基于寡核苷酸探针套painting的小麦“中国春”非整倍体高清核型及应用  

Development and Application of High Resolution Karyotypes of Wheat “Chinese Spring” Aneuploids

作　　者：王丹蕊 杜培 裴自友[2] 庄丽芳[1] 亓增军[1] WANG Dan-Rui;DU Pei;PEI Zi-You;ZHUANG Li-Fang;QI Zeng-Jun(State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China;Crop Science Institute, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China)

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,江苏南京210095 [2]山西省农业科学院作物科学研究所,山西太原030030

出　　处：《作物学报》2017年第11期1575-1587,共13页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31370385);山西省农业科学院所长青年引导专项(yydzx09)资助~~

摘　　要：基于寡核苷酸探针套painting的染色体鉴定技术简单、经济和高效,可以促进小麦品种及亲缘物种染色体识别和变异体鉴定,提高染色体工程效率。我们前期开发了寡核苷酸探针套,包含p As1-1、p As1-3、AFA-4、(GAA)10和p Sc119.2-1共5个探针。本研究通过一次荧光原位杂交(FISH),对源于17个非整倍体的18份材料分析发现,其中14个染色体组成正确,可以清晰识别相应的缺体、四体和端体。还构建了基于寡核苷酸探针套涂染的、能准确识别3个基因组和7个部分同源群染色体的高清核型,发现4个非整倍体发生变异,其中从N5BT5D中鉴定出一个可能的小片段相互易位系T6AS·6AL-6DL和T6DS·6DL-6AL。进一步对7个地方品种、10个栽培品种(系)和1个人工合成小麦分析,发现15条染色体存在多态性,涉及6条B组(除4B)、5条A组(除1A和3A)和4条D组(1D、2D、4D和7D)染色体,可以清晰识别我国小麦生产上广泛应用的3种易位类型(T1RS·1BL、T6VS·6AL及相互易位T1RS·7DL和T7DS·1BL),省去了基因组原位杂交(GISH)程序。另外,对5个亲缘物种分析发现,该探针套可以识别栽培一粒小麦、硬粒小麦Langdon、荆州黑麦、长穗偃麦草(2n=2x=14)全部和中间偃麦草30条染色体,并构建了这5个物种的核型。本研究结果证实该寡核苷酸探针套可以有效用于小麦及亲缘物种染色体鉴定,高清晰的中国春非整倍体核型为小麦染色体工程提供了参考标准。Oligonucleotide(oligo hereafter)multiplex-based chromosome painting facilitates chromosome identification of both wheat cultivars and its relatives in a simple,easy and high efficient way.In this study,an oligo multiplex containing oligos pAs1-1,pAs1-3,AFA-4,(GAA)10,and pSc119.2-1developed earlier was used for chromosome painting of18accessions from17Chinese Spring(CS)aneuploids.The high resolution karyotypes allowed to clearly distinguish individual wheat chromosomes.Fourteen aneuploids had the expected chromosome constitutions whereas the other four had chromosome variations including one with a possible small segmental reciprocal translocation T6AS·6AL-6DL and T6DS·6DL-6AL occurred in N5BT5D.The following analysis on eight landraces,nine cultivars(lines),and one synthetic hexaploid wheat,observed karyotype diversities from15chromosomes including six B-(except for4B),five A-(except for1A and3A),and four D-genome(1D,2D,4D,and7D)chromosomes.The three widely-used translocations in China,i.e.T1BL·1RS,T6AL·6VS and the reciprocal translocation T1RS·7DL and T7DS·1BL,were clearly detected after only once fluorescence in situ hybridization(FISH)using the oligo multiplex and without genomic in situ hybridization(GISH).This oligo multiplex also produced rich signals in all chromosomes of Triticum monococum,rye cultivar“Jingzhouheimai”,durum wheat“Langdon”,and Thinopyrum elongatum,and30chromosomes of Thinopyrumintermedium.The karyotypes of these five species were thus developed.These results indicate that oligo multiplex-based chromosome painting will play active roles on chromosome identifying,and provide a reference for the standard karyotypes of CS aneuploids.

关 键 词：寡核苷酸探针套 染色体painting 染色体多样性 小麦易位系 非整倍体 

分 类 号：S512.1[农业科学—作物学]