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通过限制性内切酶相关DNA(RAD)测序进行菜豆单核苷酸多态性(SNP)发现以进行遗传多样性和群体结构分析。

SNP discovery in common bean by restriction-associated DNA (RAD) sequencing for genetic diversity and population structure analysis.

作者信息

Valdisser Paula Arielle M R, Pappas Georgios J, de Menezes Ivandilson P P, Müller Bárbara S F, Pereira Wendell J, Narciso Marcelo G, Brondani Claudio, Souza Thiago L P O, Borba Tereza C O, Vianello Rosana P

机构信息

Laboratório de Biotecnologia, Embrapa Arroz e Feijão, Santo Antônio de Goiás, GO, Brazil.

Laboratório de Biologia Molecular, Departamento de Biologia Celular, Universidade de Brasília (UnB), Brasilia, DF, Brazil.

出版信息

Mol Genet Genomics. 2016 Jun;291(3):1277-91. doi: 10.1007/s00438-016-1182-3. Epub 2016 Mar 1.

Abstract

Researchers have made great advances into the development and application of genomic approaches for common beans, creating opportunities to driving more real and applicable strategies for sustainable management of the genetic resource towards plant breeding. This work provides useful polymorphic single-nucleotide polymorphisms (SNPs) for high-throughput common bean genotyping developed by RAD (restriction site-associated DNA) sequencing. The RAD tags were generated from DNA pooled from 12 common bean genotypes, including breeding lines of different gene pools and market classes. The aligned sequences identified 23,748 putative RAD-SNPs, of which 3357 were adequate for genotyping; 1032 RAD-SNPs with the highest ADT (assay design tool) score are presented in this article. The RAD-SNPs were structurally annotated in different coding (47.00 %) and non-coding (53.00 %) sequence components of genes. A subset of 384 RAD-SNPs with broad genome distribution was used to genotype a diverse panel of 95 common bean germplasms and revealed a successful amplification rate of 96.6 %, showing 73 % of polymorphic SNPs within the Andean group and 83 % in the Mesoamerican group. A slightly increased He (0.161, n = 21) value was estimated for the Andean gene pool, compared to the Mesoamerican group (0.156, n = 74). For the linkage disequilibrium (LD) analysis, from a group of 580 SNPs (289 RAD-SNPs and 291 BARC-SNPs) genotyped for the same set of genotypes, 70.2 % were in LD, decreasing to 0.10 %in the Andean group and 0.77 % in the Mesoamerican group. Haplotype patterns spanning 310 Mb of the genome (60 %) were characterized in samples from different origins. However, the haplotype frameworks were under-represented for the Andean (7.85 %) and Mesoamerican (5.55 %) gene pools separately. In conclusion, RAD sequencing allowed the discovery of hundreds of useful SNPs for broad genetic analysis of common bean germplasm. From now, this approach provides an excellent panel of molecular tools for whole genome analysis, allowing integrating and better exploring the common bean breeding practices.

摘要

研究人员在普通菜豆基因组方法的开发和应用方面取得了巨大进展,为推动更切实可行的遗传资源可持续管理策略以用于植物育种创造了机会。这项工作为通过RAD(限制性位点关联DNA)测序开发的高通量普通菜豆基因分型提供了有用的多态性单核苷酸多态性(SNP)。RAD标签来自于12种普通菜豆基因型混合的DNA,包括不同基因库和市场类别的育种系。比对后的序列鉴定出23748个推定的RAD-SNP,其中3357个适用于基因分型;本文展示了ADT(分析设计工具)得分最高的1032个RAD-SNP。RAD-SNP在基因的不同编码(47.00%)和非编码(53.00%)序列成分中进行了结构注释。使用384个具有广泛基因组分布的RAD-SNP子集对95份普通菜豆种质的多样化样本进行基因分型,成功扩增率为96.6%,安第斯组内多态性SNP占73%,中美洲组占83%。与中美洲组(0.156,n = 74)相比,安第斯基因库的He值(0.161,n = 21)略有增加。对于连锁不平衡(LD)分析,在对同一组基因型进行基因分型的580个SNP(289个RAD-SNP和291个BARC-SNP)中,70.2%处于LD状态,在安第斯组中降至0.10%,在中美洲组中降至0.77%。对来自不同来源的样本中跨越基因组310 Mb(60%)的单倍型模式进行了表征。然而,安第斯(7.85%)和中美洲(5.55%)基因库的单倍型框架分别代表性不足。总之,RAD测序能够发现数百个用于普通菜豆种质广泛遗传分析的有用SNP。从现在起,这种方法为全基因组分析提供了一组出色的分子工具,有助于整合并更好地探索普通菜豆育种实践。

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