Article(id=1276601418417046123, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276601397818814642, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2025.01.018, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1720454400000, receivedDateStr=2024-07-09, revisedDate=1722528000000, revisedDateStr=2024-08-02, acceptedDate=null, acceptedDateStr=null, onlineDate=1782295096257, onlineDateStr=2026-06-24, pubDate=1737734400000, pubDateStr=2025-01-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782295096257, onlineIssueDateStr=2026-06-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782295096257, creator=13701087609, updateTime=1782295096257, updator=13701087609, issue=Issue{id=1276601397818814642, tenantId=1146029695717560320, journalId=1235980609244409860, year='2025', volume='46', issue='1', pageStart='1', pageEnd='245', issueExtLink='null', onlineDate='null', pubDate='1737734400000', pubDateStr='2025-01-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782295091347, creator='13701087609', updateTime=1782295207335, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276601884408418422, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276601397818814642, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276601884408418423, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276601397818814642, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=171, endPage=179, ext={EN=ArticleExt(id=1276601418702258797, articleId=1276601418417046123, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Biological Characteristics and Acclimation Cultivation of Auricularia villosula, columnId=1236256434120348225, journalTitle=Chinese Journal of Tropical Crops, columnName=Plant Cultivation, Physiology & Biochemistry, runingTitle=null, highlight=null, articleAbstract=

Auricularia villosula Malysheva is a similar species of Auricularia heimuer F. Wu, B.K. Cui & Y. C. Dai, mainly distributed in high altitude areas of tropical and subtropical regions. It can adapt to the high temperature and high humidity climate in Guangxi. There are few reports on the domestication of A. villosula. In this paper, the fruiting bodies of auriculae collected in the field was used, and the molecular marker was established by sequence analysis. The effects of different temperature, carbon and nitrogen sources on the mycelia growth of A. villosula were explored through single factor tests, and the carbon and nitrogen sources of the medium were further optimized by an orthogonal test. The fruiting body of fungus was preliminarily identified as A. villosula (TY081). The results showed that the developed molecular labeling method could quickly and effectively identify A. villosula. The optimal temperature for mycelia growth of A. villosula TY081 was 30 ℃, the optimal carbon source was glucose, and the optimal nitrogen source was soybean powder. The optimized medium carbon and nitrogen combination was fructose, yeast powder, mycelium growth rate reached 0.92 mm/d. A. villosula TY081 was incubated at 28 ℃ in a culture bag mainly composed of sawdust, and the bag was full for 60 days. After puncturing, it was cultured in a greenhouse for 10 days to form the primordia. The fruity body growth temperature was 24 ℃, the humidity was 85%, and the yield of each bag was 20.943 g (dry weight).

, authors=null, authorsList=Chenhong NIE, Bingzhao ZHOU, Jing FENG, Shiyan WEI, Liangliang QI, Xiaoguo WANG, authorCompany=null, correspAuthors=Xiaoguo WANG, 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, fund=null), CN=ArticleExt(id=1276601422334526078, articleId=1276601418417046123, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=短毛木耳的生物学特性与驯化栽培研究, columnId=1236256434313286224, journalTitle=热带作物学报, columnName=作物栽培与生理生化, runingTitle=null, highlight=null, articleAbstract=

短毛木耳是黑木耳的近似种,主要分布于热带、亚热带高海拔地区,能较好地适应广西高温高湿的气候,目前有关短毛木耳驯化栽培的报道较少。本研究以野外采集的短毛木耳子实体(TY081)为材料,采用序列分析方法,建立短毛木耳鉴定的分子标记;通过单因素试验探究不同温度、碳源、氮源对短毛木耳菌丝生长的影响,用正交试验对培养基的碳、氮源进一步优化。结果表明:开发的分子标记方法可以快速有效地鉴别出短毛木耳;短毛木耳TY081菌丝生长的最适温度是30 ℃,最适碳源为葡萄糖,最适氮源为黄豆粉;优化后的培养基碳氮组合为果糖、酵母浸粉,菌丝生长速度达0.92 mm/d。短毛木耳TY081在栽培菌袋中的发菌温度为28 ℃,培养60 d满袋,刺孔后大棚培养10 d形成原基,子实体生长温度为24 ℃,湿度为85%,每袋的产量为20.943 g(干质量)。

