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Article(id=1237016044326417249, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1237016039171608726, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2025.09.005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1746633600000, receivedDateStr=2025-05-08, revisedDate=null, revisedDateStr=null, acceptedDate=1748188800000, acceptedDateStr=2025-05-26, onlineDate=1772857207613, onlineDateStr=2026-03-07, pubDate=1758729600000, pubDateStr=2025-09-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772857207613, onlineIssueDateStr=2026-03-07, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772857207613, creator=13701087609, updateTime=1772857207613, updator=13701087609, issue=Issue{id=1237016039171608726, tenantId=1146029695717560320, journalId=1235980609244409860, year='2025', volume='46', issue='9', pageStart='2031', pageEnd='2286', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1772857206385, creator=13701087609, updateTime=1773049161445, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1237821157118890427, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1237016039171608726, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1237821157118890428, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1237016039171608726, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2076, endPage=2085, ext={EN=ArticleExt(id=1237016044607435619, articleId=1237016044326417249, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Development of hAT-MITE Transposon Insertion Polymorphism Molecular Markers and Cultivars Identification in Pitaya, columnId=1236256430060261740, journalTitle=Chinese Journal of Tropical Crops, columnName=Germplasm Resources, Genetics & Breeding, runingTitle=null, highlight=null, articleAbstract=

Miniature inverted-repeat transposable elements (MITEs), as active mobile elements in eukaryotic genomes, provide novel molecular markers for germplasm genetic diversity analysis through the insertion polymorphisms. This study systematically identified hAT family MITE transposons in pitaya using bioinformatics approaches based on the whole-genome data of the Guanhua Bai cultivar. A total of 2350 candidate elements with complete terminal inverted repeats (TIRs) and target site duplications (TSDs) were screened, with an average of 213 elements per chromosome pair. Through sequence specificity and polymorphism analysis, 110 highly divergent loci (10 per chromosome pair) were selected for primer design, ultimately yielding 41 stable polymorphic hAT-MITE markers after PCR validation. Genome-wide scanning of 48 pitaya germplasms using the 41 primer pairs detected 81 polymorphic loci (polymorphism rate: 97.59%) via agarose gel electrophoresis. UPGMA cluster analysis revealed substantial genetic variation and diversity among the 48 accessions, with genetic similarity coefficients ranging from 0.57 to 0.91. At a genetic similarity threshold of 0.63, the germplasm resources were categorized into four genetically distinct groups. A digital fingerprinting system covering all germplasm resources was constructed using four core primer pairs (HU-MIT-02/06/26/75) based on the primer-band pattern combination method, achieving 100% cultivar identification accuracy. This study established the first hAT-MITE molecular marker system for pitaya, providing an efficient toolkit including a marker library and standardized identification protocols for precise germplasm classification, cultivar identification and varieties property protection of pitaya. The advancements is of significant application value for intellectual property protection in the pitaya seed industry.

, correspAuthors=Hongli LI, Qiong LI, 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=Wenbin HU, Hao WANG, Xincheng ZHOU, Wenhui PU, Yanli FENG, Hongli LI, Qiong LI), CN=ArticleExt(id=1237016045718926198, articleId=1237016044326417249, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=火龙果hAT-MITE转座子插入多态性分子标记开发与品种鉴别, columnId=1236256430219645304, journalTitle=热带作物学报, columnName=种质资源与遗传育种, runingTitle=null, highlight=null, articleAbstract=

微型反向重复转座子（MITE）作为真核基因组中活跃的可移动元件，其插入多态性为种质遗传多样性分析提供了新型分子标记。本研究基于火龙果品种莞华白全基因组数据，通过生物信息学方法系统鉴定hAT家族MITE转座子，共筛选出2350个具有完整末端反向重复（TIR）和靶位点重复（TSD）特征的候选元件，平均每对染色体213个。基于序列特异性及多态性分析，从中选出110个（每对染色体10个）高差异位点设计引物，经PCR扩增验证后成功开发出41个稳定多态性hAT-MITE标记。进一步利用41对引物对48份火龙果种质进行全基因组扫描，通过琼脂糖凝胶电泳共检测到81个多态性位点（多态率为97.59%）；UPGMA聚类分析表明，48份材料存在着丰富的变异和遗传多样性（遗传相似系数在0.57~0.91之间），在遗传相似系数为0.63时，将品种资源划分为4个遗传结构显著差异的类群。基于引物-带型组合法，精选4对核心引物（HU-MIT-02/06/26/75）构建覆盖全部种质资源的数字化指纹图谱，其品种鉴别准确率达100%。本研究首次建立火龙果hAT-MITE分子标记技术体系，所开发的标记库及标准化鉴定流程为火龙果种质资源精准分类、品种鉴定和品种权保护提供了高效工具，对火龙果种业知识产权保护具有重要应用价值。

, correspAuthors=李洪立, 李琼, authorNote=null, correspAuthorsNote=
* 李洪立（LI Hongli），E-mail：。
李琼（LI Qiong），E-mail：。
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胡文斌（1986—），男，硕士，副研究员，研究方向：种质资源学。

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3.Key Laboratory of Crop Gene Resources and Germplasm Innovation in South China, Ministry of Agriculture and Rural Affairs, Haikou, Hainan 571101, China
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3.农业农村部华南作物基因资源与种质创制重点实验室，海南海口　571101
4.海南省热带作物资源遗传改良与创新重点实验室，海南海口　571101, bio={"content":"

胡文斌（1986—），男，硕士，副研究员，研究方向：种质资源学。

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胡文斌（1986—），男，硕士，副研究员，研究方向：种质资源学。

