摘要
以不同时期不同花色的滇牡丹花瓣为试材,采用RT-PCR技术克隆得到PdANR基因的全长cDNA序列,并利用生物信息学预测PdANR蛋白的结构特征及理化性质,结合实时荧光定量PCR(qPCR)分析PdANR基因在各色组织间的表达模式,以期为滇牡丹原花青素的相关研究提供参考依据。结果表明:PdANR基因含有一个1 011 bp的完整开放阅读框(OFR),编码336个氨基酸残基,蛋白分子量为36.46×10^(3)g·mol^(-1),等电点为8.82,半衰期为30 h,不稳定指数为29.56,疏水指数为0.071,推测其为疏水性的稳定蛋白。同时,亚细胞预测显示该蛋白在细胞外中的分值最高。系统进化树表明,ANR蛋白具有明显的种属特性,其中PdANR蛋白与其它科植物的亲缘关系都较远。qPCR结果表明PdANR基因在花瓣、萼片及花药中均有表达,花瓣中的表达量会随着组织的发育而显著减少。
The full-length cDNA sequence of PdANR gene was cloned by RT-PCR from the petals of Paeonia delavayi with different flower colors at different times.The structural characteristics and physical and chemical properties of PdANR protein were predicted by bioinformatics, and the expression patterns of PdANR gene in different color tissues was analyzed by real-time fluorescent quantitative PCR(qPCR),in order to provide reference for the related research of proanthocyanidins in Paeonia delavayi.The results showed that PdANR gene contained a 1 011 bp complete open reading frame(OFR),encoding 336 amino acid residues.The molecular weight of the protein was 36.46×10^(3) g·mol^(-1),the isoelectric point was 8.82,the half-life was 30 hours, the instability index was 29.56 and the hydrophobic index was 0.071.It was speculated that it was a hydrophobic stable protein.At the same time, subcellular prediction showed that the protein had the highest score in extracellular.Phylogenetic tree showed that ANR protein had obvious species characteristics, and PdANR protein was far from other families.qPCR results showed that PdANR gene was expressed in petals, sepals and anthers, and the expression in petals decreased significantly with tissue development.
作者
杜春
平怀磊
王文治
张平芳
张怀壁
王娟
DU Chun;PING Huailei;WANG Wenzhi;ZHANG Pingfang;ZHANG Huaibi;WANG Juan(College of Forestry,Southwest Forestry University,Kunming,Yunnan 650224;Quality Standard and Testing Technology Institute,Yunnan Academy of Agricultural Sciences,Kunming,Yunnan 650205;College of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Kunming,Yunnan 650224;New Zealand Institute for Plant&Food Research Limited,Palmerston North,New Zealand 11600;Eco-Development Academy,Southwest Forestry University,Kunming,Yunnan 650224)
出处
《北方园艺》
CAS
北大核心
2022年第24期42-52,共11页
Northern Horticulture
基金
国家自然科学基金资助项目(32060089)
云南省万人计划“云岭产业技术领军人才”专项资助项目(云发改[2018]212号)
云南省重大基础专项生物资源数字化开发应用资助项目(202002AA10007)
