产。本研究通过对165份小麦-近缘植物染色体系进行抗秆锈病鉴定，筛选小麦秆锈病新抗源并建立抗病基因所在染色体特异分子标记，以发掘小麦秆锈病新抗源，培育抗病品种，有效防御Ug99导致的秆锈病。【方法】将供试的165份小麦-近缘植物染色体系、3份六倍体小麦及感病对照小密穗分别播种于直径10 cm的瓦盆中，生长到一叶一心时，用中国小麦秆锈菌流行小种34MKGQM和21C3CTHSM进行接种，当感病品种小密穗充分发病时，按照0—4级标准调查记载侵染型，对供试材料的抗秆锈病级别进行统计。同时，提取免疫、近免疫、高抗秆锈病附加系/代换系及其对应的整套染色体系、中国春等材料的基因组DNA，利用101对PLUG引物进行PCR扩增，扩增产物经限制性内切酶酶切后进行电泳检测，筛选并建立抗秆锈基因所在染色体特异分子标记。【结果】在165份小麦-近缘植物染色体系中，中国春-卵穗山羊草7M^g#1附加系、中国春-卵穗山羊草7M^g#1(7A)和7M^g#1(7B)代换系、中国春-帝国黑麦1R附加系、中国春-中间偃麦草?Ai附加系（?表示外源染色体同源群未鉴定）、中国春-单芒山羊草6N附加系、中国春-易变山羊草6S^vS端体附加系和中国春-智利大麦6H^ch附加系9份材料对秆锈病表现为免疫或近免疫；ALCD-尾状山羊草7C#1附加系、中国春-卵穗山羊草7M^g#1(7D)代换系、中国春-帝国黑麦6R附加系、中国春-高大山羊草6S^l#3附加系、中国春-高大山羊草6S^l#2(6B)代换系、中国春-希尔斯山羊草3S#1附加系和中国春-拟斯卑尔脱山羊草2S^g#3附加系7份材料对秆锈病表现为高抗；其余材料均表现为中感或高感。抗秆锈性基因定位信息比较分析发现，高大山羊草6S^l#2和6S^l#3、帝国黑麦6R、智利大麦6H^ch、卵穗山羊草7M^g#1、尾状山羊草7C、中间偃麦草?Ai染色体上可能含有抗秆锈新基因。分子标记筛选、定位及特异性验证研究共获得8个新的多态性标记，其中5个（TNAC1715、TNAC1718、TNAC1737、TNAC1739和TNAC1753）和3个（TNAC1740、TNAC1751和TNAC1756）分别被定位在6R和6S^l染色体上。【结论】筛选得到8份可能含有抗秆锈新基因的材料，建立了抗秆锈基因所在染色体特异新标记8个。

关键词: 小麦近缘植物, 染色体系, 秆锈病, 多态性

Abstract: 【Objective】Wheat stem rust, caused by Puccinia graminis Pers. f. sp. tritici Eriks. and E. Henn (Pgt) is one of the most potentially destructive wheat diseases, seriously threatening world wheat production. The emergence of new races Ug99 of stem rust caused the global wheat to be under the threat.Exploring new resistant source of wheat stem rust is one of the effective measures against Ug99. In order to explore stem rust new resistant source, the mixed dominant stem rust physiological races in China were inoculated to 165 wheat-alien species chromosome lines at the seeding stages. 【Method】All of the 165 wheat-alien species chromosome lines, 3 hexaploid wheat and the susceptible control Little Club (LC), were sown in the 10 cm diameter clay pots. When the primary leaves were fully expanded, they were inoculated using talcurediospore powder mixture of the common races 34MKGQM and 21C3CTHSM. The Infection Types (ITs) of the material tested was recorded according to the standard ‘0-4’. Meanwhile, genomic DNA was extracted from stem rust immune, nearly immune, or highly resistant additions/substitutions and their corresponding whole set of chromosome lines and Chinese Spring (CS). PCR was performed on these materials by screening 101 pairs of PLUG primer. PCR products were firstly digested by DNA restricted enzymes TaqI and HaeⅢ, and then were detected through 2.0% agarose gel electrophoresis to screen and establish chromosome-specific molecular marker where the stem rust resistance gene was located on.【Result】Among the 165 wheat-alien species chromosome lines, CS-Ae. geniculata 7M^g#1 addition, CS-Ae. geniculata 7M^g#1(7A) and 7M^g#1 (7B) substitutions, CS-Imperial rye 1R addition, CS-Th. intermedium ?Ai addition (? indicates homoeologous group of the alien chromosomes in wheat background was not identified), CS-Ae. uniaristata 6N addition, CS-Ae. variabilis 6S^vS telosomic addition, CS-Chile barley 6H^ch addition are immune or nearly immune to stem rust. ALCD-Ae. caudata 7C#1 addition, CS-Ae. geniculata 7M^g#1(7D) substitution, CS-Imperial rye 6R addition, CS-Ae. longissima 6S^l#3 addition, CS-Ae. longissima 6S^l#2(6B) substitution, CS-Ae. searsii 3S^s#1 addition, CS-Ae. speltoides 2S#3 addition are highly resistant to stem rust. Comparative analysis of chromosomal locations of stem rust resistant genes indicates that chromosomes 6S^l#2 and 6S^l#3 of Aegilops longissima, chromosome 6R of Imperial rye, chromosome 6H^chof Chile barley, chromosome 7M^g#1 of Ae. geniculata, chromosome7C of Ae. caudate and ?Ai of Th. intermedium may harbor new stem rust resistance gene (s). Molecular marker screening, localization, specificity verification showed that 8 new molecular markers have been developed. Among them, 5 (TNAC1715, TNAC1718, TNAC1737, TNAC1739 and TNAC1753) and 3 (TNAC1740, TNAC1751, and TNAC1756) have been assigned to chromosomes 6R and 6S^l, respectively. 【Conclusion】 Eight materials which probably contain new gene(s) against stem rust were obtained, and eight new chromosome specific molecular markers for stem rust resistance gene(s) were established.

Key words: wheat alien species, chromosome lines, stem rust, molecular marker

韩冉，李天亚，宫文萍，李豪圣，宋健民，刘爱峰，曹新有，程敦公，赵振东，刘成，刘建军. 小麦秆锈病新抗源及抗病基因所在染色体特异分子标记[J]. 中国农业科学, 2018, 51(7): 1223-1232.

HAN Ran, LI TianYa, GONG WenPing, LI HaoSheng, SONG JianMin, LIU AiFeng, CAO XinYou, CHENG DunGong, ZHAO ZhenDong, LIU Cheng, LIU JianJun. New Resistance Sources of Wheat Stem Rust and Molecular Markers Specific for Relative Chromosomes That the Resistance Genes are Located on[J]. Scientia Agricultura Sinica, 2018, 51(7): 1223-1232.

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小麦秆锈病新抗源及抗病基因所在染色体特异分子标记

New Resistance Sources of Wheat Stem Rust and Molecular Markers Specific for Relative Chromosomes That the Resistance Genes are Located on

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