百合类胡萝卜素代谢途径关键酶基因的分析及LoLcyB的克隆

Analysis of Key Enzyme Genes in Carotenoid Metabolism Pathway of Lilium and Cloning of LoLcyB Gene

本研究以东方百合品种‘Justina’叶片的5个生长阶段(10 d,25 d,50 d,75 d和100 d)和生长100 d的花柱、花药、花瓣、叶片、茎、鳞茎和根这7个不同部位为试验材料,采用有机溶剂法测量百合类胡萝卜素和叶绿素含量,根据转录组文库测序结果对百合类胡萝卜素代谢途径的12个关键酶基因(PSY,PDS,Z-ISO,ZDS,CRTISO,LCYB,LCYE,%HYB,VDE,ZEP,CCS和NCED)进行半定量表达分析,并克隆百合LCYB基因的ORF序列,并对其进行相关的生物信息学分析。结果显示,在百合叶片生长的5个不同阶段和百合成熟期的7个不同部位中,类胡萝卜素和叶绿素含量出现显著差异的现象,在不同生长阶段和不同部位12个关键酶基因的表达量各不相同。LCYB基因的完整开放阅读框(ORF)被克隆得到,全长1 503 bp,编码500个氨基酸,具有典型的Carotene Cycl超级家族保守区域特征,在已知全基因组植物中与小果野蕉(Musa acuminata)亲缘关系最近。本研究为百合类胡萝卜素代谢途径的进一步研究,深入了解百合花色和叶色合成的分子机理提供了坚实的理论基础。

In this study,five growth stages of Oriental lily ＇Justina＇ leaves（10 d,25 d,50 d,75 d and 100 d）,and seven different parts of style,anthers,petals,leaves,stems,bulbs and roots in 100 day growth were used as test materials to measure the content of carotenoids and chlorophyll in Lilium by organic solvent method.The semiquantitative analysis of 12 key enzyme genes（PSY,PDS,Z-ISO,ZDS,CRTISO,LCYB,LCYE,HYB,VDE,ZEP,CCS and NCED） of Lilium carotenoid metabolic pathway was carried out according to the sequencing results of transcriptome library.The study cloned the open reading frame（ORF） sequence of LiliumLCYB gene and carried out related bioinformatics analysis.The results showed that carotenoids and chlorophyll content were significantly different in the five different stages of Lilium leaf growth and seven different parts of Lilium ripening stage.The expression levels of 12 key enzyme genes were variant in different growth stages and parts.The complete ORF of the LCYB gene was cloned and the whole length was 1 503 bp,encoding 500 amino acids,which has typical conservedregion characteristics of C arotene Cycl super family.It showed closest relationship with Musa acuminata in the known genome.This study might provide a solid theoretical basis for further study of Lilium carotenoid metabolic pathways and further understanding the molecular mechanism of Lilium color and leaf color synthesis.