| 羟基红花黄色素A生物合成途径短链还原酶基因的特征及功能研究 |
| 投稿时间：2022-01-17 修订日期：2022-04-08 点此下载全文 |
| 引用本文：王璐暖,吴建辉,何贝轩,张彦洁,郭美丽.羟基红花黄色素A生物合成途径短链还原酶基因的特征及功能研究[J].药学实践杂志,2022,40(3):218~225 |
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| 基金项目:国家自然科学基金（81973421，81473300）；国家重点研发计划（2019YFC1711100） |
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| 中文摘要:目的 探究参与红花黄酮类生物合成途径特别是羟基红花黄色素A（HSYA）关键短链脱氢还原酶基因（SDRs）的功能。方法 基于红花转录组数据库以及代谢组数据库，筛选参与HSYA生物合成途径的SDRs，用qRT-PCR法分析表达模式。采用无缝克隆技术构建过表达载体，以农杆菌GV3101介导遗传转化云南巍山红花品系，对转基因T2代植株进行阳性验证，并对花冠SDRs的基因表达量进行分析，UPLC-Q-TOF/MS法测定次生代谢物的含量。结果 筛选出3个参与HSYA生物合成途径的关键短链脱氢还原酶基因CtSDR1、CtSDR2、CtSDR3，表达量从高到低依次为花冠>叶>茎>根。花冠中表达量随花冠发育逐渐升高。对转基因T2代植株进行阳性验证后的花冠进行qRT-PCR分析发现：与空白对照组相比，转CtSDR3过表达T2代阳性植株花冠中CtSDR3基因的转录水平增加了2~3倍，次生代谢物HSYA的含量提高了7.1%～16.6%（P＜0.05）。结论 CtSDR3可能参与了红花中黄酮类化合物特别是HSYA的生物合成，为阐释CtSDR3在HSYA生物合成途径中的功能提供了数据支撑。 |
| 中文关键词:短链脱氢还原酶 羟基红花黄色素A 红花 |
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| Characterization and function of short-chain dehydrogenases/reductases in hydroxysafflower yellow A biosynthesis pathway |
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| Abstract:Objective To explore the function of short-chain dehydrogenases/reductases (SDRs) in safflower flavonoid, especially hydroxysafflower yellow A (HSYA) biosynthesis. Methods SDRs involved in HSYA biosynthesis pathway were screened based on safflower transcriptome database and metabolome database. The expression pattern was analyzed by qRT-PCR. The overexpression vector was constructed by seamless cloning technology, then genetically transformed to the Yunnan Weishan safflower strain by Agrobacterium gv3101. The transgenic T2 generation plants were positively verified, and the gene expression of corolla SDRs was analyzed. The content of secondary metabolites was assayed by UPLC-Q-TOF/MS. Results Three SDRs genes named CtSDR1, CtSDR2 and CtSDR3 involved in HSYA biosynthesis pathway were screened. Their expression in safflower from high to low was corolla > leaf > stem > root. The expression level in corolla increased gradually with corolla development. qRT-PCR analysis of corolla with positive verification of genome insertion sequence showed that the transcription level of CtSDR3 in corolla of T2 positive plants increased by 2~3 times compared with the blank control group, and the content of secondary metabolite HSYA increased by 7.1%~16.6% (P< 0.05). Conclusion CtSDR3 may be involved in the biosynthesis of flavonoids, especially HSYA, in safflower. It provides the support data for explaining the function of CtSDR3 in HSYA biosynthesis pathway. |
| keywords:short-chain dehydrogenase reductase hydroxysafflor yellow A safflower |
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