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3.Key Laboratory of Crop Gene Resources and Germplasm Innovation in South China, Ministry of Agriculture and Rural Affairs, Haikou, Hainan 571101, China
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3.农业农村部华南作物基因资源与种质创制重点实验室，海南海口　571101
4.海南省热带作物资源遗传改良与创新重点实验室，海南海口　571101, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1237023447398806511, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, xref=1., ext=[AuthorCompanyExt(id=1237023447407195120, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, companyId=1237023447398806511, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China), AuthorCompanyExt(id=1237023447415583729, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, companyId=1237023447398806511, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.中国热带农业科学院热带作物品种资源研究所，海南海口　571101)]), 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3.Key Laboratory of Crop Gene Resources and Germplasm Innovation in South China, Ministry of Agriculture and Rural Affairs, Haikou, Hainan 571101, China
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3.农业农村部华南作物基因资源与种质创制重点实验室，海南海口　571101
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(in Chinese), articleTitle=Identification and analysis of repeat sequences in the genome of cactaceae family plants, refAbstract=null), Reference(id=1237023461227426399, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, doi=null, pmid=null, pmcid=null, year=2021, volume=8, issue=null, pageStart=164, pageEnd=null, url=null, language=null, rfNumber=[21], rfOrder=32, authorNames=CHEN J Y, XIE F F, CUI Y Z, CHEN C B, LU W J, HU X D, HUA Q Z, ZHAO J, WU Z J, GAO D, ZHANG Z K, JIANG W K, SUN Q M, HU G B, QIN Y H, journalName=Horticulture Research, refType=null, unstructuredReference=CHEN J Y, XIE F F, CUI Y Z, CHEN C B, LU W J, HU X D, HUA Q Z, ZHAO J, WU Z J, GAO D, ZHANG Z K, JIANG W K, SUN Q M, HU G B, QIN Y H. A chromo-some-scale genome sequence of pitaya (Hylocereus undatus) provides novel insights into the genome evolution and regulation of betalain biosynthesis[J]. 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(in Chinese), articleTitle=Accurate identification of varieties by nucleotide polymorphisms and establishment of scannable variety IDs for Soybean germplasm, refAbstract=null), Reference(id=1237023461705577093, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, doi=null, pmid=null, pmcid=null, year=2017, volume=15, issue=2, pageStart=640, pageEnd=647, url=null, language=null, rfNumber=[24], rfOrder=37, authorNames=熊发前, 刘俊仙, 贺梁琼, 韩柱强, 黄志鹏, 唐秀梅, 蒋菁, 钟瑞春, 吴海宁, 李忠, 罗赛云, 唐荣华, 何新华, journalName=分子植物育种, refType=null, unstructuredReference=熊发前, 刘俊仙, 贺梁琼, 韩柱强, 黄志鹏, 唐秀梅, 蒋菁, 钟瑞春, 吴海宁, 李忠, 罗赛云, 唐荣华, 何新华. 花生LTR和MITE转座子及其分子标记开发利用研究进展[J]. 分子植物育种, 2017, 15(2): 640-647., articleTitle=花生LTR和MITE转座子及其分子标记开发利用研究进展, refAbstract=null), Reference(id=1237023461789463181, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, doi=null, pmid=null, pmcid=null, year=2017, volume=15, issue=2, pageStart=640, pageEnd=647, url=null, language=null, rfNumber=[24], rfOrder=38, authorNames=XIONG F Q, LIU J X, HE L Q, HAN Z Q, HUANG Z P, TANG X M, JIANG J, ZHONG R C, WU H N, LI Z, LUO S Y, TANG R H, HE X H, journalName=Molecular Plant Breeding, refType=null, unstructuredReference=XIONG F Q, LIU J X, HE L Q, HAN Z Q, HUANG Z P, TANG X M, JIANG J, ZHONG R C, WU H N, LI Z, LUO S Y, TANG R H, HE X H. Recent advances on the development and utilization of molecular markers based on LTR Retrotransposons and MITE transposons from peanut (Arachis hypogaea L.)[J]. Molecular Plant Breeding, 2017, 15(2): 640-647. 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articleId=1237016044326417249, language=EN, label=Fig. 2, caption=Cluster dendrogram of tested pitaya germplasm resources based on alleles amplified by 41 pair of core primers, figureFileSmall=w57SVYTHORODQJv4rJYtow==, figureFileBig=2biRQ3YUr/mFtOthZ/PyBg==, tableContent=null), ArticleFig(id=1237023452910121112, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=CN, label=图2, caption=基于41对核心引物的火龙果品种资源聚类分析, figureFileSmall=w57SVYTHORODQJv4rJYtow==, figureFileBig=2biRQ3YUr/mFtOthZ/PyBg==, tableContent=null), ArticleFig(id=1237023453002395802, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=EN, label=Tab. 1, caption=

Names and origins of 48 pitaya germplasm resources

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编号No.名称Name来源Origin编号No.名称Name来源Origin编号No.名称Name来源Origin
H1大翼水晶中国广东H17粤红3号中国广东H33美白美国
H2桂红龙1号中国广西H18康尼梅椰美国H34富贵红中国台湾
H3蜜宝中国台湾H19越南白肉越南H35EL美国
H4紫龙中国海南H20莞华红中国广东H36无刺黄龙中国海南
H5桂族双色中国广西H21大红中国台湾H37红皮燕窝中国海南
H6紫蜜龙中国海南H22莞华白中国广东H38巴西白肉1巴西
H7柠檬中国云南H23双色1号中国广东H39黄麒麟-G哥伦比亚
H849-3中国云南H24莞华粉红中国广东H40白花双色中国海南
H9红花青龙中国海南H25BNM巴拿马H41金燕窝中国海南
H10双色锦中国海南H26大花蛇鞭柱中国海南H42双色球中国海南
H11白水晶中国广东H27BM中国海南H43红麒麟中国广西
H12无刺红龙中国海南H28巴西粉肉巴西H44丰景双色中国海南
H13红水晶中国广东H29黄锦中国海南H45黄麒麟中国广西
H14红冠2号中国广东H30秘鲁燕窝秘鲁H46S2中国海南
H15粤红中国广东H31糖龙美国H47沙漠国王以色列
H16翠西亚美国H32红宝龙中国广西H48S3中国海南
), ArticleFig(id=1237023453082087583, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=CN, label=表1, caption=

