作 者：
董晨;魏永赞;王弋;郑雪文;李伟才;石胜友
关键词：
荔枝;妃子笑;泛素结合酶;基因;实时荧光定量PCR;生物信息学;花穗发育;组织表达
摘 要：
【目的】分析荔枝泛素结合酶(E2)基因(LcUBC12)的生物学信息,并检测其组织表达特异性及烯效唑处理下花穗不同发育阶段中的表达情况,为研究E2响应烯效唑的作用机制及泛素化修饰在调控荔枝花穗发育中的功能提供理论依据。【方法】以从妃子笑荔枝花穗转录组(RNA-seq)数据中筛选出的LcUBC12基因为参考序列,利用本地BLAST从荔枝基因组中查找出该基因cDNA全长(Litchi_GLEAN_10004716),并利用实时荧光定量PCR(qRT-PCR)检测LcUBC12基因在妃子笑荔枝不同组织及烯效唑处理下花穗不同发育阶段的表达情况。【结果】LcUBC12基因c DNA全长519 bp,编码172个氨基酸,其编码的蛋白分子量19.68 kD,等电点6.52,不稳定指数35.70,亲水性的平均值-0.734,为亲水稳定蛋白,定位于细胞核,不存在信号肽和跨膜结构,包含典型的UBC12保守结构域,属于Class I家族E2蛋白,二级结构以无规则卷曲为主。LcUBC12蛋白与向日葵(XP_022009656.1)、柑橘(XP_006466720.1)、草莓(XP_004300599.1)和萝卜(XP_018466680.1)等UBC12蛋白的氨基酸序列同源性较高,分别为70.11%、68.16%、67.76%和67.03%。LcUBC12蛋白与芸香科的柑橘UBC12亲缘关系最近,其次是与同属菊科的向日葵和莴苣UBC12蛋白亲缘关系较近。LcUBC12基因在不同组织中表达量排序为雌花>雄花>种子>果皮>叶>茎>根>果肉。烯效唑处理0～21 d LcUBC12基因在花穗中的表达量与对照无明显差异,烯效唑处理28 d其表达量明显低于对照,但烯效唑处理35 d其表达量上调,且较对照高。【结论】LcUBC12基因具有组织表达特异性,在妃子笑荔枝雌花、雄花和种子中高效表达,推测其主要在花穗发育和生殖生长过程中发挥重要调控作用,但烯效唑处理后LcUBC12基因对花穗发育发挥负调控作用。
译 名：
Bioinformatics and expression characteristics analysis of Litchi chinesis Sonn ubiquitin-conjugating enzyme gene(LcUBC12)
作 者：
DONG Chen;WEI Yong-zan;WANG Yi;ZHENG Xue-wen;LI Wei-cai;SHI Sheng-you;Institute of South Subtropical Corp Research,Chinese Academy of Tropical Agricultural Science/Key Laboratory of Tropical Fruit Biology,Ministry of Agriculture and Rural Affairs;
关键词：
litchi;;Feizixiao;;ubiquitin-conjugating enzyme;;gene;;real-time fluorescence quantitative PCR;;bioinformatics;;inflorescences development;;tissue expression
摘 要：
【Objective】Biological information of ubiquitin-conjugating enzyme(E2)gene(LcUBC12)in litchi was analyzed,and the expression pattern specificity and the expression of LcUBC12 were studied in different stages of litchi inflorescences under uniconazole treatment. The results could provided a theoretical basis for studying the mechanism of E2 response to uniconazole and the function of ubiquitination modification in regulating the development of Feizixiao litchi inflorescences.【Method】With LcUBC12 gene screened from the transcriptome data of the Feizixiao litchi inflorescences(RNA-seq)as reference sequence,the full-length cDNA of LcUBC12 was obtained by local BLAST to find the litchi genome(Litchi_GLEAN_10004716). Real-time fluorescence quantitative PCR(qRT-PCR)was used to analyze the expression of Feizixiao litchi LcUBC12 in different tissues and at different developmental stages of inflorescence which were treated by uniconazole.【Result】The cDNA of LcUBC12 gene was 519 bp in length,encoding 172 amino acids with a molecular weight of 19.68 kD and an isoelectric point of 6.52,and the instability index was 35.70,the average hydrophilic value was-0.734,which was a hydrophilic stable protein. The predicted subcellular localization was in the nucleus. There was no signal peptide and transmembrane. The structure contained a typical conserved UBC domain. It belonged to Class I family E2 protein,the secondary structure of the protein was mainly random coiling. NCBI blast analysis indicated the homology of UBC12 protein amino acid sequence of LcUBC12 amino acid sequence with sunflower(XP_022009656.1),citrus(XP_006466720.1),strawberry(XP_004300599.1)and radish(XP_018466680.1)were high,which were 70.11%,68.16%,67.76% and 67.03%,respectively. Phylogenetic analysis indicated that LcUBC12 protein was the closest to citrus UBC12 of Rutaceae and also had close relationship with sunflowers and lettuce of the Compositae. qRT-PCR analysis indicated the expression of LcUBC12 gene in different tissues was female flower>male flower>seed>peel>leaf>stem>root>pulp. The expression treated by uniconazole at 28 d was lower than that in control,however the expression at 35 d up-regulated in treatment and was higher than that in control.【Conclusion】The LcUBC12 gene has tissue expression specificity and is abundantly expressed in Feizixiao litchi female flowers,male flowers and seeds. It is speculated that it mainly plays an important role in the inflorescences development and reproductive growth of litchi,but LcUBC12 gene plays a negative regulatory role in inflorescences development after uniconazole treatment.
