Article(id=1190310109248061832, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1190243275249390089, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.1475.2024.0731, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1699459200000, receivedDateStr=2023-11-09, revisedDate=null, revisedDateStr=null, acceptedDate=1705852800000, acceptedDateStr=2024-01-22, onlineDate=1761721644936, onlineDateStr=2025-10-29, pubDate=1748361600000, pubDateStr=2025-05-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1761721644936, onlineIssueDateStr=2025-10-29, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1761721644936, creator=13701087609, updateTime=1761721644936, updator=13701087609, issue=Issue{id=1190243275249390089, tenantId=1146029695717560320, journalId=1189873630562394117, year='2025', volume='50', issue='5', pageStart='505', pageEnd='640', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1761705710470, creator=13701087609, updateTime=1765784077922, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1207349188233372409, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1190243275249390089, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1207349188233372410, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1190243275249390089, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=607, endPage=611, ext={EN=ArticleExt(id=1190310109961093521, articleId=1190310109248061832, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=Research progress of transcriptomics sequencing technology in evaluating human endometrial receptivity, columnId=1190243275882729994, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Review, runingTitle=null, highlight=null, articleAbstract=

Good endometrial receptivity is an essential factor for embryo implantation, and gene expression in endometrial tissue during the window of implantation (WOI) is closely related to receptivity. Transcriptome sequencing technology enables the identification of gene expression profiles of endometrium during different menstrual phases, as well as microRNAs and long-chain non-coding RNA sequences involved in regulating gene expression. Combining this technology with bioinformatics analysis provides a better understanding of specific gene expression during the receptive period and offers technical support for studying its regulatory mechanism. Moreover, gene expression profiles of the endometrium during different menstrual phases hold significant clinical application value for accurately assessing endometrium receptivity in infertility patients and those with repeated implantation failure, thereby guiding individualized embryo transfer strategies. This review summarizes the progress of transcriptome sequencing in evaluating human endometrial receptivity and discusses future research directions. This review aims to understand the complex molecular mechanisms of endometrial receptivity formation and regulation from the transcriptional level, in order to improve the implantation rate of embryos in assisted reproductive technology and reduce the abortion rate.

, correspAuthors=Ying Qin, authorNote=null, correspAuthorsNote=
E-mail:
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良好的子宫内膜容受性是胚胎植入的关键,容受期子宫内膜组织的基因表达与容受性密切相关。通过转录组测序技术可获得不同月经时期子宫内膜的基因表达谱,以及调控基因表达的微小RNA和长链非编码RNA序列;结合生物信息学分析可更好地了解容受期子宫内膜特异基因的表达,并为其调控机制研究提供技术支持。同时,不同月经时期子宫内膜的基因表达谱对于准确判断不孕症及反复植入失败患者的子宫内膜容受性,进一步指导个体化胚胎移植具有重要的临床应用价值。本文对近年来转录组测序技术在评价人子宫内膜容受性中的应用进展进行综述,并探讨未来的研究方向,旨在从转录层面深入了解子宫内膜容受性形成和调控的复杂分子机制,以期提高辅助生殖技术的胚胎着床率,降低流产率。

, correspAuthors=秦莹, authorNote=null, correspAuthorsNote=
秦莹,E-mail:
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马丽娜,医学博士,主要从事妇产科生殖内分泌等方面的研究

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转录组测序技术在人子宫内膜容受性研究中的应用进展
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马丽娜 1, 2 , 漆海宁 1 , 刘梅 1 , 刘洋 3 , 葛航 4 , 卢凤娟 4 , 吴效科 3 , 秦莹 1, *
解放军医学杂志 | 综述 2025,50(5): 607-611
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解放军医学杂志 | 综述 2025, 50(5): 607-611
转录组测序技术在人子宫内膜容受性研究中的应用进展
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马丽娜1, 2, 漆海宁1, 刘梅1, 刘洋3, 葛航4, 卢凤娟4, 吴效科3, 秦莹1, *
作者信息
  • 1山东中医药大学附属医院产科,山东济南 250014
  • 2山东中医药大学博士后流动站,山东济南 250000
  • 3黑龙江中医药大学附属第一医院妇科,黑龙江哈尔滨 150040
  • 4黑龙江中医药大学研究生院,黑龙江哈尔滨 150040
  • 马丽娜,医学博士,主要从事妇产科生殖内分泌等方面的研究

通讯作者:

