Toward Identification of Black Lemma and Pericarp Gene Blp1 in Barley Combining Bulked Segregant Analysis and Specific-Locus Amplified Fragment Sequencing.
Toward Identification of Black Lemma and Pericarp Gene Blp1 in Barley Combining Bulked Segregant Analysis and Specific-Locus Amplified Fragment Sequencing.
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结合批量分离分析和特定位点扩增片段测序，鉴定大麦中的黑外稃和果皮基因 Blp1
DOI：
10.3389/fpls.2017.01414
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发表时间：
2017
影响因子：
5.6
通讯作者：
Liang Z
中科院分区：
文献类型：
--
作者：
Jia Q;Wang J;Zhu J;Hua W;Shang Y;Yang J;Liang Z
黑大麦是由外稃和/或果皮中合成的植物黑素引起的，该性状由一个显性基因Blp1控制。通过分子标记将该基因定位在1H染色体上，但尚未被分离出来。特异位点扩增片段测序（SLAF - seq）是一种大规模从头发现单核苷酸多态性（SNP）和基因分型的有效方法。在本研究中，采用结合混合分组分析（BSA）的SLAF - seq方法，在来自哈铁系1号×浙5819的F2群体中获取足够的标记来精细定位Blp1基因。根据SNP筛选标准，共有77542个多态性SNP符合关联分析的要求。结合两种关联分析方法，在1H染色体上获得了一个大小为32.41 Mb的重叠区域作为Blp1基因的候选区域。根据SLAF - seq数据，在目标区域开发了标记并用于Blp1基因的定位。连锁分析表明，在哈铁系1号×浙5819的172株纯合黄色籽粒F2植株中，Blp1与HZSNP34和HZSNP36共分离，并由两个标记（HZSNP35和HZSNP39）界定，跨度为8.1 cM。在缩小的目标区域筛选出更多的多态性标记并用于群体的基因分型。结果，Blp1被上游标记HZSNP63和下游标记HZSNP59界定在1H染色体上的1.66 Mb范围内。我们的研究结果证明了SLAF - seq - BSA方法在识别候选区域以及发现特定目标基因组区域的多态性标记方面的实用性。
Black barley is caused by phytomelanin synthesized in lemma and/or pericarp and the trait is controlled by one dominant gene Blp1. The gene is mapped on chromosome 1H by molecular markers, but it is yet to be isolated. Specific-locus amplified fragment sequencing (SLAF-seq) is an effective method for large-scale de novo single nucleotide polymorphism (SNP) discovery and genotyping. In the present study, SLAF-seq with bulked segregant analysis (BSA) was employed to obtain sufficient markers to fine mapping Blp1 gene in an F2 population derived from Hatiexi No.1 × Zhe5819. Based on SNP screening criteria, a total of 77,542 polymorphic SNPs met the requirements for association analysis. Combining two association analysis methods, the overlapped region with a size of 32.41 Mb on chromosome 1H was obtained as the candidate region of Blp1 gene. According to SLAF-seq data, markers were developed in the target region and were used for mapping the Blp1 gene. Linkage analysis showed that Blp1 co-segregated with HZSNP34 and HZSNP36, and was delimited by two markers (HZSNP35 and HZSNP39) spanning 8.1 cM in 172 homozygous yellow grain F2 plants of Hatiexi No.1 × Zhe5819. More polymorphic markers were screened in the reduced target region and were used to genotype the population. As a result, Blp1 was delimited within a 1.66 Mb on chromosome 1H by the upstream marker HZSNP63 and the downstream marker HZSNP59. Our results demonstrated the utility of SLAF-seq-BSA approach to identify the candidate region and discover polymorphic markers at the specific targeted genomic region.
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影响因子：
3.7
作者：
Geng X;Jiang C;Yang J;Wang L;Wu X;Wei W
通讯作者：
Wei W
影响因子：
5.6
作者：
Hu MJ;Zhang HP;Liu K;Cao JJ;Wang SX;Jiang H;Wu ZY;Lu J;Zhu XF;Xia XC;Sun GL;Ma CX;Chang C
通讯作者：
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影响因子：
5.4
作者：
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影响因子：
6.1
作者：
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通讯作者：
Chung, Ill-Min
影响因子：
3.1
作者：
Jones, Elizabeth;Chu, Wen-Chy;Smith, Stephen
通讯作者：
Smith, Stephen