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菌物学报  2017 , 36 (9): 1192-1209 https://doi.org/10.13346/j.mycosystema.170010

Orginal Article

曲霉属巢状亚属真菌研究现状及部分鉴定实例

孙炳达1, 丁刚2, 张雨森3, 赵国柱3, 周宇光1, 陈娟2*

1中国科学院微生物研究所中国普通微生物菌种保藏管理中心 北京 100101
2中国医学科学院北京协和医学院药用植物研究所 北京 100193
3北京林业大学生物科学与技术学院 北京 100083

Current taxonomy of Aspergillus subgenus Nidulantes and re-identification of several strains

SUN Bing-Da1, DING Gang2, ZHANG Yu-Sen3, ZHAO Guo-Zhu3, ZHOU Yu-Guang1, CHEN Amanda-Juan2*

1China General Microbiological Culture Collection Centre, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
2Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
3College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China

通讯作者:  *Corresponding author. E-mail: amanda_j_chen@163.com*Corresponding author. E-mail: amanda_j_chen@163.com

责任编辑:  SUN Bing-DaDING GangZHANG Yu-SenZHAO Guo-ZhuZHOU Yu-GuangCHEN Amanda-Juan

收稿日期: 2017-01-23

接受日期:  2017-05-3

网络出版日期:  2017-09-22

版权声明:  2017 中国科学院微生物研究所《菌物学报》编辑部 版权所有 

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摘要

基于“一种真菌一个名称”以及命名法的优先原则,曲霉属巢状亚属Aspergillus subgenus Nidulantes真菌的分类系统已发生很大的变化,原来属于裸胞壳属Emericella的种已全部转入曲霉属巢状亚属中,另外该亚属中种的概念也有所更新。本文基于国内外文献,对该亚属的分类现状、种的概念以及主要次生代谢产物产生情况进行了综述。同时基于钙调蛋白基因片段的分子系统发育树以及形态学观察,对收录于《中国真菌志·第五卷·曲霉属及其相关有性型》中的以及其他保存于中国普通微生物保藏中心的、来自于我国17个省及直辖市的裸胞壳属菌株进行了重新鉴定。

关键词: 曲霉属 ; 裸胞壳属 ; 有性型 ; 分子系统发育 ; 黄曲霉毒素 ; 棘白霉素

Abstract

The taxonomy of Aspergillus subgenus Nidulantes has been changed greatly because of the proposal of “one fungus, one name” and nomenclature priority rules. The teleomorphic species belonging to Emericella were transferred to Aspergillus subgenus Nidulantes and some species concepts were updated. In this paper, we reviewed the taxonomic status, species concepts and secondary metabolites of subgenus Nidulantes. In addition, the strains cited in the Flora Fungorum Sinicorum Vol. 5 Aspergillus et teleomorphi cognati or deposited in CGMCC, being identified as Emericella species, isolated from 17 provinces and cities in China, were re-identified using calmodulin gene based phylogeny and morphological methods.

Keywords: Aspergillus ; Emericella ; teleomorph ; phylogeny ; aflatoxin ; echinocandins

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孙炳达, 丁刚, 张雨森, 赵国柱, 周宇光, 陈娟. 曲霉属巢状亚属真菌研究现状及部分鉴定实例[J]. , 2017, 36(9): 1192-1209 https://doi.org/10.13346/j.mycosystema.170010

SUN Bing-Da, DING Gang, ZHANG Yu-Sen, ZHAO Guo-Zhu, ZHOU Yu-Guang, CHEN Amanda-Juan. Current taxonomy of Aspergillus subgenus Nidulantes and re-identification of several strains[J]. 菌物学报, 2017, 36(9): 1192-1209 https://doi.org/10.13346/j.mycosystema.170010

曲霉属 Aspergillus P. Micheli ex Haller 真菌具有极强的生存能力,分布非常广泛,是常见的食品和空气污染菌,有些种类还是重要的人类及动物条件致病菌。部分曲霉属真菌如黄曲霉A. flavus、赭曲霉A. ochraceus、黑曲霉A. niger等能产生黄曲霉毒素(aflatoxins)、赭曲霉毒素(ochratoxins)等真菌毒素。此外,曲霉菌还能产生各种有益的次生代谢产物,如抗生素、有机酸以及一些酶类,如纤维素酶、果胶酶等,被广泛应用于食品发酵以及工业生产中(Thom & Raper 1945;Raper & Fennell 1965;齐祖同 1997;Samson 2010)。

