Identification and Molecular Cytogenetic Analysis of Wheat-rye (Triticum aestivum-Secale cereale) 1RS-1BS/1BL Small Fragment Translocation Lines Against Fusarium Head Blight
LV Ting-Ting*, ZHANG Zhen-Yue*, YAO Xiao-Ni, ZHANG Han-Bing, LI Jia-Chuang, LIU Shu-Hui, YANG Qun-Hui, WU Jun, CHEN Xin-Hong**
Shaanxi Key Laboratory of Plant Genetic Engineering Breeding, College of Agronomy, Northwest A&F University, Yangling 712100, China
Abstract：Fusarium head blight (FHB) is one of the main diseases which caused a serious threat to the normal growth and yield formation of wheat (Triticum aestivum; 2n=6x=42, AABBDD). Rye (Secale cereal; 2n=2x=14, RR) with developed root system, strong stress resistance and disease resistance, is an important genetic resource for improving wheat traits. For the purpose of improving the FHB resistance in wheat, Mexican rye and common wheat W770B were hybridized to obtain several derived lines with genetic stability through multi-generation breeding. At the blooming stage of wheat, single flower inoculation (SFI) was used to identify the parents and progenies resistance to FHB. The results showed that the parents rye had moderate resistance to FHB whereas W770B showed high sensitivity, and the 12-5-1 derived line showed moderate resistance. Morphological and cytogenetic identification and analysis were conducted in 12-5-1 derived line by genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH) sequence amplification, sequence characterized amplified regions (SCAR), simple sequence repeats (SSR) molecular markers. The results showed that the root tip cell of 12-5-1 contained 42 chromosomes (2n=42). In the first meiotic metaphase of pollen mother cell, there is a proper chromosome pairing forming 21 bivalents(2n=42=21Ⅱ), and normal separation. GISH identification was performed on 12-5-1 using rye genome as a probe. Yellow and green signals were observed at the ends of two chromosomes of wheat, indicating that there is a translocation between wheat chromosome and rye chromosome. FISH results showed that the translocation of 1BS of wheat and 1RS of rye occurred in 12-5-1. Therefore, it is inferred that the hybridization of wheat and rye resulted in translocation of small chromosome segments. Moreover, Rye specific marker AF1/AF4 and SCAR marker of 1RS were used for identification and the corresponding bands were amplified in rye and 12-5-1 derived line. SSR analysis showed that three pairs of primers (Xgwm11、Xgwm31 and Xgwm550) on the 1BS chromosome of wheat could not be used to amplify the target bands in 12-5-1, while the remaining primers could be used to amplify the corresponding target bands, which indicated that 12-5-1 was a 1RS-1BS/1BL translocation line in Rye-Wheat. The creation of the derived line with FHB resistance provides a new germplasm resource for wheat breeding against FHB resistance.
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