Article(id=1256548243622736346, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256548241764639069, articleNumber=null, orderNo=null, doi=10.13346/j.mycosystema.250259, pmid=null, cstr=32115.14.j.mycosystema.250259, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1756396800000, receivedDateStr=2025-08-29, revisedDate=null, revisedDateStr=null, acceptedDate=1762185600000, acceptedDateStr=2025-11-04, onlineDate=1777514046817, onlineDateStr=2026-04-30, pubDate=1776787200000, pubDateStr=2026-04-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1777514046817, onlineIssueDateStr=2026-04-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1777514046817, creator=13701087609, updateTime=1777514046817, updator=13701087609, issue=Issue{id=1256548241764639069, tenantId=1146029695717560320, journalId=1255847803461844995, year='2026', volume='45', issue='4', pageStart='250187', pageEnd='250358', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1777514046373, creator=13701087609, updateTime=1777516895320, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1256560191206711468, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256548241764639069, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1256560191206711469, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256548241764639069, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=250259, endPage=, ext={EN=ArticleExt(id=1256548245006856668, articleId=1256548243622736346, tenantId=1146029695717560320, journalId=1255847803461844995, language=EN, title=Members of the LeATG gene family and their expression during mycelial colouring process of Lentinula edodes, columnId=1256263562373226548, journalTitle=Mycosystema, columnName=Research paper, runingTitle=null, highlight=null, articleAbstract=

The autophagy-related gene family members in Lentinula edodes (LeATG family members) were identified based on the whole genome data of L. edodes using homologous alignment methods, and their biological information was analyzed. The expression patterns of these family members during mycelial colouring process were analyzed by combining transcriptome sequencing data and RT-qPCR. The results showed that 29 LeATG family members belonging to 23 classes of autophagy-related genes were identified in the L. edodes genome. Chromosomal localization analysis revealed that they were mainly distributed on chromosomes 4 and 1; no distribution was detected on chromosome 9. The full length of LeATG family members ranges from 595 to 6 772 bp. The length of protein sequence encoded by the LeATG gene ranged from 127 to 1 998 aa, with relative molecular weights of 14.8-219.7 kDa and theoretical isoelectric points of 4.27-9.51. Except for LeATG9, LeATG22, and LeATG27, the other 24 LeATG proteins contained no transmembrane domains. The phylogenetic analysis showed that the 29 LeATG proteins clustered into three evolutionary branches, and proteins of the same class were dispersed across different sub-branches, suggesting significant differences in protein structure. Transcriptome data analysis and RT-qPCR revealed differential expression patterns of LeATG genes before and after mycelial colouring process: 2 LeATG genes were highly expressed before browning, 5 were highly expressed after colouring. It was speculated that these 7 genes might be involved in regulating the mycelial colouring process. This study systematically identified the LeATG gene family in L. edodes, and preliminarily revealed their expression regulation patterns during mycelial colouring process, laying an important foundation for further analysis of the molecular mechanism of the LeATG gene during mycelial colouring process of L. edodes.

, correspAuthors=Yuanyuan LIU, authorNote=null, correspAuthorsNote=
*
, 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=

ORCID: LIU Yuanyuan (0009-0000-7180-0601)

, authorsList=Mengjia HU, Qi GAO, Yangyang FAN, Dong YAN, Shouxian WANG, Yu LIU, Xianchun ZONG, Yuanyuan LIU), CN=ArticleExt(id=1256548250593669627, articleId=1256548243622736346, tenantId=1146029695717560320, journalId=1255847803461844995, language=CN, title=香菇LeATG基因家族及其在菌丝转色过程中的表达, columnId=1256263563312771301, journalTitle=菌物学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

为解析香菇Lentinula edodes自噬相关基因(autophagy-related gene, ATG)家族成员在菌丝转色过程中的功能,本研究基于香菇全基因组数据,采用同源比对的方法筛选LeATG家族成员,并对其进行生物信息学分析。同时,结合转录组测序数据和RT-qPCR,分析了该家族成员在菌丝转色过程中的表达模式。结果表明,在香菇全基因组中共鉴定出29个LeATG家族成员,属于23类自噬相关基因。染色体定位分析显示,其主要分布于4号和1号染色体,9号染色体中未发现分布。LeATG家族成员基因全长在595-6 772 bp之间,LeATG基因编码蛋白序列长度介于127-1 998 aa,蛋白相对分子量为14.8-219.7 kDa,理论等电点为4.27-9.51,除LeATG9、LeATG22和LeATG27外,其余24个LeATG蛋白均不含跨膜结构。进化分析结果显示29个LeATG蛋白聚类为3个进化分支,同一类LeATG蛋白分散于不同亚支,说明蛋白结构可能存在显著差异。转录组数据和RT-qPCR分析结果表明不同LeATG基因在菌丝转色前后的差异表达模式,其中2个LeATG基因在转色前高表达,5个在转色后高表达,推测以上7个基因可能参与调控菌丝的转色过程。本研究系统鉴定了香菇LeATG基因家族,初步揭示了该家族成员在菌丝转色过程中的表达调控模式,为深入解析LeATG基因在香菇转色过程中的分子机制奠定了重要基础。

, correspAuthors=刘媛媛, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=hy3SSjLnAcoQowLi+B+Ptw==, magXml=e5UKN3NPrsvE/2h2zAkkNw==, pdfUrl=null, pdf=GMtou22fYU/r/E6YY0Rldw==, pdfFileSize=1888390, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=LC+HEtUHiZ3IWLMjLtXAjw==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=gimqokJhBIw6kAkY8+mSTQ==, mapNumber=null, authorCompany=null, fund=null, authors=

作者贡献

胡梦佳:试验、图表制作、数据分析及论文撰写;高琪:提供实验材料、论文构思与实验指导;范阳阳:数据统计及分析;严冬:论文构思及指导;王守现:试验指导;刘宇:论文指导;宗宪春:论文审核;刘媛媛:论文构思、实验指导、论文指导和修改。