曲霉属由Micheli(1729)建立,在1768年得到有效发表(von Haller 1768)。传统的曲霉属鉴定和分类以菌落培养和微观形态特征为主要依据(Thom & Raper 1945;Raper & Fennell 1965;齐祖同 1997)。根据微观形态特征如分生孢子头的形状、颜色,顶囊的形状以及产孢细胞的排列方式等可以将曲霉属真菌分为6个亚属:曲霉亚属subgen. Aspergillus、烟色亚属subgen. Fumigati、棒状亚属subgen. Clavati、巢状亚属subgen. Nidulantes、环绕亚属subgen. Circumdati以及华丽亚属subgen. OrnatiGams et al. 1986)。曲霉属中的一些种具有有性型,在复型真菌的概念下,这些种被归在有性型属中。已报道的曲霉有性型属有11个:包括散囊菌属Eurotium Link: Fr、毛萨托菌属Chaetosartorya Subram.、裸胞壳属Emericella Berk.、芬尼菌属Fennellia B.J. Wiley & E.G. Simmons、半内果属Hemicarpenteles A.K. Sarbhoy & Elphick、新萨托菌属Neosartorya Malloch & Cain、石座菌属Petromyces Malloch & Cain、核闭壳属Sclerocleista Subram.、束梗丛霉属Stilbothamnium Henn.、新石座菌属Neopetromyces Frisvad & Samson以及新内果属Neocarpenteles Udagawa & Uchiy(Frisvad & Samson 2000;Pitt et al. 2000;Udagawa & Uchiyama 2002)。

随着分子系统学研究的深入以及“一种真菌一个名称”(one fungus,one name)概念在曲霉分类中的提出和广泛接受,现代的曲霉分类系统也发生了很大变化。首先曲霉的有性型属全部被处理为曲霉属的异名,其次根据分子系统学的聚类结果,原先的棒状亚属被合并到烟色亚属,华丽亚属被移出曲霉属,现在曲霉属中包括4个亚属,超过366个种(Houbraken et al. 2014;Samson et al. 2014;Chen et al. 2016a,2016b)。Samson et al.(2014)对曲霉属真菌的分类、鉴定、系统发育以及命名法进行了全面的综述,提供了几乎所有模式种的rDNA内转录间隔区(ITS)、钙调蛋白(calmodulin,CaM)、β-微管蛋白(β-tubulin,BenA)以及核糖核酸聚合酶II亚基(RNA polymerase II second largest subunit,RPB2)4种基因的序列信息,并推荐CaM作为曲霉鉴定的第二分子标记(secondary identification marker)。

巢状亚属目前是曲霉属中最大的亚属,包括巢状组sect. Nidulantes、杂色组sect. Versicolores、焦色组sect. Usti、土色组sect. Terrei以及黄梗组sect. Flavipedes 5个组(Gams et al. 1986)。其中巢状组sect. Nidulantes的有性型为裸胞壳属Emericella。《中国真菌志·曲霉及其相关有性型》中共收录巢状亚属17种,裸胞壳属8种(齐祖同 1997)。李冬梅等(1998)研究了我国北方地区裸胞壳属的种,报道了新记录种Emericella foeniculicola以及E. miyajii。随后又有一些巢状亚属的新种及新记录种在中国被发现(Wang 2012,2013;Zhang et al. 2013;Yu et al. 2015)。本研究基于国内外文献对曲霉属巢状亚属的分类现状进行了综述,并基于钙调蛋白基因的分子系统发育树以及形态学观察,对收录于《中国真菌志·第五卷·曲霉属及其相关有性型》中的以及其他保存于中国普通微生物保藏中心的裸胞壳属菌株进行了重新鉴定和定名。

