School of Biological Sciences and Institute of Agriculture, The University of Western Australia, Perth, WA, Australia.
Simons Center for Quantitative Biology, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, NY, 11724, USA.
BMC Plant Biol. 2020 Dec 7;20(1):546. doi: 10.1186/s12870-020-02756-y.
Brassica napus is an important oilseed crop cultivated worldwide. During domestication and breeding of B. napus, flowering time has been a target of selection because of its substantial impact on yield. Here we use double digest restriction-site associated DNA sequencing (ddRAD) to investigate the genetic basis of flowering in B. napus. An F mapping population was derived from a cross between an early-flowering spring type and a late-flowering winter type.
Flowering time in the mapping population differed by up to 25 days between individuals. High genotype error rates persisted after initial quality controls, as suggested by a genotype discordance of ~ 12% between biological sequencing replicates. After genotype error correction, a linkage map spanning 3981.31 cM and compromising 14,630 single nucleotide polymorphisms (SNPs) was constructed. A quantitative trait locus (QTL) on chromosome C2 was detected, covering eight flowering time genes including FLC.
These findings demonstrate the effectiveness of the ddRAD approach to sample the B. napus genome. Our results also suggest that ddRAD genotype error rates can be higher than expected in F populations. Quality filtering and genotype correction and imputation can substantially reduce these error rates and allow effective linkage mapping and QTL analysis.
油菜是全球广泛种植的重要油料作物。在油菜的驯化和选育过程中,开花时间一直是选择的目标,因为它对产量有很大的影响。本文利用双酶切相关 DNA 测序(ddRAD)技术研究油菜开花的遗传基础。以一个早花春性品种和一个晚花冬性品种杂交得到一个 F1 作图群体。
在该作图群体中,个体间的开花时间差异最大可达 25 天。初始质量控制后,基因型错误率仍然很高,生物测序重复之间的基因型不一致率约为 12%。经过基因型错误校正后,构建了一张包含 14630 个单核苷酸多态性(SNP)标记、全长 3981.31cM 的连锁图谱。在 C2 染色体上检测到一个包含 FLC 等八个开花时间基因的数量性状位点(QTL)。
这些结果表明 ddRAD 方法可以有效地对油菜基因组进行取样。我们的结果还表明,在 F1 群体中,ddRAD 基因型错误率可能高于预期。质量过滤、基因型校正和模拟可以显著降低这些错误率,并允许有效的连锁图谱构建和 QTL 分析。