, authors=

聂晨红(2000—),女,硕士研究生,研究方向:食用菌遗传育种。

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* 王晓国(WANG Xiaoguo),E-mail:
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3.广西壮族自治区农业科学院微生物研究所,广西南宁 530007, bio={"content":"

聂晨红(2000—),女,硕士研究生,研究方向:食用菌遗传育种。

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聂晨红(2000—),女,硕士研究生,研究方向:食用菌遗传育种。

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(in Chinese), articleTitle=Study on biological characteristics and acclimation culture of Rosinomyces rosin, refAbstract=null), Reference(id=1276601945238401947, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, doi=null, pmid=null, pmcid=null, year=2024, volume=45, issue=5, pageStart=973, pageEnd=982, url=null, language=null, rfNumber=[32], rfOrder=54, authorNames=刘潮, 田雪莲, 唐仕香, 杨才静, 韩利红, journalName=热带作物学报, refType=null, unstructuredReference=刘潮, 田雪莲, 唐仕香, 杨才静, 韩利红. 一株野生蝉花的生物学特性及驯化栽培研究[J]. 热带作物学报, 2024, 45(5): 973-982., articleTitle=一株野生蝉花的生物学特性及驯化栽培研究, refAbstract=null), Reference(id=1276601945313899420, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, doi=null, pmid=null, pmcid=null, year=2024, volume=45, issue=5, pageStart=973, pageEnd=982, url=null, language=null, rfNumber=[32], rfOrder=55, authorNames=LIU C, TIAN X L, TANG S X, YANG C J, HAN L H, journalName=Chinese Journal of Tropical Crops, refType=null, unstructuredReference=LIU C, TIAN X L, TANG S X, YANG C J, HAN L H. Biological characteristics and domestication of a wild Cordyceps chanhua[J]. Chinese Journal of Tropical Crops, 2024, 45(5): 973-982. 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1: A. heimuer, 7 and 52: A. villosula.

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1:黑木耳,7和52:短毛木耳。

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M: DL2000 DNA marker; 1, 2, 9, 12: A. heimuer; 3: A. cornea; 6, 8: A. delicata; 4, 5, 7, 10, 11: A. villosula.

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M:DL2000 DNA marker;1、2、9、12:黑木耳;3:毛木耳;6、8:皱木耳;4、5、7、10、11:短毛木耳。

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A represents the carbon source of the basic medium, B-H represent the carbon source of glucose, fructose, sucrose, maltose, lactose, starch and trehalose medium, respectively.

, figureFileSmall=hQo55S/aXeSuO9HZzIHAOg==, figureFileBig=3NmxsagNKQM4qPHqZa1JxA==, tableContent=null), ArticleFig(id=1276601933502739280, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=CN, label=图4, caption=碳源对菌丝生长的影响

A表示碳源基础培养基,B~H分别表示碳源为葡萄糖、果糖、蔗糖、麦芽糖、乳糖、淀粉和海藻糖的培养基。

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A represents the nitrogen source of the basic medium, B-H represents the nitrogen source of peptone, potassium nitrate, yeast extract powder, ammonium nitrate, urea, beef paste and soybean powder medium, respectively.

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A表示氮源基础培养基,B~H分别表示氮源为蛋白胨、硝酸钾、酵母浸粉、硝酸铵、尿素、牛肉膏和黄豆粉的培养基。

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A, B and C represent fructose, lactose and maltose respectively; X, Y, Z represent beef paste, soybean powder, yeast infusion powder respectively.