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ArticleFig(id=1256548276271198944, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Fig. 2, caption=Chromosomal distribution of the LeATG family members. Red color represents the members of the LeATG family gene., figureFileSmall=DOTIyoOjxTOqNdnEv1VPhw==, figureFileBig=3FgFCN7N0GFWTY8+EtVENw==, tableContent=null), ArticleFig(id=1256548276531245798, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=图2, caption=LeATG家族成员染色体分布情况 红色代表LeATG家族成员基因, figureFileSmall=DOTIyoOjxTOqNdnEv1VPhw==, figureFileBig=3FgFCN7N0GFWTY8+EtVENw==, tableContent=null), ArticleFig(id=1256548276904538857, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Fig. 3, caption=Prediction of conserved domains in the LeATG family., figureFileSmall=sw25LNe5z0EqHH0IU6zCrw==, figureFileBig=SyIGuTPTQ2aldoQ9HIogcw==, tableContent=null), ArticleFig(id=1256548277277831917, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=图3, caption=LeATG家族保守结构域预测, figureFileSmall=sw25LNe5z0EqHH0IU6zCrw==, figureFileBig=SyIGuTPTQ2aldoQ9HIogcw==, tableContent=null), ArticleFig(id=1256548277474964209, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Fig. 4, caption=Multiple sequence alignment and conserved motif analysis of ATG8 proteins in Lentinula edodes and other fungi. A: Multisequence alignment of ATG8 protein in L. edodes and other fungi; Sequence similarity is graded from light grey (low similarity) to black (highest similarity); Underlined ones indicate the conservative domains of ubiquitin-like (Ubl) domains; S.cerevisiae: Saccharomyces cerevisiae; L.bicolor: Laccaria bicolor; R.earlei: Russula earlei; B: Conserved motif analysis of ATG8 protein in L. edodes and other fungi., figureFileSmall=VLQ6DpxW71ERntdCF8L1Mg==, figureFileBig=FdxEyI/nAGE4XKnC2l5RQA==, tableContent=null), ArticleFig(id=1256548277705650933, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=图4, caption=香菇与其他真菌ATG8蛋白的多序列比对与保守基序分析 A:香菇与其他真菌ATG8蛋白的多序列比对,碱基的背景色越深代表该位点的保守性越高,下划线所示为ubiquitin-like (Ubl) domain的保守结构域,S.cerevisiae:Saccharomyces cerevisiae,L.bicolor:Laccaria bicolor,R.earlei:Russula earlei;B:香菇与其他真菌ATG8蛋白的保守基序分析, figureFileSmall=VLQ6DpxW71ERntdCF8L1Mg==, figureFileBig=FdxEyI/nAGE4XKnC2l5RQA==, tableContent=null), ArticleFig(id=1256548277902783223, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Fig. 5, caption=Phylogenetic analysis of LeATG family members., figureFileSmall=i2uxYX8oSBd42n3/nI+7iw==, figureFileBig=ZvFlh8Fix3yIEDB6MH1LIQ==, tableContent=null), ArticleFig(id=1256548278137664252, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=图5, caption=LeATG家族成员系统进化分析, figureFileSmall=i2uxYX8oSBd42n3/nI+7iw==, figureFileBig=ZvFlh8Fix3yIEDB6MH1LIQ==, tableContent=null), ArticleFig(id=1256548278594843392, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Fig. 6, caption=Transcriptomic analysis of LeATG family genes. A: Correlation analysis plot; B: Heatmap of transcriptomic expression levels of LeATG family genes; C: Bar chart of some differentially expressed genes (DEGs) within the LeATG family. LeHW-3×30: Uncoloured mycelia of BIPP21020482; LeHB-3×30: Coloured mycelia of BIPP21020482. ***and **** represent significance at P<0.001 and P<0.000 1, respectively (n=3)., figureFileSmall=/wNEeZyghCPHK40KwoMvPA==, figureFileBig=RIenZy+cRW2AeSTyyEYYPQ==, tableContent=null), ArticleFig(id=1256548280352256772, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=图6, caption=LeATG家族成员基因的转录组分析结果 A:相关性分析图;B:LeATG家族基因的转录组表达量热图;C:LeATG家族中部分差异表达基因的柱状图;LeHW-3×30:BIPP21020482未转色菌丝;LeHB-3×30:BIPP21020482转色菌丝;***和****分别表示在P<0.001、P<0.000 1水平上差异显著(n=3), figureFileSmall=/wNEeZyghCPHK40KwoMvPA==, figureFileBig=RIenZy+cRW2AeSTyyEYYPQ==, tableContent=null), ArticleFig(id=1256548280977208071, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Fig. 7, caption=Expression analysis and conserved structure analysis of LeATG15 family members. A: Bar chart of differential expression of the LeATG15 gene, LeHW-3×30: Uncoloured mycelia of BIPP21020482; LeHB-3×30: Coloured mycelia of BIPP21020482; **and *** represent significance at P<0.01 and P<0.001, respectively (n=3). B: Multiple sequence alignment of LeATG15 protein; Sequence similarity is graded from light grey (low similarity) to black (highest similarity). C: Conserved motif analysis of LeATG15., figureFileSmall=nI2dZpzZq3grcXqfANNdgA==, figureFileBig=tCygW0xaY4Ub76NAPQilRw==, tableContent=null), ArticleFig(id=1256548281228866314, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=图7, caption=LeATG15家族成员的表达分析及保守结构分析 A:LeATG15基因的差异表达柱状图,LeHW-3×30:BIPP21020482未转色菌丝;LeHB-3×30:BIPP21020482转色菌丝,**和***分别表示在P<0.01、P<0.001水平上差异显著(n=3);B:LeATG15的蛋白多序列比对图,碱基的背景色越深代表该位点的保守性越高;C:LeATG15的保守基序分析图, figureFileSmall=nI2dZpzZq3grcXqfANNdgA==, figureFileBig=tCygW0xaY4Ub76NAPQilRw==, tableContent=null), ArticleFig(id=1256548281392444172, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Table 1, caption=

Primers sequences

, figureFileSmall=null, figureFileBig=null, tableContent=
引物名称
Primer
引物序列
Primer sequence (5ʹ→3ʹ)
用途
Function
Btu-1 F: CAGTTCACGGCCATGTTCA 荧光定量PCR检测
R: CGACGGTGGCATCCTGGTA RT-qPCR
LeATG8-1Q F: ATCGTATTCCTGTAATCTGCG 荧光定量PCR检测
R: TAGACGAACTGCCCCACA RT-qPCR
LeATG27-1Q F: TGATGGCAGCGACAAT 荧光定量PCR检测
R: TGCACCGAGCACAAAG RT-qPCR
), ArticleFig(id=1256548281518273295, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=表1, caption=

引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
引物名称
Primer
引物序列
Primer sequence (5ʹ→3ʹ)
用途
Function
Btu-1 F: CAGTTCACGGCCATGTTCA 荧光定量PCR检测
R: CGACGGTGGCATCCTGGTA RT-qPCR
LeATG8-1Q F: ATCGTATTCCTGTAATCTGCG 荧光定量PCR检测
R: TAGACGAACTGCCCCACA RT-qPCR
LeATG27-1Q F: TGATGGCAGCGACAAT 荧光定量PCR检测
R: TGCACCGAGCACAAAG RT-qPCR
), ArticleFig(id=1256548281597965073, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Table 2, caption=

