百合花香合成相关基因LiMCS的克隆、定位和表达特性研究 |
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引用本文:张 茜,罗景琳,王浩楠,李迎迎,冷平生,胡增辉.百合花香合成相关基因LiMCS的克隆、定位和表达特性研究[J].西北农业学报,2022,(8):981~989 |
DOI:10.7606/j.issn.1004-1389.2022.08.005 |
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基金项目:国家重点研发计划(2018YFD1000400);国家自然科学基金(31201645);北京市自然科学基金(6172006)。 |
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中文摘要:百合花香中含有大量的萜类化合物,2-C-甲基-D-赤藻糖醇-2,4-环二磷酸合成酶(MCS)是合成单萜的甲基赤藓糖醇磷酸(MEP)途径中的关键酶之一。为了进一步探究MCS在百合花香合成中的作用,克隆了‘西伯利亚’百合LiMCS基因,进行生物信息学分析,使用亚细胞定位确定该基因编码蛋白的位置,根据荧光定量PCR确定了该基因的时空表达模式。结果表明,LiMCS基因开放阅读框为669 bp,编码222个氨基酸,与盾叶薯蓣的相似度最高。LiMCS蛋白定位于烟草叶表皮细胞的叶绿体中。LiMCS基因在‘西伯利亚’百合花的半开期表达量最高,花蕾期最低;在盛开期,该基因在花瓣中表达量最高,在其他部位表达量较低。结果证明LiMCS可能参与百合花香单萜类物质合成,这为后续利用基因工程手段调控百合花香释放奠定了理论基础。 |
中文关键词:‘西伯利亚’百合 LiMCS 基因克隆 表达分析 |
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Cloning,Localization and Expression Characteristics of Flower Fragance Synthesis Related Gene LiMCS in Lilium |
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Abstract:The floral volatiles of Lilium are composed of a large number of terpenoids,among which 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate synthase (MCS) is one of the key enzymes in the monoterpene synthesis 2-C-methyl-D-erythritol 4-phosphate pathway. In order to further explore theregulation mechanism of MCS on lily floral fragrancesynthesis,we cloned the LiMCS gene of Lilium ‘Siberia’. Subsequently,the bioinformatics analysis was carried out,and the subcellular location was used to determine the location of the protein encoded by the gene,and finally the spatiotemporal expression pattern of the gene was determined according to the quantitative real-time PCR. The results showed that the open reading frame of the LiMCS gene was 669 bp,encoding 222 amino acids,and had the highest similarity with Dioscorea zingiberensis. LiMCS protein was located in the chloroplasts of tobacco leaf epidermal cells. The expression level of LiMCS gene was highest in the early flowering stage and lowest in the bud stage. During flowering,the expression of this gene was highest mainly in petals,but low in other parts. The results proved that LiMCS may be involved in the synthesis of monoterpenoids of lilium fragrance,which laid a theoretical foundation for the subsequent use of genetic engineering methods to regulate the release of lily fragrance. |
keywords:Lilium ‘Siberia’ LiMCS Gene cloning Expression analysis |
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