1 材料与方法

1.1 供试菌株

本研究的菌株来源于中国普通微生物菌种保藏中心(CGMCC)(表1)。

表1   本研究中使用的菌株

Table 1   Strains used in this study

菌株号
Strain No.
(CGMCC)
分离基质
Substrate
采样地点
Location
原鉴定名称
Previous identification
本研究鉴定结果
Current identification
CaM基因
GenBank登录号
CaM GenBank
No.
3.00727电器
Household appliances
-Emericella nidulansAspergillus nidulansKY472609
3.01322--E. nidulansA. nidulansKY472610
3.01323--E. nidulansA. nidulansKY472611
3.01324--E. nidulansA. nidulansKY472612
3.01325--E. nidulansA. nidulansKY472613
3.01326--E. nidulansA. nidulansKY472614
3.01327--E. nidulansA. nidulansKY472615
3.01328--E. nidulansA. nidulansKY472616
3.03915PVC塑料
PVC plastic
-E. nidulansA. nidulansKY472617
3.03916发霉小米
Moldy millet
-E. nidulansA. nidulansKY472618
3.03917空气
Air
-E. nidulans var.
acristata
A. quadrilineatus KY472619
3.03960皮料
Leather
-E. nidulansA. nidulansKY472620
3.03964--E. nidulansA. nidulansKY472621
3.03965猪油
Lard
-E. nidulansA. nidulansKY472622
3.04510T土壤
Soil
湖北神农架
Shennongjia, Hubei
E. undulatusA. undulatusKY472623
3.05270土壤
Soil
四川茂县
Mao, Sichuan
E. nidulansA. nidulansKY472624
3.05272土壤
Soil
四川茂县
Mao, Sichuan
E. nidulansA. nidulansKY472625
3.05277发霉猪肉
Moldy pork
四川茂县
Mao, Sichuan
E. echinulataA. spinulosporus KY472626
3.05363发霉纸盒
Moldy paper box
贵州铜仁市
Tongren, Guizhou
E. nidulansA. nidulansKY472627
3.05445内衣
Underwear
上海市
Shanghai
E. nidulansA. nidulansKY472628
待续
表1
3.05446空气
Air
上海市
Shanghai
E. nidulansA. nidulansKY472629
3.06262 = MQ142-云南宾川
Binchuan, Yunnan
E. nidulans var.
acristata
A. quadrilineatusKY472630
3.06263 =
MQ6396
-宁夏贺兰山
Helan Mountain, Ningxia
E. nidulans var.
acristata
A. quadrilineatusKY472631
3.06264 =
MQ5277
-浙江金华
Jinhua, Zhejiang
E. nidulansA. latusKY472632
3.06265 =
MQ5480
土壤
Soil
山东泰安市
Tai’an, Shandong
E. nidulansA. nidulansKY472633
3.06266 =
MQ5636
-河南潢川县
Huangchuan, Henan
E. nidulansA. nidulansKY472634
3.06267 =
MQ5697
-辽宁铁岭市
Tieling, Liaoning
E. nidulansA. nidulansKY472635
3.06268 =
MQ6397
-宁夏贺兰山
Helan Mountain, Ningxia
E. nidulansA. nidulansKY472636
3.06269 =
MQ6460
-宁夏贺兰山
Helan Mountain, Ningxia
E. nidulansA. nidulansKY472637
3.06270-福建福州市
Fuzhou, Fujian
E. nidulansA. nidulansKY472638
3.06271-江苏扬州市
Yangzhou, Jiangsu
E. nidulansA. nidulansKY472639
3.06272 = MQ20-北京市
Beijing
E. nidulansA. nidulansKY472640
3.06287 =
MQ6525
-宁夏陶乐
Taole, Ningxia
E. nidulansA. nidulansKY472641
3.06288 =
MQ8438
-山西阳泉
Yangquan, Shanxi
E. nidulansA. nidulansKY472642
3.06289 =
MQ8755
-北京市
Beijing
E. nidulansA. nidulansKY472643
3.06290 =
MQ8797
-河南郑州市
Zhengzhou, Henan
E. nidulansA. nidulansKY472644
3.06292 =
MQ8507
玻璃板
Glass pane
辽宁通化
Tonghua, Liaoning
E. variecolorA. stellatusKY472645
待续
表1
3.06295 =
MQ7839
土壤
Soil
湖北神农架
Shennongjia, Hubei