Gene sequence information of the LeATG family members

, figureFileSmall=null, figureFileBig=null, tableContent=
基因名称
Gene
外显子个数
Number of
exons
内含子个数
Number of
introns
开放阅读框长度
ORF length/bp
5ʹ端非翻译区长度
Length of the
5ʹ-untranslated region
(5ʹ-UTR)/bp
3ʹ端非翻译区长度
Length of the
3ʹ-untranslated
region (3ʹ-UTR)/bp
基因全长
Gene length
/bp
LeATG1-1 10 9 2 652 213 305 3 152
LeATG2-1 16 15 5 997 140 71 6 772
LeATG2-2 11 17 1 617 1 339 56 2 248
LeATG3-1 7 6 1 116 40 156 1 458
LeATG4-1 5 9 3 405 577 377 3 630
LeATG5-1 4 3 1 101 75 112 1 257
LeATG6-1 6 5 1 455 17 94 1 710
LeATG7-1 10 9 2 127 34 36 2 614
LeATG8-1 5 4 384 122 221 603
LeATG9-1 10 9 2 865 65 208 3 355
LeATG10-1 3 2 636 8 80 741
LeATG11-1 16 15 4 020 63 271 4 803
LeATG12-1 5 4 375 19 76 595
LeATG13-1 3 2 2 877 152 406 3 009
LeATG15-1 5 4 1 584 155 132 1 996
LeATG15-2 3 2 681 332 151 876
LeATG15-3 6 5 1 254 23 136 1 544
LeATG15-4 4 3 1 269 34 333 1 438
LeATG16-1 4 3 822 17 134 1 006
LeATG17-1 9 8 1 449 21 165 1 881
LeATG18-1 7 6 1 275 20 214 1 587
LeATG20-1 8 7 1 728 40 216 2 150
LeATG22-1 8 7 1 674 1 119 2 041
LeATG22-2 14 13 1 608 60 87 2 305
LeATG22-3 19 18 1 617 58 200 2 626
LeATG24-1 8 7 1 401 25 214 1 768
LeATG26-1 19 18 4 425 64 144 5 415
LeATG27-1 5 4 681 204 123 931
LeATG29-1 5 4 1 299 78 88 1 508
), ArticleFig(id=1256548281925120789, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=表2, caption=

LeATG家族成员基因序列信息

, figureFileSmall=null, figureFileBig=null, tableContent=
基因名称
Gene
外显子个数
Number of
exons
内含子个数
Number of
introns
开放阅读框长度
ORF length/bp
5ʹ端非翻译区长度
Length of the
5ʹ-untranslated region
(5ʹ-UTR)/bp
3ʹ端非翻译区长度
Length of the
3ʹ-untranslated
region (3ʹ-UTR)/bp
基因全长
Gene length
/bp
LeATG1-1 10 9 2 652 213 305 3 152
LeATG2-1 16 15 5 997 140 71 6 772
LeATG2-2 11 17 1 617 1 339 56 2 248
LeATG3-1 7 6 1 116 40 156 1 458
LeATG4-1 5 9 3 405 577 377 3 630
LeATG5-1 4 3 1 101 75 112 1 257
LeATG6-1 6 5 1 455 17 94 1 710
LeATG7-1 10 9 2 127 34 36 2 614
LeATG8-1 5 4 384 122 221 603
LeATG9-1 10 9 2 865 65 208 3 355
LeATG10-1 3 2 636 8 80 741
LeATG11-1 16 15 4 020 63 271 4 803
LeATG12-1 5 4 375 19 76 595
LeATG13-1 3 2 2 877 152 406 3 009
LeATG15-1 5 4 1 584 155 132 1 996
LeATG15-2 3 2 681 332 151 876
LeATG15-3 6 5 1 254 23 136 1 544
LeATG15-4 4 3 1 269 34 333 1 438
LeATG16-1 4 3 822 17 134 1 006
LeATG17-1 9 8 1 449 21 165 1 881
LeATG18-1 7 6 1 275 20 214 1 587
LeATG20-1 8 7 1 728 40 216 2 150
LeATG22-1 8 7 1 674 1 119 2 041
LeATG22-2 14 13 1 608 60 87 2 305
LeATG22-3 19 18 1 617 58 200 2 626
LeATG24-1 8 7 1 401 25 214 1 768
LeATG26-1 19 18 4 425 64 144 5 415
LeATG27-1 5 4 681 204 123 931
LeATG29-1 5 4 1 299 78 88 1 508
), ArticleFig(id=1256548282327773976, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=EN, label=Table 3, caption=

Physicochemical properties of LeATG family members

, figureFileSmall=null, figureFileBig=null, tableContent=
基因名称
Gene
蛋白序列长度
Protein length/aa
分子量
Molecular
weight/kDa
等电点
Isoelectric
point
跨膜结构个数
Transmembrane
domain
信号肽
Signal peptide
亚细胞定位
Subcellular localization
LeATG1-1 883 98.62 8.89 0 No 细胞核 Nuclear
LeATG2-1 1 998 219.70 5.43 0 No 细胞核 Nuclear
LeATG2-2 538 58.83 6.99 0 No 细胞质 Cytoplasmic
LeATG3-1 371 40.86 4.62 0 No 细胞质 Cytoplasmic
LeATG4-1 1 134 120.63 6.75 0 No 细胞核 Nuclear
LeATG5-1 366 40.94 6.46 0 No 细胞核 Nuclear
LeATG6-1 484 54.24 6.96 0 No 细胞核 Nuclear
LeATG7-1 708 78.23 5.31 0 No 细胞质 Cytoplasmic
LeATG8-1 127 14.8 5.76 0 No 细胞质 Cytoplasmic
LeATG9-1 954 107.87 6.64 4 No 质膜 Plasma Membrane
LeATG10-1 211 24.11 4.66 0 No 质膜 Plasma Membrane
LeATG11-1 1 339 149.36 6.22 0 No 细胞核 Nuclear
LeATG12-1 772 86.58 5.89 0 No 线粒体 Mitochondrial
LeATG13-1 958 100.10 9.45 0 No 细胞核 Nuclear
LeATG15-1 527 58.31 5.46 0 No 细胞外 Extracellular
LeATG15-2 226 24.81 4.76 0 Yes 细胞外 Extracellular
LeATG15-3 417 46.10 4.88 0 No 细胞外 Extracellular
LeATG15-4 422 46.67 5.55 0 Yes 细胞外 Extracellular
LeATG16-1 273 30.71 5.45 0 No 细胞核 Nuclear
LeATG17-1 482 54.93 4.56 0 No 细胞核 Nuclear
LeATG18-1 424 45.62 7.58 0 No 质膜 Plasma membrane
LeATG20-1 575 64.49 5.99 0 No 细胞核 Nuclear
LeATG22-1 557 61.63 7.58 12 No 质膜 Plasma membrane
LeATG22-2 535 59.72 8.31 12 No 质膜 Plasma membrane
LeATG22-3 538 59.94 5.91 12 No 质膜 Plasma membrane
LeATG24-1 466 53.03 5.41 0 No 细胞质 Cytoplasmic
LeATG26-1 1 474 162.88 6.39 0 No 细胞核 Nuclear
LeATG27-1 226 24.59 4.27 1 No 细胞核 Nuclear
LeATG29-1 432 46.46 9.51 0 No 细胞核 Nuclear
), ArticleFig(id=1256548282524906270, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256548243622736346, language=CN, label=表3, caption=

