Article(id=1236318334518612276, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1236318325282763627, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2023.02.001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1650729600000, receivedDateStr=2022-04-24, revisedDate=1654531200000, revisedDateStr=2022-06-07, acceptedDate=null, acceptedDateStr=null, onlineDate=1772690860632, onlineDateStr=2026-03-05, pubDate=1677254400000, pubDateStr=2023-02-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772690860632, onlineIssueDateStr=2026-03-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772690860632, creator=13701087609, updateTime=1772690860632, 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=225, endPage=232, ext={EN=ArticleExt(id=1236318334787047756, articleId=1236318334518612276, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Establishment of PARMS Reaction System for Pineapple (Ananas comosus. L), columnId=1236256430337085821, journalTitle=Chinese Journal of Tropical Crops, columnName=Omics & Biotechnology, runingTitle=null, highlight=null, articleAbstract=

Single nucleotide polymorphism (SNP) is widely distributed in plant genomes, which is one of the most abundant form of DNA variation. The molecular markers based on single nucleotide polymorphism are considered to be of great application prospect. Penta-primer amplification re-fractory mutation system (PARMS) is a kind of new genotyping system based on single nucleotide polymorphism (SNP) which has the advantages of high throughput, high accuracy, low cost and short time consuming. The establishment of the PARMS system of pineapple is significant in germplasm identification, gene mapping and marker-assisted selection of pineapple. In this study, three germplasms of significant difference in phenotypes were used as the materials. A specific primer SNP31 was designed based on the resequencing data of 130 germplasm resources. The result revealed SNP31 could effectively group the pineapple germplasm resources, and be used for the subsequent optimization of the system. Reaction volume,primer concentration, method of DNA extracting and template DNA amount were optimized. The results showed that reaction volume, primer concentration and template DNA amount could affect the fluorescence signal value of the genotype signal point. The fluorescence signal value of genotypic signal point decreased when the reaction volume was larger or smaller. The optimal reaction volume was 6 μL. When the concentration of primer and template DNA increased, the fluorescence signal value of genotype signal point increased. The optimal concentration of primer and template DNA was 100 μmol/L and 25 ng/μL, respectively. In addition, different methods of genomic DNA extraction could group well the PARMS-SNP for the three germplasms of pineapple. The optimal PARMS reaction system was as follows:total volume 6 μL, containing 1 μL template DNA (25 ng), 3 μL PARMS mix (2×), 0.45 μL primer mix (100 μmol/L) and 1.55 μL ddH₂O. With the optimal PARMS reaction system, high quality results of PARMS-SNP genotyping was produced on sixty-five pineapple germplasm resources, which indicated that the reaction system was accurate and stable. The establishment of the optimized PARMS genotyping system could provide a basis for the genetic diversity analysis, genetic linkage map construction, gene mapping and marker-assisted selection of pineapple in this study.

, correspAuthors=Wenqiu LIN, 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=Yunfei GAO, Wenqiu LIN, Qingsong WU, Xiumei ZHANG, Weisheng SUN, Shenghui LIU, Yanli YAO), CN=ArticleExt(id=1236318335034511709, articleId=1236318334518612276, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=菠萝PARMS反应体系的建立, columnId=1236256430517440904, journalTitle=热带作物学报, columnName=组学与生物技术, runingTitle=null, highlight=null, articleAbstract=

单核苷酸多态性（single nucleotide polymorphism, SNP）广泛分布在植物的基因组中，具有丰富的DNA变异形式。基于单核苷酸多态性开发的标记被认为是极具应用前景的分子标记。五引物扩增受阻突变体系（Penta-primer amplification re-fractory mutation system, PARMS）是一种基于单核苷酸多态性的新型基因分型技术，具有通量高、准确性高、成本低和耗时短等优点。因此，建立高效、简便可行的PARMS分型技术对菠萝种质资源的鉴定、基因定位的开展以及分子辅助选择育种体系的建立具有重要意义。本研究以3份表型差异显著的菠萝种质资源为材料，基于130份菠萝种质资源的重测序数据分析结果，设计特异引物SNP31。结果显示，SNP31可将菠萝种质资源进行较好地分型，可用于后续反应体系的优化。为了建立适用于菠萝的PARMS-SNP分型体系，对PARMS的反应体积、引物浓度、DNA的提取方法和DNA浓度等参数进行优化。研究结果表明，反应体积、引物浓度以及DNA的浓度均能影响基因型信号点的荧光信号值。在较大或较小的反应体积下基因型信号点的荧光信号值均降低，最适的反应体积为6 μL。随着引物和DNA浓度的增加，基因型信号点的荧光信号值增加，最佳的引物和DNA浓度分别为100 μmol/L和25 ng/μL。此外，不同基因组DNA的提取方法均能对3份种质实现较好的PARMS-SNP分型。因此，PARMS-SNP分型体系的最佳反应体系为：反应总体积6 μL，DNA（25 ng）1 μL，PARMS mix 3 μL，Primer mix（100 μmol/L）0.45 μL，ddH₂O 1.55 μL。利用65份菠萝种质资源验证该体系的准确性和稳定性，获得了较好的PARMS-SNP分型结果。本研究建立的PARMS-SNP分型体系为开展菠萝种质资源遗传多样性分析、遗传图谱的构建、基因定位及分子标记辅助选择提供基础。

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^3.云南农业大学热带作物学院，云南普洱　665099, bio={"content":"

高云飞（1999—），男，本科生，研究方向：菠萝生物技术；

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高云飞（1999—），男，本科生，研究方向：菠萝生物技术；

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