E. undulataA. undulatusKY472646
3.06370 = C44-福建福州
Fuzhou, Fujian
E. nidulansA. nidulansKY472647
3.06371 = C4100蔬菜地土壤
Soil from vegetable field
云南昆明市
Kunming, Yunnan
E. nidulansA. nidulansKY472648
3.06374 = C5002茄子地土壤
Soil from eggplant field
云南丽江
Lijiang, Yunnan
E. nidulansA. nidulansKY472649
3.06375 = C5314玉米地土壤
Soil from maize field
云南保山
Baoshan, Yunnan
E. nidulansA. nidulansKY472650
3.06376 = C5755草地土壤
Grassland
云南丽江
Lijiang, Yunnan
E. nidulansA. nidulansKY472651
3.06377 = C5798白菜地土壤
Soil from cabbage field
云南丽江
Lijiang, Yunnan
E. nidulansA. nidulansKY472652
3.06378 = C5372玉米地土壤
Soil from maize field
云南丽江
Lijiang, Yunnan
E. nidulansA. nidulansKY472653
3.06379 = C5399霉橘皮
Moldy orange peel
云南大理
Dali, Yunnan
E. nidulansA. nidulansKY472654
3.06380 = C6314红薯地土壤
Soil from sweet potato field
云南保山市
Baoshan, Yunnan
E. nidulansA. nidulansKY472655
3.06381 = C6460土壤
Soil
云南芒市
Mangshi, Yunnan
E. nidulansA. nidulansKY472656
3.06382 = C6565霉扁豆
Moldy bean
云南芒市
Mangshi, Yunnan
E. nidulansA. nidulansKY472657
3.06383 = C6599红薯地土壤
Soil from sweet potato field
云南芒市
Mangshi, Yunnan
E. nidulansA. nidulansKY472658
3.06384 = C6676芭蕉地土壤
Soil growing banana
云南芒市
Mangshi, Yunnan
E. nidulansA. nidulansKY472659
3.06385 = C6867发霉竹子
Moldy bamboo
云南瑞丽
Ruili, Yunnan
E. nidulansA. nidulansKY472660
3.06386 = C7568菠萝蜜下土壤
Soil under Artocarpus
heterophyllus
云南西双版纳
Xishuangbanna, Yunnan
E. nidulansA. nidulansKY472661
3.06387 = C7596土壤
Soil
云南西双版纳
Xishuangbanna, Yunnan
E. nidulansA. nidulansKY472662
3.06388 = C8424霉黄豆壳
Moldy shell of soybean
云南思茅
Simao, Yunnan
E. nidulansA. nidulansKY472663
待续
表1
3.06390菜地土壤
Soil from vegetable field
云南昆明
Kunming, Yunnan
E. nidulansA. nidulansKY472664
3.06393 = C6692芭蕉地土壤
Soil growing banana
云南芒市
Mangshi, Yunnan
E. nidulans var.
acristata
A. quadrilineatusKY472665
3.06394 = C5237玉米粉
Corn flour
云南丽江
Lijiang, Yunnan
E. rugulosaA. rugulosusKY472666
3.06395 = C5997发霉红木果
Moldy annatto fruit
云南
Yunnan
E. rugulosaA. rugulosusKY472667
3.06404 = C8619土壤
Soil
北京市
Beijing
E. unguisA. unguisKY472668
3.06546 = C1350树皮
Bark
河北小五台山
Xiaowutai Mountain, Hebei
E. nidulansA. nidulansKY472669
3.11526土壤
Soil
新疆阿克苏
Akesu, Xinjiang
E. corrugataA. nidulansKY472670
3.11528土壤
Soil
新疆阿克苏
Akesu, Xinjiang
E. nidulansA. quadrilineatusKY472671
3.11530土壤
Soil
新疆阿克苏
Akesu, Xinjiang
E. nidulansA. quadrilineatusKY472672
3.11566土壤
Soil
新疆乌鲁木齐
Urumchi, Xinjiang
E. corrugataA. protuberus KY472673
3.11573土壤
Soil
新疆乌鲁木齐
Urumchi, Xinjiang
E. corrugataA. nidulansKY472674
3.14984T狭叶圆穗蓼
Polygonum
macrophyllum var.
stenophyllum
西藏米拉山
Mira Mountain, Tibet
E. miraensiA. miraensisKY472675
3.15313土壤
Soil
山东
Shandong
E. heterothallicaA. heterothallicusKY472676