LeATG家族成员理化性质

, figureFileSmall=null, figureFileBig=null, tableContent=
基因名称
Gene
蛋白序列长度
Protein length/aa
分子量
Molecular
weight/kDa
等电点
Isoelectric
point
跨膜结构个数
Transmembrane
domain
信号肽
Signal peptide
亚细胞定位
Subcellular localization
LeATG1-1 883 98.62 8.89 0 No 细胞核 Nuclear
LeATG2-1 1 998 219.70 5.43 0 No 细胞核 Nuclear
LeATG2-2 538 58.83 6.99 0 No 细胞质 Cytoplasmic
LeATG3-1 371 40.86 4.62 0 No 细胞质 Cytoplasmic
LeATG4-1 1 134 120.63 6.75 0 No 细胞核 Nuclear
LeATG5-1 366 40.94 6.46 0 No 细胞核 Nuclear
LeATG6-1 484 54.24 6.96 0 No 细胞核 Nuclear
LeATG7-1 708 78.23 5.31 0 No 细胞质 Cytoplasmic
LeATG8-1 127 14.8 5.76 0 No 细胞质 Cytoplasmic
LeATG9-1 954 107.87 6.64 4 No 质膜 Plasma Membrane
LeATG10-1 211 24.11 4.66 0 No 质膜 Plasma Membrane
LeATG11-1 1 339 149.36 6.22 0 No 细胞核 Nuclear
LeATG12-1 772 86.58 5.89 0 No 线粒体 Mitochondrial
LeATG13-1 958 100.10 9.45 0 No 细胞核 Nuclear
LeATG15-1 527 58.31 5.46 0 No 细胞外 Extracellular
LeATG15-2 226 24.81 4.76 0 Yes 细胞外 Extracellular
LeATG15-3 417 46.10 4.88 0 No 细胞外 Extracellular
LeATG15-4 422 46.67 5.55 0 Yes 细胞外 Extracellular
LeATG16-1 273 30.71 5.45 0 No 细胞核 Nuclear
LeATG17-1 482 54.93 4.56 0 No 细胞核 Nuclear
LeATG18-1 424 45.62 7.58 0 No 质膜 Plasma membrane
LeATG20-1 575 64.49 5.99 0 No 细胞核 Nuclear
LeATG22-1 557 61.63 7.58 12 No 质膜 Plasma membrane
LeATG22-2 535 59.72 8.31 12 No 质膜 Plasma membrane
LeATG22-3 538 59.94 5.91 12 No 质膜 Plasma membrane
LeATG24-1 466 53.03 5.41 0 No 细胞质 Cytoplasmic
LeATG26-1 1 474 162.88 6.39 0 No 细胞核 Nuclear
LeATG27-1 226 24.59 4.27 1 No 细胞核 Nuclear
LeATG29-1 432 46.46 9.51 0 No 细胞核 Nuclear
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香菇LeATG基因家族及其在菌丝转色过程中的表达
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胡梦佳 1, 2 , 高琪 1 , 范阳阳 1 , 严冬 1 , 王守现 1 , 刘宇 1 , 宗宪春 2 , 刘媛媛 1, *
菌物学报 | 研究论文 2026,45(4): 250259
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菌物学报 | 研究论文 2026, 45(4): 250259
香菇LeATG基因家族及其在菌丝转色过程中的表达
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胡梦佳1, 2, 高琪1, 范阳阳1, 严冬1, 王守现1, 刘宇1, 宗宪春2, 刘媛媛1, *
作者信息
  • 1 北京市农林科学院植物保护研究所 北京市食用菌工程技术研究中心, 北京 100097
  • 2 牡丹江师范学院生命科学与技术学院, 黑龙江 牡丹江 157011
Members of the LeATG gene family and their expression during mycelial colouring process of Lentinula edodes
Mengjia HU1, 2, Qi GAO1, Yangyang FAN1, Dong YAN1, Shouxian WANG1, Yu LIU1, Xianchun ZONG2, Yuanyuan LIU1, *
Affiliations
  • 1 Beijing Engineering Research Center for Edible Mushroom, Institute of Plant Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
  • 2 College of Life Sciences and Technology, Mudanjiang Normal University, Mudanjiang 157011, Heilongjiang, China
出版时间: 2026-04-22 doi: 10.13346/j.mycosystema.250259
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为解析香菇Lentinula edodes自噬相关基因(autophagy-related gene, ATG)家族成员在菌丝转色过程中的功能,本研究基于香菇全基因组数据,采用同源比对的方法筛选LeATG家族成员,并对其进行生物信息学分析。同时,结合转录组测序数据和RT-qPCR,分析了该家族成员在菌丝转色过程中的表达模式。结果表明,在香菇全基因组中共鉴定出29个LeATG家族成员,属于23类自噬相关基因。染色体定位分析显示,其主要分布于4号和1号染色体,9号染色体中未发现分布。LeATG家族成员基因全长在595-6 772 bp之间,LeATG基因编码蛋白序列长度介于127-1 998 aa,蛋白相对分子量为14.8-219.7 kDa,理论等电点为4.27-9.51,除LeATG9、LeATG22和LeATG27外,其余24个LeATG蛋白均不含跨膜结构。进化分析结果显示29个LeATG蛋白聚类为3个进化分支,同一类LeATG蛋白分散于不同亚支,说明蛋白结构可能存在显著差异。转录组数据和RT-qPCR分析结果表明不同LeATG基因在菌丝转色前后的差异表达模式,其中2个LeATG基因在转色前高表达,5个在转色后高表达,推测以上7个基因可能参与调控菌丝的转色过程。本研究系统鉴定了香菇LeATG基因家族,初步揭示了该家族成员在菌丝转色过程中的表达调控模式,为深入解析LeATG基因在香菇转色过程中的分子机制奠定了重要基础。

香菇  /  自噬相关基因  /  菌丝转色  /  基因表达

The autophagy-related gene family members in Lentinula edodes (LeATG family members) were identified based on the whole genome data of L. edodes using homologous alignment methods, and their biological information was analyzed. The expression patterns of these family members during mycelial colouring process were analyzed by combining transcriptome sequencing data and RT-qPCR. The results showed that 29 LeATG family members belonging to 23 classes of autophagy-related genes were identified in the L. edodes genome. Chromosomal localization analysis revealed that they were mainly distributed on chromosomes 4 and 1; no distribution was detected on chromosome 9. The full length of LeATG family members ranges from 595 to 6 772 bp. The length of protein sequence encoded by the LeATG gene ranged from 127 to 1 998 aa, with relative molecular weights of 14.8-219.7 kDa and theoretical isoelectric points of 4.27-9.51. Except for LeATG9, LeATG22, and LeATG27, the other 24 LeATG proteins contained no transmembrane domains. The phylogenetic analysis showed that the 29 LeATG proteins clustered into three evolutionary branches, and proteins of the same class were dispersed across different sub-branches, suggesting significant differences in protein structure. Transcriptome data analysis and RT-qPCR revealed differential expression patterns of LeATG genes before and after mycelial colouring process: 2 LeATG genes were highly expressed before browning, 5 were highly expressed after colouring. It was speculated that these 7 genes might be involved in regulating the mycelial colouring process. This study systematically identified the LeATG gene family in L. edodes, and preliminarily revealed their expression regulation patterns during mycelial colouring process, laying an important foundation for further analysis of the molecular mechanism of the LeATG gene during mycelial colouring process of L. edodes.

