Article(id=1266801753946211267, tenantId=1146029695717560320, journalId=1266358746360856629, issueId=1266801713492153305, articleNumber=null, orderNo=null, doi=10.13324/j.cnki.jfcf.202505002, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1746720000000, receivedDateStr=2025-05-09, revisedDate=1753545600000, revisedDateStr=2025-07-27, acceptedDate=null, acceptedDateStr=null, onlineDate=1779958674179, onlineDateStr=2026-05-28, pubDate=1763136000000, pubDateStr=2025-11-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1779958674179, onlineIssueDateStr=2026-05-28, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1779958674179, creator=13701087609, updateTime=1779958674179, updator=13701087609, issue=Issue{id=1266801713492153305, tenantId=1146029695717560320, journalId=1266358746360856629, year='2025', volume='45', issue='6', pageStart='570', pageEnd='672', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1779958664534, creator=13701087609, updateTime=1779959653473, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1266805862552199994, tenantId=1146029695717560320, journalId=1266358746360856629, issueId=1266801713492153305, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1266805862552199995, tenantId=1146029695717560320, journalId=1266358746360856629, issueId=1266801713492153305, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=664, endPage=672, ext={EN=ArticleExt(id=1266801754235618246, articleId=1266801753946211267, tenantId=1146029695717560320, journalId=1266358746360856629, language=EN, title=Identification of the PEBP gene family and functional prediction of FT gene in Prunus campanulata, columnId=1266801754164315077, journalTitle=Journal of Forest and Environment, columnName=Forest genetics, runingTitle=null, highlight=null, articleAbstract=

To elucidate the genetic basis of early flowering and the molecular mechanisms underlying floral regulation in plants of Rosaceae, this study selected Prunus campanulata, an early-flowering ornamental plant endemic to East Asia, as the target species. Genome-wide comparison, phylogenetic analysis, conserved motif and cis-acting element prediction, weighted gene co-expression network analysis (WGCNA) were employed to characterize the PEBP gene family and its role in floral induction. The results revealed that the member composition and motif structure of the PEBP gene family in P. campanulata were highly conserved compared with those in related species, and no significant positive selection was detected based on branch-site models, indicating a lack of functional divergence among family members. Expression profiling showed that PcamFT gene exhibited high tissue-specific expression in floral organs. Promoter analysis indicated an enrichment of light-responsive G-box elements and the absence of AE-box and GT1 repressive elements, suggesting its transcriptional regulation may be highly responsive to photoperiodic signals. WGCNA identified 33 co-expression modules, among which the module 25 showed the strongest positive correlation with floral tissue. PcamFT gene was assigned to this module and was co-expressed with several core MADS-box floral regulators, including AP1, SEP3, and AGL1. Protein-protein interaction analysis indicated that PcamFT gene may interact with these transcription factors to form regulatory complexes that promoted floral meristem identity establishment and organ differentiation.

, correspAuthors=Xianzhen ZHOU, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2025 Journal of Forest and Environment. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Yanhua YU, Chengjie FU, Jianyong CHEN, Xianzhen ZHOU), CN=ArticleExt(id=1266801756227912663, articleId=1266801753946211267, tenantId=1146029695717560320, journalId=1266358746360856629, language=CN, title=福建山樱花PEBP基因家族鉴定及FT基因功能预测, columnId=1266801754365641671, journalTitle=森林与环境学报, columnName=森林遗传学, runingTitle=null, highlight=null, articleAbstract=

为探明蔷薇科植物早花性状的遗传基础及开花调控机制, 以东亚特有的早花观赏树种福建山樱花为研究对象, 采用全基因组比对、系统发育分析、保守基序与顺式作用元件分析、加权基因共表达网络分析等方法对其PEBP基因家族进行系统研究。结果表明, 福建山樱花PEBP基因家族在成员组成和基序结构上与近缘物种高度保守, 且枝-位点模型未检测到正选择信号, 推测其家族成员未发生明显功能分化。表达分析结果显示, PcamFT基因在花组织中特异性高表达, 其启动子区域富含G-box光响应元件, 缺失AE-box和GT1等负调控元件, 提示该基因易受光周期调控。共表达网络分析鉴定出33个表达模块, 其中模块25与花组织高度相关, PcamFT基因正属于该模块, 并与AP1、SEP3、AGL1等关键开花调控因子共表达。蛋白互作网络预测进一步显示, PcamFT基因可能通过与这些MADS-box因子形成转录复合体, 参与花分生组织的身份确立及花器官发育。

