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7Mg#1附加系、中国春-卵穗山羊草7Mg#1(7A)和7Mg#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6SvS端体附加系和中国春-智利大麦6Hch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7Mg#1(7D)代换系、中国春-帝国黑麦6R附加系、中国春-高大山羊草6Sl#3附加系、中国春-高大山羊草6Sl#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2Sg#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6Sl#2和6Sl#3、帝国黑麦6R、智利大麦6Hch、卵穗山羊草7Mg#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6Sl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。
【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 34 MKGQM and 21 C3 CTHSM. 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 Taq I 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 7 Mg#1 addition, CS-Ae. geniculata 7 Mg#1(7 A) and 7 Mg#1(7 B) substitutions, CS-Imperial rye 1 R addition, CS-Th. intermedium ?Ai addition(? indicates homoeologous group of the alien chromosomes in wheat background was not identified), CS-Ae. uniaristata 6 N addition, CS-Ae. variabilis 6 SvS telosomic addition, CS-Chile barley 6 Hch addition are immune or nearly immune to stem rust. ALCD-Ae. caudata 7 C#1 addition, CS-Ae. geniculata 7 Mg#1(7 D) substitution, CS-Imperial rye 6 R addition, CS-Ae. longissima 6 Sl#3 addition, CS-Ae. longissima 6 Sl#2(6 B) substitution, CS-Ae. searsii 3 Ss#1 addition, CS-Ae. speltoides 2 S#3 addition are highly resistant to stem rust. Comparative analysis of chromosomal locations of stem rust resistant genes indicates that chromosomes 6 Sl#2 and 6 Sl#3 of Aegilops longissima, chromosome 6 R of Imperial rye, chromosome 6 Hch of Chile barley, chromosome 7 Mg#1 of Ae. geniculata, chromosome7 C 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 6 R and 6 Sl, 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.
作者
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
HAN Ran;LI TianYa;GONG WenPing;LI HaoSheng;SONG JianMin;LIU AiFeng;CAO XinYou;CHENG DunGong;ZHAO ZhenDong;LIU Cheng;LIU JianJun(Crop 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, Ji 'nan 250100;College of Plant Protection, Shenyang Agricultural University, Shenyang 110866)
出处
《中国农业科学》
CAS
CSCD
北大核心
2018年第7期1223-1232,共10页
Scientia Agricultura Sinica
基金
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)
关键词
小麦近缘植物
染色体系
秆锈病
多态性
wheat alien species
chromosome lines
stem rust
molecular marker