摘要
生长调节因子(growth regulating factor,GRF)是植物体内特有的一种转录因子,在植物生长发育、渗透胁迫中发挥重要的作用.基于龙眼基因组和转录组数据,采用生物信息学分析的方法对龙眼GRF(DlGRF)家族成员进行鉴定命名;分析其基本理化性质、基因结构、蛋白结构域、启动子序列顺式作用元件;构建系统进化树、预测可能直接调控DlGRF的miRNAs;分析DlGRF家族在龙眼非胚性及胚性培养物、组织器官、光质与激素处理下的表达水平;采用实时荧光定量PCR(qRT-PCR)检测DlGRF的表达模式.结果显示,龙眼DlGRF家族包含9个成员,均含有QLQ和WRC保守结构域;包含2-5个外显子,以3个为主;分布于四大分支上,与甜橙、拟南芥GRF亲缘关系较近;DlGRF蛋白包含7个motif;启动子序列中包含众多光响应、激素应答、厌氧感应、胁迫响应及其他与生长发育相关的作用元件.转录组结果显示,DlGRF家族8个成员在球形胚阶段表达量高于其他阶段;各成员均在种子中有较高表达量,对蓝、白光及激素响应明显. qRT-PCR结果显示,DlGRF1-2、DlGRF2-1、DlGRF2-2、DlGRF4、DlGRF5-1及DlGRF7在球形胚阶段表达量最高;DlGRF1-1、DlGRF1-2、DlGRF2-2及DlGRF5-2在脱落酸(ABA)、茉莉酸甲酯(MeJA)和赤霉素(GA3)等激素处理中表达上调.此外,DlGRF可能受miR396a调控,调控方式为裂解靶标.本研究表明DlGRF在进化过程中保守性较高,且可能受到蓝光和激素的诱导,参与龙眼非胚性及胚性培养物尤其是球形胚及种子发育形成的过程.(图6表3参42)
Growth regulating factor(GRF),a specific transcription factor in plants,plays an important role in plant growth and osmotic stress responses.Based on the longan genome and transcriptome dataset,this study used bioinformatics and quantitative real-time polymerase chain reaction(qRT-PCR)analyses to identify Dimocarpus longan Lour.GRF(DlGRF)family members.Firstly,we analyzed the basic physicochemical properties,gene structure,protein domain,and cis-acting elements of the promoter sequence.Then,we constructed a phylogenetic tree and predicted miRNAs that may directly regulate the DlGRF genes.In addition,we used the transcriptome dataset to analyze the expression levels of the DlGRF gene family in longan non-embryonic and embryogenic cultures,different tissues,different light conditions,and phytohormone treatments.qRT-PCR analysis was also used to detect the expression patterns of DlGRF family members in non-embryonic and embryogenic cultures and different phytohormone treatments.The results showed that the DlGRF family contains 9 members,which contain QLQ and WRC conservative structural domains.The DlGRF family members contained two to five exons and mainly contained three exons.The DlGRF protein contained 7 conservative motifs.The phylogenetic tree analysis found that the DlGRF family distributed among four branches and they were genetically close to Citrus sinensis and Arabidopsis thaliana.The cis-acting element analysis found that the DlGRF family presented massive light response,hormone response,anaerobic induction,stress response,and growth-related elements.An expression pattern analysis(FPKM values)revealed that eight members of the DlGRF family showed the highest expression levels in the global embryo(GE)stage.Additionally,the DlGRF genes were significantly expressed in seeds,and they also responded to light and phytohormone treatments.The qRT-PCR results showed that DlGRF1-2,DlGRF2-1,DlGRF2-2,DlGRF4,DlGRF5-1,and DlGRF7 had the highest expression levels in the GE stage.The expression levels of DlGRF1-1,DlGRF1-
作者
刘蒲东
张舒婷
陈晓慧
苏立遥
姚德恒
李汉生
张梓浩
陈裕坤
林玉玲
赖钟雄
LIU Pudong;ZHANG Shuting;CHEN Xiaohui;SU Liyao;YAO Deheng;LI Hansheng;ZHANGZihao;CHEN Yukun;LIN Yuling;LAI Zhongxiong(Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2020年第2期236-245,共10页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金项目(31572088,31672127)
福建省高原学科建设经费(102/71201801101)
福建农林大学科技创新专项基金(CXZX2017189、CXZX2017314、CXZX2018076、KF2015108)资助。
