摘要
【目的】检测果实甜菜红素含量,克隆HpCYP76AD1基因并分析其表达模式,分析基因表达与甜菜红素含量之间的关系,为研究红肉火龙果果实甜菜红素积累的分子机制和改良果实营养价值提供理论依据。【方法】利用比色法测量果实成熟期间甜菜红素的相对含量,基于果实转录组测序数据结合RT-PCR克隆获得HpCYP76AD1基因,分析该基因编码蛋白质的理化性质和系统进化关系,通过qRT-PCR检测HpCYP76AD1基因在果实成熟期间的表达模式。【结果】红肉火龙果果实成熟期间甜菜红素快速积累,成熟果实(授粉30 d)中的含量约是授粉20 d的150倍。HpCYP76AD1基因的开放读码框长度为1521 bp,编码长度为506个氨基酸的蛋白质序列。HpCYP76AD1蛋白的相对分子量为57.5 kDa,预测等电点为8.86。HpCYP76AD1氨基酸序列与梨果仙人掌OfCYP76AD8相似度最高,达89.96%。HpCYP76AD1与甜菜BvCYP76AD1、紫茉莉MjCYP76AD3和紫色苋菜花AcCYP76AD2属于CYP76AD1家族α类,HpCYP76AD1在果实甜菜红素积累的两个羟基化反应中均具有催化活性。HpCYP76AD1基因在果实成熟期间显著上调表达,与果实甜菜红素的积累模式正相关。【结论】红肉火龙果细胞色素P450家族成员HpCYP76AD1基因属于α类,可能在甜菜红素积累的两步羟基化反应中均具有催化活性;在果实成熟期间大幅上调表达,参与果实甜菜红素积累。
【Objective】The present paper aimed to provide theoretical basis for studying the molecular mechanism of betacyanin accumulation and improving the nutritional value of Hylocereus polyrhizus,the betacyanin content in fruits,cloned HpCYP76 AD1 gene are detected,and the relationship between gene expression and betacyanin content are analyzed.【Method】The betacyanin content during fruit maturation was measured using the colorimetric method.HpCYP76 AD1 gene was obtained based on fruit transcriptome sequencing data and RT-PCR clone.The physicochemical properties and phylogenetic relationship of protein encoding by HpCYP76 AD1 were analyzed and the gene expression pattern during fruit maturation was detected by qRT-PCR.【Result】The betacyanin accumulated rapidly during fruit ripening,whose content in mature fruit(30 days after pollination)was about 150 times higher than the one 20 days after pollination.The open reading frame of HpCYP76 AD1 gene was 1521 bp,which encoded a protein sequence of 506 amino acids.The relative molecular weight of HpCYP76 AD1 protein was 57.5 kDa and the predicted isoelectric point was 8.86.The HpCYP76 AD1 amino acid sequence had a highest similarity with pear cactus OfCYP76 AD8,which up to 89.96%.HpCYP76 AD1 and Beta vulgaris BvCYP76 AD1,Mirabilis jalapa MjCYP76 AD3 and Amaranthus cruentus belonged to theαclass of CYP76 AD1 family,which suggested that HpCYP76 AD1 had a catalytic activity in the two hydroxylation reactions of betacyanin accumulation in fruits.The expression of HpCYP76 AD1 gene was significantly up-regulated during fruit ripening,which was positively correlated with the betacyanin accumulation pattern in fruits.【Conclusion】HpCYP76 AD1 gene from cytochrome P450 family member of H.polyrhizus belonged to theαclass,which indicated that it may have a catalytic activity in the two-step hydroxylation reaction of betacyanin accumulation.The expression of HpCYP76 AD1 gene was greatly up-regulated during fruit ripening,which was involved in betacyanin accumulation.
作者
郑乾明
王小柯
马玉华
ZHENG Qian-ming;WANG Xiao-ke;MA Yu-hua(Institute of Pomology Science,Guizhou Academy of Agricultural Science,Guiyang 550006,China)
出处
《西南农业学报》
CSCD
北大核心
2022年第6期1269-1273,共5页
Southwest China Journal of Agricultural Sciences
基金
贵州省科学技术基金项目[黔科合基础(2016)1146]。