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A、B、C分别代表果糖、乳糖、麦芽糖;X、Y、Z分别代表牛肉膏、黄豆粉、酵母浸粉。

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Orthogonal experiment design of mycelia growth of A. villosula

, figureFileSmall=null, figureFileBig=null, tableContent=
水平Level因素Factor
碳源Carbon source氮源Nitrogen source
1果糖A牛肉膏X
2葡萄糖B黄豆粉Y
3麦芽糖C酵母浸粉Z
), ArticleFig(id=1276601936291951448, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=CN, label=表1, caption=

短毛木耳菌丝生长的正交试验设计

, figureFileSmall=null, figureFileBig=null, tableContent=
水平Level因素Factor
碳源Carbon source氮源Nitrogen source
1果糖A牛肉膏X
2葡萄糖B黄豆粉Y
3麦芽糖C酵母浸粉Z
), ArticleFig(id=1276601936405197657, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=EN, label=Tab. 2, caption=

Effects of different temperatures on mycelium

, figureFileSmall=null, figureFileBig=null, tableContent=
温度Temperature/℃菌丝生长速率Mycelial growth rate/(mm·d–1)菌丝长势Mycelium growth state
350.34±0.01d+
301.12±0.03a+++
250.88±0.03b+++
200.70±0.09c++
150.44±0.06d++
), ArticleFig(id=1276601936510055258, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=CN, label=表2, caption=

不同温度对菌丝体的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
温度Temperature/℃菌丝生长速率Mycelial growth rate/(mm·d–1)菌丝长势Mycelium growth state
350.34±0.01d+
301.12±0.03a+++
250.88±0.03b+++
200.70±0.09c++
150.44±0.06d++
), ArticleFig(id=1276601936581358427, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=EN, label=Tab. 3, caption=

Effects of different carbon sources on mycelium growth

, figureFileSmall=null, figureFileBig=null, tableContent=
碳源Carbon source菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state碳源Carbon source菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state
果糖3.30±0.02a+++淀粉3.05±0.01ab+++
麦芽糖3.27±0.02ab+++蔗糖2.94±0.00ab+++
葡萄糖3.25±0.01ab+++对照2.76±0.05b++
海藻糖3.14±0.05ab+++乳糖2.02±0.52c+
), ArticleFig(id=1276601936661050204, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=CN, label=表3, caption=

不同碳源对菌丝生长的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
碳源Carbon source菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state碳源Carbon source菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state
果糖3.30±0.02a+++淀粉3.05±0.01ab+++
麦芽糖3.27±0.02ab+++蔗糖2.94±0.00ab+++
葡萄糖3.25±0.01ab+++对照2.76±0.05b++
海藻糖3.14±0.05ab+++乳糖2.02±0.52c+
), ArticleFig(id=1276601936723964765, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=EN, label=Tab. 4, caption=

Effects of different nitrogen sources on mycelium growth

, figureFileSmall=null, figureFileBig=null, tableContent=
氮源Nitrogen source菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state
CK2.62±0.03c++
黄豆粉3.21±0.05a+++
酵母浸粉2.97±0.04b+++
牛肉膏2.13±0.04d++
硝酸钾2.11±0.01d++
硝酸铵1.71±0.18e++
蛋白胨1.41±0.09f+
尿素0.60±0.03g+
), ArticleFig(id=1276601936786879326, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=CN, label=表4, caption=

不同氮源对菌丝生长的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
氮源Nitrogen source菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state
CK2.62±0.03c++
黄豆粉3.21±0.05a+++
酵母浸粉2.97±0.04b+++
牛肉膏2.13±0.04d++
硝酸钾2.11±0.01d++
硝酸铵1.71±0.18e++
蛋白胨1.41±0.09f+
尿素0.60±0.03g+
), ArticleFig(id=1276601936849793887, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=EN, label=Tab. 5, caption=

Growth rate of mycelia in orthogonal experiment

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正交组合Orthogonal combination菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state
AZ0.92±0.03a+++
CZ0.85±0.12ab+++
CY0.80±0.19ab+++
BY0.72±0.06abc+++
BZ0.65±0.05abcd+++
AY0.61±0.03bcd+++
BX0.51±0.03cd++
CX0.45±0.14cd+
AX0.37±0.08d+
), ArticleFig(id=1276601936925291360, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=CN, label=表5, caption=

