摘要
为明确蓝星睡莲(Nymphaea colorata)蓝色、白色两种花色花瓣在不同发育时期的类黄酮物质含量、成分及其代谢途径中的关键基因,本研究采用高效液相色谱法(HPLC)和超高效液相色谱-串联四极杆飞行时间高分辨质谱(UPLC-Triple TOF-MS/MS)技术,分析了蓝星睡莲5个发育时期花瓣的花青苷和黄酮醇苷含量和成分.结果表明,蓝、白花瓣中类黄酮含量差异明显,白花花瓣花青苷和黄酮醇苷含量均保持在较低水平,尤其是在S4,S5时期与蓝花达到了显著性差异,可能是造成蓝白花色差异的物质成因.在蓝星睡莲花瓣中,两种花色花瓣类黄酮成分相同,共鉴定出3种花青苷和11种黄酮醇苷.花青苷为飞燕草素-3-O-β-半乳糖苷、飞燕草素-3-O-(2″-O-没食子酰-6″-O-乙酰-β-半乳糖苷)和飞燕草素-3′-O-(2″-O-没食子酰-6″-O-乙酰-β-半乳糖苷);黄酮醇苷主要有槲皮素3-O-半乳糖苷、杨梅素3-O-α-L-(3″-O-丙二酰)-鼠李糖苷、杨梅素3-O-α-L-鼠李糖甘、杨梅素3-O-α-L-(3″-O-乙酰)-鼠李糖苷、槲皮素3-O-α-L-(3″-O-丙二酰)-鼠李糖苷等.通过实时荧光定量PCR(RT-qPCR)技术进一步分析了类黄酮代谢途径相关基因在两种花色花瓣中的转录表达情况,结果表明,在蓝花花瓣5个发育时期,类黄酮3′5′-羟化酶基因(F3′5′H)、黄酮醇合成酶基因(FLS)、二氢黄酮醇4-还原酶基因(DFR)的转录表达变化与花青苷积累趋势一致,推测其可能是蓝色花瓣花青苷积累的关键结构基因;F3′5′H和糖基转移酶基因(GT)在蓝花中的表达量均明显高于白花,推测其是造成蓝白花色差异的重要基因.本研究为进一步解析蓝星睡莲花色形成分子机理提供了新的证据,为睡莲蓝色花分子育种提供了参考.
In order to determine the flavonoid content,components and key genes in the flavonoid metabolic pathway of waterlily(Nymphaea colorata)petals at different developmental stages,high performance liquid chromatography(HPLC)and ultra-performance liquid chromatography-tandem quadrupolar time of flight high resolution mass spectrometry(UPLC-Triple-TOF-MS/MS)were used to analyze the content of anthocyanin and flavonol glycosides in the petals of N.colorata in five developmental stages(S1-S5).The results showed that the difference on flavonoid content between blue and white petals was obvious.The content of anthocyanin and flavonol glycosides in white petals remained at a low level,especially in S4 and S5 stages,while the flavonoids of blue petals were significantly higher than that of white.This may be the cause of white and blue color of petal.Three anthocyanins and 11 flavonol glycosides were identified from the petals of N.colorata.The flavonoid compositions of blue petals and white petals were the same.The three anthocyanins were delphinidin 3-O-β-galactopyranoside,delphinidin 3-O-(2″-O-galloyl-6″-O-acetyl-β)-galactopyranoside,and delphinidin 3'-O-(2″-O-galloyl)-6″-O-acetyl-β-galactopyranoside.The flavonol glycosides mainly included quercetin 3-O-galactoside,myricetin 3-O-α-L-(3″-O-malonyl)-rhamnopyranoside,myricetin 3-O-α-L-rhamnopyranoside,myricetin 3-O-α-L-(3″-O-acetyl)-rhamnopyranoside,and quercetin 3-O-α-L-(3″-O-malonyl)-rhamnopyranoside.Besides,Real-Time quantitative PCR(RT-qPCR)was used to further analyze the expression of genes related to flavonoid biosynthetic pathway.We found that during the five developmental stages of blue flower petals,the genes’s transcriptional expression patterns of flavonoid 3'5'-hydroxylase(F3'5'H),flavonol synthetase(FLS)and dihydroflavonol 4-reductase(DFR)were consistent with the trend of anthocyanin accumulation,indicating that these genes might be responsible for anthocyanin accumulation in blue petals.Moreover,the expression levels of F3'5'H and glycosyl
作者
陈凯利
班文卓
杜灵娟
周兴华
张雄飞
CHEN Kaili;BAN Wenzhuo;DU Lingjuan;ZHOU Xinghua;ZHANG Xiongfei(College of Fishery,Southwest University,Rongchang Chongqing 402460,China;College of Landscape Architecture and Arts,Northwest A&F University,Yangling Shaanxi 712100,China;Management Committee of Rongchang Campus,Southwest University,Rongchang Chongqing 402460,China)
出处
《西南大学学报（自然科学版）》
CSCD
北大核心
2024年第2期71-80,共10页
Journal of Southwest University(Natural Science Edition)
基金
国家自然科学基金面上项目(32171863)
重庆市“大学生创新创业训练计划”(重点支持领域)项目(S202110635232)。
关键词
睡莲
花青苷
黄酮醇苷
高效液相色谱
液质联用
基因表达
waterlily
anthocyanin
flavonol glycosides
high performance liquid chromatography
LCMS/MS
gene expression
