芒属植物叶绿体InDel标记的开发与应用

研究利用芒属植物(芒02381和荻04005)叶绿体全基因组测序的结果,开发12对InDel标记。采用正交设计法优化cpInDelPCR体系,得到模板DNA、dNTPs、Mg2+、引物、TaqDNA聚合酶5个关键因素在体系中的最佳组合。12对InDel标记对43份芒属和2份甘蔗材料进行扩增,共扩增出538条条带,其中3对引物扩增出13条多态性条带,占总数的2.4%。将所有位点读带转化为数值矩阵并用UPGMA法聚类,在遗传相似系数1.4的水平上,和南荻聚为一类,而芒和与五节芒、双药芒、红山茅、尼泊尔芒、甘蔗聚为一类,4个具有杂交种特征的材料单独聚为一类。试验开发的芒属植物cpInDel标记可用于芒与荻的区分,指导芒属植物的杂交育种。     

基于 RAD-seq 的菜心 InDel 标记开发及应用

【目的】开发多态性丰富的的InDel分子标记,为菜心育种提供研究基础。【方法】以Chiifu-401-42的基因组序列为模板,利用4份菜心材料的RAD-seq重测序数据,在全基因组范围内鉴定InDel位点。利用生物信息学方法筛选4份菜心材料间具有潜在多态性的InDel位点,挑选分布于10条染色体的80个InDel位点设计引物,对55份菜心种质材料进行遗传多样评价。【结果】通过与参考基因组序列比对,在4份菜心材料的重测序数据中共鉴定出84 510个InDel位点,其中插入/缺失长度大于5 bp的3 609个InDel位点在4份菜心材料间具有潜在多态性。挑选80个InDel位点进行PCR扩增验证,58个（72.5%）在4份测序样品中具有多态性,在55份菜心种质材料中检测到133个等位位点,多态性信息含量（PIC）为0.263～0.618,平均值为0.443。基于InDel标记的检测结果可知,55份菜心种质的遗传相似系数在0.366～0.756,平均值为0.570;在遗传相似系数为0.564时55份菜心种质被分为4个类群,但各类群间的耐热性没有明显差异。【结论】本研究开发的InDel标记具有...     

樱桃番茄种质资源亲缘关系的InDel和SSR标记分析

选用18对InDel引物和15对SSR引物对30份樱桃番茄高代自交系育种材料进行遗传多样性分析。多态性分析结果表明,InDel标记比SSR标记的多态性低;遗传距离分析结果表明,30份樱桃番茄种质材料两两间的遗传距离(GD)在0.052~0.993之间,平均遗传距离仅为0.391,遗传距离小于0.5的种质材料占到了77.24%。在相似系数为0.65的水平上,可将30份樱桃番茄材料分为3类。     

Development ofNew InDel Marker to Detect Genotypes of Conferring Fertility Restoration of BT-Type Cytoplasmic Male Sterility inRice简

Restorer line breeding is an important approach to enhance the heterosis and improve the yields of japonica hybrid rice. To improve the selection efficiency of restorer lines for BT-type cytoplasmic male sterility (CMS) in japonica rice, a functional marker InDel-Rf-1a based on the difference of nucleotide sequence in Rf-1a locus between BT-type CMS lines and restorer lines was developed to detect the genotypes of different rice materials. Conventional indica rice varieties, restorer and maintainer lines without 574 bp deletion could restore the fertility for BT-type CMS in japonica rice. By contrast, most conventional japonica rice varieties except Aichi 106 and Yijing 12, with genotype of rf-1arf-1a showed the 574 bp deletion maintained sterility for BT-type CMS lines. To further verify the effect of genotyping detection in Rf-1a locus, this marker was also used to amplify the genomic DNA in different japonica rice restorer lines, CMS lines, hybrids and F₂ segregation population, and three genotypes in Rf-1a locus could be distinguished distinctly. Therefore, the marker InDel-Rf-1a could be widely used for genetic identification and marker-assisted selection (MAS) in breeding japonica restorer lines.     

InDel分子标记WC656的开发及其在鉴别小麦-簇毛麦5VS纯合易位中的应用

簇毛麦是小麦品种改良重要的遗传资源,其5VS上含有硬度基因Dina/Dinb、抗白粉病基因Pm55和抗条锈病基因Yr5V,创制普通小麦-簇毛麦5VS易位系新种质,对于高效利用5VS上的有益基因进行小麦品种遗传改良具有重要意义。为了便于鉴别普通小麦-簇毛麦5VS纯合易位,本研究以mInDel软件分析普通小麦中国春基因组序列,开发了小麦第5同源群短臂的InDel特异分子标记WC656,对小麦品种以及普通小麦-簇毛麦5VS.5AL和5VS.5DL易位系及其衍生后代进行PCR扩增,根据扩增片段大小来区分5AS、5BS、5DS染色体臂以及5VS易位系。结果表明,在21个普通小麦品种、小麦-簇毛麦5VS回交F₂代中杂合易位单株,均扩增出379 bp（5AS）、471 bp（5BS）、422 bp（5DS）3条带。在T5VS.5AL高代品系、回交F₂代中纯合易位单株扩增出471 bp（5BS）、422 bp（5DS）2条带,379 bp（5AS）条带缺失。T5VS.5DL高代品系、回交F₂代中纯合易位单株扩增出379 bp（5AS）、471 bp（5BS）2条带,422 bp（5DS）条带缺失。WC656鉴别的5VS纯合易位结果与顺序FISH-GISH分析结果一致。T5VS.5AL、T5VS.5DL具有抗白粉病、籽粒硬度指数低等优良特性。因此,利用InDel分子标记WC656可以鉴别普通小麦-簇毛麦T5VS.5AL、T5VS.5DL衍生后代中的纯合易位,PCR扩增实验操作相对简便,可以为分子标记辅助选育携有5VS纯合易位的软质弱筋小麦提供帮助。     