正交试验菌丝的生长速度

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正交组合Orthogonal combination菌丝生长速率Mycelial growth rate/(mm·d-1)菌丝长势Mycelium growth state
AZ0.92±0.03a+++
CZ0.85±0.12ab+++
CY0.80±0.19ab+++
BY0.72±0.06abc+++
BZ0.65±0.05abcd+++
AY0.61±0.03bcd+++
BX0.51±0.03cd++
CX0.45±0.14cd+
AX0.37±0.08d+
), ArticleFig(id=1276601936996594529, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=EN, label=Tab. 6, caption=

Comparison of yields of A. heimuer and A. villosula

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种类Species编号Code每袋干质量Dry weight of each bag/g每袋鲜质量Wet weight of each bag/g干质量/鲜质量Dry weight/wet weight
黑木耳TY91623.319155.5356.67
黑木耳TY02610.60083.8467.91
短毛木耳TY08120.943171.5238.19
), ArticleFig(id=1276601937088869218, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276601418417046123, language=CN, label=表6, caption=

黑木耳与短毛木耳的产量对比

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种类Species编号Code每袋干质量Dry weight of each bag/g每袋鲜质量Wet weight of each bag/g干质量/鲜质量Dry weight/wet weight
黑木耳TY91623.319155.5356.67
黑木耳TY02610.60083.8467.91
短毛木耳TY08120.943171.5238.19
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短毛木耳的生物学特性与驯化栽培研究
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聂晨红 1, 3 , 周炳沼 2, 3 , 丰景 1 , 韦仕岩 3 , 祁亮亮 3 , 王晓国 3, *
热带作物学报 | 作物栽培与生理生化 2025,46(1): 171-179
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热带作物学报 |作物栽培与生理生化 2025 , 46 (1) : 171 -179
短毛木耳的生物学特性与驯化栽培研究
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聂晨红1, 3, 周炳沼2, 3, 丰景1, 韦仕岩3, 祁亮亮3, 王晓国3, *
作者信息
  • 1.广西民族大学海洋生物与技术学院,广西南宁 530006
  • 2.贺州学院食品与生物工程学院,广西贺州 542899
  • 3.广西壮族自治区农业科学院微生物研究所,广西南宁 530007
通讯作者:
* 王晓国(WANG Xiaoguo),E-mail:
Biological Characteristics and Acclimation Cultivation of Auricularia villosula
Chenhong NIE1, 3, Bingzhao ZHOU2, 3, Jing FENG1, Shiyan WEI3, Liangliang QI3, Xiaoguo WANG3, *
Affiliations
  • 1.College of Marine Biology and Technology, Guangxi University of Nationalities, Nanning, Guangxi 530006, China
  • 2.College of Food and Bioengineering, Hezhou University, Hezhou, Guangxi 542899, China
  • 3.Institute of Microbiology, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi 530007, China
出版时间: 2025-01-25 doi: 10.3969/j.issn.1000-2561.2025.01.018
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短毛木耳是黑木耳的近似种,主要分布于热带、亚热带高海拔地区,能较好地适应广西高温高湿的气候,目前有关短毛木耳驯化栽培的报道较少。本研究以野外采集的短毛木耳子实体(TY081)为材料,采用序列分析方法,建立短毛木耳鉴定的分子标记;通过单因素试验探究不同温度、碳源、氮源对短毛木耳菌丝生长的影响,用正交试验对培养基的碳、氮源进一步优化。结果表明:开发的分子标记方法可以快速有效地鉴别出短毛木耳;短毛木耳TY081菌丝生长的最适温度是30 ℃,最适碳源为葡萄糖,最适氮源为黄豆粉;优化后的培养基碳氮组合为果糖、酵母浸粉,菌丝生长速度达0.92 mm/d。短毛木耳TY081在栽培菌袋中的发菌温度为28 ℃,培养60 d满袋,刺孔后大棚培养10 d形成原基,子实体生长温度为24 ℃,湿度为85%,每袋的产量为20.943 g(干质量)。