48份火龙果种质的名称和来源

, figureFileSmall=null, figureFileBig=null, tableContent=
编号No.名称Name来源Origin编号No.名称Name来源Origin编号No.名称Name来源Origin
H1大翼水晶中国广东H17粤红3号中国广东H33美白美国
H2桂红龙1号中国广西H18康尼梅椰美国H34富贵红中国台湾
H3蜜宝中国台湾H19越南白肉越南H35EL美国
H4紫龙中国海南H20莞华红中国广东H36无刺黄龙中国海南
H5桂族双色中国广西H21大红中国台湾H37红皮燕窝中国海南
H6紫蜜龙中国海南H22莞华白中国广东H38巴西白肉1巴西
H7柠檬中国云南H23双色1号中国广东H39黄麒麟-G哥伦比亚
H849-3中国云南H24莞华粉红中国广东H40白花双色中国海南
H9红花青龙中国海南H25BNM巴拿马H41金燕窝中国海南
H10双色锦中国海南H26大花蛇鞭柱中国海南H42双色球中国海南
H11白水晶中国广东H27BM中国海南H43红麒麟中国广西
H12无刺红龙中国海南H28巴西粉肉巴西H44丰景双色中国海南
H13红水晶中国广东H29黄锦中国海南H45黄麒麟中国广西
H14红冠2号中国广东H30秘鲁燕窝秘鲁H46S2中国海南
H15粤红中国广东H31糖龙美国H47沙漠国王以色列
H16翠西亚美国H32红宝龙中国广西H48S3中国海南
), ArticleFig(id=1237023453203722403, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=EN, label=Tab. 2, caption=

Distribution of hAT-MITE elements in genic regions, intergenic regions, and promoter regions of pitaya genome

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区域
Region		拷贝数
Copies		长度
Length/bp		平均拷贝数
Average copies
基因区225154 187 0151.459
启动子区24355 470 0004.381
基因间隔区18821 232 580 6321.527
), ArticleFig(id=1237023453308580007, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=CN, label=表2, caption=

火龙果基因组hAT-MITE在基因区、间隔区和启动子区的分布

, figureFileSmall=null, figureFileBig=null, tableContent=
区域
Region		拷贝数
Copies		长度
Length/bp		平均拷贝数
Average copies
基因区225154 187 0151.459
启动子区24355 470 0004.381
基因间隔区18821 232 580 6321.527
), ArticleFig(id=1237023453442797741, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=EN, label=Tab. 3, caption=

Chromosomal distribution of hAT-MITE elements in genic regions, intergenic regions, and promoter regions of pitaya genome

, figureFileSmall=null, figureFileBig=null, tableContent=
类型
Types		启动子
Promoter		外显子
Exon		内含子
Intron		基因间隔区
Intergenic region		总数
Total
chr0113115210239
chr0221117165204
chr0328115175219
chr0415118194228
chr0524018191233
chr0623527134189
chr0722219168211
chr0819535161220
chr0922017152191
chr1029113135178
chr1126113188228
scaffold858100910
), ArticleFig(id=1237023453618958515, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=CN, label=表3, caption=

火龙果基因组hAT-MITE在不同染色体基因区、基因间隔区和启动子区的分布

, figureFileSmall=null, figureFileBig=null, tableContent=
类型
Types		启动子
Promoter		外显子
Exon		内含子
Intron		基因间隔区
Intergenic region		总数
Total
chr0113115210239
chr0221117165204
chr0328115175219
chr0415118194228
chr0524018191233
chr0623527134189
chr0722219168211
chr0819535161220
chr0922017152191
chr1029113135178
chr1126113188228
scaffold858100910
), ArticleFig(id=1237023453711233209, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=EN, label=Tab. 4, caption=

Information of hAT-MITE primers

, figureFileSmall=null, figureFileBig=null, tableContent=
引物名称
Primer name		正向引物（5′–3′）
Forward primer（5′–3′）		反向引物（5′–3′）
Reverse primer（5′–3′）		目标片段长度
Target fragment length/bp
HU-MIT1CCTCCAATTTCTGACGCCCTACTCGTGGAGGAGACAGGAA740
HU-MIT2TTGACCTGGTCTTTTTGGCCTGGGAAGTCACACGGAGTTG622
HU-MIT6ACCGTGCAAGGTAGTTCCAGGTTGTGGGCCTTTCGTCAAC700
HU-MIT10GGTCCACATGCAAAAACATCCAAGAGTGACAGTTACAGCGGC964
HU-MIT12CGGGAGTTGATCGGAGGATGCGTAATACACCTAGGCTGGCA762
HU-MIT16ATTCTTCGCCCAAGCCTACCACCGAATCACACCCAATCCA1098
HU-MIT17TCCTGATCCTCCCTGCGTTAGAGAGAGAGAGATGCACCGC885
HU-MIT18GGCAAGGCACTGACATGTTCTGCACTGAAGACAGCCTCTG971
HU-MIT22ATGCTTGTCGGCCCTCTATGTATGTTCGGGCTCGGCATTT640
HU-MIT26CACCGACCTGAATCCAAGCTCGGGATCAGCAAACCAGACT761
HU-MIT27GGTTTGGTGGAAATTGGGCCGTTACAGCAATGGAGCCCCT928
HU-MIT30GAATCTGGGGGTCGCTAAGGCCACGCAAACTGACTCGAAC635
HU-MIT33CAAACCGCCCCTATCCAGTTATACGTCTGCTGGTTGCCTC558
HU-MIT36AGCATCCCATTGTCGTCACATCAACTCAGTGGCAATCCCC975
HU-MIT38CCCACCGGAAAAGAACTCGAGCTTTTGGGGTTTCTGCAGG575
HU-MIT40TCTCCTGTCGGATCTGGGTTATCAGCCATGTTGCGTCTGA673
HU-MIT45TGAAGCACGGGATTCACATTAGTTTGCGTACCTTCACCGT869
HU-MIT46TGAGGGGGTTGTTGGAGGTAAACCCAAAACCCACCTACGG1004
HU-MIT48CGAAGTCAACCGGAACCAGAAACAAAACAACAGGCCAGCC772
HU-MIT50TGCTACAATCGAAGGGCGTTACTTGCACGCTGACTGAGAA771
HU-MIT52TGGAAGGAAGAGAGTGGGGTCACACCCCCATAGTCACGAC590
HU-MIT53CCAGTGCTCAGCTCATCCAAAATCCTCTCCGGCCACTAGT541
HU-MIT60TACTCTGCCCTTTTCTGCCGTCTCGGTCGTGGTCAGTTTG781
HU-MIT67CTGCACCAGTTCAACAAGGCAACCACTACGGAAGCATGGG606
HU-MIT68TGCACATGTTTGGGTTTGGCGCAGCAGCTTTCTTTGCAGT1008
HU-MIT70ACTCCAACAAGAATCAAGCCAATGGCGAGGAAGGACACAAG738
HU-MIT72TGCAGATTCGCACATTGAGGTTTCCGCCATCATCGTGGAA724
HU-MIT75AGATGCGCTTGTCATGCTCTACATGGTGGAAGTGAGCCAG997
HU-MIT76AGTGTGGCATTCCATCCCTGTGAGCAGGTTCTTACCCATCT632
HU-MIT77GCAAGACTCGAATCCAAGGCCACGTTGCTTTGGGATGCAA909
HU-MIT84AGGACTCTCATGCGGGTAGTACATGGAGGGAAGCTTGTCG672
HU-MIT86AAGAAGTTCGGCCCAACCAAGAGGCCCAAGTGCAAGGTAT829
HU-MIT88TCCTCGCAAAGCCTCACATTAAGACATGTCGGTGCTTCGT771
HU-MIT91GCACTGAAAGGAGGCAGGAACAGGAGCTGTTGGGGGAAAA1065
HU-MIT94CTGAAGCGTTCATGGGACCTAATGCGGGAGACAGAAAGGG1197
HU-MIT97AGTCACACCCACCACTTTACAAGCGCGCATCTACCATACTG938
HU-MIT99CATGTAGCTCCGCCCATGATATTCGCACTTAGCCAAGCCT679
HU-MIT107TCCATGTGTGTGCCCCTTTTCACCACCCCACAAATGCATG638
HU-MIT108GGGTGTAGTGACTTGCTCCCACATCCACCAGAGGATTGGC968
HU-MIT109AGGTGCAAGTTGACGTTGGTATCAAGCCTTTGGACCTGGG604
HU-MIT110ACAGAGCAAGCCGACGTTAATTCCTTGCTACTGCTGCTCC613
), ArticleFig(id=1237023453858033858, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=CN, label=表4, caption=

