叶用芥菜叶缘裂刻的功能优势探究及其候选基因验证

Functional Advantages Study and Identification of Candidate Genes for Margin Lobes Traits in Leaf Mustard

利用2个叶用芥菜叶缘全缘DH系‘MN001’和叶缘裂刻‘MU056’构建F;代为试材,通过测定七叶期叶片的光响应和CO_(2)响应曲线,及利用生物信息学分析QTL初定位区间内的候选基因和RT-qPCR验证不同时期不同组织器官中候选基因BjuA040054的表达,对叶用芥菜叶缘裂刻的功能优势及其形成机制进行了研究。结果表明:裂刻叶能更充分地利用光能,且Pn值与叶片裂刻程度有一定相关性,裂刻程度在一定范围内越大越有利于进行光合作用。且生物信息学分析筛选到1个与叶片形态发生相关的候选基因BjuA040054,开放阅读框660 bp,编码220个氨基酸,该基因编码的蛋白与甘蓝型油菜BnaA10g26320D编码的蛋白同源。RT-qPCR分析推测该基因主要在营养生长期四叶和五叶期的叶片部位发生作用,且在各组织器官中均有表达。研究为进一步克隆叶用芥菜裂刻基因和揭示其裂刻性状的调控机理提供了一定的生物信息学基础,并为裂刻芥菜品种的选育提供了理论依据。

Two leaf mustard doubled haploid(DH)lines with leaf margin entire’MN001’and margin lobes’MU-056’were used as materials,and the F;generation was constructed also used as materials.In order to study the functional advantages and formation mechanisms of leaf margin lobes traits in leaf mustardr,the Light Curve and Aci Curve were determined of leaves at seven-leaf,and QTL analysis of the candidate genes were analyzed by bioinformatics method in initial region,and RT-qPCR analysis of the expression of BjuA040054 in different stages and tissues or organs.The results showed that lobes leaves get the utmost out of light energy,and there was certain correlation between Pn value and level of margin lobes,the deeper of the margin lobes within a certain range,the more photosynthesis.And one candidate gene BjuA040054 related to leaf morphogenesis was selected by bioinformatics method,and the open reading frame of BjuA040054 contained 660 bp and encoded 220 amino acids,the homology analysis showed that BjuA040054 protein had the closest relationship with the homologous protein BnaA10g26320D of Brassica napus.By RT-q PCR analyzed the results showed that BjuA040054 might mainly acts on the leaf parts of the four-leaf and five-leaf stage in vegetative stages and was expressed in all tissues and organs.This study provided a certain bioinformatics foundation for further cloning the lobe-leafed gene of leaf mustard and revealing the regulation mechanism of margin lobes traits,and provided theoretical basis for the breeding selection of margin lobes mustard varieties.