秦莹,E-mail:
Research progress of transcriptomics sequencing technology in evaluating human endometrial receptivity
Li-Na Ma1, 2, Hai-Ning Qi1, Mei Liu1, Yang Liu3, Hang Ge4, Feng-Juan Lu4, Xiao-Ke Wu3, Ying Qin1, *
Affiliations
  • 1Department of Obsterics, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong 250014, China
  • 2Post Doctoral Station of Shandong University of Traditional Chinese Medicine, Jinan, Shandong 250000, China
  • 3Department of Gynecology, First Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang 150040, China
  • 4Graduate School of Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang 150040, China
出版时间: 2025-05-28 doi: 10.11855/j.issn.0577-7402.1475.2024.0731
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良好的子宫内膜容受性是胚胎植入的关键,容受期子宫内膜组织的基因表达与容受性密切相关。通过转录组测序技术可获得不同月经时期子宫内膜的基因表达谱,以及调控基因表达的微小RNA和长链非编码RNA序列;结合生物信息学分析可更好地了解容受期子宫内膜特异基因的表达,并为其调控机制研究提供技术支持。同时,不同月经时期子宫内膜的基因表达谱对于准确判断不孕症及反复植入失败患者的子宫内膜容受性,进一步指导个体化胚胎移植具有重要的临床应用价值。本文对近年来转录组测序技术在评价人子宫内膜容受性中的应用进展进行综述,并探讨未来的研究方向,旨在从转录层面深入了解子宫内膜容受性形成和调控的复杂分子机制,以期提高辅助生殖技术的胚胎着床率,降低流产率。

子宫内膜  /  RNA测序技术  /  胚胎移植  /  子宫内膜容受性

Good endometrial receptivity is an essential factor for embryo implantation, and gene expression in endometrial tissue during the window of implantation (WOI) is closely related to receptivity. Transcriptome sequencing technology enables the identification of gene expression profiles of endometrium during different menstrual phases, as well as microRNAs and long-chain non-coding RNA sequences involved in regulating gene expression. Combining this technology with bioinformatics analysis provides a better understanding of specific gene expression during the receptive period and offers technical support for studying its regulatory mechanism. Moreover, gene expression profiles of the endometrium during different menstrual phases hold significant clinical application value for accurately assessing endometrium receptivity in infertility patients and those with repeated implantation failure, thereby guiding individualized embryo transfer strategies. This review summarizes the progress of transcriptome sequencing in evaluating human endometrial receptivity and discusses future research directions. This review aims to understand the complex molecular mechanisms of endometrial receptivity formation and regulation from the transcriptional level, in order to improve the implantation rate of embryos in assisted reproductive technology and reduce the abortion rate.