hAT-MITE引物信息

, figureFileSmall=null, figureFileBig=null, tableContent=
引物名称
Primer name		正向引物（5′–3′）
Forward primer（5′–3′）		反向引物（5′–3′）
Reverse primer（5′–3′）		目标片段长度
Target fragment length/bp
HU-MIT1CCTCCAATTTCTGACGCCCTACTCGTGGAGGAGACAGGAA740
HU-MIT2TTGACCTGGTCTTTTTGGCCTGGGAAGTCACACGGAGTTG622
HU-MIT6ACCGTGCAAGGTAGTTCCAGGTTGTGGGCCTTTCGTCAAC700
HU-MIT10GGTCCACATGCAAAAACATCCAAGAGTGACAGTTACAGCGGC964
HU-MIT12CGGGAGTTGATCGGAGGATGCGTAATACACCTAGGCTGGCA762
HU-MIT16ATTCTTCGCCCAAGCCTACCACCGAATCACACCCAATCCA1098
HU-MIT17TCCTGATCCTCCCTGCGTTAGAGAGAGAGAGATGCACCGC885
HU-MIT18GGCAAGGCACTGACATGTTCTGCACTGAAGACAGCCTCTG971
HU-MIT22ATGCTTGTCGGCCCTCTATGTATGTTCGGGCTCGGCATTT640
HU-MIT26CACCGACCTGAATCCAAGCTCGGGATCAGCAAACCAGACT761
HU-MIT27GGTTTGGTGGAAATTGGGCCGTTACAGCAATGGAGCCCCT928
HU-MIT30GAATCTGGGGGTCGCTAAGGCCACGCAAACTGACTCGAAC635
HU-MIT33CAAACCGCCCCTATCCAGTTATACGTCTGCTGGTTGCCTC558
HU-MIT36AGCATCCCATTGTCGTCACATCAACTCAGTGGCAATCCCC975
HU-MIT38CCCACCGGAAAAGAACTCGAGCTTTTGGGGTTTCTGCAGG575
HU-MIT40TCTCCTGTCGGATCTGGGTTATCAGCCATGTTGCGTCTGA673
HU-MIT45TGAAGCACGGGATTCACATTAGTTTGCGTACCTTCACCGT869
HU-MIT46TGAGGGGGTTGTTGGAGGTAAACCCAAAACCCACCTACGG1004
HU-MIT48CGAAGTCAACCGGAACCAGAAACAAAACAACAGGCCAGCC772
HU-MIT50TGCTACAATCGAAGGGCGTTACTTGCACGCTGACTGAGAA771
HU-MIT52TGGAAGGAAGAGAGTGGGGTCACACCCCCATAGTCACGAC590
HU-MIT53CCAGTGCTCAGCTCATCCAAAATCCTCTCCGGCCACTAGT541
HU-MIT60TACTCTGCCCTTTTCTGCCGTCTCGGTCGTGGTCAGTTTG781
HU-MIT67CTGCACCAGTTCAACAAGGCAACCACTACGGAAGCATGGG606
HU-MIT68TGCACATGTTTGGGTTTGGCGCAGCAGCTTTCTTTGCAGT1008
HU-MIT70ACTCCAACAAGAATCAAGCCAATGGCGAGGAAGGACACAAG738
HU-MIT72TGCAGATTCGCACATTGAGGTTTCCGCCATCATCGTGGAA724
HU-MIT75AGATGCGCTTGTCATGCTCTACATGGTGGAAGTGAGCCAG997
HU-MIT76AGTGTGGCATTCCATCCCTGTGAGCAGGTTCTTACCCATCT632
HU-MIT77GCAAGACTCGAATCCAAGGCCACGTTGCTTTGGGATGCAA909
HU-MIT84AGGACTCTCATGCGGGTAGTACATGGAGGGAAGCTTGTCG672
HU-MIT86AAGAAGTTCGGCCCAACCAAGAGGCCCAAGTGCAAGGTAT829
HU-MIT88TCCTCGCAAAGCCTCACATTAAGACATGTCGGTGCTTCGT771
HU-MIT91GCACTGAAAGGAGGCAGGAACAGGAGCTGTTGGGGGAAAA1065
HU-MIT94CTGAAGCGTTCATGGGACCTAATGCGGGAGACAGAAAGGG1197
HU-MIT97AGTCACACCCACCACTTTACAAGCGCGCATCTACCATACTG938
HU-MIT99CATGTAGCTCCGCCCATGATATTCGCACTTAGCCAAGCCT679
HU-MIT107TCCATGTGTGTGCCCCTTTTCACCACCCCACAAATGCATG638
HU-MIT108GGGTGTAGTGACTTGCTCCCACATCCACCAGAGGATTGGC968
HU-MIT109AGGTGCAAGTTGACGTTGGTATCAAGCCTTTGGACCTGGG604
HU-MIT110ACAGAGCAAGCCGACGTTAATTCCTTGCTACTGCTGCTCC613
), ArticleFig(id=1237023453950308549, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=EN, label=Tab. 5, caption=