短毛木耳  /  分子验证  /  生物学特性  /  驯化栽培

Auricularia villosula Malysheva is a similar species of Auricularia heimuer F. Wu, B.K. Cui & Y. C. Dai, mainly distributed in high altitude areas of tropical and subtropical regions. It can adapt to the high temperature and high humidity climate in Guangxi. There are few reports on the domestication of A. villosula. In this paper, the fruiting bodies of auriculae collected in the field was used, and the molecular marker was established by sequence analysis. The effects of different temperature, carbon and nitrogen sources on the mycelia growth of A. villosula were explored through single factor tests, and the carbon and nitrogen sources of the medium were further optimized by an orthogonal test. The fruiting body of fungus was preliminarily identified as A. villosula (TY081). The results showed that the developed molecular labeling method could quickly and effectively identify A. villosula. The optimal temperature for mycelia growth of A. villosula TY081 was 30 ℃, the optimal carbon source was glucose, and the optimal nitrogen source was soybean powder. The optimized medium carbon and nitrogen combination was fructose, yeast powder, mycelium growth rate reached 0.92 mm/d. A. villosula TY081 was incubated at 28 ℃ in a culture bag mainly composed of sawdust, and the bag was full for 60 days. After puncturing, it was cultured in a greenhouse for 10 days to form the primordia. The fruity body growth temperature was 24 ℃, the humidity was 85%, and the yield of each bag was 20.943 g (dry weight).

