甘蓝花色性状QTL定位及候选基因变异分析
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西南大学农学与生物科技学院

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国家重点研发计划项目(2018YFD0100500);国家油菜产业体系(CARS-12);重庆市社会事业与民生保障科技创新项目 (cstc2017shms-xdny80050)


QTL Mapping and Variation Analysis of Candidate Gene for Flower Color in Brassica oleracea L.
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College of Agronomy and Biotechnology, Southwest University

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    摘要:

    花色性状是甘蓝的一个重要性状,在吸引和指示传粉者、保护花器官、维持花组织能量平衡、测定品种异交率及 纯度、检测性状转移等方面有重要作用。为了鉴定控制和影响甘蓝花色的遗传位点和候选基因,本研究利用芥蓝(白花)与 野生甘蓝(黄花)构建了 F2分离群体,并分别利用基于分子标记遗传连锁图谱和基于 SNP 芯片分析的 QTL 扫描技术,对甘 蓝花色性状进行了 QTL 定位。具体研究结果如下:利用分子标记遗传连锁图谱对花色表型进行 QTL 扫描,将花色 QTL 初步 定位在甘蓝的 C02 和 C03 染色体上,其中 C03 上的 QTL 为主效位点,解释 78%的表型变异,表现部分显性效应,C02 上的 位点为微效 QTL,解释 8%的表型变异,表现加性效应;进一步采用油菜 60K SNP 芯片分析了 63 份白花和 31 份黄花 F2材料, 采用 QTL-seq 分析策略在 C03 染色体 48.08~48.92Mb 的位置鉴定到显著的峰值区间,该区间位于初定位鉴定到的 C03 QTL 区 间内部,跨度 0.84 Mb,且覆盖一个与油菜已知花色基因同源的基因 BoCCD4;RT-PCR 检测结果显示 BoCCD4 在黄色花瓣中 基本不表达,而在白色花瓣中存在不同程度的表达;双亲重测序结果发现 BoCCD4 的 CDS 第 778~780bp 处存在 3 bp 的 indel, 以此设计的 indel 标记 LJ04 在 133 份甘蓝 F2材料中与花色共分离,并在 101 份油菜 DH 系(白花性状来源于白花芥蓝)中与 花色性状共分离。本研究结论如下:甘蓝花色性状由 C03 染色体上一个部分显性主效 QTL 位点控制,并受到 C02 上一个微 效加性 QTL 的影响;主效 QTL 候选基因 BoCCD4 编码区 778-780 bp 处 3 个碱基的插入极可能导致 BoCCD4 功能丧失,从而呈 现黄色表型。本研究确认了控制甘蓝白花性状的主效 QTL 位置,鉴定到了候选基因并发现了与文献报道不同的新变异位点, 为进一步了解芸薹属物种花色的遗传和变异提供了新的数据。

    Abstract:

    Flower color is an important trait in Brassica oleracea which can attract and guide the insect pollinators, protect flower organs, maintain the energy homeostasis in flower, reveal the rate and purity of out-crossing and indicate the introgressions of traits. In order to identify the genetic loci and candidate genes controlling flower color in B. oleracea, we constructed an F2 population from the cross between a B. alboglabra line (white flower) and a wild B. oleracea accession (yellow flower), and conducted QTL analysis using a previous molecular-marker-based genetic linkage map and a SNP microarray. By using a previously developed genetic linkage map, four QTL for flower color were detected from chromosome C02 and C03, respectively. The QTL on C03 was identified as a major QTL (explaining 78% of the phenotypic variation) with a partial dominant effect, and the QTL on C02 was a minor QTL (explaining 8% of the phenotypic variation) with an additive effect. To confirm and delimit the QTL, a QTL-seq analysis was conducted by using the B. napus 60K SNP microarray among 63 white and 31 yellow F2 genotypes. A significant peak was detected from 48.08 to 48.92Mb (0.84 Mb interval) on chromosome C03, within the confidence interval of the C03-QTL. One candidate gene BoCCD4 which is homologous to a known gene controlling flower color in B. napus was found from this QTL region. Nearly no expression of BoCCD4 was found by QT-PCR in yellow-flower B. oleracea genotypes, while high expression was detected in white-flower genotypes. A 3-bp insertion was detected at 778-780 bp of the coding region of BoCCD4 between two parental lines, and an indel marker LJ04 was developed accordingly, exhibiting co-segregation with flower color in 133 F2 genotypes of B. oleracea and 101 DH lines of B. napus (derived from B. alboglabra with white-flower). Our results suggest that the white flower in B. oleracea is controlled by a partial dominant locus on chromosome C03 and possibly regulated by a minor additive QTL on C02. Furthermore, the 3-bp insertion in the candidate gene BoCCD4 was assumed to result in the loss of function of BoCCD4, leading to a yellow flower. Thus, this study identified a novel allele of the candidate gene controlling the flower color in B. oleracea, and this result might contribute to understand the variation and evolution of flower color in Brassica species.

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杨茹涵,刘进,李加纳,等.甘蓝花色性状QTL定位及候选基因变异分析[J].植物遗传资源学报,2019,20(5):1271-1277.

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  • 收稿日期:2018-12-28
  • 最后修改日期:2019-07-06
  • 录用日期:2019-04-04
  • 在线发布日期: 2019-09-17
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