TePPO基因克隆及表达分析

Cloning and expression analysis of TePPO gene in Tagetes erecta

万寿菊(Tagetes erecta L.)作为一种兼具观赏和经济价值的优质植物资源,在世界范围内广泛种植。文章依据万寿菊转录组数据,通过RNA提取、反转录克隆了万寿菊原卟啉原氧化酶(Tagetes erecta protoporphyrinogen oxidase,TePPO)基因。PPO作为催化叶绿素合成的关键酶,在植物叶绿素形成中发挥重要作用。TePPO三维结构预测及系统进化树结果表明,其编码蛋白三维结构较为稳定,与向日葵和紫花苜蓿同源基因的一致性最高。进一步利用不同花色、不同发育时期万寿菊花组织样品,对TePPO基因表达水平进行定量分析,结果表明:TePPO在万寿菊的花组织中表达,且在叶绿素量较高的花发育早期丰度更高;在花色中绿色更明显、叶绿素量更高的万寿菊品种中,TePPO基因表达水平也明显升高,提示TePPO基因与万寿菊花中叶绿素的合成及花色形成相关。

Marigold(Tagetes erecta L.),as a high-quality plant resource with both ornamental and economic value,is widely cultivated around the world.In this study,the Tagetes erecta protoporphyrinogen oxidase(TePPO)gene was cloned by RNA extraction and reverse transcription based on transcriptome data from marigold.PPO is a key enzyme in catalyzing the synthesis of chlorophyll and plays an important role in the plant chlorophyll formation.Protein three-dimensional structure prediction and phylogenetic tree analysis of TePPO indicated relatively stable coding protein and the highest identity with homologous genes from Helianthus annuus and Medicago sativa.Furthermore,TePPO expression levels in marigold flowers in different colors and different developmental stages were analyzed.The results showed that TePPO was expressed in flower tissues and contained more abundance in the early development stages with higher chlorophyll content.Moreover,the expression of TePPO significantly increased in the marigold variety with more obvious green color and higher chlorophyll content,suggesting that TePPO is related to chlorophyll synthesis and flower color formation in Tagetes erecta.