利用单片段代换系研究水稻抽穗期等位基因变异

doi:10.7668/hbnxb.20192258

摘要/Abstract

摘要： 为了研究水稻单片段代换系（Single segment substitution line，SSSL）与受体亲本HJX74（Huajingxian 74）抽穗期差异的原因，以HJX74和5个具有抽穗期差异显著性的SSSLs WY18（W06-15-18-3-11）、WY24（W08-16-3-2）、WY26（W11-15-7-1-1）、WY60（W15-3-1-38）、WY61（W17-10-5-5-35-9-2）为试验材料，对其代换片段上已知的抽穗期等位基因进行克隆，获取其CDS序列，用多种生物信息学方法进行序列比对，包括CDS序列、氨基酸序列，比较其氨基酸理化性质的异同及表达的差异。田间抽穗期表型性状调查结果表明，WY18、WY60、WY61相对于HJX74表现为晚抽穗，WY24、WY26表现为早抽穗。与HJX74相比，WY18代换片段上抽穗期等位基因DTH8存在4个SNP的差异、WY26中OsDof12存在7个SNP的差异、WY60 DTH8存在6个SNP的差异，但以上SNP的突变均未改变其氨基酸序列的亲水性及理化性质，表明这些抽穗期等位基因的突变可能并不是引起其抽穗期变化的原因。进一步通过qRT-PCR技术分析发现，WY18中DTH8、WY26中OsDof12的表达水平整体高于HJX74中的相应基因，表明该等位基因上游调节区域对其基因的表达进行了调控，可能是导致其抽穗期变异的原因。WY24中se14、WY61中Hd1存在明显的序列片段缺失，造成se14和Hd1等位基因功能缺失，可能是WY24、WY61抽穗期变异的主要原因。

关键词: 水稻, 抽穗期, 单片段代换系, 等位基因, 遗传分化

Abstract: In order to study the reasons for the difference of rice heading date between SSSL (Single segment substitution line) and acceptor parent HJX74 (Huajingxian 74), used HJX74 and five SSSLs WY18 (W06-15-18-3-11), WY24 (W08-16-3-2), WY26 (W11-15-7-1-1), WY60 (W15-3-1-38) and WY61 (W17-10-5-5-35-9-2) with significant difference in heading date as materials, cloned the known heading date alleles on their substitution fragments, obtained their CDS sequence, and used a variety of bioinformatics methods to perform sequence alignment, including CDS sequences and amino acid sequences, compared the similarities and differences of amino acid physicochemical properties and expression differences. According to the phenotype of heading date in the field, WY18, WY60 and WY61 showed late heading date compared with HJX74, while WY24 and WY26 showed early heading date. Compared with HJX74, there were 4 SNP differences in the heading date allele DTH8 on the WY18 substitution fragment, 7 SNP differences in WY26 OsDof12, and 6 SNP differences in WY60 DTH8. However, none of the above SNP mutations changed the hydrophilicity and physicochemical properties of its amino acid sequence, indicating that the mutations of these heading date alleles might not be the cause of the heading date changes. Further expression analysis by qRT-PCR founded that the expression level of heading date alleles DTH8 of WY18 and OsDof12 of WY26 were higher than that of HJX74, indicating that their expression was regulated by the upstream regulatory region. The regulation of this expression might be the reason for the variation of heading date. The WY24 se14 and WY61 Hd1 had obvious sequence deletions, leading to loss function of se14 and Hd1 alleles which probably were the main reason for heading date variations of WY24 and WY61.

Key words: Rice, Heading date, Single segment substitution line, Allelic gene, Genetic differentiation

中图分类号:

S511.03

高瑞钰, 张华, 宣宁, 柳絮, 张浩, 张梦琦, 姚方印. 利用单片段代换系研究水稻抽穗期等位基因变异[J]. 华北农学报, 2021, 36(5): 29-34. doi: 10.7668/hbnxb.20192258.

GAO Ruiyu, ZHANG Hua, XUAN Ning, LIU Xu, ZHANG Hao, ZHANG Mengqi, YAO Fangyin. Variation Analysis of Heading Date Alleles Using Rice Single Segment Substitution Lines[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(5): 29-34. doi: 10.7668/hbnxb.20192258.

http://www.hbnxb.net/CN/Y2021/V36/I5/29

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