Fingerprint of 48 pitaya germplasm resources

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编号No.HU-MIT2HU-MIT6HU-MIT26HU-MIT75编号No.HU-MIT2HU-MIT6HU-MIT26HU-MIT75
H10110110111100111H250100010000000111
H20010010011100000H260010110101011101
H30110111110000111H270100110011000110
H40110111111100101H280010011000110111
H50100110011010111H290110110010100011
H60111111111110111H300110010000111011
H71000011001010011H310010010001001010
H80110010011100111H320001110011010111
H90011010011010111H330110010110000000
H100110110110100011H340111010011010111
H110111010010100100H350010010000100011
H120100110111110111H360110110011000111
H130100110000100100H370110001000101011
H140110110011000000H380000010010000111
H150111110011110111H391111110000100111
H160110010000000000H400111110011010111
H170010110010000110H410110010000100000
H180110110100011111H420000010111000010
H190110000011100000H430110000000110111
H200010010011100110H440111111001010111
H210100000010100000H450010010100111001
H220110010010000111H460110010001000111
H230110010110100111H470010011000111011
H240010110001010111H480110010001111110
), ArticleFig(id=1237023454042583244, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016044326417249, language=CN, label=表5, caption=

48个火龙果品种资源的指纹图谱

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编号No.HU-MIT2HU-MIT6HU-MIT26HU-MIT75编号No.HU-MIT2HU-MIT6HU-MIT26HU-MIT75
H10110110111100111H250100010000000111
H20010010011100000H260010110101011101
H30110111110000111H270100110011000110
H40110111111100101H280010011000110111
H50100110011010111H290110110010100011
H60111111111110111H300110010000111011
H71000011001010011H310010010001001010
H80110010011100111H320001110011010111
H90011010011010111H330110010110000000
H100110110110100011H340111010011010111
H110111010010100100H350010010000100011
H120100110111110111H360110110011000111
H130100110000100100H370110001000101011
H140110110011000000H380000010010000111
H150111110011110111H391111110000100111
H160110010000000000H400111110011010111
H170010110010000110H410110010000100000
H180110110100011111H420000010111000010
H190110000011100000H430110000000110111
H200010010011100110H440111111001010111
H210100000010100000H450010010100111001
H220110010010000111H460110010001000111
H230110010110100111H470010011000111011
H240010110001010111H480110010001111110
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火龙果hAT-MITE转座子插入多态性分子标记开发与品种鉴别
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胡文斌 1, 3, 4 , 王好 1, 3, 4 , 周新城 2 , 濮文辉 1, 3, 4 , 冯艳丽 1, 3, 4 , 李洪立 1, 3, 4, * , 李琼 1, 3, 4, *
热带作物学报 | 种质资源与遗传育种 2025,46(9): 2076-2085
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热带作物学报 | 种质资源与遗传育种 2025, 46(9): 2076-2085
火龙果hAT-MITE转座子插入多态性分子标记开发与品种鉴别
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胡文斌1, 3, 4, 王好1, 3, 4, 周新城2, 濮文辉1, 3, 4, 冯艳丽1, 3, 4, 李洪立1, 3, 4, * , 李琼1, 3, 4, *
作者信息
  • 1.中国热带农业科学院热带作物品种资源研究所，海南海口　571101
  • 2.中国热带农业科学院热带生物技术研究所，海南海口　571101
  • 3.农业农村部华南作物基因资源与种质创制重点实验室，海南海口　571101
  • 4.海南省热带作物资源遗传改良与创新重点实验室，海南海口　571101

    • 胡文斌（1986—），男，硕士，副研究员，研究方向：种质资源学。

通讯作者:

* 李洪立（LI Hongli），E-mail：。
李琼（LI Qiong），E-mail：。
Development of hAT-MITE Transposon Insertion Polymorphism Molecular Markers and Cultivars Identification in Pitaya
Wenbin HU1, 3, 4, Hao WANG1, 3, 4, Xincheng ZHOU2, Wenhui PU1, 3, 4, Yanli FENG1, 3, 4, Hongli LI1, 3, 4, * , Qiong LI1, 3, 4, *
Affiliations
  • 1.Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
  • 2.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
  • 3.Key Laboratory of Crop Gene Resources and Germplasm Innovation in South China, Ministry of Agriculture and Rural Affairs, Haikou, Hainan 571101, China
  • 4.Key Laboratory of Genetic Improvement and Innovation of Tropical Crops, Haikou, Hainan 571101, China
出版时间: 2025-09-25 doi: 10.3969/j.issn.1000-2561.2025.09.005
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摘要
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微型反向重复转座子（MITE）作为真核基因组中活跃的可移动元件，其插入多态性为种质遗传多样性分析提供了新型分子标记。本研究基于火龙果品种莞华白全基因组数据，通过生物信息学方法系统鉴定hAT家族MITE转座子，共筛选出2350个具有完整末端反向重复（TIR）和靶位点重复（TSD）特征的候选元件，平均每对染色体213个。基于序列特异性及多态性分析，从中选出110个（每对染色体10个）高差异位点设计引物，经PCR扩增验证后成功开发出41个稳定多态性hAT-MITE标记。进一步利用41对引物对48份火龙果种质进行全基因组扫描，通过琼脂糖凝胶电泳共检测到81个多态性位点（多态率为97.59%）；UPGMA聚类分析表明，48份材料存在着丰富的变异和遗传多样性（遗传相似系数在0.57~0.91之间），在遗传相似系数为0.63时，将品种资源划分为4个遗传结构显著差异的类群。基于引物-带型组合法，精选4对核心引物（HU-MIT-02/06/26/75）构建覆盖全部种质资源的数字化指纹图谱，其品种鉴别准确率达100%。本研究首次建立火龙果hAT-MITE分子标记技术体系，所开发的标记库及标准化鉴定流程为火龙果种质资源精准分类、品种鉴定和品种权保护提供了高效工具，对火龙果种业知识产权保护具有重要应用价值。