, correspAuthors=周显臻, authorNote=null, correspAuthorsNote=
周显臻(1992-), 男, 讲师, 从事生物信息学研究。Email:
, copyrightStatement=版权所有©《森林与环境学报》编辑部2025, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=TO9LyvIkjaZ/9JUWIQNIhg==, magXml=hAMEWpcTZD9/JACpbdkHhQ==, pdfUrl=null, pdf=yOxNb0xQf6XkF21aTCwCcQ==, pdfFileSize=6735400, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=p+Y1ucGmUzFdKtXt1MCkLQ==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=VBvlHolWoCqo5BmhEPj4Zg==, mapNumber=null, authorCompany=null, fund=null, authors=

余燕华(1984-), 女, 副教授, 从事种苗繁育研究。Email:

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余燕华(1984-), 女, 副教授, 从事种苗繁育研究。Email:

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余燕华(1984-), 女, 副教授, 从事种苗繁育研究。Email:

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journalId=1266358746360856629, articleId=1266801753946211267, language=CN, orderNo=4, keyword=早花性状), Keyword(id=1266801762653586426, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801753946211267, language=CN, orderNo=5, keyword=花期调控)], refs=[Reference(id=1266801764863983628, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801753946211267, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=1, rfOrder=0, authorNames=null, journalName=null, refType=null, unstructuredReference=杨小凤, 李小蒙, 廖万金. 植物开花时间的遗传调控通路研究进展[J]. 生物多样性, 2021, 29(6): 825-842., articleTitle=null, refAbstract=null), Reference(id=1266801764943675405, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801753946211267, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=2, rfOrder=1, 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注:*表示P < 0.05;* *表示P < 0.01;* * *表示P < 0.001。

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注:黑线表明存在互作关系。

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Data sources of tested plants

, figureFileSmall=null, figureFileBig=null, tableContent=
参试植物
Tested plant
数据库
Database
网址
Web address
福建山樱花PcamPrunus campanulata Genome v1.0https://www.rosaceae.org/Analysis/17650653
山桃PdaPrunus davidiana Whole Genome v2.0https://www.rosaceae.org/Analysis/11857924
扁桃PruduPrunus dulcis Lauranne Genome v1.0https://www.rosaceae.org/species/prunus/prunus_dulsis/lauranne/genome_v1.0
新疆桃PfePrunus ferganensis Whole Genome v2.0https://www.rosaceae.org/Analysis/12080705
杏PARGPrunus armeniaca Genome v1.0https://www.rosaceae.org/species/prunus_armeniaca/genome_v1.0
光核桃PmiPrunus mira Whole Genome v2.0https://www.rosaceae.org/Analysis/12080707
甘肃桃PkaPrunus kansuensis Whole Genome v2.0https://www.rosaceae.org/Analysis/12080706
欧洲甜樱桃PavPrunus avium Tieton Genome v2.0https://www.rosaceae.org/Analysis/9262820
桃PpPrunus persica Whole Genome Assembly v2.0https://www.rosaceae.org/species/prunus_persica/genome_v2.0.a1
), ArticleFig(id=1266801764582965257, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801753946211267, language=CN, label=表1, caption=

