Adhikari T B, Yang X, Cavaletto J R, Hu X, Buechley G, Ohm H W, Shaner G, Goodwin S B
Crop Production and Pest Control Research, US Department of Agriculture-Agricultural Research Service, Department of Botany and Plant Pathology, Purdue University, 915 W. State Street, West Lafayette, IN 47907-2054, USA.
Theor Appl Genet. 2004 Sep;109(5):944-53. doi: 10.1007/s00122-004-1709-6. Epub 2004 May 15.
Septoria tritici blotch (STB), caused by the ascomycete Mycosphaerella graminicola (anamorph Septoria tritici), was the most destructive disease of wheat in Indiana and adjacent states before deployment of the resistance gene Stb1 during the early 1970s. Since then, Stb1 has provided durable protection against STB in widely grown wheat cultivars. However, its chromosomal location and allelic relationships to most other STB genes are not known, so the molecular mapping of Stb1 is of great interest. Genetic analyses and molecular mapping were performed for two mapping populations. A total of 148 F1 plants (mapping population I) were derived from a three-way cross between the resistant line P881072-75-1 and the susceptible lines P881072-75-2 and Monon, and 106 F6 recombinant-inbred lines (mapping population II) were developed from a cross between the resistant line 72626E2-12-9-1 and the susceptible cultivar Arthur. Bulked-segregant analysis with random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and microsatellite or simple-sequence repeat (SSR) markers was conducted to identify those that were putatively linked to the Stb1 gene. Segregation analyses confirmed that a single dominant gene controls the resistance to M. graminicola in each mapping population. Two RAPD markers, G7(1200) and H19(520), were tightly linked to Stb1 in wheat line P881072-75-1 at distances of less than 0.68 cM and 1.4 cM, respectively. In mapping population II, the most closely linked marker was SSR Xbarc74, which was 2.8 cM proximal to Stb1 on chromosome 5BL. Microsatellite loci Xgwm335 and Xgwm213 also were proximal to Stb1 at distances of 7.4 cM and 8.3 cM, respectively. The flanking AFLP marker, EcoRI-AGC/ MseI-CTA-1, was 8.4 cM distal to Stb1. The two RAPD markers, G7(1200) and H19(520), and AFLP EcoRI-AGC/ MseI-CTA-1, were cloned and sequenced for conversion into sequence-characterized amplified region (SCAR) markers. Only RAPD allele H19(520) could be converted successfully, and none of the SCAR markers was diagnostic for the Stb1 locus. Analysis of SSR and the original RAPD primers on several 5BL deletion stocks positioned the Stb1 locus in the region delineated by chromosome breakpoints at fraction lengths 0.59 and 0.75. The molecular markers tightly linked to Stb1 could be useful for marker-assisted selection and for pyramiding of Stb1 with other genes for resistance to M. graminicola in wheat.
小麦黄斑叶枯病(STB)由子囊菌小麦球腔菌(无性型为小麦壳针孢)引起,在20世纪70年代初抗性基因Stb1应用之前,是印第安纳州及邻近各州最具破坏性的小麦病害。从那时起,Stb1在广泛种植的小麦品种中提供了对STB的持久保护。然而,其染色体定位以及与大多数其他STB基因的等位基因关系尚不清楚,因此Stb1的分子定位备受关注。对两个作图群体进行了遗传分析和分子定位。共有148株F1植株(作图群体I)来自抗性品系P881072 - 75 - 1与感病品系P881072 - 75 - 2和莫农之间的三交,106个F6重组自交系(作图群体II)由抗性品系72626E2 - 12 - 9 - 1与感病品种亚瑟杂交培育而成。利用随机扩增多态性DNA(RAPD)、扩增片段长度多态性(AFLP)和微卫星或简单序列重复(SSR)标记进行混合分组分析法,以鉴定那些可能与Stb1基因连锁的标记。分离分析证实,每个作图群体中对小麦球腔菌的抗性由单个显性基因控制。两个RAPD标记G7(1200)和H19(520)在小麦品系P881072 - 75 - 1中与Stb1紧密连锁,距离分别小于0.68 cM和1.4 cM。在作图群体II中,最紧密连锁的标记是SSR Xbarc74,它在5BL染色体上位于Stb1近端2.8 cM处。微卫星位点Xgwm335和Xgwm213也分别在距离Stb1 7.4 cM和8.3 cM处位于近端。侧翼AFLP标记EcoRI - AGC / MseI - CTA - 1在Stb1远端8.4 cM处。克隆并测序了两个RAPD标记G7(1200)和H19(520)以及AFLP EcoRI - AGC / MseI - CTA - 1,以便转化为序列特征化扩增区域(SCAR)标记。只有RAPD等位基因H19(520)能够成功转化,并且没有一个SCAR标记对Stb1位点具有诊断性。在几个5BL缺失系上对SSR和原始RAPD引物进行分析,将Stb1位点定位在由染色体断裂点界定的、片段长度为0.59和0.75的区域内。与Stb1紧密连锁的分子标记可用于标记辅助选择,以及将Stb1与其他抗小麦球腔菌的基因进行聚合。