Lentinula edodes  /  autophagy-related genes  /  mycelial colouring  /  gene expression
胡梦佳, 高琪, 范阳阳, 严冬, 王守现, 刘宇, 宗宪春, 刘媛媛. 香菇LeATG基因家族及其在菌丝转色过程中的表达. 菌物学报, 2026 , 45 (4) : 250259 - . DOI: 10.13346/j.mycosystema.250259
Mengjia HU, Qi GAO, Yangyang FAN, Dong YAN, Shouxian WANG, Yu LIU, Xianchun ZONG, Yuanyuan LIU. Members of the LeATG gene family and their expression during mycelial colouring process of Lentinula edodes[J]. Mycosystema, 2026 , 45 (4) : 250259 - . DOI: 10.13346/j.mycosystema.250259
香菇Lentinula edodes (Berk.) Pegler是一种食药用菌(Wu et al. 2019),因其丰富的营养成分和显著的生物活性在全球受到广泛欢迎 (Meng et al. 2025;Ren et al. 2025)。据中国食用菌协会统计,2023年食用菌年产量为4 334.17万t (谷莉等 2025),其中香菇的总产量达1 303.75万t,占全国30%,稳居我国食用菌总产量的第一位。作为重要的经济作物,香菇转色能力会直接影响子实体的产量和质量(管婉等 2022a),在经济效益中起到关键作用。因此,探索香菇转色的分子机制对香菇高品质的性状改良具有重要意义。
自噬是真核生物中高度保守的细胞过程,包括自噬诱导、脂质输送及囊泡成核、吞噬泡的扩张与闭合、自噬体的运输、自噬体与液泡膜的融合、物质降解等步骤(Yang et al. 2021),主要用于降解不必要或功能失调的细胞成分(Kanayama & Shinohara 2016)。正常条件下,低水平自噬可以清除细胞损伤,维持代谢稳态(刘晓良等 2025);当受到干旱、极端温度和营养亏缺等非生物胁迫时,自噬过程增强,从而提高生物抗逆性(任云儿等 2025)。Takeshige et al. (1992)在酿酒酵母中发现营养缺乏会诱导自噬降解,后续又通过筛选自噬缺陷突变体鉴定了一系列Apg自噬相关基因。迄今为止,在真核生物中共鉴定出42个负责自噬体形成和选择性调节自噬的Atg自噬相关基因(Sheng & Qin 2019)。自噬相关基因能够直接或间接调控生物的生长发育和抗胁迫能力。在水稻中,自噬相关基因OsATG7能响应黑暗胁迫,且胁迫时间与基因表达量呈正相关,敲除该基因后会致使菌株高度不育,衰老相关基因SGR表达显著上调,叶绿素含量降低,呈现早衰表型(孟盼盼等 2017);在酵母中,过表达自噬相关基因ATG8后会导致活性氧含量降低,氧化应激相关的基因表达上调(杨晓迪等 2025);在桃褐腐病菌中,基于转录组及基因组数据,鉴定了1个自噬相关基因MfATG1,通过敲除技术发现突变株的生长速度和致病力下降,提高了对NaCl的敏感性(黄松等 2025)。对于食用菌来说,在蛹虫草中,Wang et al. (2025)分别构建了自噬相关基因CmATG1CmATG13敲除菌株,表型结果发现两种突变株与野生型相比无法形成正常子实体,且菌丝形态异常,分生孢子产量和萌发率显著降低。在香菇中,房丽丽等(2017)通过western blot技术发现菌丝转色后Atg8蛋白的表达量比转色前高,管婉等(2022a, 2022b)通过检测转色前后该基因的表达量及自噬特征进一步证明自噬与转色过程紧密相关。目前大量研究均集中在对单个自噬相关基因功能的探究,而关于香菇自噬家族基因成员的系统性研究以及其是否参与菌丝转色过程尚不明确。
本研究基于香菇全基因组数据,利用同源比对筛选获得自噬相关基因LeATG家族成员,并确定其基因定位。采用生物信息学的方法,预测并分析了各家族成员基因结构、蛋白理化性质,初步解析其序列特征,同时构建系统进化树以分析其进化关系。结合菌丝转色前后的转录组数据和RT-qPCR,分析了LeATG家族成员不同的表达模式。本研究结果将为解析LeATG家族成员的生物学功能奠定基础。
北方香菇主栽品种BIPP21020482及其原生质体单核化菌株SP3均保藏于北京市食用菌种质资源库。
本实验室已采用Illumina HiSeq X Ten和PacBio SequelⅡ平台对SP3的基因组进行测序,并通过Hi-C数据辅助组装,获得10条染色体。采用Illumina双端测序法对BIPP21020482的白色菌丝阶段、转色菌丝阶段、小原基、大原基菌柄、大原基菌盖、菌盖表皮、菌盖菌肉、菌盖菌柄连接处、菌褶、菌柄表皮、菌柄外侧菌肉、菌柄中心菌肉样品进行二代转录组测序,并结合Genescan软件(http://hollywood.mit.edu/GENSCAN.html)、Augustus软件(https://bioinf.uni-greifswald.de/augustus/)和Exonerate软件(http://ftp.ebi.ac.uk/pub/software/vertebrategenomics/exonerate/exonerate-2.2.0-x86_64.tar.gz)等对SP3基因组进行基因预测,获得15 864个转录本。
从NCBI数据库(https://www.ncbi.nlm.nih.gov/)中下载酿酒酵母中已报道的ATG的氨基酸序列,利用BioEdit软件将其与香菇单核菌株SP3蛋白序列进行本地BlastP比对,设置e-value为1e-5,获取相似性较高的香菇ATG蛋白序列。基于香菇SP3基因组的预测信息,以鉴定的香菇LeATG基因序列为参考,采用BWA软件(https://github.com/lh3/bwa)将转录组reads与之进行比对,结合GT-AG的内含子保守结构判断规则,获取基因的结构信息。将内含子剪切后的基因序列提交至NCBI数据库中的ORF finder (https://www.ncbi.nlm.nih.gov/orffinder/)在线网站预测开放阅读框,并获取校正后的氨基酸序列。采用GSDS 2.0 (http://gsds.gao-lab.org/index. php)在线网站绘制基因结构图。
基于香菇SP3基因组的注释文件及校正后的基因序列。采用MapGene2Chromosome软件(http://mg2c.iask.in/mg2c_v2.0/)绘制LeATG家族成员在基因组中的定位图。