Auricularia villosula  /  molecular verification  /  biological characteristics  /  acclimation culture
聂晨红, 周炳沼, 丰景, 韦仕岩, 祁亮亮, 王晓国. 短毛木耳的生物学特性与驯化栽培研究. 热带作物学报, 2025 , 46 (1) : 171 -179 . DOI: 10.3969/j.issn.1000-2561.2025.01.018
Chenhong NIE, Bingzhao ZHOU, Jing FENG, Shiyan WEI, Liangliang QI, Xiaoguo WANG. Biological Characteristics and Acclimation Cultivation of Auricularia villosula[J]. Chinese Journal of Tropical Crops, 2025 , 46 (1) : 171 -179 . DOI: 10.3969/j.issn.1000-2561.2025.01.018
木耳属(Auricularia)是世界广泛分布的木生真菌,目前中国已有的木耳属种类为15个,而世界范围内的木耳种类有27个,其中主要包括黑木耳(A. heimuer)、毛木耳[A. polytricha(Mont.)Sacc.]、皱木耳[A. delicata(Mont. ex Fr.)Henn.]、脆木耳(A. fibrillifera Kobayasi,Bull.)和短毛木耳(A. villosula[1]。木耳属主要生长在热带、亚热带、温带以及海拔500~1000 m的山区森林中的阔叶树、死树、树桩、倒木上、枯枝或腐烂木,偶尔也生长在竹子上[2-3]。毛木耳和黑木耳是木耳属中我国主要栽培的品种,黑木耳的人工栽培历史已有1300多年[4],据中国食用菌协会统计,2021年全国黑木耳产量约为712.3万t,毛木耳产业年产量近190万t,其中仅广西占总产量的17.14%[5]
木耳属中的大多数均可食用,其质地滑嫩营养丰富,含有多种人体必需的氨基酸、矿物质和膳食纤维等,并且可以分离出多种活性物质[6-8]。木耳不但有重要的食用价值,而且具有多种药用功能,随着现代药理学研究的不断深入,木耳被证实了具有免疫调节、降血脂血糖、抗凝血、抗氧化的作用,以及具有清肺解毒的功效[9-12],因此在开发保健品等方面具有重要的研究价值。
随着近年对木耳属其他种类的驯化,如黑木耳、皱木耳、脆木耳、短毛木耳亦被驯化成功并开始规模化生产[11,13-14]。但南方如广西的湿热气候使栽培存在挑战,限制了生产规模和经济效益[15-17]。短毛木耳是黑木耳的近似种,研究发现栽培黑木耳中混进了短毛木耳,说明短毛木耳也可以人工种植[1]。短毛木耳主要分布在亚热带高海拔地区,在生态适应上能够适应高温高湿的环境,因此能更好地适应广西高温高湿的气候。目前有关短毛木耳的研究相对较少,驯化栽培方面更是未见报道。本研究通过深入研究短毛木耳的生物学特性,优化栽培技术,以期推动广西木耳产业的发展,为木耳产业的多样化和可持续发展提供支持。
本研究所用菌株黑木耳916、黑木耳TY026、毛木耳TY193、短毛木耳TY012、皱木耳TY504、短毛木耳TE011、皱木耳TY503、黑木耳黑TY003、短毛木耳1413、短毛木耳1416、黑木耳新科4号保藏于广西农业科学院微生物研究所。短毛木耳TY081保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO.15278。
碳源基础培养基:20 g琼脂,2 g酵母浸粉,1000 mL水。碳源:葡萄糖、果糖、蔗糖、麦芽糖、乳糖、淀粉、海藻糖。氮源基础培养基:20 g琼脂,20 g葡萄糖,1000 mL水。氮源:蛋白胨、酵母浸粉、硝酸钾、硝酸铵、尿素、牛肉膏、黄豆粉。栽培料:70%杂木屑、10%麦麸、0.5%石膏、0.5%石灰、10%棉籽壳。
用试剂盒法提取供试子实体和对应菌丝体的DNA并进行PCR扩增和ITS序列测定,测定的结果在NCBI中进行序列比对。PCR用的引物为ITS1/ITS4,ITS1(5'-TCCGTAGGTGAACCTGCGG-3'),ITS4(5'-TCCTCCGCT TATTGATATGC-3')。根据短毛木耳与黑木耳碱基序列的不同区域,并以此为模板,反向设计引物,设计的引物为:TY081-F(5ʹ-CGAAAGTCCAGAATGTGATCTA-3ʹ),TY081-R(5ʹ-AGCACTCTAAAAGCGCCAGCTAATG-3ʹ)共293 bp,通过特异引物鉴定以达到快速鉴定出短毛木耳的目的。在PCR反应过程中,PCR扩增反应采用30 μL体系,反应条件为:95 ℃预变性3 min,95 ℃变性30 s;56 ℃退火30 s;72 ℃延伸30 s;设29个循环,72 ℃终延伸7 min。反应结束后用1%的琼脂糖凝胶电泳检测PCR产物。