火龙果  /  hAT-MITE转座子  /  指纹图谱  /  品种鉴定  /  遗传多样性

Miniature inverted-repeat transposable elements (MITEs), as active mobile elements in eukaryotic genomes, provide novel molecular markers for germplasm genetic diversity analysis through the insertion polymorphisms. This study systematically identified hAT family MITE transposons in pitaya using bioinformatics approaches based on the whole-genome data of the Guanhua Bai cultivar. A total of 2350 candidate elements with complete terminal inverted repeats (TIRs) and target site duplications (TSDs) were screened, with an average of 213 elements per chromosome pair. Through sequence specificity and polymorphism analysis, 110 highly divergent loci (10 per chromosome pair) were selected for primer design, ultimately yielding 41 stable polymorphic hAT-MITE markers after PCR validation. Genome-wide scanning of 48 pitaya germplasms using the 41 primer pairs detected 81 polymorphic loci (polymorphism rate: 97.59%) via agarose gel electrophoresis. UPGMA cluster analysis revealed substantial genetic variation and diversity among the 48 accessions, with genetic similarity coefficients ranging from 0.57 to 0.91. At a genetic similarity threshold of 0.63, the germplasm resources were categorized into four genetically distinct groups. A digital fingerprinting system covering all germplasm resources was constructed using four core primer pairs (HU-MIT-02/06/26/75) based on the primer-band pattern combination method, achieving 100% cultivar identification accuracy. This study established the first hAT-MITE molecular marker system for pitaya, providing an efficient toolkit including a marker library and standardized identification protocols for precise germplasm classification, cultivar identification and varieties property protection of pitaya. The advancements is of significant application value for intellectual property protection in the pitaya seed industry.