endometrium  /  RNA sequencing  /  embryo transfer  /  endometrial receptivity
马丽娜, 漆海宁, 刘梅, 刘洋, 葛航, 卢凤娟, 吴效科, 秦莹. 转录组测序技术在人子宫内膜容受性研究中的应用进展. 解放军医学杂志, 2025 , 50 (5) : 607 -611 . DOI: 10.11855/j.issn.0577-7402.1475.2024.0731
Li-Na Ma, Hai-Ning Qi, Mei Liu, Yang Liu, Hang Ge, Feng-Juan Lu, Xiao-Ke Wu, Ying Qin. Research progress of transcriptomics sequencing technology in evaluating human endometrial receptivity[J]. Medical Journal of Chinese People’s Liberation Army, 2025 , 50 (5) : 607 -611 . DOI: 10.11855/j.issn.0577-7402.1475.2024.0731
人子宫内膜在月经周期中被分为增殖期、分泌期及月经期3个阶段[1]。分泌期可再分为分泌早、中、晚期,其中分泌中期是非常短暂的特定时期,此时的子宫内膜允许囊胚的黏附、侵入,这种能够接纳胚胎植入的能力称为子宫内膜容受性(endometrial receptivity,ER)[2]。ER缺陷或异常容易导致着床失败、流产甚至不孕症的发生。据报道,在体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET)助孕者中,约2/3的患者胚胎着床失败与ER低下有关,而胚胎质量问题仅占1/3[3-4]。因此,如何通过改善ER来提高胚胎着床率,以获得满意的妊娠结局一直是生殖医学等领域亟待解决的核心问题。尽管当前可采用子宫内膜组织学检测、超声检查、白血病抑制因子等分子检测等手段来评估ER[5],但其准确性及可重复性饱受争议。分子生物学技术已为生理、病理条件下的子宫内膜功能特征研究提供了便捷有效的分析方法,而转录组学数据在一定程度上可为研究群体、个体、器官、组织、细胞、亚细胞等层面的生物学功能及分子调控途径提供可靠的数据支持。通过对分泌期子宫内膜基因差异表达谱的研究,得到与ER相关的编码基因及非编码基因,可更深入地了解ER形成和调控的分子机制,并提高ER异常诊断的准确性及可靠性。本文就ER及其影响因素、转录组学及其技术在生理性和病理性子宫内膜中的应用研究进展进行阐述,旨在从转录层面深入了解ER形成及调控的复杂分子机制,为临床上提高IVF-ET的胚胎着床率、降低流产率提供新的思路和方法。
子宫内膜仅在分泌中期一段极短的时间内允许胚胎植入,这一时期被称为“着床窗”(window of implantation,WOI)[6]。在人类,WOI一般在排卵后的第6~10天,即月经周期的第20~24天,且仅持续12~48 h,但WOI的开放、关闭及持续时间因人而异[7]。对于ER的形成过程,目前较为统一的说法是雌孕激素结合其同源受体调控下游靶基因,同时协同转录因子、生长因子、细胞因子等对子宫基质及上皮细胞有序的增殖和(或)分化进行调控,诱导子宫内膜发生一系列形态、生化及功能转化,进而为胚胎成功植入创造条件[8]。目前,干扰ER的因素主要包括以下几个方面:(1)子宫解剖形态异常,包括子宫肌瘤、子宫腺肌症、子宫内膜异位症(endometriosis,EMT)、子宫内膜息肉及宫腔粘连等;(2)母体免疫异常,包括自身免疫性疾病、反复着床失败等;(3)内分泌因素,包括多囊卵巢综合征、糖尿病等;(4)医源性因素,包括控制性超促排卵过程中超生理剂量雌孕激素引起的内膜容受性损伤;(5)微生物群紊乱,包括肠道、阴道及子宫内膜微生物群紊乱导致的ER下降,进而影响胚胎着床[9-12]。因此,深入全面研究ER的形成机制及影响因素,识别与ER相关的靶分子,对于揭示胚胎着床的生殖机制,提高妊娠率有重要的临床意义。
组学是指运用高通量技术同时检测不同分子区域的变化,对生物系统进行全面解读的学科,包括基因组学、转录组学、蛋白质组学等[13-14]。同一个基因组如何产生不同的细胞类型,以及基因表达如何被调控是后基因组时代困扰科学家们的难题。转录水平调控是最常见的调控方式,因此转录组学是功能基因组学研究的重要组成部分,可从整体水平上研究细胞中所有基因表达情况及转录调控的规律[15-16]。早期因技术、成本因素,转录组学的研究只能针对少数特异基因进行比对分析[17]。现在常用的测序技术包括微阵列技术、基因芯片技术、大规模平行测序技术及RNA测序技术(RNA Sequencing,RNA-Seq)等[18-20]。与其他技术相比,RNA-Seq技术具有独特的优势:(1)不受物种的基因组数据是否完整的限制;(2)分辨率高,既能对同一组织或细胞在不同功能状态下进行研究,也可研究小RNA及长链非编码RNA等基因表达水平的差异[21];(3)敏感度高,可发现稀有或未知的转录本等[22]
目前报道的不同月经周期子宫内膜转录组学研究主要集中在容受前期及容受期生理性子宫内膜的差异表达基因,以及疾病状态下子宫内膜的转录组学变化[23-37]。通过构建不同月经周期的基因表达图谱,对比ER正常、减退患者的基因表达差异,有助于研究者了解ER及胚胎植入的分子机制,对进一步指导临床诊疗具有重要意义。