Note: “-”: Unknown. Numbers titled with MQ/C were cited in the Flora Fungorum Sinicorum Vol. 5 Aspergillus et Teleomorphi Cognati.注:“-”:采集信息不详;MQ/C编号:《中国真菌志·第五卷·曲霉属及其相关有性型》中的编号

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1.2 菌株DNA提取以及目标基因片段的PCR扩增及测序

菌株在麦芽汁琼脂培养基(MEA)上培养1周后,刮取约1g菌丝,根据DNA提取试剂盒(UltracleanTM Microbial DNA Isolation Kit,MoBio,Solana Beach,U.S.A.)的说明进行DNA的提取,提取的DNA置于-20℃冰箱保存。根据Samson et al.(2014)的方法,扩增CaM基因,扩增好的PCR产物送北京诺赛基因组研究中心有限公司测序。测序后拼接完毕的序列递交GenBank。

1.3 系统发育树构建

以曲霉属巢状亚属117个种的模式菌株的序列作为参考序列,以Aspergillus flavipes(NRRL 302T)的序列作为外群(Chen et al. 2016a),先用软件MAFFT v. 7进行序列比对(Katoh & Standley 2013),并两端切齐。采用软件Find Model(Posada & Crandall 1998)计算系统发育树的模型,随后采用最大似然法(RA x ML,Stamatakis et al. 2008)以及贝叶斯分析(MrBayes v. 3.1.2,Ronquist & Huelsenbeck 2003)建立系统发育树。

1.4 形态学研究

将菌株接种于察氏酵母膏琼脂(Czapek yeast autolysate agar,CYA),以及麦芽汁琼脂(Malt extract agar,MEA)上,在25℃黑暗培养7d后,观察菌落的颜色、质地、产孢程度、色素分泌等情况。使用在MEA培养基上生长7d的菌落进行显微玻片的制备,观察分生孢子梗、产孢细胞以及分生孢子等的特征。使用在OA培养基上生长14d后的菌落观察子囊孢子的微观形态(Chen et al. 2016a)。

2 结果与分析

2.1 曲霉属巢状亚属及其有性型裸胞壳属在中国的报道情况

我国已报道该类群36种,其中23种属于巢状亚属的5个组,其余13种属于裸胞壳属(表2)。根据目前分子系统学的研究以及命名法的优先原则,部分种名以及其分类地位已经发生了变化。其中17个种名被修订:4个变种(Aspergillus terreus var. aureusA. terreus var. terreusA. versicolor var. protuberus以及A. versicolor var. versicolor)升级为种;13个裸胞壳属的种被归到曲霉属中,其属名及部分种加词也发生了相应的改变(Samson et al. 2011a,2014;Jurjevic et al. 2012;Chen et al. 2016a)。

表2   中国已报道的曲霉属巢状亚属及其有性型裸胞壳属的记录

Table 2   Reported species in Aspergillus subgenus Nidulantes and Emericella in China

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2.2 基于CaM基因序列的系统发育分析

用于建立系统发育树的CaM基因序列全长为645bp,根据软件Find Model的计算,采用GTR+G模型,基于最大似然法以及贝叶斯分析两种算法分别计算系统进化树。两种算法获得的系统发育树具有相同的拓扑结构,采用贝叶斯分析的系统发育树形作图1,两种算法的自展支持率(bootstrap support)及后验概率(posterior probability)分别标注于分支上。本研究中的68株真菌分别聚类到11个种,分别是A. heterothallicusA. latusA. miraensisA. nidulansA. protuberusA. quadrilineatusA. rugulosusA. spinulosporusA. stellatusA. undulatus以及A. unguis图1)。这些菌株来自中国15个省、2个直辖市,分离基质多为土壤,此外还有空气、发霉食品等(表1)。这68株真菌的系统发育鉴定的结果,与前人基于形态学鉴定的结果基本吻合,仅有4个菌株分别被重新鉴定为A. latus(“E. nidulans” CGMCC 3.06264),A. nidulans (“E. corrugata” CGMCC 3.11526,“E. corrugata” CGMCC 3.11573)以及 A. protuberus(“E. corrugata” CGMCC 3.11566)。随后对这4株菌进行的形态学观察进一步证实了分子系统发育树的鉴定结果:A. latus(CGMCC 3.06264)与A. nidulans在形态上非常相似,仅有子囊孢子的赤道冠有微小差别:A. latus(CGMCC 3.06264)子囊孢子的赤道冠(1-1.5µm)较A. nidulans的赤道冠(0.5-1µm)宽;A. corrugatus子囊孢子体表面具有皱褶,而A. nidulans(CGMCC 3.11526,CGMCC 3.11573)子囊孢子体表面光滑;A. corrugatus在OA培养基上培养2-4周后能够产生子囊孢子,而A. protuberus(CGMCC 3.11566)不产生子囊孢子。4个菌株(CGMCC 3.06264、CGMCC 3.11526、CGMCC 3.11573、CGMCC 3.11566)在CYA 25℃培养7d的生长速率分别为45mm、35mm、38mm、30mm,符合相应种已报道的生长速率范围(Jurjević et al. 2012;Chen et al. 2016a)。

图1   基于钙调蛋白基因(CaM)构建的曲霉属巢状亚属种的系统发育树 加粗的分支表示后验概率高于1pp及自展支持率高于95% bs. 菌株Aspergillus flavipes(NRRL 302T)作为外群. 星号★表示本研究使用的CGMCC菌株

Fig. 1   Phylogenetic tree of subgenus Nidulantes inferred from CaM. Branches with values more than 1 posterior probability (pp) and 95% bootstrap support (bs) are thickened. The phylogram is rooted with Aspergillus flavipes (NRRL 302T). CGMCC strains used in this study were marked with star.

