农业生物技术学报2013，Vol.21Issue(3)：263-271,9.DOI:10.3969/j.issn.1674-7968.2013.03.002
抗条锈病的小麦-非洲黑麦异代换系的分子细胞学鉴定
Molecular Cytogenetic Characterization of a Triticum durum-Secale africanum Substitution Line for Resistance to Sripe Rust (Puccinia striiformis Eriks.f.sp.tritici)
摘要
Abstract
Species in genus Secale have provided novel rust resistance genes to protect wheat from this fungal disease. In order to transfer new stripe rust resistance gene (s) from wild Secale species, Secale qfricanum, we obtained a line HH41 from the advanced generation of Triticum durum-Secde qfricanum amphiploid (AABBRaRa) crossed with wheat (T. aestivum). Line HH41 was immune to stripe rust isolates from China, and had 42 chromosomes. Fluorescence in situ hybridization (FISH) using D-chromosome specific probe pAsl and sequential genomic in situ hybridization (GISH) analysis using 5. cereale cv. Qinglingheimai (QH) genomic DNA as probe demonstrated that a pair of wheat chromosomes 6D were absent, but a pair of S. qfricanum chromosomes presented in HH41. Expressed sequence tags based molecular markers also confirmed that HH41 lost 6D specific bands and amplified 6Ra specific bands, indicating that HH41 was a 6Ra (6D) substitution line. GISH analysis using genomic DNA of S. cereale cv. Qinglingheimai(QH) as a probe showed that hybridization signal covered along the chromosomes 6Ra except the telomere region of the short arm. However, the whole 6Ra chromosomes possessed hybridization signal if using genomic DNA of S. qfricanum for GISH analysis. FISH using Secale telomere or subtelomere tandem repetitive sequences pSc200 as a probe, no hybridization signal was detected on HH41. However, apparent signal was observed on chromosomes 6R from CS-S. cereale cv. Imperial disomic addition lines with strong signal on the telomere of the short arms were also scattered signal on the subtelomere of the long arms. It was revealed that chromosomes 6Ra of S. qfricanum were different from the S. cereale chromosomes 6R, especially on the telomere regions, with lost or reduced telomere or subtelomere repetitive sequences. Nine of sixteen 6R/6Ra specific markers developed by our study represented polymorphic amplification on chromosomes 6Ra of 5. africanum distinct from chromosomes 6R of S. cereale, further demonstrated polymorphism exited between the two chromosome types. Based on stripe rust races evaluation, it was deduced that the stripe rust resistance in HH41 was derived from chromosomes 6Ra of S. qfricanum. In conclusion, the 6Ra (6D) substitution line with excellent stripe rust resistance and apparent polymorphism originated from ancient species S. qfricanum will serve as an important resource for translocation lines creation, valuable gene introgression and common wheat improvement.关键词
原位杂交/分子标记/6Ra(6D)代换系/抗条锈病/非洲黑麦Key words
Fluorescence in situ hybridization/ Molecular markers/ 6Ra (6D) substitution line/ Stripe rust resistance/ Secale qfricanum引用本文复制引用
雷孟平,李光蓉,刘成,杨足君..抗条锈病的小麦-非洲黑麦异代换系的分子细胞学鉴定[J].农业生物技术学报,2013,21(3):263-271,9.基金项目
国家自然科学基金(No.31171542和31101143)和中国博士后基金(No.Y02006023601261) （No.31171542和31101143）
