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为了探究AUX/IAA家族基因在结球甘蓝叶片与茎尖发育过程中的功能,筛选出对甘蓝叶片与茎尖发育具重要影响的相关AUX/IAA基因。试验以结球甘蓝品系“519”为材料,通过对莲座期叶片与茎尖,结球期叶片与茎尖的转录组比较分析,筛选出在两个组织中均表达差异显著的2个AUX/IAA基因,分别为BoIAA2与BoIAA19。采用PCR技术分别获得基因BoIAA2与BoIAA19的编码序列,其CDS(coding sequence)全长分别为519 bp、585 bp,编码172、194个氨基酸;进一步序列分析发现BoIAA2、BoIAA19分别与大白菜BrIAA2、油菜BnIAA19氨基酸同源相似性均达99%,且均具有AUX/IAA蛋白的DomainⅠ、Ⅱ、Ⅲ和Ⅳ特异性结构域。利用荧光定量PCR分析表明BoIAA2和BoIAA19在叶片、茎尖中的表达量均呈现出结球期高于莲座期的变化趋势,与转录组分析结果相符;对其结合蛋白基因BoTIR1和BoARF8进行表达分析,结果显示二者与BoIAA2、BoIAA19在叶片、茎尖中不同时期的表达量变化趋势一致;由此,推测BoIAA2和BoIAA19可能通过与BoTIR1和BoARF8的结合影响生长素信号转导,进而调控结球甘蓝叶片及茎尖的生长发育。
In order to explore the function of AUX / IAA family genes in the development of leaf cabbage and shoot tip, we screened the related AUX / IAA genes that have important influence on the development of leaf and shoot tip of cabbage. In this study, the cabbage variety “519 ” was used as materials. The transcriptome of leaves and shoot tips at the rosette stage, AUX / IAA genes, BoIAA2 and BoIAA19 respectively. The coding sequences of BoIAA2 and BoIAA19 were obtained by PCR. The full-length coding sequence of CDS (coding sequence) was 519 bp and 585 bp, encoding 172,194 amino acids. Further analysis showed that BoIAA2 and BoIAA19 were respectively associated with BrIAA2, The amino acid identities of BnIAA19 reached 99%, and all of them had Domain Ⅰ, Ⅱ, Ⅲ and Ⅳ specific domains of AUX / IAA protein. Fluorescence quantitative PCR analysis showed that the expression level of BoIAA2 and BoIAA19 in leaves and shoot tips showed a trend of higher than that in rosette stage at the stage of coturnum, which was consistent with the result of transcriptome analysis. The expression of BoTIR1 and BoARF8 The results showed that the expression level of BoIAA2 and BoIAA19 in leaves and shoot tips were consistent at different stages. Therefore, it was speculated that BoIAA2 and BoIAA19 might affect auxin signal transduction through the combination with BoTIR1 and BoARF8, Growth and development of cabbage leaves and shoot tips.