有研究对比、筛选了健康人不同月经周期的子宫内膜差异基因,以期获得与胚胎种植及着床相关的关键基因[23-25]。Otsuka等[23]通过寡核苷酸三维微阵列分析了人增殖期与分泌期子宫内膜基因表达的差异,结果显示,在增殖期明显高表达的基因主要编码参与细胞增殖的蛋白质,而在分泌期,参与免疫反应、钙代谢及甲状腺激素反应的大量基因明显高表达。Altmäe等[24]应用微阵列技术对8名健康女性的增殖期及分泌中期(容受期)子宫内膜进行转录组学分析,结果显示共2177个基因表达存在差异,其中920个基因表达上调,1257个基因表达下调,进一步行功能富集分析发现,JAK/STAT信号通路、补体及凝血级联反应、黏着斑、黏附连接、炎症反应信号通路在胚胎植入过程中具有重要作用。另一项使用RNA-Seq技术的转录组学研究发现,与增殖期相比,可生育女性的子宫内膜分泌中期有2154个差异表达的mRNA,主要涉及色氨酸代谢、代谢途径及FoxO信号通路[25]
早在2002年,学者就已利用cDNA芯片检测了分泌早期及容受期子宫内膜,筛选出693个DEG,其中以编码细胞表面受体、参与细胞黏附功能的基因表达差异尤为明显[26]。随后,Hu等[27]通过子宫内膜活检收集人子宫内膜,并采用RNA-seq技术对自然月经周期容受前及容受期阶段内膜的转录组进行全面分析,结果发现,与容受前相比,容受期有1099个基因表达上调,1273个基因表达下调;利用生物信息学技术对基因功能进行功能注释及深度解析,发现上调的DEG富集到矿物质吸收通路,而下调的DEG主要在细胞周期通路富集;此外,通过基因共表达网络分析发现5个核心调控因子(GLI2、CDC25A、TLR9、MT1G、SLC5A1)参与了着床期间ER的建立。另一项研究利用RNA-Seq技术对容受前期与容受期子宫内膜的转录组进行比较发现,共有865个DEG,这些DEG在G蛋白耦联受体、糖皮质激素、胰岛素样生长因子1等信号转导通路中明显上调,而在Wnt/β-连环蛋白信号转导通路中明显下调[28]。Wang等[38]利用单细胞转录组测序(single-cell RNA sequencing,scRNA-seq)技术对不同月经周期的子宫内膜进行单细胞分离并测序,描绘了子宫内膜组织的细胞转录组图谱,发现随着月经期的改变,不同细胞的数量及功能特征发生了转变;该研究还比较了分泌中期及分泌晚期的子宫内膜基因图谱,揭示了上皮细胞基因转录活性在子宫内膜窗口期的变化。目前运用转录组学测序技术比较分泌中期与分泌晚期子宫内膜基因图谱的研究较少,这也是未来研究的重要方向。
除了自然月经周期阶段,还有其他一些病理因素也可影响子宫内膜转录组,尤其是在生殖功能衰竭的女性中。即便是同一种生殖疾病,也有不同的分期及表型,可能导致子宫内膜发生不同类型的改变或损伤。因此,需要对病理条件下的子宫内膜转录组进行无偏倚分析,以全面深入地了解子宫内膜的相关作用机制。
IVF-ET是一种治疗不孕症的高效方法。部分患者通过IVF获得优质胚胎,但是ET后存在胚胎反复种植失败(repeated implantation failure,RIF)的情况[39-41]。ER的病理性改变是导致RIF的原因之一。2009年,Koler等[29]应用微阵列技术比对了RIF女性与正常对照者容受期(月经周期第21天)子宫内膜的基因图谱,鉴定出313个DEG,其中288个(92%)在RIF女性中表达下调,证实RIF患者存在基因表达的异常。Shi等[30]发现,与非RIF女性比较,RIF患者容受期子宫内膜组织中有357个DEG,其中253个上调,104个下调;功能富集分析发现,明显下调的DEG主要涉及细胞因子-细胞因子受体相互作用、p53信号通路及补体凝血级联反应通路,而上调的DEG与过氧化物酶体增殖物激活受体(peroxisome proliferators-activated receptor,PPAR)信号、造血细胞谱系、细胞外基质(extracellular matrix,ECM)受体相互作用等信号通路相关;该研究还证实,RIF患者容受期子宫内膜组织中水通道蛋白3(aquaporin,AQP3)、二肽基肽酶Ⅳ(dipeptidylpeptidase-Ⅳ,DPP4)及组织金属蛋白酶抑制因子3(tissue inhibitor of metalloproteinase 3,TIMP3)明显下调,是其容受性受损的标志性基因。此外,Guo等[31]的转录组学研究发现,ICAM2ITGB2PECAM1SELPTEK是与RIF胚胎着床失败有关的基因,提示子宫内膜自身异常可能在RIF女性生殖障碍中发挥了重要作用。
