| 红花昼夜节律相关基因CtPRR1的特征及功能研究 |
| 投稿时间：2021-02-07 修订日期：2021-05-25 点此下载全文 |
| 引用本文：吴建辉,何贝轩,贾鑫磊,郭美丽.红花昼夜节律相关基因CtPRR1的特征及功能研究[J].药学实践杂志,2022,40(1):38~43 |
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| 中文摘要:目的 探究昼夜节律基因对红花黄酮类物质生物合成的影响及机制。方法 基于红花花冠转录组及代谢组数据库筛选潜在调控红花黄酮类化合物生物合成的昼夜节律基因；用qPCR测定红花各部位以及花冠单日不同时间点昼夜节律基因的表达量，液质联用测定黄酮类化合物的积累量，并分析二者的相关性；酵母双杂交实验验证昼夜节律基因的互作蛋白。结果 获得7个昼夜节律基因PRR1、PRR2、ELF3、FT、PHYB、GI、ZTL，其中PRR1基因与黄酮类化合物积累量呈正相关（r≥0.7）。PRR1全长3 201 bp，编码421个氨基酸，与水稻OsPRR73基因高度同源，将其命名为CtPRR1（GenBank登录号：MW492035）。CtPRR1主要在花中表达，表达量在日间逐渐升高，晚间逐渐下降；黄酮类化合物芹菜素、槲皮素、HSYA、山奈酚、Carthamin、山奈酚-3-O-葡萄糖苷以及野黄芩素的含量为白天逐渐降低，晚间逐渐升高，二者都有昼夜节律性且呈负相关（r≥-0.7）。酵母双杂实验得到2个热休克蛋白、3个AP2转录因子。结论 CtPRR1对红花黄酮类成分的昼夜节律性积累起负调节作用；CtPRR1可能受这些互作蛋白的影响调控红花黄酮类成分的昼夜节律性积累。 |
| 中文关键词:红花 昼夜节律基因 黄酮类化合物 互作蛋白 |
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| Characterization and function study of circadian rhythm gene CtPRR1 in Carthamus tinctorius L. |
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| Abstract:Objective To explore the effect of circadian rhythm genes on flavonoids biosynthesis in safflower and its molecular mechanism.Methods Based on the transcriptome and metabolomic database of safflower corolla, we screened the circadian rhythm genes that correlate with biosynthesis of flavonoids in safflower. qPCR was used to quantify the expressions of circadian rhythm genes in different flowering stages at different time points in a single day. LC-MS was performed to determine the accumulation of flavonoids. The correlation between them was analyzed as well. Yeast Two-Hybrid experiment was used to verify the interactive proteins of these genes.Results Seven circadian rhythm genes PRR1, PRR2, ELF3, FT, PHYB, GI and ZTL were obtained. PRR1 gene was positively correlated with flavonoids accumulation (r≥0.7). The full length of PRR1 is 3 201 bp, encoding 421 amino acids, which is highly homologous with rice OsPRR73 gene and named as CtPRR1 (GenBank accession number: MW492035). CtPRR1 was mainly expressed in flowers, and the expression level increased in the daytime and declined in the evening gradually. Correspondingly, the content of flavonoids showed an opposite variation. Both of them displayed a circadian rhythm with a negative correlation (r≥-0.7). In addition, 2 heat shock proteins along with 3 AP2 transcription factors interacting with CtPRR1 protein were obtained via Yeast Two-Hybrid experiment.Conclusion CtPRR1 negatively regulated the safflower flavonoids accumulation in a circadian rhythm way, which may be affected by these interacting proteins. |
| keywords:Carthami flos circadian gene flavonoids interaction protein |
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