小麦背景中披碱草部分染色体的FISH鉴定  

Fluorescence in Situ Hybridization (FISH) Identification of Partial Elymus Chromosomes in a Wheat Background

作　　者：宫文萍[1] 汪晓璐 韩冉[1] 李光蓉[2] 李豪圣[1] 刘建军[1] 刘成[1] 杨足君[2] GONG Wenping;WANG Xiaolu;HAN Ran;LI Guangrong;LI Haosheng;LIU Jianjun;LIU Cheng;YANG Zujun(Crop Research Institute,Shandong Academy of Agriculural Sciences/Key Laboratory of Wheat Biology and Genetic Improvement in the North Yellow&Huai River Valley,Ministry of Agriculture and Rural Affairs/Engineering Laboratory for Wheat&Maize,Jinan,Shandong 250100;School of Life Science and Technology,University of Electronic Science and Technology of China,Chengdu,Sichuan 610054)

机构地区：[1]山东省农业科学院作物研究所/农业农村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,山东济南250100 [2]电子科技大学生命科学与技术学院,四川成都610054

出　　处：《核农学报》2021年第4期789-795,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金(31971874);泰山学者工程专项经费(tsqn201812123);山东省农业良种工程(2019LZGC016)。

摘　　要：为了建立小麦背景中粗穗披碱草(Elymus trachycaulus)和纤毛披碱草(Elymus ciliaris)染色体的跟踪鉴定方法,本研究利用寡聚核苷酸Oligo-pSc119.2-1和Oligo-pTa-535-1为探针的双色荧光原位杂交(FISH)和以(GAA)8为探针的单色FISH,分别对4个小麦-粗穗披碱草附加系和5个小麦-纤毛披碱草附加系染色体进行分析。结果发现,经与小麦FISH核型比对后,建立了可用于追踪小麦背景中粗穗披碱草1St、5Ht、6Ht和7Ht染色体的标准FISH核型;而纤毛披碱草Sc和Yc染色体FISH信号较弱。FISH检测发现,小麦-粗穗披碱草1St附加系自交自发变成了1St(1BS·3BL)代换系,且在其染色体组中检测到了一对T1BL·3BS易位染色体,同时发现5AS端部寡聚核苷酸Oligo-pSc119.2-1序列发生了删除。另外,在小麦-粗穗披碱草5Ht附加系中发现2B染色体短臂末端删除现象,形成了2B-del和2BS-4AS·4AL易位染色体,而另一条2B和4A为正常的完整染色体。这种染色体结构重排事件表明,部分小麦-远缘物种附加系细胞学并不稳定,因此,繁种前后应对材料进行单株细胞学鉴定。上述小麦-粗穗披碱草染色体结构变异体的获得,为研究染色体结构重排与基因转录表达和表型变化的关系提供了研究基础。To explore new approaches for the detection of Elymus trachycaulus and E. ciliaris chromosomes in a wheat background, four wheat-E. trachycaulus addition lines and five wheat-E. ciliaris addition lines were characterized by double-color and Fluorescence in situ hybridization(FISH) using oligonucleotides Oligo-pSc119.2-1 and Oligo-pTa-535-1 as probes, and one-color FISH using oligonucleotide(GAA)8 as a probe. After comparing with the FISH pattern of wheat, the standard FISH karyptypes of E. trachycaulus 1 St, 5 Ht, 6 Ht and 7 Ht chromosomes were established, while the FISH signal of E. ciliaris Sc and Yc chromosomes were very weak. FISH result showed that the wheat-E. trachycaulus 1 St addition line has already turned spantaneously to a wheat-E. trachycaulus 1 St(1 BS-3 BL) substitution line, and a pair of T1 BL·3 BS translocation chromosomes was detected in its genome. Meanwhile, it was also found that the oligonucleotide Oligo-pSc119.2-1 sequence at the end of chromosome 5 AS was deleted. In addition, a deletion of chromosome 2 B short arm end was also detected in 5 Ht addition line, formed a 2 B-del chromosome and a spontaneous 2 BS-4 AS·4 AL translocation chromosome, while the other one 2 B and 4 A chromosomes were normal and complete. These chromosomal rearrangement events indicate that the cytology of some wheat-wild species addition lines were unstable. Therefore, individual plants of these addition lines should be cytogenetically identified before and after propagating seed. The wheat-E. trachycaulus chromosomal structure variants provided materials for studying the relationships among chromosomal rearrangement, gene transcription, gene expression and phenotypic changes.

关 键 词：粗穗披碱草 纤毛披碱草 荧光原位杂交 染色体重排 

分 类 号：S512.1[农业科学—作物学]