pitaya  /  hAT-MITE transposon  /  fingerprint  /  variety identification  /  genetic diversity
胡文斌, 王好, 周新城, 濮文辉, 冯艳丽, 李洪立, 李琼. 火龙果hAT-MITE转座子插入多态性分子标记开发与品种鉴别. 热带作物学报, 2025 , 46 (9) : 2076 -2085 . DOI: 10.3969/j.issn.1000-2561.2025.09.005
Wenbin HU, Hao WANG, Xincheng ZHOU, Wenhui PU, Yanli FENG, Hongli LI, Qiong LI. Development of hAT-MITE Transposon Insertion Polymorphism Molecular Markers and Cultivars Identification in Pitaya[J]. Chinese Journal of Tropical Crops, 2025 , 46 (9) : 2076 -2085 . DOI: 10.3969/j.issn.1000-2561.2025.09.005
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火龙果是新兴的重要的热带果树，在中国、越南、哥伦比亚、厄瓜多尔、澳大利亚等国家有大规模种植。目前中国是全球最大的火龙果生产国，种植面积约6.8万hm2，主要分布在广东、广西、海南、贵州、云南、福建等省（区），在我国脱贫攻坚、农业产业结构调整中发挥重要作用。火龙果在长期的自然演化、突变和人工选育过程中形成了丰富的种质资源，但种质资源缺乏系统研究，存在遗传基础不清问题。火龙果现有主栽品种外形特征极为相似，传统形态学分类手段难以有效鉴定，存在品种混淆问题。利用分子标记技术可为火龙果种质资源准确鉴定提供依据，而转座子标记是开展种质资源遗传多样性分析、品种鉴别的重要标记之一。
转座子的转座是植物基因组进化的重要动力，是植物遗传多样性的重要来源，在植物基因组进化、基因表达调控、系统发育和遗传多样性评价方面具有重要作用[1]。基于转座子的分子标记可以用于品种及物种鉴定、遗传多样性分析和功能标记。在植物中，数量较多的DNA类型转座子包括hAT（hobo，Activator and Tam3）、CACTA以及Mutator超家族，基因组上常见的MITE（miniature inverted-repeat transposon element，微型倒置重复转座元件）是属于非自主型DNA类型的转座子家族。一些MITE元件在基因组上可达几千到上万个拷贝[2]。同一作物不同品种间在相同位点常有MITE插入与缺失形成的多态性，即有的品种在某位点有MITE转座元件，有的品种则没有[3]。插入缺失标记（InDel）可以用于作物的指纹图谱[4]，MITE转座元件是否插入基因组上某位点，本质上也属于InDel标记，因此其也可以作为指纹图谱用于鉴别作物品种[5-6]。另外，在物种演化方面，MITE的这种多态性也可以用于种质资源遗传多样性分析，常被用于属内不同物种亲缘关系的研究[7-9]。与单核苷酸多态性（SNP）、简单序列重复（SSR）等分子标记一样，MITE等转座子在植物基因组中数量大、分布广，但是相比于SNP、SSR等标记，MITE产生的插入缺失标记多态性明确、易识别、开发方便、试验成本低且操作简单（琼脂糖电泳足以分辨），是种质资源遗传多样性分析、构建品种指纹图谱的良好标记之一。MITE标记在花生[10-11]、油菜[12]、水稻[13]等农作物品种鉴定、杂交种真伪鉴定上得到应用，目前ISSR[14-15]、SSR[16-18]等标记在火龙果种质资源遗传多样性分析、指纹图谱构建中已得到应用。陶金等[19]开发了基于反转录转座子的IRAP分子标记，用于火龙果遗传多样性分析，但尚未有基于MITE标记的分子身份证构建相关报道。
前人研究表明，在火龙果等仙人掌科植物基因组中，DNA转座子的占比均大幅高于反转录转座子，且绝大多数DNA转座子属于TIR类型，在TIR类型的DNA转座子中，hAT超家族在转座子代表性数据集中占比最大[20]。本研究从火龙果基因组中发掘出hAT-MITE转座子序列，基于转座子插入多态性开发适于火龙果品种的分子标记，并以48份火龙果主要商业品种为材料，开展遗传多样性分析和分子身份证构建研究，以期为火龙果种质分类、品种鉴定和品种权保护提供重要的理论依据与技术支撑。
1　材料与方法
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1.1　材料
供试的48份火龙果品种资源（表1）主要包括国内主要审认定品种、地方品种和特色资源。其中H5、H18、H31、H37、H40、H41、H45、H46作为核心引物筛选的材料。以上材料均保存于海南儋州中国热带农业科学院热带作物品种资源研究所省级火龙果种质资源圃。
1.2　方法
1.2.1　火龙果hAT类MITEs的鉴定
从火龙果基因组数据库（http://www.pitayagenomic.com/）[21]和NCBI（https://www.ncbi.nlm.nih.gov/bioproject/PRJNA691451）数据库中下载火龙果基因组序列和基因注释信息，利用MITE-Hunter等软件对火龙果基因组进行分析，主要是查找适合做分子标记的hAT家族MITE等DNA类型转座元件，对其类型、分布、偏好性进行分析。利用primer 3软件在位于基因间隔区、基因区和启动子区转座子外侧设计引物。
1.2.2　DNA提取
使用天根生化科技（北京）有限公司的多糖多酚植物基因组DNA提取试剂盒（DP360）提取48份火龙果品种资源嫩茎的总DNA，操作参照产品说明书进行。样品统一稀释至20 ng/μL后，–20 ℃保存备用。
1.2.3　核心引物筛选
每条染色体分别选10个以上的多态性位点，在其两端一定位置处分别设计PCR引物。如果因为序列不合适等原因无法设计引物，可选择一端引物位于转座子内部，另一端在转座子外。合成引物后，对8个代表性火龙果种质进行扩增，明确引物扩增片段大小、PCR退火温度等信息。每条染色体上至少获得2个在群体内呈现多态性的标记。
1.2.4　hAT-MITE程序及PCR产物检测
引物委托生工生物工程（上海）股份有限公司合成。MITE-PCR反应体系总体积为20 μL，含2×Taq PCR Master Mix 10.0 μL，F-primer（10 μmol/L）0.8 μL，R-primer（10 μmol/L）0.8 μL，模板DNA（20 ng/μL）3.0 μL，ddH2O 5.4 μL。扩增程序为：94 ℃预变性5 min；94 ℃变性30 s，复性30 s（根据引物Tm值设计退火温度），72 ℃延伸30 s，共35个循环；72 ℃延伸10 min。4 ℃保存。PCR扩增结束后用2%琼脂糖凝胶对PCR扩增产物进行电泳检测。
1.2.5　数据处理与遗传多样性分析
根据电泳结果，选择条带清晰、多态性高的进行统计，将同一位点的所有等位基因从小到大排列，有条带记为“1”，空白记为“0”，统计所得的数据利用Excel软件转换成NTSYS 2.10所需要的格式，利用软件NTSYSpc 2.10计算遗传相似系数，采用非加权组平均法（UPGMA），利用筛选出的hAT-MITE核心引物对48份火龙果种质资源进行聚类分析。
1.2.6　指纹图谱构建
从上述结果中选择鉴定力强的hAT-MITE引物构建火龙果品种资源的指纹图谱，以所分析的hAT-MITE引物名称为前缀，该标记在某样品上扩增带的分子量为后缀，得到每个品种在某个标记的带型编号，按照固定的引物排序结合不同引物分析结果，串联各带型编号，形成该品种的hAT-MITE指纹图谱。根据指纹图谱出现的概率公式P=1/2n（n为多态位点数，2n则为检测n个位点涉及的所有可能的试验材料个数），统计图谱的置信概率。
2　结果与分析
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2.1　 hAT-MITEs的鉴定与特征分析
利用MITE-Hunter软件对栽培种莞华白火龙果基因组进行分析，鉴定和筛选出hAT家族的MITE转座子，同时统计hAT-MITE元件在基因区、基因间隔区和启动子区的数量，并对其插入偏好性进行分析。根据火龙果基因组基因结构注释结果计算基因区的总长度（基因区包括内含子区、外显子区），启动子区为基因区上游2000 bp，基因组上其他区域则为非基因区，统计分析结果见表2。栽培种莞华白的基因组存在2350个完整结构的hAT-MITE转座子，其中243个位于基因启动子区，1882个位于基因间隔区，225个位于基因区，207个位于内含子区，18个位于外显子区和UTR区。基因区平均每兆碱基（平均拷贝数）有1.459个，略低于基因间隔区1.527个。而启动子区每兆碱基含4.381个转座元件，表明hAT-MITE转座子更倾向于分布在基因启动子区。