参试植物数据来源

, figureFileSmall=null, figureFileBig=null, tableContent=
参试植物
Tested plant
数据库
Database
网址
Web address
福建山樱花PcamPrunus campanulata Genome v1.0https://www.rosaceae.org/Analysis/17650653
山桃PdaPrunus davidiana Whole Genome v2.0https://www.rosaceae.org/Analysis/11857924
扁桃PruduPrunus dulcis Lauranne Genome v1.0https://www.rosaceae.org/species/prunus/prunus_dulsis/lauranne/genome_v1.0
新疆桃PfePrunus ferganensis Whole Genome v2.0https://www.rosaceae.org/Analysis/12080705
杏PARGPrunus armeniaca Genome v1.0https://www.rosaceae.org/species/prunus_armeniaca/genome_v1.0
光核桃PmiPrunus mira Whole Genome v2.0https://www.rosaceae.org/Analysis/12080707
甘肃桃PkaPrunus kansuensis Whole Genome v2.0https://www.rosaceae.org/Analysis/12080706
欧洲甜樱桃PavPrunus avium Tieton Genome v2.0https://www.rosaceae.org/Analysis/9262820
桃PpPrunus persica Whole Genome Assembly v2.0https://www.rosaceae.org/species/prunus_persica/genome_v2.0.a1
), ArticleFig(id=1266801764658462730, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801753946211267, language=EN, label=Tab.2, caption=

Types and copy numbers of cis-acting elements in FT genes of tested plants

, figureFileSmall=null, figureFileBig=null, tableContent=
参试植物
Tested plant
光响应元件
Light-responsive element
激素响应元件
Plant hormone- responsive element
应激响应元件
Stress- responsive element
转录因子结合元件
Transcription factor-binding element
基本元件
Basic promoter element
G-boxAE-boxGT1-motifTCT-motifABREGARE-motifP-boxLRTW-boxMYBMYCCCAAT-box
福建山樱花Pcam300330101235
新疆桃Pfe112310101625
光核桃Pmi112310101717
山桃Pda113310101526
甘肃桃Pka113310101727
欧洲甜樱桃Pav101311111537
桃Pp112310101516
杏PARG102420101626
), ArticleFig(id=1266801764742348811, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801753946211267, language=CN, label=表2, caption=

参试植物FT基因的顺式作用元件类型及数量

, figureFileSmall=null, figureFileBig=null, tableContent=
参试植物
Tested plant
光响应元件
Light-responsive element
激素响应元件
Plant hormone- responsive element
应激响应元件
Stress- responsive element
转录因子结合元件
Transcription factor-binding element
基本元件
Basic promoter element
G-boxAE-boxGT1-motifTCT-motifABREGARE-motifP-boxLRTW-boxMYBMYCCCAAT-box
福建山樱花Pcam300330101235
新疆桃Pfe112310101625
光核桃Pmi112310101717
山桃Pda113310101526
甘肃桃Pka113310101727
欧洲甜樱桃Pav101311111537
桃Pp112310101516
杏PARG102420101626
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福建山樱花PEBP基因家族鉴定及FT基因功能预测
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余燕华 , 傅成杰 , 陈剑勇 , 周显臻 *
森林与环境学报 | 森林遗传学 2025,45(6): 664-672
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森林与环境学报 | 森林遗传学 2025, 45(6): 664-672
福建山樱花PEBP基因家族鉴定及FT基因功能预测
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余燕华 , 傅成杰, 陈剑勇, 周显臻*
作者信息
  • 福建林业职业技术学院林学系, 福建 南平 353000
  • 余燕华(1984-), 女, 副教授, 从事种苗繁育研究。Email:

通讯作者:

周显臻(1992-), 男, 讲师, 从事生物信息学研究。Email:
Identification of the PEBP gene family and functional prediction of FT gene in Prunus campanulata
Yanhua YU , Chengjie FU, Jianyong CHEN, Xianzhen ZHOU*
Affiliations
  • Department of Forestry, Fujian Forestry Vocational and Technical College, Nanping, Fujian 353000, China
出版时间: 2025-11-15 doi: 10.13324/j.cnki.jfcf.202505002
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为探明蔷薇科植物早花性状的遗传基础及开花调控机制, 以东亚特有的早花观赏树种福建山樱花为研究对象, 采用全基因组比对、系统发育分析、保守基序与顺式作用元件分析、加权基因共表达网络分析等方法对其PEBP基因家族进行系统研究。结果表明, 福建山樱花PEBP基因家族在成员组成和基序结构上与近缘物种高度保守, 且枝-位点模型未检测到正选择信号, 推测其家族成员未发生明显功能分化。表达分析结果显示, PcamFT基因在花组织中特异性高表达, 其启动子区域富含G-box光响应元件, 缺失AE-box和GT1等负调控元件, 提示该基因易受光周期调控。共表达网络分析鉴定出33个表达模块, 其中模块25与花组织高度相关, PcamFT基因正属于该模块, 并与AP1、SEP3、AGL1等关键开花调控因子共表达。蛋白互作网络预测进一步显示, PcamFT基因可能通过与这些MADS-box因子形成转录复合体, 参与花分生组织的身份确立及花器官发育。

