摘要
[目的]为进一步探明LsABF5参与叶用莴苣高温诱导抽薹过程中的分子机制.[方法]以易抽薹叶用莴苣品种'GB-30'为材料,采用同源克隆的方法克隆出LsABF5基因,运用生物信息学软件对其cDNA序列进行分析,并通过病毒诱导基因沉默技术分析LsABF5基因在叶用莴苣抽薹中的作用.[结果]LsABF5基因的全长CDS序列为1 035 bp,共编码着344个氨基酸,理论等电点值为7.59,蛋白相对分子量为38.8 kDa;该转录因子含有bZIP保守结构域,并且其高级结构大多由α螺旋和无规则卷曲组成;通过与其他植物的ABF家族成员进行同源比对和进化分析,叶用莴苣LsABF5与刺苞菜蓟亲缘关系最接近;瞬时沉默LsABF5后,在茎长测量观察中发现,沉默LsABF5后的植株茎长明显高于空白对照组、阴性对照组.对其茎尖花芽分化情况观察分析,发现空白对照组、阴性对照组植株茎尖生长点呈锥形,花芽尚未分化,沉默组植株茎尖生长点呈扁平状,已进入花芽分化期.病毒诱导LsABF5沉默可以促进叶用莴苣抽薹.[结论]LsABF5基因是典型的ABF转录因子,具有ABF蛋白家族的功能,在叶用莴苣抽薹中起抑制作用.
Abstract
[Objective]To further investigate the molecular mechanism of LsABF5 involved in high temperature induced bolting of leaf lettuce.[Methods]The LsABF5 gene was cloned by homologous cloning from lettuce'GB-30'and its cDNA sequence was analyzed by bioinformatics software.The role of LsABF5 gene in leaf lettuce bolting was analyzed by virus induced gene si-lencing technique.[Results]The full-length CDS sequence of LsABF5 gene is 1 035 bp,encoding 344 amino acids,the theoretical isoelectric point value is 7.59,and the relative molecular weight of the protein is 38.8 kDa.The transcription factor contains a bZIP conserved domain,and its higher structure is mainly composed of a helix and random coil.Through homology comparison and evolutionary analysis with ABF family members of other plants,leaf lactuca LsABF5 was the closest relative to Spinithistle.After instantaneous silencing of LsABF5,the stem length of plants silenced by LsABF5 was significantly higher than that of blank control group and negative control group.The flower bud differentiation was observed and analyzed.It was found that the growth point of the flower bud was conical in the blank control group and the negative control group,and the growth point of the flower bud was flat in the silent group,and the flower bud differentiation had entered the stage.Virus in-duced LsABF5 silencing can promote bolting of leaf lettuce.[Conclusion]LsABF5 gene is a typical ABF transcription factor,which has the function of ABF protein family and plays an inhibitory role in leaf lettuce bolting.
基金项目
国家自然科学基金面上资助项目(32072560)
北京市自然科学基金-教委联合重点资助项目(KZ20231002034)
北京市种质创制和品种选育联合攻关项目(G20220628003)
维普
万方数据