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Screening of Candidate Leaf Morphology Genes by Integration of QTL Mapping and RNA Sequencing Technologies in Oilseed Rape (Brassica napus L.)

机译:QTL定位和RNA测序技术在油菜中候选叶片形态基因的筛选

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摘要

Leaf size and shape play important roles in agronomic traits, such as yield, quality and stress responses. Wide variations in leaf morphological traits exist in cultivated varieties of many plant species. By now, the genetics of leaf shape and size have not been characterized in Brassica napus. In this study, a population of 172 recombinant inbred lines (RILs) was used for quantitative trait locus (QTL) analysis of leaf morphology traits. Furthermore, fresh young leaves of extreme lines with more leaf lobes (referred to as ‘A’) and extreme lines with fewer lobes (referred to as ‘B’) selected from the RIL population and leaves of dissected lines (referred to as ‘P’) were used for transcriptional analysis. A total of 31 QTLs for the leaf morphological traits tested in this study were identified on 12 chromosomes, explaining 5.32–39.34% of the phenotypic variation. There were 8, 6, 2, 5, 8, and 2 QTLs for PL (petiole length), PN (lobe number), LW (lamina width), LL (Lamina length), LL/LTL (the lamina size ratio) and LTL (leaf total length), respectively. In addition, 74, 1,166 and 1,272 differentially expressed genes (DEGs) were identified in ‘A vs B’, ‘A vs P’ and ‘B vs P’ comparisons, respectively. The Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were used to predict the functions of these DEGs. Gene regulators of leaf shape and size, such as ASYMMETRIC LEAVES 2, gibberellin 20-oxidase 3, genes encoding gibberellin-regulated family protein, genes encoding growth-regulating factor and KNOTTED1-like homeobox were also detected in DEGs. After integrating the QTL mapping and RNA sequencing data, 33 genes, including a gene encoding auxin-responsive GH3 family protein and a gene encoding sphere organelles protein-related gene, were selected as candidates that may control leaf shape. Our findings should be valuable for studies of the genetic control of leaf morphological trait regulation in B. napus.
机译:叶的大小和形状在农艺性状(如产量,品质和胁迫响应)中起着重要作用。许多植物品种的栽培品种叶片形态特征均存在很大差异。到目前为止,在甘蓝型油菜中尚未鉴定叶片形状和大小的遗传学。在这项研究中,将172个重组自交系(RIL)的种群用于叶片形态性状的定量性状基因座(QTL)分析。此外,从RIL种群中选择具有更多叶裂片(称为“ A”)的极品系的新鲜幼叶和具有较少叶裂片(称为“ B”)的极品系和解剖系的叶片(称为“ P”) ')被用于转录分析。在这项研究中,共鉴定了31个针对12个染色体的叶片形态性状的QTL,解释了表型变异的5.32–39.34%。 PL(叶柄长度),PN(叶数),LW(薄片宽度),LL(薄片长度),LL / LTL(薄片尺寸比)和8个,6、2、5、8和2个QTL LTL(叶总长)分别。此外,在“ A vs B”,“ A vs P”和“ B vs P”比较中分别鉴定出74、1,166和1,272个差异表达基因(DEG)。基因本体论(GO)和京都基因与基因组百科全书(KEGG)数据库被用来预测这些DEG的功能。在DEGs中也检测到了叶片形状和大小的基因调节剂,例如不对称叶2,赤霉素20-氧化酶3,编码赤霉素调节的家族蛋白的基因,编码生长调节因子的基因和类似KNOTTED1的同源盒。在整合了QTL定位和RNA测序数据后,选择了33个基因(包括编码植物生长素应答性GH3家族蛋白的基因和编码球形细胞器蛋白相关基因的基因)作为可以控制叶片形状的候选基因。我们的发现对于研究甘蓝型油菜叶片形态性状调控的遗传控制具有重要的参考价值。

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