用于黄瓜白粉病抗性鉴定的InDel标记

根据黄瓜白粉病抗性主效基因/QTL的插入/缺失突变,将其设计成共显性的In Del分子标记。通过群体连锁分析,发现In Del-MLO1标记与黄瓜白粉病抗性主效基因共分离。应用该标记对24份来自世界各地且白粉病抗性已知的黄瓜材料进行抗性验证,21份材料的抗性表型与标记的带型一致,说明在黄瓜进化和驯化过程中,造成抗病表型的该插入/缺失突变发生普遍。因此,该In Del标记适合于大多数黄瓜材料的白粉病抗性鉴定和分子标记辅助育种,从而加快黄瓜白粉病抗病育种的进程。     

基于SNP和InDel标记的巴西木薯遗传多样性与群体遗传结构分析

As a typical tropical crop, cassava (Manihot esculenta Crantz) has the characteristics of drought resistance, barren resistance, high biomass and so on. In addition to being used for food and forage, it can also be used for production, processing and starch extraction. Due to highly heterozygous cassava genome, breeding is more difficult. Enriching the genetic diversity of cassava germplasm, comprehensively evaluating its genetic background and traits, and discovering superior alleles that control excellent traits are of great significance for cassava breeding in the future. In order to analyze the genetic diversity, genetic relationship and population structure of cassava germplasm in Brazil, 7946 SNPs and 1997 InDels molecular markers were used. Population structure analysis was performed by ADMIXTURE software, and principal component analysis was performed by GCTA software. Brazilian cassava was divided into nine subgroups, and was roughly consistent with the results of cluster analysis using PHYLIP. Among them, subgroup 1, subgroup 2, subgroup 4, subgroup 6, and subgroup 8 could be clustered together respectively, while the samples of other subgroups could be roughly clustered, and there was a certain cross between the samples. The genetic diversity of cassava germplasm in Brazil (0.274) was higher than the genetic diversity level of cassava germplasm in China and Nigeria. Subgroup 5 of Brazil cassava had a relatively high genetic diversity (0.29). The genetic differentiation of subgroups was low (the genetic differentiation vary from 0.03 to 0.15), but higher than domestic cassava germplasm. The genetic distance between cassava accessions varied from 0.084 to 0.297, with the average of 0.228. The results of this study can provide a basis for subsequent association analysis to identify great alleles and introduction.     

白菜基因组导入芥菜的远缘杂交及后代分离偏向性

[目的]研究远缘杂交将结球白菜的A基因组导入包心芥菜,利用高分辨率熔解曲线(high-resolution melting,HRM)结合InDel标记辅助选择技术,筛选出保留白菜基因较多的回交后代,改良包心芥的结球性,丰富包心芥菜的种质资源.[方法]利用幼胚培养技术辅助6份包心芥菜与2份结球白菜进行远缘杂交和回交,选取...     

Localization of a Dwarfing Mutation in Upland Cotton Using InDel Markers Based on Genome Re-Sequencing Data

[Objective] InDel markers that were developed by comparing whole genome re-sequencing data were used to map the AS98 gene of a dwarf mutant of upland cotton (Gossypium hirsutum L.). [Method] An F2 segregation population was constructed from an upland-cotton dwarf mutant (AS98) and its wild type (LHF10), and InDel markers were developed based on genome re-sequencing data from the two parents. The InDel markers and the F2 generation were then used to map the AS98 gene of the dwarf mutant. [Result] A total of 114 InDel markers located in a previous mapping region were screened and used to genotype 223 F2 individuals. Ultimately, phenotypic data and 20 polymorphic primer pairs were used to construct a genetic linkage map. Molecular marker analysis mapped AS98 within a 6.9 cM distance of the marker InDel-50 on chromosome D12. [Conclusion] This study has demonstrated the feasibility of using InDel markers developed from re-sequencing data for gene localization in isolated populations derived from upland cotton dwarf materials. The resulting gene map provides a basis for future fine mapping and molecular-assisted selection breeding.     

北方地区白菜型冬油菜与春油菜的SSR和InDel遗传多样性分析

为了合理评价北方地区白菜型油菜种质资源,为抗寒育种及组合配制提供依据,从252对标记中筛选出9对SSR和36对In Del标记对感温性、抗寒性、品质等性状差异明显的19份白菜型油菜品种的遗传多样性和亲缘关系进行了分析。结果表明,共检测到95个等位变异,平均每个标记2.15个;有效等位基因变幅为1.05~3.27个,平均为1.70个。Shannon指数的变幅范围在0.121 7~1.269 5间,平均值为0.580 3;多态性信息量PIC变幅范围为0.049 9~0.637 7,平均值为0.308 1。通过NTSYS计算遗传相似系数GS并按加权配对法(UPGMA)聚类,结果表明19份材料的GS变异在0.52~0.86之间。在GS为0.605水平上可将19份材料按两室与多室性划分为I-1和I-2两大类群,I-1类群在GS为0.655水平上可按冬春性分为II-1、II-2两个亚类群。在GS为0.71水平上,可将冬性材料划分为4个小群。聚类结果表明北方地区白菜型油菜的遗传多样性丰富,冬、春性品种间及春性品种间亲缘关系较远,存在较大遗传差异。