(1)温度对菌丝体生长的影响。试验用培养基为PDA培养基,共设5个培养条件(15、20、25、30、35 ℃)。接入菌种后于28 ℃培养1 d,然后置于5个温度进行培养,每个处理重复5次。
(2)碳源对菌丝体生长的影响。以碳源基础培养基为空白对照组(CK),分别设置葡萄糖、果糖、蔗糖、麦芽糖、乳糖、淀粉、海藻浓度均为20 g/L,每个处理重复5次。接种后于28 ℃进行恒温培养。
(3)氮源对菌丝体生长的影响。以氮源基础培养基为空白对照组(CK),分别设置蛋白胨、酵母浸粉、硝酸钾、硝酸铵、尿素、黄豆粉、牛肉膏,浓度均为2 g/L。
(4)正交试验优化。采用L9(32)正交试验在以上的碳源、氮源2个因素中各选出3个水平,进行正交试验。选用的碳源为果糖、葡萄糖、麦芽糖,氮源为牛肉膏、黄豆粉、酵母浸粉(表1)。每个处理重复6次。
测量方法:用“米”字划线法测量菌落的直径,每隔2 d测量1次并做好标记,下一次测量的起始点为上一次测量的标记点,连续测量10 d,并观察菌丝长势,直至菌丝长满培养基。
将组织分离得到的菌种接入斜面PDA培养基中,28 ℃暗培养至菌丝长满,得到生产母种。木屑提前24 h预湿,依次加入麦麸、石膏、石灰、棉籽壳、水,混匀至含水量为58%~60%,每袋装3/4。灭菌冷却至28 ℃以下,于接种室打孔接种。每袋打4个孔,按“井”字形堆放,相对湿度低于65%,25~29 ℃遮光培养,60 d即可长满菌丝。菌丝长满后刺孔,培养7~14 d。原基形成后可摆地出耳,成熟即可采收。
利用SPSS 27软件对试验数据进行统计分析,在单因素和正交试验方差分析中,P<0.05代表差异显著。
使用引物ITS1/ITS4进行ITS序列鉴定,通过NCBI中的Blast对比样品的ITS和GenBank中的短毛木耳(Auricularia villosula),一致性为525/528(99%),初步推断采集到的野生木耳子实体为短毛木耳。短毛木耳ITS序列的121为C,131~134为TCTA,这与黑木耳、毛木耳等其他木耳属种类的序列不同(图1)。以此为标记反向设计引物可以快速鉴别出短毛木耳。将短毛木耳与黑木耳、毛木耳等12个不同的DNA进行ITS检测和特异引物检测,结果表明不同品种的木耳均可以用ITS测序法测出,而在使用特异引物的情况下能鉴别出来的只有短毛木耳(图2)。
菌丝在15~30 ℃内均能生长,且随着温度的升高菌丝生长速度逐渐加快。15 ℃时菌丝生长缓慢;20 ℃时菌丝生长相对缓慢;25 ℃时菌丝的生长快且菌丝洁白浓密;30 ℃时菌丝生长最快,但菌丝长势较差;30 ℃之后,随着培养温度的升高菌丝生长速度逐渐减慢,甚至死亡;在35 ℃时菌丝几乎不生长,推测高温度可导致菌丝死亡(图3)。其生长速度表现为30 ℃>25 ℃>20 ℃>15 ℃>35 ℃,5组数据中,15 ℃和35 ℃无显著差异,其他均存在显著差异(表2)。
短毛木耳菌丝在不同的碳源上均能生长,菌丝洁白且浓密,但未加入碳源的培养皿菌丝长势相对稀疏,且只有最外面的一圈菌丝是毛绒状的。短毛木耳菌丝对不同碳源的喜好程度为果糖>麦芽糖>葡萄糖>海藻糖>淀粉>蔗糖>乳糖,其中,麦芽糖、葡萄糖、海藻糖、淀粉、蔗糖无显著差异,果糖、乳糖存在显著差异(图4表3)。
短毛木耳在不同氮源培养基上均能生长,其中在含有黄豆粉的培养基上长势最好,菌丝洁白且浓密;其次是酵母浸粉,在含有尿素的培养基上生长速度最慢。
在未加入氮源的培养基菌丝长势稀疏,培养基上只有薄薄一层菌丝,加入蛋白胨的培养基菌丝长势不平整且菌丝相对稀疏,加入酵母浸粉的培养基菌丝生长较快且菌丝浓密洁白,加入硝酸钾的培养基菌丝相对稀疏也只有薄薄一层,加入硝酸铵的培养基菌丝生长速度较慢,加入尿素的培养基菌丝不生长;加入牛肉膏的培养皿菌丝与CK相比生长较慢,但菌丝长势较好;加入黄豆粉的培养基菌丝生长速度最快但菌丝长势较差。短毛木耳菌丝在含有无机氮源和动物氮源的培养基上生长速度较慢,在无机氮源上菌丝稀疏,在含有植物氮源的培养基上菌丝生长较快且菌丝洁白浓密(图5)。短毛木耳菌丝对氮源的喜好程度为黄豆粉>酵母浸粉>硝酸钾>牛肉膏>硝酸铵>蛋白胨>尿素(表4)。
在2因素3水平的正交试验中,短毛木耳菌丝在含有果糖、酵母浸粉的组合中生长状况最好,菌丝洁白且生长旺盛(图6)。结合菌丝的生长速度和长势,最适合短毛木耳的正交组合是果糖、酵母浸粉。在试验中不同碳氮源组合的长势表现为AZ>CZ>CY>BY>BZ>AY>BX>CX>AX,其中CZ、CY不存在显著差异,BX、CX不存在显著差异,其他均存在显著差异(表5)。