3 讨论

曲霉属巢状亚属的主要形态特征为具有双层的产孢结构,分生孢子梗多为淡褐色(Raper & Fennell 1965;Gams et al. 1986)。其有性型裸胞壳属的主要形态特征为具有由壳细胞包被的闭囊壳;子囊不规则排列,球形或近球形,壁易消解;子囊孢子双凸镜形,通常具有鸡冠状突起,橙红色至紫红色,表面光滑或具有不同纹饰(Benjamin 1955;Raper & Fennell 1965)。在传统分类系统中,子囊孢子的颜色、形状、大小以及纹饰是该类群鉴定种的重要依据(Thom & Raper 1939;Christensen & Raper 1978;Horie 1980;Christensen & States 1982;Ismail et al. 1995;齐祖同 1997;Zalar et al. 2008;Guarro et al. 2012;Matsuzawa et al. 2012)。

近20年来诸多学者从分子系统发育角度对该亚属的分类系统进行了重新整理。Peterson(2008)分析了该亚属所有模式种的4个基因片段(ITS、BenACaM以及RPB2),将该亚属分为7个组:sect. Bispori、sect. Nidulantes、sect. Ochraceorosei、sect. Raperi、sect. Silvati、sect. Sparsi以及sect. Usti。随后,Varga et al.(2010a)基于BenACaM以及ITS的多基因分析,提出了一个新组sect. Aenei。关于有性型裸胞壳属的分子系统学研究开展的较晚,Matsuzawa et al.(2012)首次使用ActinBenA以及CaM 3个基因片段研究了裸胞壳属各个种之间的系统发育关系,发现子囊孢子的形态在种下水平有一定的变异范围。根据“一种真菌一个名称”以及命名法的优先原则,Samson et al.(2014)将裸胞壳属的所有种都转入了曲霉属巢状亚属。此后,曲霉属巢状亚属的分类学研究进入一个新的时代,无性世代以及有性世代作为同一种真菌的不同生活史,不再被孤立地分别命名。Hubka et al.(2016) 采用4个基因片段(ITS、BenA、CaM以及RPB2)对巢状亚属内所有种(包括无性型以及有性型种)进行了分子系统学的研究,报道了2个新种Aspergillus askiburgiensis以及A. croceus。Chen et al.(2016a)采用多相分类的方法,综合分子系统学、形态学、生理学以及次生代谢产物分析,对巢状亚属进行了系统整理,重新界定了该亚属中种的概念:即曲霉属巢状亚属中所接受的种符合系统发生种(phylogenetic species)的概念,并且系统发育的聚类结果与形态学、生理学以及次生代谢产物的结果吻合。在此基础上,他们提出一个新组sect. Cavernicolus,报道新种10个,并针对所有种都提供了详细的形态、多基因序列、生长温度以及主要次生代谢产物谱的数据,目前曲霉属巢状亚属共接受9个组117个种。由于ITS片段在曲霉属各种间的分辨率不高,Samson et al.(2014)建议使用CaM作为曲霉属鉴定的第二分子标记,Chen et al.(2016a)发现CaM在曲霉属巢状亚属中也具有较好的分辨能力,除了Aspergillus qinqixianii以及A. filifer 2个种以外,其余种都具有特异的CaM基因。

《中国真菌志·第五卷·曲霉属及其相关有性型》对中国的巢状亚属及其有性型裸胞壳属Emericella进行了描述(齐祖同 1997)。近年来我国也有一些新种被陆续报道,如Emericella miraensisAspergillus hongkongensisA. keveioidesWang 2013;Zhang et al. 2013;Tsang et al. 2016)。还有一些新种如Emericella foeniculicolaAspergillus qinqixianiiA. pachycristatus由日本学者在我国分离到并发表(Udagawa & Muroi 1979;Horie et al. 2000;Matsuzawa 2012)。本研究依据最新的分类系统对收录于《中国真菌志·第五卷·曲霉属及其相关有性型》中的以及其他保存于CGMCC的裸胞壳属菌株进行了重新的鉴定,分子鉴定的结果大部分与原形态学鉴定结果相吻合。子囊孢子的形态特征是这一类群真菌鉴定的重要依据,其中子囊孢子的形状、大小以及颜色较容易观察,而纹饰需要借助扫描电镜进行准确观察。我们在进行电镜观察时发现过于剧烈的预处理方法可能会在光滑的子囊孢子表面造成皱褶,从而引起鉴定的误判。对形态上非常相近的种,建议采用目前广泛使用的单基因(CaM)或多基因(ITS、BenACaMRPB2)分子系统学手段进行准确的鉴定(Chen et al. 2016a)。随着曲霉现代分类系统的建立和模式种多基因序列数据库的完善,曲霉属的分类学已进入一个新的时代。我国具有丰富的物种多样性,但是目前报道的曲霉物种数仅占全世界已报道种的约1/4至1/5,因此我国还有大量的曲霉属物种资源有待发掘。