EMT是导致育龄期女性盆腔疼痛及不孕的常见原因,发病机制未明。识别EMT患者子宫内膜的基因表达异常是了解EMT发病机制及开发相关不孕不育和疼痛治疗新策略的重要一步。目前,转录组学在EMT内膜研究中的应用较为广泛,TIMP1、半椎蛋白(hemicentin1,HMCN1)、接触蛋白-1(contactin-1,CNTN1)等几个基因已被鉴定为EMT的诊断标志物[32-35]。如田鑫[32]基于RNA-seq测序技术对EMT患者的异位病灶、在位内膜及健康女性子宫内膜的RNA进行全面分析,发现DEG中的TIMP1HMCN1与EMT细胞迁移、侵袭及浸润高度相关,提示这些DEG可能成为EMT新的分子标志物及治疗靶基因。此外,赵旭旭[33]借助全基因组表达谱分析比较了卵巢EMT与非EMT患者的异位内膜组织的转录差异,发现异位内膜的黏附功能发生了明显变化,其中变化最明显的黏附分子是CNTN1,可作为卵巢EMT潜在的诊断标志物及药物治疗靶点。Chen等[34]采用RNA-seq分析在位及异位子宫内膜间质细胞中的DEG,发现共有1309个基因上调,663个基因下调,这些DEG主要涉及磷脂酰肌醇-3-激酶/蛋白激酶B(phosphatidyli-nositol-3-kinase-protein kinase B,PI3K/AKT)、细胞因子-细胞因子受体相互作用及MAPK信号通路。以上转录组学研究筛选的分子标志物并不一致,分析原因可能与EMT疾病的异质性较大有关。因此,建立统一的表型分类方法、规范的标本提取及检测流程,对评估、筛选公认的生物标志物具有重要意义。目前,对EMT的研究依然采取整体组织转录组测序的方法,缺乏对异位病灶组织异质性及单细胞水平基因表达情况的认知。黄志雄[42]通过scRNA-seq绘制了EMT在位与异位内膜的细胞图谱,揭示了异位病灶细胞数量及生理功能的改变,从单细胞层面展示了异位病灶中各类细胞生理功能的异常转变。
CE是一种可引起子宫内膜结构及功能破坏的持续性炎症过程,局部内膜炎性细胞浸润及炎症介质渗出会改变子宫内膜微环境,从而影响子宫内膜的功能。研究发现,CE可造成子宫内膜的容受性受损[43],导致自然妊娠及辅助生殖技术助孕的受孕率明显降低。Chen等[36]发现,CE患者子宫内膜的一些炎性相关基因发生了变化,影响了子宫内膜的容受性,进而影响了女性的生殖功能。近期,Oshina等[37]通过scRNA-seq技术获得了CE与非CE女性分泌期的内膜基因组,发现有8个基因表达上调,包括2个免疫球蛋白基因(IGHG4JCHAIN),6个非免疫球蛋白基因(OVGP1MTUS2CLIC6ACSM1MESP1LTF),与CE的主要特征是浆细胞浸润一致。目前尚未有系统的研究来描述CE在整个月经周期中ER的基因表达特征。
良好的ER是胚胎着床的必要前提,对于ER的影响因素、评估手段及治疗方法仍在不断探索中,如何准确评价及有效改善ER是临床亟须解决的问题。转录组测序技术广泛应用于人生理、病理状态下ER的机制研究中,一些与ER相关的关键基因相继被挖掘。然而,既往研究报道的与ER相关的DEG重复性较低,其原因除研究的样本量、样本可变性、异质性等以外,还与测序技术的局限性有关。相信随着转录组测序技术的不断革新,其在ER及胚胎植入机制研究领域中的应用必将更加广泛,能更好地为女性的生殖健康服务。
  • 国家重点研发计划(2019YFC1709500)
  • 国家重大疑难疾病中西医临床协作试点项目(国中医药办医政发[2018]3号)
  • 山东省自然科学基金面上项目(ZR2022MH321)
  • 山东省中医药科技项目重点项目(Z-2023075)
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doi: 10.11855/j.issn.0577-7402.1475.2024.0731
  • 接收时间:2023-11-09
  • 首发时间:2025-10-29
  • 出版时间:2025-05-28
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  • 收稿日期:2023-11-09
  • 录用日期:2024-01-22
基金
National Key Research and Development Project of China(2019YFC1709500)
国家重点研发计划(2019YFC1709500)
National Pilot Project of Clinical Cooperation between Traditional Chinese and Western Medicine for Major Difficult Diseases ([2018]No.3 of the State Office of Traditional Chinese Medicine)
国家重大疑难疾病中西医临床协作试点项目(国中医药办医政发[2018]3号)
Natural Science Foundation of Shandong Province(ZR2022MH321)
山东省自然科学基金面上项目(ZR2022MH321)
Key Project of TCM Science and Technology Project of Shandong Province(Z-2023075)
山东省中医药科技项目重点项目(Z-2023075)
作者信息
    1山东中医药大学附属医院产科,山东济南 250014
    2山东中医药大学博士后流动站,山东济南 250000
    3黑龙江中医药大学附属第一医院妇科,黑龙江哈尔滨 150040
    4黑龙江中医药大学研究生院,黑龙江哈尔滨 150040

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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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