福建山樱花  /  PEBP基因家族  /  FT基因  /  早花性状  /  花期调控

To elucidate the genetic basis of early flowering and the molecular mechanisms underlying floral regulation in plants of Rosaceae, this study selected Prunus campanulata, an early-flowering ornamental plant endemic to East Asia, as the target species. Genome-wide comparison, phylogenetic analysis, conserved motif and cis-acting element prediction, weighted gene co-expression network analysis (WGCNA) were employed to characterize the PEBP gene family and its role in floral induction. The results revealed that the member composition and motif structure of the PEBP gene family in P. campanulata were highly conserved compared with those in related species, and no significant positive selection was detected based on branch-site models, indicating a lack of functional divergence among family members. Expression profiling showed that PcamFT gene exhibited high tissue-specific expression in floral organs. Promoter analysis indicated an enrichment of light-responsive G-box elements and the absence of AE-box and GT1 repressive elements, suggesting its transcriptional regulation may be highly responsive to photoperiodic signals. WGCNA identified 33 co-expression modules, among which the module 25 showed the strongest positive correlation with floral tissue. PcamFT gene was assigned to this module and was co-expressed with several core MADS-box floral regulators, including AP1, SEP3, and AGL1. Protein-protein interaction analysis indicated that PcamFT gene may interact with these transcription factors to form regulatory complexes that promoted floral meristem identity establishment and organ differentiation.