曲霉属巢状亚属真菌能产生丰富的次生代谢产物,例如黄曲霉毒素等真菌毒素类和一些具有潜在价值的药物先导化合物。目前巢状亚属中有4个种:Aspergillus astellatusA. miraensisA. olivicola以及A. venezuelensis能产生黄曲霉毒素B1(Aflatoxin B1)(Frisvad & Samson 2004;Frisvad et al. 2004;Zalar et al. 2008;Chen et al. 2016a);有包括A. nidulans在内的40个种能产生杂色曲霉素(sterigmatocystin)(Horie & Yamazaki 1985;Frisvad 1986;Rabie et al. 1977;Frisvad & Samson 2004;Frisvad et al. 2004;Varga et al. 2009,2010a,2010b;Rank et al. 2011;Jurjević et al. 2013;Hubka et al. 2016)。该亚属产生的药物先导化合物类包括棘白霉素echinocandins、calbistrins、mulundocandins、terrein、varitriols以及variecolins等(Chen et al. 2016a)。已上市的阿尼芬净(anidulafungin),是从A. spinulosporus(= A. nidulans var. echinulatus)发酵产物提取的半合成脂肽,属于棘白霉素。已报道能产生该化合物的种还包括巢状亚属的A. navahoensisA. pachycristatusA. parvatheciusA. quadrilineatus以及A. rugulosusKlich et al. 2001;de la Cruz et al. 2012;Matsuzawa et al. 2012;Bills et al. 2014;Yue et al. 2015)。在今后的工作中进一步丰富我国该类群真菌资源,理清其分类及系统发育关系,对于发现和利用该类群真菌产生的有用次生代谢产物,规避害处,具有重要意义。

致谢:作者向毕生致力于青曲霉及其有性型研究的分类学家齐祖同、孔华忠先生致敬,感谢中国科学院微生物研究所王龙在菌株资源收集方面所做的工作。


参考文献

[1] Benjamin CR, 1955.

Ascocarps of Aspergillus and Penicillium

.Mycologia, 47(5): 669-687

[本文引用: 1]     

[2] Bills G, Li Y, Chen L, Yue Q, Niu XM, An ZQ, 2014.

New insights into the echinocandins and other fungal non-ribosomal peptides and peptaibiotics

.Natural Product Reports, 31(10): 1348-1375

[本文引用: 1]     

[3] Chen AJ, Frisvad JC, Sun BD, Varga J, Kocsubé S, Dijksterhuis J, Kim DH, Hong SB, Houbraken J, Samson RA, 2016a.

Aspergillus section Nidulantes (formerly Emericella): polyphasic taxonomy, chemistry and biology

.Studies in Mycology, 84: 1-118

[本文引用: 8]     

[4] Chen AJ, Varga J, Frisvad JC, Jiang XZ, Samson RA, 2016b.

Polyphasic taxonomy of Aspergillus section Cervini

.Studies in Mycology, 85: 65-89

[本文引用: 1]     

[5] Christensen M, Raper KB, 1978.

Synoptic key to Aspergillus nidulans group species and related Emericella species

.Transactions of the British Mycological Society, 71(2): 177-191

[本文引用: 1]     

[6] Christensen M, States JS, 1982.

Aspergillus nidulans group: Aspergillus navahoensis, and a revised synoptic key

.Mycologia, 74(2): 226-235

[本文引用: 1]     

[7] de la Cruz M, Martín J, González-Menéndez V, Pérez-Victoria I, Moreno C, Tormo JR, El Aouad N, Guarro J, Vicente F, Reyes F, Bills GF, 2012.

