北美鹅掌楸YABBY转录因子家族的鉴定与表达分析  

Identification and expression analysis of the YABBY family in Liriodendron tulipifera

作　　者：刘换换 杨立春 张成阁 郝自远 李火根 LIU Huanhuan;YANG Lichun;ZHANG Chengge;HAO Ziyuan;LI Huogen(College of Forestry,Nanjing Forestry University,Nanjing 210037,Jiangsu,China;College of Landscape Architecture,Jiangsu Vocational College of Agriculture and Forestry,Jurong 212400,Jiangsu,China)

机构地区：[1]南京林业大学林学院,江苏南京210037 [2]江苏农林职业技术学院风景园林学院,江苏句容212400

出　　处：《中南林业科技大学学报》2023年第2期133-143,165,共12页Journal of Central South University of Forestry & Technology

基　　金：国家自然科学基金项目(31770718,31470660);江苏省高校优势学科项目(PAPD)。

摘　　要：【目的】YABBY转录因子家族仅存在于种子植物中,对植物的早期胚胎发育、果实发育、次生代谢物合成、逆境应答、侧生器官分化和背腹极性建立等方面均具有重要作用,而目前对鹅掌楸属植物的YABBY家族基因的研究较少。为了解北美鹅掌楸YABBY家族的成员和特征,对其成员进行了鉴定、亚细胞定位及表达模式分析等研究工作。【方法】从北美鹅掌楸花器官转录组中筛选出YABBY家族基因序列,采用RACE扩增技术获得了其中2个差异表达基因的全长,分别命名为LtYABBY1,5,并对他们进行生物信息学分析。随后,利用RT-qPCR检测LtYABBY1,5在北美鹅掌楸的根、茎、叶、花芽、萼片、花瓣、雄蕊、雌蕊等8个组织中的表达水平。最后,通过烟草瞬时转化体系对LtYABBY1,5蛋白进行进一步亚细胞定位分析。【结果】在北美鹅掌楸中共鉴定出6个YABBY成员,分别属于YABBY1/FILAMENTOUS FLOWER(FIL)、YABBY2、YABBY5亚族,外显子数量均为7个。克隆所得其中2个基因LtYABBY1,5,全长分别为1255、1078 bp,编码了210、188 aa,生物信息学分析结果显示,两者均为亲水蛋白与非跨膜蛋白。RT-qPCR试验发现LtYABBY1整体表达水平较高,尤其是在北美鹅掌楸的花芽和雌蕊中高表达,而LtYABBY5整体表达水平较低,仅在叶片、花芽、雌蕊和萼片中有少量表达。亚细胞定位分析结果表明,LtYABBY1,5蛋白定位于细胞核,与预测结果基本一致。【结论】北美鹅掌楸YABBY成员较少,其中LtYABBY1在花芽中特异表达,基于上述结果,推测该基因可能参与了北美鹅掌楸花芽的早期分生组织的发育调控。【Objective】The transcription factor YABBY family is unique to seed plants and plays an important role in the early stages of embryo development,fruit development,secondary metabolite synthesis,stress response,lateral organ differentiation and dorsoventral polarity establishment.However,there are few studies about the YABBY family in Liriodendron.Thus,identification,subcellular localization and expression pattern assays were conducted to analyze the members and characteristics of the YABBY family in L.tulipifera.【Method】The YABBY family homologous sequences were screened from the floral organ RNA-seq data of L.tulipifera.The full length of two differentially expressed genes,LtYABBY1 and 5,were obtained by RACE technology,and they were analyzed by bioinformatics approaches.Then,their expression levels in roots,stems,leaves,flower buds,sepals,petals,stamens and gynoecia of L.tulipifera were detected by RT-qPCR assay.Subsequently,the subcellular localization analyses of LtYABBY1 and LtYABBY5 proteins were conducted by the transient transformation system of tobacco.【Result】A total of six YABBY members of L.tulipifera were identified,which belonged to YABBY1/FILAMENTOUS FLOWER(FIL),YABBY2 and YABBY5 subfamilies with seven exons each.The full length of LtYABBY1 and LtYABBY5 genes were 1255 bp and 1078 bp,encoding 210 aa and 188 aa,respectively.LtYABBY1 and LtYABBY5 were both hydrophilic proteins without transmembrane domains.The expression of LtYABBY1 was high,especially in flower buds and gynoecia.However,the expression level of LtYABBY5 was lower,with only a small amount of transcripts in leaves,flower buds,gynoecia and sepals.Subcellular localization analysis showed that LtYABBY1 and LtYABBY5 proteins were located in the nucleus,which was consistent with the prediction.【Conclusion】The YABBY family members of L.tulipiferahas are few,and LtYABBY1 was specifically expressed in flower buds.Based on the above results,we speculate that LtYABBY1 may be involved in the early floral meristem development of L.t

关 键 词：北美鹅掌楸 YABBY 转录因子 表达分析 花芽 

分 类 号：S792.21[农业科学—林木遗传育种]