Article(id=1236340103610692372, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1236340101991691008, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2023.03.004, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1653408000000, receivedDateStr=2022-05-25, revisedDate=1655136000000, revisedDateStr=2022-06-14, acceptedDate=null, acceptedDateStr=null, onlineDate=1772696050789, onlineDateStr=2026-03-05, pubDate=1679673600000, pubDateStr=2023-03-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772696050789, onlineIssueDateStr=2026-03-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772696050789, creator=13701087609, updateTime=1772696050789, updator=13701087609, issue=Issue{id=1236340101991691008, tenantId=1146029695717560320, journalId=1235980609244409860, year='2023', volume='44', issue='3', pageStart='447', pageEnd='660', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1772696050403, creator=13701087609, updateTime=1772696379070, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1236341480579715870, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1236340101991691008, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1236341480579715871, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1236340101991691008, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=476, endPage=483, ext={EN=ArticleExt(id=1236340103900099364, articleId=1236340103610692372, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Identification of the Di-Mon Mating Heterozygote in Lentinula edodes Using a New ISSR Technique, columnId=1236256430337085821, journalTitle=Chinese Journal of Tropical Crops, columnName=Omics & Biotechnology, runingTitle=null, highlight=null, articleAbstract=

Identification of heterozygote is a key step in the cross breeding of Lentinula edodes. A new ISSR technique (N-ISSR) was designed in this study based on discrimination between the two nucleus of the donor strain to identify quickly and accurately the authenticity of hybrid strains of L. edodes. The new ISSR was used to identify 40 hybrid strains which were from the Di-mon mating between dikaryon wild strain YX7 and spore monokaryons of cultivated strain 808, as well as microscopy observation combined with antagonistic test (MOCAT) and the traditional ISSR technique (T-ISSR) as the control. N-ISSR results showed 27 strains were confirmed to be true heterozygote as they possessed specific DNA bands from one of the nucleus of donor strain YX7, and the other 13 strains were confirmed to not be heterozygote. Among them, 11 strains were strain YX7 itself as they possessed both specific DNA bands from the two nucleus of strain YX7, and 2 strains were spore monocaryons of strain 808 as they had no any specific DNA bands of donor strain YX7. Compared with MOCAT, N-ISSR could thorough identify all the 27 heterozygote strains, which included the 19 strains identified as heterozygote, the 6 strains unable to be identified and 2 strains incorrectly identified as non-heterozygote by MOCAT. N-ISSR could identify all the 27 heterozyote stains and 13 non-heterozyote stains with simple primer, and could classify the heterozygote strains into two groups according to the type of nucleus received from the donor. On contrast, T-ISSR needed 4 primers to identify all the 27 heterozygote strains, and was unable to con firm the other 13 stains as non-heterozygote stains, and was unable to classify the heterozygote strains into two groups. In conclusion, N-ISSR was an effective tool to be used for fast, extensive and accurate identification of heterozygote and non-heterozygote strains of L. edodes, and would be a technical support for further study on the cross breeding and genetic analysis of L. edodes.

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杂合子的鉴别是香菇杂交育种中的关键步骤，为了快速准确地鉴别香菇双单杂交后代，本研究设计一种基于区分供体亲本双核的新型ISSR分子标记技术，采用该技术对香菇野生双核菌株YX7为供体和菌株808孢子单核体为受体杂交获得的40个杂交菌株进行鉴别，并以传统的显微观察与拮抗试验和传统ISSR分子标记技术为对照。新型ISSR分析结果表明：仅拥有供体菌株YX7一个核的特异性条带的菌株有27个，可判定为杂合子；其余13个菌株均判定为非杂合子，其中有11个菌株同时拥有供体菌株YX7两个核的特异性条带，是菌株YX7自身；有2个菌株不含供体菌株YX7两个核的任何特异性条带，属于菌株808的孢子单核体。与传统显微观察和拮抗试验方法相比，新型ISSR技术鉴定出的杂合子菌株更全面，不仅包含前者鉴定出的全部19个杂合子，还包括其无法判断的菌株6个和错判为非杂合子的菌株2个。新型ISSR技术采用单个引物就可确定全部27个杂合子菌株和13个非杂合子菌株，并能根据导入的供体细胞核类别将杂合子归为2类，而传统ISSR技术需采用4个引物才能鉴别出27个杂合子，并且不能确定剩余13个菌株为非杂合子，也不能对杂合子进行分类。综上，新型ISSR技术能更快速、全面和精准鉴别香菇双单杂交菌株的杂合子和非杂合子，可为香菇杂交选育和遗传分析提供技术支撑。

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吴圣进（1973—），男，博士，研究员，研究方向：食用菌育种与栽培，E-mail:gxwusj@126.com。

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吴圣进（1973—），男，博士，研究员，研究方向：食用菌育种与栽培，E-mail:gxwusj@126.com。

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