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[1]刘蒲东 张舒婷 陈晓慧 苏立遥 姚德恒 李汉生 张梓浩.龙眼GRF家族全基因组鉴定及表达模式*[J].应用与环境生物学报,2020,26(02):1-15.[doi:10.19675/j.cnki.1006-687x.2019.06028]
　LIU Pudong,ZHANG Shuting,CHEN Xiaohui,et al.Genomic identification and expression patterns of longan GRF family*[J].Chinese Journal of Applied & Environmental Biology,2020,26(02):1-15.[doi:10.19675/j.cnki.1006-687x.2019.06028]
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龙眼GRF家族全基因组鉴定及表达模式*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
26卷
期数:
2020年02期
页码:
1-15
栏目:
研究论文
出版日期:
2020-03-20

文章信息/Info

Title:
Genomic identification and expression patterns of longan GRF family*
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** 福建农林大学园艺植物生物工程研究所 福州 350002
Author(s):
LIU Pudong;? ZHANG Shuting;? CHEN Xiaohui;? SU Liyao;? YAO Deheng;? LI Hansheng;? ZHANG Zihao;? CHEN Yukun;? LIN Yuling & LAI Zhongxiong**
Institute of horticultural plant biological engineering, fujian agriculture and forestry university , fuzhou 350002, China
关键词:
龙眼;?GRF转录因子;?成员鉴定;?光质;?激素;?实时荧光定量PCR
Keywords:
Dimocarpus longan Lour.;? GRF transcription factor;? Members identification;? Light quality;? Hormone;? qRT-PCR
DOI:
10.19675/j.cnki.1006-687x.2019.06028
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4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 参43）
Abstract:
Growth regulating factor (GRF), a specific transcription factor in plants, plays an important role in plant growth and osmotic stress. Based on longan genome and transcriptome dataset, this study used bioinformatics and quantitative real-time polymerase chain reaction (qRT-PCR) analyses to identified Dimocarpus longan Lour.GRF (DlGRF) family members. Firstly, we analyzed the basic physicochemical property, gene structure, protein domain and cis-acting elements of promoter sequence. Then we constructed phylogenetic tree, predicted miRNAs that may directly regulate the DlGRF genes. In addition, we used the transcriptome dataset to analyzed the expression levels of DlGRF gene family in longan non-embryonic and embryogenic cultures, different tissues, different light quality and phytohormone treatments. qRT-PCR analysis also used to detected the expression pattern of DlGRF family members in non-embryonic and embryogenic cultures and different phytohormone treatments. The results showed that DlGRF family contain 9 members, which contain QLQ and WRC conservative structural domains. The DlGRF family members contained two to five exons, which mainly contained three exons. DlGRF protein contained 7 conservative motifs. Phylogenetic tree analysis found that the DlGRF family distributed in four branches, and they were genetically close to Citrus sinensis and Arabidopsis thaliana. Cis-acting element analysis found that DlGRF family contained massive light response, hormone response, anaerobic induction, stress response and growth-related elements. Expression patterns 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 response to light and phytohormone treatments. 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, DlGRF1-2, DlGRF2-2 and DlGRF5-2 were upregulated in the ABA, MeJA and GA 3 treatments. In addition, miR396a may regulates DlGRFs by cleavaged their mRNAs. This study indicated that DlGRF is highly conserved in the evolutionary process, and may be induced by blue light and phytohormone, thus they may participate in the formation and development of non-embryogenic and embryogenic longan cultures, especially in the GE stage and seeds.

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备注/Memo

备注/Memo:
收稿日期 Received: 2019-06-23 接受日期 Accepted: 2019-08-30
*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）和福建农林大学科技创新专项基金（CXZX2017189、CXZX2017314、CXZX2018076、KF2015108）资助
**通讯作者 Corresponding author (E-mail : Laizx01@163.com)
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更新日期/Last Update: 2019-09-04