Prunus campanulata  /  PEBP gene family  /  FT gene  /  early flowering trait  /  regulation of flowering time
余燕华, 傅成杰, 陈剑勇, 周显臻. 福建山樱花PEBP基因家族鉴定及FT基因功能预测. 森林与环境学报, 2025 , 45 (6) : 664 -672 . DOI: 10.13324/j.cnki.jfcf.202505002
Yanhua YU, Chengjie FU, Jianyong CHEN, Xianzhen ZHOU. Identification of the PEBP gene family and functional prediction of FT gene in Prunus campanulata[J]. Journal of Forest and Environment, 2025 , 45 (6) : 664 -672 . DOI: 10.13324/j.cnki.jfcf.202505002
开花时间的精准调控是植物应对环境变化、实现生殖成功的重要策略,不仅直接影响物种的生态适应性,还为农业性状的改良提供了重要潜力[1-2]。磷脂酰乙醇胺结合蛋白(phosphatidyl ethanolamine-binding proteins,PEBP) 高度保守,广泛参与植物从营养生长向生殖生长的转变及器官形态建成[3],主要分为3个功能分支,分别与开花位点T (FT)、终花基因1 (TFL1)、FTTFL1母源基因(MFT) 的形成和表达有关。FT基因是成花信号通路的核心元件,能整合光周期、春化、植物年龄及自主发育等多种信号,驱动茎尖分生组织向花分生组织的转化,从而启动开花进程[4]。已有研究[5-7]在多种蔷薇科(Rosaceae) 植物中解析了PEBP基因家族的进化特征与功能分化,并证实FT基因在花芽分化和开花调控中发挥关键的正向调节作用,但福建山樱花(Prunus campanulata) PEBP基因家族的成员组成、表达特征及其参与开花调控的机制尚缺乏系统研究。
福建山樱花是东亚特有的早花观赏树种,其花期普遍早于同属其他物种14~28 d,是探索蔷薇科植物开花时间调控及其适应性进化机制的理想模型[8]。尽管其早花性状具有重要的生态和园艺价值,但目前其形成机制尚未被充分阐明[9],制约了其在分子育种与花期精准调控等方面的实际应用潜力。基因家族的扩张、编码序列的多态性以及启动子区顺式作用元件的变异,是驱动基因功能多样化与表达调控重塑、进而影响植物关键性状进化的重要机制[10-12]。因此,系统解析福建山樱花PEBP基因家族的基因组特征与表达调控模式,对于揭示这些特征与其早花表型形成之间的密切关系至关重要。
本研究以福建山樱花为研究对象,系统鉴定其PEBP基因家族成员,并结合蔷薇科8个代表物种的PEBP基因家族数据,综合开展系统发育分析、保守基序与结构特征比对、启动子区顺式调控元件预测、选择压力分析及共表达网络构建等研究工作,重点探究福建山樱花PEBP基因家族是否通过序列变异实现了功能分化,其关键成员FT基因的启动子区顺式元件组成是否存在特异性变异,FT基因如何与其他基因形成调控网络协同驱动开花过程。本研究旨在揭示福建山樱花早花表型的分子基础,以期为蔷薇科植物花期调控机制研究提供新见解,并为其花期改良与分子育种提供理论依据与基因资源。
在蔷薇科基因组数据库(genome database for rosaceae,GDR) 中获取9种参试植物[福建山樱花(Pcam)、山桃(P. davidiana,Pda)、扁桃(P. dulcis,Prudu)、新疆桃(P. ferganensis,Pfe)、杏(P. armeniaca,PARG)、光核桃(P. mira,Pmi)、甘肃桃(P. kansuensis,Pka)、欧洲甜樱桃(P. avium,Pav) 和桃(P. persica,Pp)] 的全部编码蛋白的氨基酸序列、编码序列(coding dna sequence,CDS) 以及基因组结构注释文件(GFF3格式),参试植物数据来源如表 1所示。从蛋白质家族(protein families,Pfam) 数据库(http://pfam.xfam.org) 中获得PEBP基因的典型结构域特征文件(编号PF01161),通过HMMER 3.0软件[13]隐马尔可夫模型(hidden Markov model,HMM) 算法筛选出参试植物PEBP的蛋白序列。为进一步确定预测的PEBP基因家族成员,通过Pfam数据库检测所有候选蛋白所含的结构域,去除不含PEBP保守结构域的蛋白序列。
