Rui Xu, Junxian Wu, Yazhong Zhang, Lu Jiang, Jinchen Yao, Liangping Zha, Jin Xie
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引用次数: 0
摘要
1-脱氧-d-木醛糖-5-磷酸合成酶和1-脱氧-d-木醛糖-5-磷酸还原异构酶被认为是萜类生物合成2- c -甲基-d-赤藓糖-4-磷酸途径中的两个关键酶,与倍半萜类的合成和积累有关。从苍术中克隆了两个DXS和DXR基因,分析了它们在不同组织中的表达和对茉莉酸甲酯(MeJA)的响应。亚细胞定位分析表明,AlDXS和AlDXR1蛋白位于叶绿体和细胞质中,而AlDXR2蛋白仅位于叶绿体中。pET-AlDXS-28a和pGEX-AlDXR-4T-1分别在大肠杆菌BL21(DE3)和BL21中表达。基于非生物胁迫分析,重组pGEX-AlDXR-4T-1在HCl和NaOH条件下的生长速度高于对照。AlDXS基因在湖北忍冬根茎中表达量最高,在河南忍冬叶片中表达量最高。相比之下,AlDXR在湖北和河南的刺楠叶片中表达量最大。此外,MeJA对DXS和DXR基因表达、酶活性和抗氧化酶活性有一定的影响,表达峰值出现在不同的时间点。本研究结果表明,AlDXS和AlDXR基因的特征及其功能可为进一步阐明仙丹DXR和DXR基因的功能提供依据。
Isolation, characterisation, and expression profiling of DXS and DXR genes in Atractylodes lancea.
1-Deoxy-d-xylulose-5-phosphate synthase and 1-deoxy-d-xylulose-5-phosphate reductoismerase are considered two key enzymes in the 2-C-methyl-d-erythritol-4-phosphate pathway of terpenoid biosynthesis and are related to the synthesis and accumulation of sesquiterpenoids. We cloned two DXS and DXR genes from Atractylodes lancea and analysed their expression in different tissues and in response to methyl jasmonate (MeJA). Subcellular localisation analysis revealed that the AlDXS and AlDXR1 proteins are located in the chloroplasts and cytoplasm, whereas AlDXR2 is only located in the chloroplasts. pET-AlDXS-28a and pGEX-AlDXR-4T-1 were expressed in Escherichia coli BL21(DE3) and BL21, respectively. Based on the abiotic stress analysis, the growth rate of the recombinant pGEX-AlDXR-4T-1 was higher than that of the control in HCl and NaOH. AlDXS exhibited the highest expression level in rhizomes of A. lancea from Hubei but was highest in leaves from Henan. In contrast, AlDXR showed maximum expression in the leaves of A. lancea from Hubei and Henan. Moreover, DXS and DXR gene expression, enzyme activities, and antioxidant enzyme activities oscillated in response to MeJA, with expression peaks appearing at different time points. Our findings indicated that the characterisation and function of AlDXS and AlDXR could be useful for further elucidating the functions of DXR and DXR genes in A. lancea.
期刊介绍:
Genome is a monthly journal, established in 1959, that publishes original research articles, reviews, mini-reviews, current opinions, and commentaries. Areas of interest include general genetics and genomics, cytogenetics, molecular and evolutionary genetics, developmental genetics, population genetics, phylogenomics, molecular identification, as well as emerging areas such as ecological, comparative, and functional genomics.