目的 对铁皮石斛Dendrobium officinale中糖基转移酶基因DoUGT83A1进行克隆，并对其进行生物信息学及表达模式分析。方法 从铁皮石斛“雁荡山1号”叶组织中获得DoUGT83A1基因，利用软件进行生物信息学分析，利用MEGA 11构建系统发育树，采用实时荧光定量PCR（qRT-PCR）检测基因表达模式。结果 通过PCR扩增得到1个DoUGT83A1基因（GI：1622023669），该基因的CDS序列为1 347 bp，编码448个氨基酸。DoUGT83A1蛋白相对分子质量为54 330，理论等电点为5.75，为亲水性不稳定蛋白，无信号肽和跨膜区域，含有41个磷酸化位点，具有UDPGT和PSPG 2个保守结构域。氨基酸序列分析表明DoUGT83A1蛋白与金钗石斛Dendrobium nobile的UGT83A1蛋白的同源性最高，相似度为96.36%。分子对接结果证明DoUGT83A1蛋白对山柰酚具有较好的催化活性。组织特异性表达分析结果显示，DoUGT83A1在“雁荡山1号”不同组织相对表达量为：叶＞茎＞花＞根（白根、绿根尖）。顺式作用元件分析结果表明，DoUGT83A1基因启动子序列含有多个与非生物胁迫响应相关的元件，且在低温、缺磷和菌根共生诱导下显著上调表达，推测该基因可能参与调控铁皮石斛菌根共生及非生物胁迫响应过程。结论 成功克隆获得铁皮石斛DoUGT83A1基因，并进行生物信息学分析和基因表达模式分析。为进一步探究DoUGT83A1基因功能及其在菌根共生和非生物胁迫响应中的作用机制提供理论依据。

Objective To clone one glycosyl transferase gene named DoUGT83A1 from Dendrobium officinale and analyze its bioinformatics information and expression patterns. Methods The DoUGT83A1 gene was obtained from the leaf tissue of D. officinale ‘Yandangshan 1’. Online softwares were used for bioinformatics analysis. Phylogenetic tree was constructed using MEGA 11. Gene expression patterns were analyzed by qRT-PCR. ResultsThe DoUGT83A1 gene (GI: 1622023669) was cloned by PCR. The CDS sequence of the DoUGT83A1 gene was 1347 bp, encoding 448 amino acids. The DoUGT83A1 protein was a unstable hydrophilic protein, with no signal peptide or transmembrane region. The relative molecular weight of DoUGT83A1 was 54 330, and its theoretical isoelectric point (pI) was 5.75. DoUGT83A1 contained 41 phosphorylation sites and two conserved domains (UDPGT, PSPG). Amino acid sequence analysis showed that DoUGT83A1 had the highest homology with UGT83A1 protein of D. nobile, with a similarity of 96.36%. Molecular docking demonstrated the catalytic activity of DoUGT83A1 protein on kaempferol. Tissue-specific expression analysis showed that the relative expression levels of DoUGT83A1 varied across different tissues of ‘Yandangshan 1’ as follows: leaves > stems > flowers > roots (white roots, green root tips). The results of cis-acting element analysis showed that the promoter sequence of DoUGT83A1 contained multiple elements, which were related to abiotic stress response. And the expression was significantly up-regulated under the induction of low temperature, phosphorus deficiency and mycorrhizal symbiosis, which suggested that this gene may regulate the mycorrhizal symbiosis and abiotic stress response process of D. officinale. Conclusion The cloning, bioinformatics analysis and expression patterns analysis of DoUGT83A1 of D. officinale were completed, which provided a theoretical basis for further exploring the function of DoUGT83A1 and its mechanism in mycorrhizal symbiosis and abiotic stress response.

R286.12

国家自然科学基金项目(32102485);温州市科技特派员专项项目(X2023018);乐清市农业科技计划项目(2023N002);浙江省农业科学院乐清共同富裕产业研究院项目