摘要
对香豆酸-3-羟基化酶(coumarate-3-hydroxylase,C3H)是木质素合成途径的关键酶。本研究采用RACE方法从‘突尼斯软籽’(Punica granatum cv.Tunisiruanzi)中获得C3H的同源基因PgC3H。该基因开放阅读框为1527bp,编码508个氨基酸。相对分子量为57759.0,等电点为8.37,推测该蛋白定位于叶绿体中。系统进化树分析显示,PgC3H与其他物种起源相同,而与寇阿相思树AkF5H的亲缘关系最近。荧光定量PCR分析表明:PgC3H在花、嫩茎、种皮、叶片四个组织中均有表达,其中嫩茎中的表达量最高,而花中的表达量最低;PgC3H基因在‘突尼斯软籽’籽粒不同发育时期均有表达,呈现先降低、再升高再降低的趋势。30~60d相对表达量逐步下降,后在第75天时表达量升至最高,第90天时急剧下降至最低值,至第120天时维持较低水平表达。该研究对于后续研究PgC3H的功能提供了理论依据。
Coumarate-3-hydroxylase is a key enzyme in lignin biosynthesis pathway. In this study, PgC3H, the homologous gene of C3H, was obtained from Punica granatum cv. Tunisiruanzi by using RACE method. The open reading frame of PgC3H was 1 527 bp, encoding 508 amino acids. The relative molecular weight was 57 759.0 and the isoelectric point was 8.37. It was speculated that the protein was localized in the chloroplast. Phylogenetic tree analysis indicated that PgC3H shared the same origin with other species, and had the closest relationship with Eucalyptus camaldulensis AkF5H. The fluorescent quantitative PCR analysis showed that expression of PgC3H was detected in flowers, young stems, seed coats and leaves. The expression level in young stems was the highest, while the lowest in flowers. The expression of PgC3H was detected in different development stages of 'P. granatum cv. Tunisiruanzi' grain. The relative expression showed a tendency of decreasing first, then increasing, and finally decreasing. The relative expression level of PgC3H gradually decreased from 30~ 60 d, then reached the highest level on the 75th day, later dropped to the lowest value on the 90th day, and remained lower level until the 120th day. This study might provide the theory basis for the subsequent studies on the function of PgC3H.
作者
熊枫
陈磊
刘同瑞
王琦
刘娜
张水明
董丽丽
Xiong Feng;Chen Lei;Liu Tongrui;Wang Qi;Liu Na;Zhang Shuiming;Dong Lili(College of Horticulture, Anhui Agricultural University, Hefei,230036)
出处
《分子植物育种》
CAS
CSCD
北大核心
2018年第23期7628-7633,共6页
Molecular Plant Breeding
基金
安徽省高等学校自然科学研究重点项目(KJ2017A154)资助
关键词
石榴
C3H
基因克隆
表达分析
Pomegranate
C3H
Gene cloning
Expression analysis
