| 缪娜娜,丁明全，杨思晴，戎均康.转录组测序技术在鉴定小麦染色体臂置换系染色体组成上的应用[J].麦类作物学报,2019,(10):1146 |
| 转录组测序技术在鉴定小麦染色体臂置换系染色体组成上的应用 |
| Application of RNA-seq Technique in Identifying the Chromosome Constitution of Chromosome Arm Substitution Lines（CASLs） |
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| DOI：10.7606/j.issn.1009-1041.2019.10.02 |
| 中文关键词: 小麦 RNA-seq 染色体臂置换系 SNP 染色体组成 |
| 英文关键词:Wheat RNA-seq CASL SNP Chromosome constitution |
| 基金项目:浙江省农业（粮食）新品种选育重大科技专项（子课题：小麦高蛋白质含量材料的创建与品系选育，2016C02050-9-9）;国家重点研发计划项目（2016YFD0102002）;国家自然科学基金项目（31671684） |
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| 中文摘要: |
| 为给染色体片段置换系的快速鉴定提供新的技术手段，用3个以普通小麦品种中国春（Chinese Spring，CS）为背景的野生二粒小麦（TTD140）染色体臂置换系（chromosome arm substitution line,CASL）进行转录组测序，并结合SNP分析，获得纯合的SNP，用于鉴定CASL材料中TTD140的置换区段。结果发现，CASL3AL的SNP主要分布在染色体3A的108~750 Mb区段；CASL7BS的SNP分布于染色体7B的0~536 Mb和5A的22~482 Mb区段；CASL4AL的SNP分布比较复杂，除集中在4A的40~745 Mb、7B的0~570 Mb以及5B的410~675 Mb外，还在1A、2A、3A、7A、2B、2D、5D、6D和7D染色体上成簇分布，表明该材料除在预期的染色体4A上保留TTD140的大片段外，还在其他染色体上残留许多TTD140小片段。通过97对多态性SSR标记验证CASL3AL的染色体组成，发现84对标记能检测到TTD带型，其分布范围与转录组数据鉴定结果基本一致。对SNP富集区域的320 bp PCR产物直接测序，发现CASL3AL与CS之间存在7个SNP位点，但与TTD140以及Zavitan参考序列一致。因此，本研究利用转录组测序技术能够有效鉴定小麦染色体片段置换系材料的染色体组成，且比传统的分子标记技术准确、方便、快捷。 |
| 英文摘要: |
| Inorder to develop new and effective method for quickly identifying the chromosome constitution of these genetic materials,three chromosome arm substitution lines(CASL3AL,4AL and 7BS) of wild emmer accession TTD140 in the genetic background of common wheat variety Chinese Spring(CS) were used for transcriptome sequencing and SNP analysis. The results of homozygous SNP were used to identify the TTD140 substitution segment of CASLs.The results showed that homozygous SNPs of CASL3AL were mainly distributed on 108-750 Mb segment of chromosome 3A. SNPs of CASL7BS had two main locations:one on the segments of 0-510 Mb of chromosome 7B,and the other one on 30-460 Mb of chromosome 5A. Distributions of the homozygous SNPs of CASL4AL were complicated,besides the locations on 40-745 Mb segments of chromosome 4A,0-570 Mb of chromosome 7B and 410-675 Mb of chromosome 5B,the remaining SNPs were clustered on the end of chromosomes 1A,2A,3A,7A,2B,2D,5D,6D and 7D,suggesting that CASL4AL retained large TTD140 fragments on chromosome 4A and many small TTD140 fragments on other chromosomes. The chromosome composition of CASL3AL was further verified with 97 polymorphic SSR markers,of which 84 amplified bands from TTD,and the distributions of these SSR markers were consistent with our transcriptome results.320 bp PCR fragment in SNP rich regions of CASL3AL,TTD140 and CS were directly sequenced.CASL3AL showed seven SNPs with CS,which were consistent with TTD140 and Zavitan reference sequences. Therefore,our method combining homozygous SNP analysis with transcriptome data is an accurate,feasible and convenient way to verify chromosome constitution in CASL. |
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