寡聚核苷酸FISH在鉴定小麦-单芒山羊草种质中的应用

Application of Oligo-Nucleotide Fluorescence in situ Hybridization( FISH) in Identifying Wheat-Aegilops uniaristata Germplasm

在小麦-单芒山羊草杂交种质鉴定研究中,尚缺乏可同时鉴定小麦和单芒山羊草染色体的细胞遗传学方法。本研究用寡聚核苷酸Oligo-p Sc119.2-1和Oligo-p Ta-535-1为探针的双色FISH和以(GAA)8为探针的单色FISH分别对小麦-单芒山羊草双二倍体、中国春-单芒山羊草附加系进行分析,建立了可以同时鉴定小麦和单芒山羊草全部染色体的FISH标准核型。同时,利用建立的FISH标准核型在1N附加系自交后代材料中发现了5BS.3BS和5BL.3BL易位,在5N附加系自交后代中发现5NL端部寡聚核苷酸序列删除现象,这为研究染色体结构变异与表型变化的关系等提供了研究材料。此外,还在4N附加系自交后代中发现4B染色体丢失现象,造成该附加系自发形成4N(4B)代换系,为诱导产生涉及4N染色体的小麦-单芒山羊草罗伯逊易位系奠定了基础。

So far,the cytogenetic approaches for the identification of wheat-Aegilops uniaristata germplasm are lacking. In the present research,Fluorescence in situ hybridization（ FISH） using Oligo-p Ta535-1 and Oligo-p Sc119. 2-1 as probes,and FISH using（ GAA）8 as probe are performed on wheat-Ae. uniaristata amphiploid and additions. As a result,a standard FISH pattern was established for the identification ofall chromosomes of wheat and Ae. uniaristata simultaneously. Meanwhile,chromosome translocations 5BS.3BS and 5BL. 3BL were detected in the self-pollinated 1N addition progenies,and oligo-nucleotide sequence deletion was also detected on chromosome arm 5NL of Ae. uniaristata in the self-pollinated 5N addition progenies,providing important material for researching the relationships between chromosome structure variation and phenotypic changes. Moreover,a spontaneously formed wheat-Ae. uniaristata 4N（ 4B） substitution line was identified by the established FISH karyotype from the self-pollinated 4N addition progenies,making a solid foundation for further inducing wheat-Ae. uniaristata chromosome translocations with chromosome 4N involved.