利用Mafft程序对上一步获得的PEBP蛋白序列进行比对[14],之后用Gblock程序提取序列中的保守区域。将比对好的序列,利用ModelTest-NG程序选择最佳模型构建系统发育树[15]。最终,利用RAxML-NG程序,以“JTTDCMut+G4”为替换模型,构建最大似然法(maximum likelihood, ML) 系统发育树,Bootstrap值设置为1 000。
将所筛选到的9种参试植物的PEBP基因对应的蛋白序列提交到在线分析工具MEME (http://meme-suite.org/tools/meme),对蛋白序列中含有的保守基序(motif) 类型和排列进行分析。氨基酸序列检索设置最大基序数值为10,其他参数选择默认参数。从参试植物基因组序列中提取PEBP基因家族成员转录起始位点上游2 000 bp的序列,选用PlantCARE在线工具(http://bioinformatics.psb.ugent.be/webtools/plantcare/html/) 分析启动子区的顺式作用元件。
采用PAML 4.9软件[16]通过Branch-site模型进行选择压力分析,将PEBP基因家族各亚群的祖先枝设为前景枝以分析该亚群分化形成时是否受到选择压力的影响;将亚群中福建山樱花的序列设为前景枝, 则可分析福建山樱花在演化过程中是否受到选择压力的影响。分析时选用Model A (Model=2,NSsites=2) 作为备择假设模型,选用Null Model A (Model=2,NSsites=2,fix_ omega=1,omega=1) 作为零假设模型,最后通过计算两两嵌套模型2ΔL似然值之间卡方值的显著情况来判断备择假设模型和零假设模型间的差异性。
福建山樱花根、叶、花、叶柄和茎的转录组测序数据下载自美国国家生物信息中心(National Center for Biotechnology Information,NCBI) 的SRA数据库(编号SRR23365082~SRR23365096)。转录组数据分析流程如下:使用FastQC软件对FASTQ文件进行质量评估,识别并去除低质量的序列、接头序列以及污染序列;利用HISAT2软件比对工具将高质量的测序读段比对到福建山樱花参考基因组上,使用Feature Counts工具根据基因注释文件计算每个基因的读数数目,生成基因表达量矩阵[17]PBEP基因家族成员在各时间点的平均表达量经过log2转换后用于绘制可视化热图。应用R包WGCNA (v1.70) 程序分析基因表达数据[18]
为分析候选基因的组织表达模式,选择在植物组织中广泛存在且稳定表达的肌动蛋白基因ACTIN作为内参基因,于2025年2月分别采集福建山樱花的根、叶、花、叶柄和茎组织进行检测,每个样品设置3个生物学重复。以花组织为对照,采用2-ΔΔCt法计算候选基因在各组织中的相对表达量。实时荧光定量-聚合酶链式反应(real-time fluorescent quantitative-polymerase chain reaction,RT-qPCR) 使用SYBR Select Master Mix (Applied Biosystems, Madrid, CA, USA) 试剂盒,在ABI PRISM 7500 FAST Sequence Detection System (Applied Biosystems, Madrid, CA, USA) 平台上进行。反应体系总量为20 μL,包括1 μL模板cDNA、10 μL SYBR Select Master Mix、1 μL上游引物、1 μL下游引物,其余体积用无RNA酶的超纯水补足。PCR扩增程序如下:50 ℃预变性2 min,95 ℃变性2 min;接着进行40个循环,每个循环包括95 ℃ 3 s和60 ℃ 30 s。
通过比对分析,从福建山樱花基因组中共鉴定出5个PEBP家族基因,其基因号分别为Pcam_evm.TU. ctg19.136、Pcam_ evm. TU. ctg16.728、Pcam_ evm. TU. ctg4.1327、Pcam_ evm. TU. ctg10.616和Pcam_evm. TU. ctg18.304。利用从9种参试植物中鉴定到的42条PEBP基因对应的蛋白序列构建系统进化树(图 1),根据桃中已报道的5条PEBP基因序列(PpTFL1,PpFTPpMFTPpBFTPpCEN),PEBP基因家族可分为5个亚群[19]。根据系统发育分析结果,可确定Pcam_ evm. TU. ctg19.136、Pcam_evm. TU. ctg16.728、Pcam_ evm. TU. ctg4.1327、Pcam_ evm. TU. ctg10.616、Pcam_ evm. TU. ctg18.304对应的基因分别为PcamMFTPcamFTPcamBFTPcamTFL1、PcamCEN