将校正正确的LeATG氨基酸序列分别提交至Batch CD-Search (https://www.ncbi.nlm.nih.gov/Structure/bwrpsb/bwrpsb.cgi)和InterProScan (https://www.ebi.ac.uk/interpro/)数据库进行保守结构域预测和功能注释;采用ProtParam (https://web.expasy.org/protparam/)在线网站对LeATG氨基酸序列进行理化性质分析;采用CELLO v.2.5 (https://cello.life.nctu.edu.tw/)在线网站对LeATG蛋白进行亚细胞定位分析;采用DeepTMHMM- 1.0 (https://services.healthtech.dtu.dk/services/DeepTMHMM-1.0/)在线网站对LeATG蛋白进行跨膜结构分析;采用SignalP-4.1 (https://services.healthtech.dtu.dk/services/SignalP-4.1/)在线网站对蛋白进行信号肽预测。
从NCBI数据库中下载担子菌和子囊菌的ATG8氨基酸序列,采用Muscle方法分别对LeATG15成员及LeATG8家族成员蛋白进行多序列比对,并利用GeneDoc软件分析保守性。同时,采用MEME在线软件(https://meme-suite.org/meme/)预测其保守基序。
采用Muscle方法对鉴定的LeATG家族成员蛋白进行多序列比对,并利用MEGA11.0软件基于邻接法(neighbor-joining, NJ)构建成系统发育树,bootstrap设置为1 000。
将BIPP21020482菌株转接至90 mm的PDA平板中进行活化,25 ℃避光静置培养8 d后转接至装有1 200 g栽培料(78%细木屑、20%麦麸、1%石灰、1%绵白糖、55%含水量)的菌袋中进行栽培种制种,45 d后菌丝体长满菌袋。后续将等量栽培种接种至2 500 g菌棒(78%木屑、20%麦麸、1%石膏、1%轻钙,55%含水量)上,参照胡建平和陈青(2025)的方法进行出菇试验。待菌丝长满菌棒时,收集未转色菌丝;待菌丝完全转色后,收集褐色转色菌丝。每组样品共收集3个生物重复,并用液氮速冻保存,-80 ℃备用。
将上述两组样品委托武汉菲沙基因信息有限公司采用Illumina Hiseq平台进行RNA双端测序。原始测序数据Raw data经FastQC软件(https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)和fastap软件(https://github.com/OpenGene/fastp)进行质量评估与质控处理,获得高质量的Clean data。利用HISAT2软件(https://github. com/DaehwanKimLab/hisat2.git)将Clean data比对至SP3参考基因组。采用Samtools软件(https://github.com/lh3/samtools)将SAM文件转换为BAM文件并重新排序后,利用Featurecounts软件(http://subread.sourceforge.net/)对获得的每个基因在各个样本中的reads计数,采用TBtools软件(https://github.com/CJ-Chen/TBtools)将count值转换为FPKM值。基于百迈客云平台(https://international.biocloud.net/zh/dashboard)对样本进行相关性分析。抽提鉴定的LeATG家族成员基因表达量的FPKM值,采用基迪奥云平台(http://www.omicshare.com/tools/)绘制基因表达热图,分析LeATG家族成员基因的表达水平。
采用RNAsimple总RNA提取试剂盒(TIANGEN)提取菌株BIPP21020482在菌丝转色前后样品的总RNA,并采用PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time)试剂盒(TaKaRa)将RNA反转录为cDNA,以上操作方法均严格按照说明书进行。采用TB Green® Premix Ex Taq Ⅱ (Tli RNaseH Plus)定量试剂盒,在Biosystems® QuantStudio™ 7 Flex实时荧光定量PCR仪(赛默飞)上检测基因相对表达量,退火温度为60 ℃,循环40个。采用Primer Premier 5软件设计定量引物并委托生工生物工程(上海)股份有限公司合成,以β-微管蛋白基因(tubulin)为内参基因,引物序列见表1。基因的相对表达量采用2-ΔΔct方法进行计算(Livak & Schmittgen 2001)。
采用GraphPad Prism软件进行作图,基于Student’s t-test比较两组之间的差异显著性。两组间以概率值表示(*P<0.05, **P<0.01, ***P< 0.001)。
以NCBI数据库中下载的各亚家族酿酒酵母自噬相关蛋白序列为参考,与香菇SP3的全部蛋白序列进行同源比对,共鉴定出29个LeATG家族成员蛋白。转录组reads校正基因结构(表2),该家族成员序列全长介于595-6 772 bp,其中,LeATG2-1基因长度最长,LeATG12-1基因长度最短。外显子结构分析表明,6个家族成员基因LeATG4-1LeATG8-1LeATG12-1LeATG15-1LeATG27-1LeATG29-1分别包含 5个外显子,含有3、4、8和10个外显子的家族成员均有3个。经ORF finder预测后发现29个家族成员基因的完整开放阅读框长度为375-5 997 bp,5ʹ端非翻译区及3ʹ端非翻译区长度范围分别为1-1 339 bp和36-406 bp。基因序列结构示意图见图1
基于香菇SP3基因组的GFF注释文件,用MapGene2Chromosome软件分析LeATG家族成员基因在染色体中的位置。28个LeATG家族成员基因非均匀分布于9条染色体,1个定位于未组装成染色体的contig上,Chr09未见分布;Chr04、Chr01、Chr03和Chr08分布的基因数量较多,分别具有6、5、4和4个基因,而Chr06、Chr10、Chr05和Chr07分布的基因数量较少,分别具有2、2、1和1个基因(图2)。亚家族分析显示,3个LeATG15成员分布于2条染色体,其中LeATG15-1LeATG15-4位于Chr01,LeATG15-3位于Chr08;3个LeATG22成员分布于Chr03、Chr04和Chr08;2个LeATG2类型成员分布于Chr01和Chr08,说明同一亚家族成员无染色体区域化集中分布特征。
采用ProtParam等系列在线网站分析LeATG家族成员蛋白相关特征(表3),LeATG家族成员蛋白序列长度范围为127-1 998 aa,蛋白相对分子量为14.8-219.7 kDa,表明该家族成员间存在较大的结构差异。理论等电点范围为4.27-9.51,其中LeATG1-1、LeATG13-1、LeATG18-1、LeATG22-1、LeATG22-2和LeATG29-1蛋白的PI值大于7,分别为8.89、9.45、7.58、7.58、8.31和9.51,均为碱性蛋白。跨膜结构域结果显示,LeATG9、LeATG22和LeATG27亚家族成员均包含跨膜结构,分别为4、12和1个,其余24个LeATG蛋白不含跨膜结构。信号肽结果显示,LeATG15-2和LeATG15-4分别在第17-18位和第18-19位氨基酸残基间有信号肽,其余 27个LeATG蛋白均不含信号肽。亚细胞定位结果显示,有13个成员分布在细胞核中、有6个成员分布在质膜中、有5个成员分布在细胞质中、有4个成员分布在细胞外,还有1个成员分布在线粒体中。
