利用叶绿体 DNA 完整测序和基因单倍型分析对六倍体小麦（L.）进行遗传分析。
Genetic Analysis of Hexaploid Wheat ( L.) Using the Complete Sequencing of Chloroplast DNA and Haplotype Analysis of the Gene.
机构信息
Institute of Molecular Genetics, Agricultural University of Georgia, #240 D. Aghmashenebeli Av, Tbilisi 0131, Georgia.
Graduate School of Agricultural Science, Kobe University, Rokko, Nada, Kobe 657-8501, Japan.
出版信息
Int J Mol Sci. 2021 Nov 24;22(23):12723. doi: 10.3390/ijms222312723.
The aim of the presented study is a genetic characterization of the hexaploid wheat L. Two approaches were used for the genealogical study of hexaploid wheats-the complete sequencing of chloroplast DNA and PCR-based haplotype analysis of the fourth intron of and of the fifth-to-sixth-exon region of The complete chloroplast DNA sequences of 13 hexaploid wheat samples were determined: Free-threshing- subsp. , one sample; subsp. , two samples; subsp. , one sample; subsp. four samples. Hulled- subsp. , three samples; subsp. jakubz two samples. The comparative analysis of complete cpDNA sequences of 20 hexaploid wheat samples (13 samples in this article plus 7 samples sequenced in this laboratory in 2018) was carried out. PCR-based haplotype analysis of the fourth intron of and of the fifth-to-sixth exon region of of all 20 hexaploid wheat samples was carried out. The 20 hexaploid wheat samples (13 samples in this article plus 7 samples in 2018) can be divided into two groups- subsp. three samples and subsp. collected in Armenia, and the remaining 16 samples, including subsp. collected in Europe (Sweden). If we take the cpDNA of Chinese Spring as a reference, 25 SNPs can be identified. Furthermore, 13-14 SNPs can be identified in subsp. and subsp. (Vav1). In the other samples up to 11 SNPs were detected. 22 SNPs are found in the intergenic regions, 2 found in introns, and 10 SNPs were found in the genes, of which seven are synonymous. PCR-based haplotype analysis of the fourth intron of and the fifth-to-sixth-exon region of provides an opportunity to make an assumption that hexaploid wheats subsp. var. and var. differ from other macha samples by the absence of a 42 bp insertion in the fourth intron of . One possible explanation for this observation would be that two Coss. (A) and (B) participated in the formation of hexaploids through the D genome: (A)-macha (1-5, 7, 8, 10-12), and (B)-macha M6, M9, subsp. cv. 'Chinese Spring' and cv. 'Red Doly'.
本研究的目的是对六倍体小麦 L.进行遗传特征分析。为了对六倍体小麦进行系统发育研究，我们采用了两种方法——叶绿体 DNA 全序列测序和第四内含子 PCR 分析和第五到第六外显子区域的 PCR 分析。我们确定了 13 个六倍体小麦样本的完整叶绿体 DNA 序列：自由脱粒亚种，1 个样本；硬粒亚种，2 个样本；波兰亚种，1 个样本；普通小麦亚种，4 个样本。皮壳亚种，3 个样本；密穗亚种，2 个样本。我们对 20 个六倍体小麦样本（本文中的 13 个样本加上 2018 年本实验室测序的 7 个样本）的完整 cpDNA 序列进行了比较分析。对所有 20 个六倍体小麦样本的第四内含子 PCR 分析和第五到第六外显子区域的 PCR 分析。这 20 个六倍体小麦样本（本文中的 13 个样本加上 2018 年的 7 个样本）可以分为两组——亚组。收集于亚美尼亚的 3 个样本和收集于亚美尼亚的 3 个样本，其余 16 个样本，包括欧洲（瑞典）收集的亚组。如果我们以中国春的 cpDNA 为参考，可以鉴定出 25 个 SNP。此外，还可以在亚组和亚组中鉴定出 13-14 个 SNP（Vav1）。在其他样本中，检测到多达 11 个 SNP。在基因间区发现了 22 个 SNP，在内含子中发现了 2 个 SNP，在基因中发现了 10 个 SNP，其中 7 个是同义的。第四内含子 PCR 分析和第五到第六外显子区域的 PCR 分析提供了一种假设，即六倍体小麦亚种。硬粒亚种 var. 和 var. 与其他 macha 样本不同，第四内含子中不存在 42 bp 的插入。这种观察结果的一种可能解释是，两个 Coss.（A）和（B）通过 D 基因组参与了六倍体的形成：（A）-macha（1-5、7、8、10-12），和（B）-macha M6、M9、普通小麦亚种 cv。‘中国春’和 cv。‘红多利’。
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