将从9种参试植物中鉴定到的42条PEBP基因对应的蛋白序列进行保守基序分析,设定共有10种基序类型存在于家族成员中,结果如图 2所示。福建山樱花PEBP基因家族5个成员与其所属亚群的其余序列在基序类型上不存在差别。通过枝-位点模型分析后发现,福建山樱花PEBP基因家族5个成员相比于亚群中的其余序列,均未能检测到显著的正选择压力信号。根据正选择压力分析结果能判断福建山樱花PEBP基因家族5个成员与其所属亚群的其余序列相比是否出现功能分化,保守基序分析和正选择压力分析结果均支持福建山樱花PEBP基因家族成员与其直系同源基因之间未出现功能分化。
为进一步了解福建山樱花PEBP基因家族成员的功能,对其在不同组织部位的表达情况进行分析,结果(图 3) 表明,只有属于FT亚群的PcamFT基因在花中有较高表达,其余4个PEBP基因在花中几乎没有表达。
参试植物扁桃缺失FT基因,其余8个参试植物的顺式作用元件的类型和拷贝数如表 2所示。对于光响应元件,各参试植物的FT基因在数量上有较大差异,只有TCT-motif数在所有FT基因中相近;福建山樱花的G-box元件比其他参试植物多,但是其AE-box和GT1-motif则缺失。关于激素响应元件,所有FT基因启动子中均包含乙烯响应转录因子和脱落酸响应元件(abscisic acid-responsive element,ABRE) 和赤霉素响应元件P-box,但只有欧洲甜樱桃中存在赤霉素响应元件基序(gibberellin-responsive element motif,GARE-motif)。对于应激响应元件,低温响应元件(low-temperature responsive element,LTR) 仅存在于欧洲甜樱桃的FT基因启动子中,与响应胁迫反应的WRKY转录因子的结合位W-box在8个FT基因启动子区域中均有检测到。此外,所有参试植物的FT基因启动子中都存在髓母细胞瘤转录因子(v-myb avian myeloblastosis viral oncogene homolog,MYB)、髓细胞瘤转录因子(myelocytomatosis transcription factor,MYC) 的结合位点。在FT基因的启动子区域中,8个FT基因中鉴定出5~7个基本元件即CCAAT-box。
利用福建山樱花各组织中的基因表达量数据,进行加权共表达网络分析,寻找能与FT基因互作的基因,结果鉴定出33个具有不同表达模式的模块。为了进一步探索这些模块与组织特异性性状之间的潜在关联性,对模块特征值与不同组织类型进行相关性分析,结果(图 4) 表明,有10个模块与花组织的形成呈显著正相关(P < 0.05),其中,模块25与花组织形成的相关性最强。
模块25基因的京都基因和基因组数据库富集分析结果如图 5所示,模块25的基因在04131膜运输、04144内吞、00020柠檬酸循环(TCA cycle)、04031 GTP结合蛋白以及00040戊糖-葡萄糖醛酸相互转化等通路中显著富集。由于FT基因正属于模块25,进一步鉴定了该模块中与PcamFT基因关联度最高的前50个基因,发现包含多个开花相关基因。在string数据库中以拟南芥(Arabidopsis thaliana) 蛋白序列及其互作关系为基础,预测这些基因的互作关系,以FT基因作为互作网络中的核心基因,发现其与拟南芥AGAMOUS基因1号(AGL1)、花瓣1基因(AP1) 以及花萼3基因(SEP3) 可能存在互作关系。此外,虽然E2F靶基因1 (ETG1)、胚胎发育缺陷基因2813号(EMB2813) 和DNA聚合酶δ4号亚基(POLD4) 之间存在互作关系,但与FT基因均无互作关系(图 6)。
AGL1、AP1、SEP3基因与FT基因在福建山樱花中表现出高度相似的表达模式,均在花组织中相对表达量最高,而在根、叶、叶柄和茎等营养器官中的相对表达量较低(图 7)。COL5基因是与光周期响应相关的重要调控因子[20],但其表达趋势与FT基因不同,其在叶、叶柄和茎的相对表达量较高,在根中的相对表达量最低,而在花中的相对表达量较低。这一结果与共表达网络预测一致,支持FT基因可能与AGL1、AP1和SEP3基因在花发育中存在功能协同,而与COL5基因之间则可能缺乏直接的互作关联。为解析PcamFT与共表达基因(AP1、SEP3) 的调控关系,分析了PcamAP1和PcamSEP3的启动子区(转录起始位点上游2 000 bp),结果显示:PcamAP1启动子含2个G-box元件(CACGTG,位置-707/-1 215),但是PcamSEP3启动子没有G-box元件的存在。