利用InterProScan数据库和NCBI数据库中的Batch CD-Search在线网站对香菇29个LeATG的蛋白序列进行保守结构域分析,结果表明LeATG蛋白均具有亚家族的保守结构域,Batch CD-Search的预测结果见图3。LeATG1-1的N末端含激酶结构域;2个LeATG2的C端均包含自噬相关蛋白C末端结构域,在细胞质到液泡靶向和自噬中发挥作用;LeATG3-1和LeATG10-1均具有自噬相关蛋白活性位点结构域;LeATG4-1具肽酶催化结构域;LeATG5-1含“APG5结构域”,参与自噬囊泡形成;LeATG6-1包含ATG6卷曲螺旋区和“Apg6 BARA结构域”,在自噬中起到重要作用;LeATG7-1具有非典型的E1酶结构域;LeATG8-1和LeATG12-1均包含泛素样(Ubl)结构域;LeATG9-1、ATG13-1、ATG16-1和ATG27-1分别属于相应自噬相关蛋白ATG9、ATG13、ATG16和ATG27超家族;LeATG11-1和LeATG17-1均包含自噬蛋白ATG17样结构域;4个LeATG15均包含脂肪酶催化结构域;LeATG18-1具有WD40结构域;LeATG20-1和LeATG24-1均包含PX和BAR结构域;3个LeATG22均具有主要促进因子MFS结构域;LeATG26-1包含PH-GRAM1_AGT26结构域;LeATG29-1具有ATG29的N端结构域,可以与ATG31相结合。
LeATG8作为自噬过程中的核心蛋白,我们对其进行解析。将香菇LeATG8与酿酒酵母Saccharomyces cerevisiae、双色蜡蘑Laccaria bicolor和红菇Russula earlei的ATG8蛋白进行多序列比对,预测其保守基序(图 4)。所有比对序列的N端及中间区域保守性较高,而C端区域变异较强。其中,属于“ubiquitin-like (Ubl) domain”的序列在所有物种中均高度保守。保守基序分析显示,4条序列均包含Motif1、Motif2和Motif3这3个核心基序。此外,所有担子菌(香菇、双色蜡蘑和红菇)均含有Motif4基序,而香菇和双色蜡蘑还共同具有特有的Motif5基序,表示二者在ATG8蛋白结构上更为接近,可能反映了功能上的细化。
为进一步探究香菇内LeATG家族成员蛋白的进化关系,采用NJ法构建29个LeATG蛋白序列的系统发育树(图5),该家族可分为3个进化分支,其中LeATG2、LeATG5、LeATG12、LeATG13和LeATG24聚集在CladeⅠ分支中;LeATG4、LeATG9、LeATG10、LeATG17、LeATG18、LeATG22和LeATG27聚集在CladeⅡ分支;剩余10类亚家族聚集在Clade Ⅲ分支,包含14个LeATG蛋白。值得注意的是,LeATG2、LeATG15和LeATG22为多拷贝蛋白亚家族,呈现差异化进化模式,其中4个LeATG15家族成员亲缘关系较近,在Clade Ⅲ内形成高支持度的小分支,置信度在98以上;2个LeATG2和3个LeATG22家族成员虽分别同属CladeⅠ和CladeⅡ分支,却分散于不同亚支,表明同一亚家族的蛋白结构可能存在显著差异。
采用百迈客云平台对未转色菌丝和转色菌丝样品获得的FPKM值进行相关性分析(图6A),组内样品的相关性在0.923-0.965之间,说明样品组内相关性较强,重复性较好;组间样品的相关性在0.673-0.87之间,表明两组之间差异较大,为后续探索样品间的差异性提供数据支撑。抽提LeATG家族成员基因在菌丝转色前后FPKM值,分析其表达情况(图6B6C),29个家族成员在两组样品中的表达模式不同,其中22个基因无显著差异;LeATG22-3LeATG27-1基因在菌丝转色前高表达,表达量分别是菌丝未转色样品的13.46倍和43.03倍;LeATG7-1LeATG8-1LeATG15-1LeATG15-3LeATG29-1在菌丝转色后高表达,相比于未转色样品表达量分别提高了47.02%、58.09%、57.94%、320.27%和34.72%,且LeATG15-3差异最显著。推测以上7个基因可能参与调控菌丝的转色过程。此外,选取在菌丝转色前高表达的LeATG27-1基因与在菌丝转色后高表达的LeATG8-1基因进行了RT-qPCR验证(图6C),二者的表达趋势与转录组数据分析结果一致,从而证实了本研究转录组数据的可靠性,可用于后续深入分析。
自噬是一种重要的细胞过程,它通过降解和回收细胞内成分,在维持细胞稳态、免疫防御、代谢平衡和调控细胞死亡等方面起到重要作用(郑祖国和张评浒 2016;Bozhkov 2018)。随着对酵母(Sheng & Qin 2019)、小鼠(王晗等 2019)、病原真菌(刘伟和杜春梅 2021)和果蝇(Katz et al. 2025)等多种物种中自噬相关基因的深入研究,发现不同物种中自噬相关基因的数量及种类存在差异。在草地贪夜蛾中共鉴定了18个ATG家族成员,其中16个为参与自噬体膜形成的核心自噬相关基因,不包含ATG29ATG31 (马秋琴等 2021);王晗等(2019)基于小鼠精子发生的3个时期转录组测序数据,共鉴定了636个自噬相关基因,且其在精子发生过程中存在3种主要表达模式;Young et al. (2019)通过拟南芥突变体库正向筛选了21个ATG基因,包含1个ATG3、2个ATG2、2个ATG5、2个ATG18a、3个ATG16和11个ATG7;在禾谷镰孢菌中共鉴定到28个ATG家族成员,其中18个参与其生长过程,研究还进一步表明除了FgATG17基因外,其余任何ATG基因的缺失均导致该真菌无法引发镰孢菌头枯病(Lv et al. 2017);Kershaw & Talbot (2009)在稻瘟病菌中构建了22个核心自噬基因突变体,发现其中16个为非选择性巨噬细胞必需基因,缺失其中任何一个均会导致自噬过程受阻,影响分生孢子程序性细胞死亡和附着胞成熟。本研究中基于香菇SP3单核菌株的全基因组数据,通过同源比对及生物信息学分析共鉴定到29个LeATG家族成员,属于23类自噬相关基因,在自噬体形成过程中起关键作用的18种自噬相关基因中(Suzuki et al. 2017),我们成功注释到17种,但未发现LeATG14的存在。香菇中LeATG14的缺失是否会影响自噬体的正常形成过程,仍需进一步地深入研究。
