Yi Yingjin, Zheng Ke, Ning Shunzong, Zhao Laibin, Xu Kai, Hao Ming, Zhang Lianquan, Yuan Zhongwei, Liu Dengcai
Triticeae Research Institute, Sichuan Agricultural University, Chengdu, 611130 Sichuan China.
Mol Cytogenet. 2019 Apr 2;12:15. doi: 10.1186/s13039-019-0428-2. eCollection 2019.
The annual allotetraploid species harbors a number of traits relevant for wheat improvement. An effective cytogenetic method has yet to be developed to distinguish between each of its 14 chromosomes.
A fluorescence in situ hybridization (FISH) approach was adopted to describe the karyotype of Each of its 14 chromosomes was unequivocally recognized using a cocktail of three probes, namely pTa-713, (AAC) and pTa71. FISH karyotyping was then used to detect and characterize selections from an × bread wheat wide cross of a chromosome 1M disomic addition line and three 4M(4B) substitution lines. The identity of the addition line was confirmed by the presence of , detected both using an SDS-PAGE separation of endosperm proteins and by applying a PCR assay directed at the locus. The status of the substitution lines was validated by genotyping using a wheat single nucleotide polymorphism chip.
FISH karyotyping based on pTa-713, (AAC) and pTa71 will be useful for determining the contribution of to derivatives of an × wheat wide cross. SNP chip-based genotyping is effective for confirming the status of whole chromosome wheat/alien substitution lines.
一年生异源四倍体物种具有许多与小麦改良相关的性状。尚未开发出一种有效的细胞遗传学方法来区分其14条染色体中的每一条。
采用荧光原位杂交（FISH）方法来描述其核型。使用三种探针（即pTa-713、(AAC)和pTa71）的混合物明确识别了其14条染色体中的每一条。然后，利用FISH核型分析来检测和鉴定来自提莫菲维小麦×普通小麦远缘杂交的1M二体附加系和三个4M(4B)代换系的选择材料。通过胚乳蛋白的SDS-PAGE分离以及针对该位点的PCR分析，均检测到了1M染色体，从而证实了附加系的身份。利用小麦单核苷酸多态性芯片进行基因分型，验证了代换系的状态。
基于pTa-713、(AAC)和pTa71的FISH核型分析将有助于确定提莫菲维小麦对提莫菲维小麦×普通小麦远缘杂交后代的贡献。基于单核苷酸多态性芯片的基因分型对于确认整条染色体小麦/外源代换系的状态是有效的。