保守的基序组成与排列模式通常预示相似的功能,而正选择压力是驱动蛋白功能分化的重要力量[21-22],福建山樱花PEBP基因家族在成员组成、基序保守性及选择压力上与参试的近缘物种无显著差异,这说明福建山樱花PEBP基因家族成员与近缘物种直系同源基因间未发生功能分化。PEBP基因家族中FT基因编码蛋白是开花素的主要成分之一,能促进植物的生殖生长,而其他成员(如TFL1) 通常抑制开花[23]。异源超量表达蔷薇科物种的FT同源基因可导致转基因植物早花,而超量表达TFL1基因则导致晚花[24]。本研究中,福建山樱花FT基因在花中的高表达提示其在局部组织(如花原基) 中的快速积累,可能加速了花器官分化的进程。同时,其他PEBP基因家族成员在花中表达的普遍缺失,可能共同促成了其早花表型。
植物开花的早晚由成花素FT的表达水平直接决定,而成花素FT的表达水平则直接由FT基因的转录水平决定[25]。本研究发现,福建山樱花FT基因启动子区展现出独特的顺式作用元件组成:富含光响应核心元件G-box,却缺失了抑制性元件AE-box和GT1-motif。G-box是光周期信号通路的核心元件,可与光响应转录因子(如HY5、PIFs) 结合,正向调控FT基因表达[26]。因此,PcamFT启动子区G-box数量的显著增加,可能增强其对光信号的敏感性,使其在较短的日照条件下即可激活开花程序。与此同时,AE-box (参与光响应负调控) 和GT1 (与逆境信号偶联) 的缺失可能进一步解除对FT基因的转录抑制。这种“增强激活”与“削弱抑制”的双重作用,很可能使PcamFT基因在较低强度光照或较短时间的光周期诱导下即可达到表达阈值,从而成为福建山樱花早花的关键调控因素之一。
虽然植物开花整合子基因(FTSOC1和LFY) 的功能保守,但是开花时间调控机制存在物种特异性[27-29]。本研究通过共表达网络预测并结合RT-qPCR验证,发现PcamFT基因与MADS-box家族成员AGL1、AP1和SEP3在花组织中呈现高度同步的表达模式,这强烈提示它们在花的发育过程中存在功能协同。FT蛋白在叶片维管束中合成后,运输至茎顶端分生组织,与bZIP类转录因子开花基因D (FD) 蛋白结合形成异源二聚体复合物(FT-FD)。该复合物通过FD蛋白的DNA结合结构域识别并结合下游基因(如花分生组织特性基因AP1等) 启动子区的G-box元件,从而激活开花相关基因的表达。AP1基因可以促进花形成相关分生组织标记基因LFY基因的表达[30],而LFY基因又可直接激活FD基因表达[31],这种相互激活表达放大了开花信号,保证开花相关基因的高水平表达水平。AGL1作为MADS-box转录因子家族成员,通常参与花分生组织维持与花被发育[32]。本研究中AGL1与FT基因的共表达及同步高表达模式,可能暗示其通过物理互作或遗传上位性调控FT基因的局部活性,但具体机制需进一步验证。
本研究首次在全基因组水平系统鉴定了福建山樱花PEBP基因家族,揭示其在成员组成、保守基序和选择压力方面与近缘物种高度保守,功能未发生显著分化。福建山樱花PEBP基因家族核心成花素基因(PcamFT) 在花组织中呈现特异性高表达,其启动子区发生显著变异——富集光响应G-box元件并缺失AE-box和GT1-motif抑制元件,这可能是其对光周期信号敏感度增高、响应提前进而促成早花的重要分子基础;进一步通过共表达网络分析及RT-qPCR验证,揭示PcamFTAP1、SEP3、AGL1等MADS-box核心开花调控因子在花发育过程中高度协同表达,提示它们可能通过形成转录复合体或参与正反馈网络协同驱动花分生组织身份确立与器官分化。本研究系统阐明了福建山樱花FT基因的表达特征、独特的转录调控机制及其与关键开花因子的协同网络,为深入解析其早花分子机制及蔷薇科植物花期分子设计育种提供了重要理论基础与候选基因资源。
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2025年第45卷第6期
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doi: 10.13324/j.cnki.jfcf.202505002
  • 接收时间:2025-05-09
  • 首发时间:2026-05-28
  • 出版时间:2025-11-15
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  • 收稿日期:2025-05-09
  • 修回日期:2025-07-27
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    福建林业职业技术学院林学系, 福建 南平 353000

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周显臻(1992-), 男, 讲师, 从事生物信息学研究。Email:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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