以木屑为主要原料,短毛木耳菌丝洁白且生长浓密,培养60 d即可满袋,刺孔培养10 d,待有原基即可摆地出耳(图7)。成熟的短毛木耳形状好,多为簇生,且具有明显的皱褶,子实体具有弹性、胶质、半透明、盘状,黄褐色,干后革质、薄,呈灰褐色或者深褐色。不孕面具柔毛;横切面无髓层。每袋的产量为20.943 g(干质量)(表6)。
分子标记可以快速鉴别金针菇[18-19]、香菇[20-22]、双孢蘑菇[23],目前基本没有用于短毛木耳快速鉴别的分子标记方法,而本研究通过设计的分子标记引物可以准确鉴别出短毛木耳,这不仅为食用菌领域的遗传多样性研究提供了重要支持,同时也为相关产业的发展提供有力的技术支持。
通过生物学特性试验进一步了解短毛木耳菌丝的最适生长条件。温度是影响短毛木耳生长速度的重要因素之一,黑木耳最适温度为25~30 ℃[24],脆木耳菌丝生长的最适温度为33 ℃[25],毛木耳最适温度为25~30 ℃[26],本研究发现低温(15 ℃)会抑制短毛木耳菌丝的生长,偏高温(35 ℃)会严重抑制菌丝的生长甚至使菌丝死亡,这与上述黑木耳与毛木耳的最适温度结果基本一致。
碳源是大型真菌最重要的生命元素和营养成分之一。它不仅是糖类等碳水化合物和蛋白质以及核酸的基本组成部分,也是一种重要的能量原料[27]。在加入的碳源中,最适碳源为果糖,二者的促进效果相差较小,这与黑木耳的最适碳源为果糖的结果[28]一致。其中,短毛木耳利用乳糖为碳源生长较差,而利用海藻糖为碳源时生长情况与利用葡萄糖为碳源相差不大,其原因可能与糖的结构与组成有关。
短毛木耳菌丝的最适氮源是黄豆粉,其次是是酵母浸粉,而有研究表明黑木耳的最适氮源为酵母粉[29]。短毛木耳在利用黄豆粉和酵母浸粉时,菌丝浓密且长速快,但以硝酸钾和硝酸铵为氮源时,菌丝长势差,说明有机氮源比无机氮源更适宜短毛木耳的生长,这可能是因为有机氮提供了核苷酸、维生素和微量元素等非氮源成分,被菌丝生长所利用,而无机氮源需要在酶的作用下转换成有机氮源才能被菌丝利用[30-32]。此外,在以尿素为无机氮源时,菌丝几乎不生长,可能是因为尿素中的氨态氮挥发致使培养基中氮源不足菌丝无法正常生长,或菌丝将其转化成抑制自身生长的成分。
综上,短毛木耳最佳生长条件为果糖、酵母浸粉于30 ℃下培养,这时的菌丝生长较快。在栽培过程中,短毛木耳适宜的培菌温度为28 ℃,培养60 d即可满袋,出耳温度为24 ℃,湿度在85%以上。与黑木耳和毛木耳相比,经过驯化栽培的短毛木耳泡发率高,产量更可观,可以在一定程度上提高经济效益。
  • 广西农科院基本业务费专项(桂农科2021YT094; 桂农科2025YP075)
  • 国家现代农业产业技术体系建设项目(CARS-20)
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doi: 10.3969/j.issn.1000-2561.2025.01.018
  • 接收时间:2024-07-09
  • 首发时间:2026-06-24
  • 出版时间:2025-01-25
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  • 收稿日期:2024-07-09
  • 修回日期:2024-08-02
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广西农科院基本业务费专项(桂农科2021YT094; 桂农科2025YP075)
国家现代农业产业技术体系建设项目(CARS-20)
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    1.广西民族大学海洋生物与技术学院,广西南宁 530006
    2.贺州学院食品与生物工程学院,广西贺州 542899
    3.广西壮族自治区农业科学院微生物研究所,广西南宁 530007

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2种不同金属材料的力学参数

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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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