Article(id=1236318325786080107, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1236318325282763627, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2023.02.003, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1652112000000, receivedDateStr=2022-05-10, revisedDate=1656000000000, revisedDateStr=2022-06-24, acceptedDate=null, acceptedDateStr=null, onlineDate=1772690858551, onlineDateStr=2026-03-05, pubDate=1677254400000, pubDateStr=2023-02-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772690858551, onlineIssueDateStr=2026-03-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772690858551, creator=13701087609, updateTime=1772690858551, updator=13701087609, issue=Issue{id=1236318325282763627, tenantId=1146029695717560320, journalId=1235980609244409860, year='2023', volume='44', issue='2', pageStart='225', pageEnd='445', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1772690858431, creator=13701087609, updateTime=1772695119289, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1236336196671033969, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1236318325282763627, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1236336196675228274, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1236318325282763627, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=246, endPage=253, ext={EN=ArticleExt(id=1236318326033544046, articleId=1236318325786080107, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Construction of Yeast Two-hybrid Normalized Library and Screening of Interaction Proteins of PnRXLR5863 in Agave hybrid No.11648, columnId=1236256430337085821, journalTitle=Chinese Journal of Tropical Crops, columnName=Omics & Biotechnology, runingTitle=null, highlight=null, articleAbstract=

Agave spp. is main resource of hard fiber. Zebra disease, caused by the pathogen Phytophthora nicotianae, is one of the most devastating sisal oomycete diseases. RXLR effector proteins are important cytoplasmic effectors of oomycete and more than 300 RXLR effector genes were identified in the genomes of P. nicotianae. RXLR effectors have the ability to manipulate or suppress host immunity by directly binding host proteins. A typical RXLR effector (named PnRXLR5863) that induced programmed cell death (PCD) was identified from P. nicotianae, but the targets in Agave were still unknown. This study aims to obtain the host plant proteins that are interacting with PnRXLR5863 in A. hybrid No.11648. A normalized cDNA library was constructed by combining Switching Mechanism at 5′ end of RNA Transcript (SMART) and Duplex-Specific Nuclease (DSN) technology. The capacity of cDNA library was 5.32×10⁷ CFU/ml, the independent clones was 1.064×10⁸ CFU, and the lengths of inserts ranged from 0.5 kb to 3.0 kb, with the recombination rate of 100%. A total of 23 proteins interacting with PnRXLR5863 were obtained by sequencing and homology analysis. Some of target proteins involve several physiological events, such as ubiquitination, protein synthesis, protein translation and protein translation, suggesting that PnRXLR5863 effector may act on protein metabolism of Agave spp. This research would lay a foundation for further study of the pathogenic mechanism of PnRXLR5863.

, correspAuthors=Wenzhao ZHOU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, 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=Ziping YANG, Qian YANG, Xun WANG, Zhi KE, Zhiwei LU, Yanmei ZHANG, Huibang SHEN, Wenzhao ZHOU), CN=ArticleExt(id=1236318326251647857, articleId=1236318325786080107, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=剑麻均一化文库构建及PnRXLR5863靶蛋白筛选, columnId=1236256430517440904, journalTitle=热带作物学报, columnName=组学与生物技术, runingTitle=null, highlight=null, articleAbstract=

剑麻是硬质纤维的主要来源，由烟草疫霉（Phytophthora nicotianae）引起的斑马纹病是剑麻生产的重要病害。RXLR效应蛋白是卵菌第一大类效应蛋白，能通过直接的蛋白质相互作用，靶向寄主防卫相关蛋白，干扰和抑制寄主防卫。利用转录组分析不同侵染阶段的烟草疫霉，发现PnRXLR5863表达量较高，并且其编码的蛋白含有信号肽、RXLR-EER和WY结构域，瞬时表达发现PnRXLR5863能引起烟草细胞死亡，推测PnRXLR5863在烟草疫霉侵染剑麻过程发挥重要作用，但是它作用于剑麻的靶蛋白和致病机制还未知。为了筛选PnRXLR5863的剑麻靶蛋白，应用Switching Mechanism at 5′ end of RNA Transcript（SMART）和Duplex-Specific Nuclease（DSN）技术构建了剑麻均一化酵母cDNA表达文库，并筛选了与PnRXLR5863相互作用的蛋白。结果显示，构建的文库库容为5.32×10⁷ CFU/mL，文库总克隆数为1.064×10⁸ CFU，文库片段平均大小为1000 bp，重组率为100%；构建的pGBKT7-PnRXLR5863诱饵载体无自激活性；利用酵母双杂交系统，以pGBKT7-PnRXLR5863诱饵载体，从构建的文库中筛选获得23个与PnRXLR5863相互作用的蛋白。获得的候选靶蛋白主要参与分子功能和生物过程，涉及泛素化过程、蛋白合成、蛋白翻译和蛋白修饰等生理事件，暗示PnRXLR5863效应蛋白可能作用于剑麻的蛋白代谢。剑麻均一化酵母表达文库的构建和PnRXLR5863靶蛋白的筛选，为进一步深入研究PnRXLR5863的致病机理奠定基础。

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杨子平（1984—），男，博士，助理研究员，研究方向：植物分子生物学。

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杨子平（1984—），男，博士，助理研究员，研究方向：植物分子生物学。

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