菌丝转色是香菇生长发育过程中特有的关键生理环节,也是由其营养生长转向为子实体发育的重要标志(Hong et al. 2020)。该过程是指白色菌丝长满菌棒表面后,在适宜的环境条件下,气生菌丝和瘤状物发生细胞死亡,同时伴随着色素的合成与分泌,最终在菌棒表面形成一层棕褐色菌膜(刘阳 2025)。菌丝转色在保护菌丝、内部支撑和催蕾促菇等方面起到重要作用(王贺祥和刘庆洪 2014)。Sano et al. (2009)研究指出香菇转色是色素及醌类物质积累的过程,本团队前期研究发现在转色后的菌丝超微结构中存在4种类黑素体和自噬体类似结构,推测细胞自噬可能参与黑素体形成,黑色素以黑素体的形式在细胞内运输并积累,最终导致菌丝转色发生(Gao et al. 2019;Yan et al. 2020)。多项研究表明自噬相关基因与蛋白在香菇菌丝转色过程中发挥重要调控作用。Tang et al. (2022)通过蛋白质组学和代谢组学分析发现,在菌丝转色样品中自噬相关的蛋白质(RPD3、TOR1、VAC8、VPS1 和 VPS27)表达显著上调。Yan et al. (2023)通过外源H2O2诱导菌丝氧化应激实验证实高渗透压甘油蛋白LeHOG1的活化/磷酸化和自噬相关蛋白LeATG8的表达协同调节ROS诱导的自噬过程,进而参与调控香菇的老化和转色;此外,荧光定量PCR和western blot分析结果显示转色菌丝中ATG8基因和蛋白表达水平均显著高于未转色菌丝,进一步证实ATG8在香菇菌丝转色过程中的关键作用(房丽丽等 2017;管婉等 2022a, 2022b)。本研究通过对菌丝转色前后的转录组分析发现,多数自噬相关基因表达稳定,可能主要维持细胞内基本的自噬过程;7个基因呈现差异表达模式,其中LeATG22-3LeATG27-1基因在菌丝转色前表达较高,LeATG7-1LeATG8-1LeATG15-1LeATG15-3LeATG29-1则在菌丝转色后显著上调。推测以上7个自噬相关基因可能共同调控菌丝的转色过程。ATG7基因编码E1样泛素激活酶,参与ATG8和ATG12的共价结合过程,Yu et al. (2022)敲除稻绿核菌中的UvAtg7基因后会导致自噬降解受阻,菌丝生长变慢,气生菌丝稀疏,分生孢子数量减少,但回补UvAtg7基因后菌丝表型恢复;ATG8基因编码的家族蛋白是自噬过程的关键参与者,其不仅与磷脂酰乙醇胺(PE)结合定位于自噬体膜上,参与调控自噬发生的各个环节(Bu et al. 2020),还能应用于自噬体标记和细胞自噬程度评价研究(赵修明等 2021),在黄曲霉中,ATG8基因缺失会导致菌丝生长和分生孢子形成显著减少(Geng et al. 2024);Yu et al. (2023)敲除荔枝霜疫霉中的PlAtg8基因后,菌落生长速率、孢子囊数量和游动孢子释放率显著降低;ATG15是一种脂酶,参与自噬体降解,赫可伟等(2021)沉默拟轮枝镰孢的Atg15基因后,孢子的萌发率和菌丝生长速度明显受到抑制。综上研究表明,自噬相关基因ATG7ATG8ATG15在多种真菌的菌丝生长、发育、细胞分化和次生代谢中发挥了关键作用,而在本研究中,属于这3类基因中的4个同源基因在香菇菌丝转色后表达显著上调,该结果进一步说明其可能共同参与调控菌丝转色过程,但其具体的功能机制还需进一步深入研究。
本研究对香菇29个LeATG家族成员进行了系统性分析,发现LeATG15为多蛋白亚家族。进一步对该亚家族4个成员的分析表明,其亚细胞定位与表达模式存在显著差异。ATG15是一种液泡磷脂酶,对于细胞质至液泡靶向(Cvt)体和自噬体的降解至关重要(Watanabe et al. 2023;Watanabe & Suzuki 2024)。Epple et al. (2001)和Marquardt et al. (2023)通过构建带有荧光标记的酿酒酵母突变株证实ATG15位于液泡,这与本研究基于香菇的预测结果在细胞外存在差异。我们推测ATG15不属于自噬过程中的核心蛋白(Suzuki et al. 2017),其在不同物种间可能存在功能分化,从而导致亚细胞定位的差异。转录组数据显示LeATG15-1LeATG15-3基因在菌丝转色后表达量显著上升,而LeATG15-2LeATG15-4基因在转色前后表达均无显著差异(图 7A)。为探索其表达差异的潜在机制,我们开展了序列比对分析,结果显示4个LeATG15成员在核酸水平的一致性为42.27%,在氨基酸水平上的一致性为44.97%,表明该家族成员间序列差异较大,推测其结构差异可能是导致表达模式不同的重要原因。蛋白多序列比对进一步发现(图 7B),这些LeATG15蛋白的C端保守性较高,而N端具有较强的变异性。保守基序分析表明(图7C),4个成员共包含4个共有的保守Motif基序(Motif1、Motif2、Motif3、Motif4),且均靠近C端。值得注意的是,除LeATG15-2外,其余3个成员均含有Motif5,据此推测LeATG15-2基因在转色前后表达量较低且无显著变化,可能与缺乏Motif5基序有关。而LeATG15-4基因在转色过程中表达未发生明显变化,是否也与其结构特征相关还需进一步的实验验证。本研究结果与植物中自噬相关基因的研究相似,Wang et al. (2024)通过对茄子全基因数据分析,共鉴定到7个SmATG8基因,其氨基酸序列长度介于118-166 aa之间,当受到低温、高温及盐胁迫等非生物胁迫时,不同的SmATG8基因会表现出明显的表达差异。本研究结果进一步支持自噬相关基因家族在进化过程中可能存在功能分化现象,后续研究可通过基因功能验证实验进一步深入探索。
  • 国家自然科学基金(32202568)
  • 北京市农林科学院植物保护研究所改革与发展项目(GGYFZ202512)
  • 现代农业产业技术体系北京市食用菌创新团队(BAIC03)
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2026年第45卷第4期
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doi: 10.13346/j.mycosystema.250259
  • 接收时间:2025-08-29
  • 首发时间:2026-04-30
  • 出版时间:2026-04-22
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  • 收稿日期:2025-08-29
  • 录用日期:2025-11-04
基金
National Natural Science Foundation of China(32202568)
国家自然科学基金(32202568)
Reform and Development Project, Institute of Plant Protection, Beijing Academy of Agriculture and Forestry Sciences(GGYFZ202512)
北京市农林科学院植物保护研究所改革与发展项目(GGYFZ202512)
Beijing Innovation Consortium of Agriculture Research System(BAIC03)
现代农业产业技术体系北京市食用菌创新团队(BAIC03)
作者信息
    1 北京市农林科学院植物保护研究所 北京市食用菌工程技术研究中心, 北京 100097
    2 牡丹江师范学院生命科学与技术学院, 黑龙江 牡丹江 157011

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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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