根据火龙果基因组序列信息和基因结构注释结果信息分析hAT-MITE元件在栽培种莞华白火龙果基因组各染色体上的数量，以及在每一条染色体基因区、基因间隔区和启动子区的数量，结果见表3。hAT-MITE在染色体上分布最多的是1号染色体，为239个；最少的是10号染色体，有178个；平均每对染色体有213个。大部分转座子分布在基因间区、启动子区，而基因区主要存在于内含子区，外显子区很少。
2.2　 hAT-MITE引物多态性的筛选
从每条染色体基因区、基因间隔区和启动子区选取代表性位点，每条染色体选10个多态性位点，共110个，设计引物。用形态、遗传背景差异较大的8份火龙果种质资源对110对hAT-MITE引物进行引物初步筛选。利用这110对hAT-MITE引物在8份火龙果种质资源中共检测出193个条带，平均每对引物扩增出1.7个，多态性条带数为121个，多态位点百分率达62.69%，条带大小在250~2000 bp之间。经2轮筛选后，选出41对具有明显多态性、条带清晰、带型稳定引物（表4）。
2.3　 hAT-MITE标记多态性引物在不同火龙果品种资源间的遗传多样性分析
利用筛选出来的41对火龙果hAT-MITE标记核心引物，对48份样品进行遗传多样性分析（部分检测结果见图1）。琼脂糖凝胶电泳结果显示41对hAT-MITE引物在48份火龙果种质资源中共检测出83个等位变异，多态性条带数为81个，多态位点百分率达97.59%，等位基因数变化范围为2~5。
根据41对核心引物扩增数据，利用NTSYS-pc 2.1软件，依据遗传相似系数采用UPGMA算法对48个火龙果品种资源进行聚类分析（图2）。结果表明，48份材料遗传相似系数在0.57~0.91之间，说明这些火龙果品种资源之间存在着丰富的变异和遗传多样性。在遗传相似系数为0.63时，48份火龙果材料总体上被分为4个类群，其中第Ⅰ类只包含了H39（黄麒麟-G）；第Ⅱ类包括H37（红皮燕窝）；第Ⅲ类H26（大花蛇鞭柱）、H48（S3）蛇鞭柱属野生资源；第Ⅳ类为常见的火龙果主栽品种和地方品种，共计44份，包括桂红龙1号、莞华粉红、莞华红、粤红、粤红3号、双色1号、红冠2号、紫龙等广东、广西、海南审认定品种，也包括大翼水晶、白水晶、红水晶、紫蜜龙、红花青龙、柠檬、黄锦、桂族双色、红宝龙等地方特色品种，还包括巴西粉肉、康尼梅椰、越南白肉、翠西亚、EL、秘鲁燕窝、BNM等来自越南、澳大利亚、巴西等国的栽培品种。这些品种的遗传相似系数基本在0.80以上，说明现有火龙果品种资源多样性较好，但是现有栽培品种差异较小，遗传背景较狭窄，其中H3（蜜宝）与H4（紫龙）之间的相似系数接近0.91，说明其亲缘关系较近。
2.4　指纹图谱构建
根据41对核心引物扩增结果可以将所有品种区分开，但从41对核心引物中选取最优的4对引物组合（HU-MIT2、HU-MIT6、HU-MIT26、HU-MIT75）可以将48份品种资源完全区分。HU-MIT2、HU-MIT6、HU-MIT26、HU-MIT75的多态性好，分别有3、5、3、5个等位变异，对应的基因型有6、12、8、16个，有很高的鉴别能力，可以将48份材料鉴别出来，可以作为指纹图谱构建的优选引物。将代表每个样品等位基因位点按顺序组成数字指纹，构建了48份火龙果品种资源的指纹图谱（表5）。这些引物的多态性位点有16个，根据概率公式1/2n可知，理论上在216（65536）份品种中才有可能存在2个品种的电泳图谱完全相同，置信概率几乎为100%，该指纹图谱可准确检测其中任何品种。
3　讨论
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前人研究表明，在火龙果等8个仙人掌科植物基因组TIR类型的转座子中，hAT超家族在转座子代表性数据集中占比最大，其比例为21.07%~ 31.86%[20]。因此hAT类MITE是分析火龙果遗传多样性和构建指纹图谱最适合的转座子标记。而本课题组前期开展火龙果栽培种David Bowie的基因组hAT类MITE转座子分析发现，2191个完整结构的hAT类MITE转座子，其中1714个位于基因间隔区，231个位于基因启动子区，基因区则为246个，平均每对染色体199个；而栽培种莞华白的基因组有2350个完整结构的hAT类MITE转座子，其中基因间隔区含有1882个，243个位于基因启动子区，基因区共含225个，平均每对染色体213个，高于栽培种David Bowie火龙果基因组。本研究中选择从莞华白火龙果的基因组上开发转座子标记，每条染色体分别选10个多态性位点，从110个位点中筛选出41个核心多态性位点，平均每条染色体3.7个，每个染色体至少有3个，既有多态性，又有代表性。
本研究鉴定开发的41个hAT-MITE标记，对48份样品进行遗传多样性分析结果中，共检测出83个等位变异，多态性条带数为81个，多态位点百分率达97.59%。等位基因数变化范围为2~5，平均每个位点等位基因数2.2个，多态性较好。能有效揭示48份火龙果种质资源的遗传多样性，也能很好地鉴别现有主要商业品种。聚类分析结果表明，除了其中第Ⅰ类只包含了H39（黄麒麟-G）；第Ⅱ类包括H37（红皮燕窝）；第Ⅲ类H26（大花蛇鞭柱）、H48（S3）蛇鞭柱属野生资源；其他的大部分材料遗传相似系数基本在0.80以上，说明现有品种资源遗传背景较狭窄，相似度较高，但也存在明显的多样性。本研究对hAT-MITE转座子在国内外火龙果品种资源中的多态性分布情况做出了初步的判断，为后期的火龙果种质资源收集、分类以及品种选育提供一定的理论参考依据。
目前品种鉴定主要使用SSR、SNP标记。前期本课题组利用SSR标记构建了现有主要商业品种的分子身份证，对完成现有主要商业品种分类、品种鉴定和品种权保护提供了技术支撑[16-17]。而利用SNP（单核苷酸序列多态性）标记进行品种鉴定在玉米[22]、大豆[23]等主要农作物均得到广泛利用。但是SSR标记法存在SSR位点差异小、检测方法成本高、容易出现错误基因型等问题；SNP标记法存在单个SNP位点鉴定能力有限、SNP位点多、检测成本高等。而MITE标记产生的插入缺失标记多态性明确、易识别、开发方便、试验成本低且操作简单利用，实验条件简单，能满足大部分研究人员开展种质资源遗传多样性分析、构建品种指纹图谱研究。有研究表明，MITE转座子分子标记揭示栽培种花生的DNA多态性要明显高于SSR分子标记，展示了其潜在的应用前景[24]。另外，MITE较高的转座活性和转座倾向于插入基因或基因附近区域的特性，常分布在基因的启动子区或内含子区，对基因的表达起到调控作用[1]，可以用于QTL和功能基因定位和功能标记开发。hAT超家族的MITE转座子非常适合开发为新一代的分子标记。
本课题组将结合MITE与SSR、SNP、MNP标记法开展火龙果种质资源鉴定与遗传多样性研究，建立品种鉴定技术规程，开发功能标记，更系统更准确地对火龙果种质资源鉴定评价，提高火龙果育种的效率，为火龙果分类、育种、品种鉴定、品种权保护提供重要的技术支撑和理论依据，加快培育具有自主知识产权的优异品种，推动我国火龙果产业健康可持续发展。
基金
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  • 海南省自然科学基金面上项目(323MS080)
  • 热带作物生物育种全国重点实验室创新团队项目(NKLTCBCXTD14)
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2025年第46卷第9期
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doi: 10.3969/j.issn.1000-2561.2025.09.005
  • 接收时间：2025-05-08
  • 首发时间：2026-03-07
  • 出版时间：2025-09-25
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  • 收稿日期：2025-05-08
  • 录用日期：2025-05-26
基金
海南省自然科学基金面上项目(323MS080)
热带作物生物育种全国重点实验室创新团队项目(NKLTCBCXTD14)
作者信息
    1.中国热带农业科学院热带作物品种资源研究所，海南海口　571101
    2.中国热带农业科学院热带生物技术研究所，海南海口　571101
    3.农业农村部华南作物基因资源与种质创制重点实验室，海南海口　571101
    4.海南省热带作物资源遗传改良与创新重点实验室，海南海口　571101

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* 李洪立（LI Hongli），E-mail：。
李琼（LI Qiong），E-mail：。
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
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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