Chemical and physical modulation of antibiotic activity in Emericella species

.Chemistry & Biodiversity, 9(6): 1095-1113

[本文引用: 1]     

[8] Frisvad JC, 1986.

Secondary metabolites as an aid to Emericella classification. In: Samson RA, Pitt JI (eds.) Advances in Penicillium and Aspergillus systematics. NATO ASI Series

. Series A: life sciences. Vol. 102. Plenum Press, New York. 437-444

[本文引用: 1]     

[9] Frisvad JC, Samson RA, 2000.

Neopetromyces gen. nov. and an overview of teleomorphs of Aspergillus subgenus Circumdati

.Studies in Mycology, 45: 201-207

[本文引用: 1]     

[10] Frisvad JC, Samson RA, 2004.

Emericella venezuelensis, a new species with stellate ascospores producing sterigmatocystin and aflatoxin B1

.Systematic and Applied Microbiology, 27(6): 672-680

[本文引用: 2]     

[11] Frisvad JC, Samson RA, Smedsgaard J, 2004.

Emericella astellata, a new producer of aflatoxin B1, B2 and sterigmatocystin

.Letters in Applied Microbiology, 38(5): 440-445

[本文引用: 2]     

[12] Gams W, Christensen M, Onions AH, Pitt JI, Samson RA, 1986.

Infrageneric taxa of Aspergillus. In: Samson RA, Pitt JI (eds.) Advances in Penicillium and Aspergillus systematics

. NATO ASI Series. Series A: life sciences. Vol. 102. Plenum Press, New York. 55-62

[本文引用: 3]     

[13] Guarro J, Gené J, Stchigel AM, Figueras MJ, 2012.

Atlas of soil Ascomycetes. CBS Biodiversity Series 10

. CBS-KNAW Fungal Biodiversity Centre, Utrecht. 169-183

[本文引用: 1]     

[14] Haller AV, 1768.

Historia Stirpium Indigenarum Helvetiae Inchoata

. Sumptibus Societatis Typographicae, Bernae.

[本文引用: 1]     

[15] Horie Y, 1980.

Ascospore ornamentation and its application to the taxonomic re-evaluation in Emericella

.Transactions of the Mycological Society of Japan, 21: 483-493

[本文引用: 1]     

[16] Horie Y, Abliz P, Hui Y, Fukiharu T, Nishimura K, Li DM, Li RY, 2000.

Emericella qinqixianii, a new species from desert soil in China

.Mycoscience, 41(2): 183-187

[本文引用: 1]     

[17] Horie Y, Yamazaki M, 1985.

Production of carcinogenic mycotoxins, sterigmatocystin and its allied compounds, by Emericella species

.Transactions of the Mycological Society of Japan, 26: 411-419

[本文引用: 1]     

[18] Houbraken J, de Vries RP, Samson RA, 2014.

Chapter four—modern taxonomy of biotechnologically important Aspergillus and Penicillium species

.Advances in Applied Microbiology, 86: 199-249

[本文引用: 1]     

[19] Hubka V, Nováková A, Kolařík M, Jurjević Ž, Peterson SW, 2015.

Revision of Aspergillus section Flavipedes: seven new species and proposal of section Jani sect. nov

.Mycologia, 107(1): 169-208

[20] Hubka V, Nováková A, Peterson SW, Frisvad JC, Sklenář F, Matsuzawa T, Kubátová A, Kolařík M, 2016.

A reappraisal of Aspergillus section Nidulantes with descriptions of two new sterigmatocystin-producing species

.Plant Systematics and Evolution, 302(9): 1267-1299

[本文引用: 1]     

[21] Ismail MA, Abdel-Sater MA, Zohri AA, 1995.

A synoptic key to species of the Aspergillus nidulellus-Emericella assemblage common to Egypt

.Mycotaxon, 53: 391-405

[本文引用: 1]     

[22] Jurjević Ž, Peterson SW, Horn BW, 2012.

Aspergillus section Versicolores: nine new species and multilocus DNA sequence based phylogeny

.IMA Fungus, 3(1): 59-79

[本文引用: 2]     

[23] Jurjević Ž, Peterson SW, Solfrizzo M, Peraica M, 2013.

Sterigmatocystin production by nine newly described Aspergillus species in section Versicolores grown on two different media

.Mycotoxin Research, 29(3): 141-145

[本文引用: 1]     

[24] Katoh K, Standley DM, 2013.

MAFFT multiple sequence alignment software version 7: improvements in performance and usability

.Molecular Biology and Evolution, 30(4): 772-780

[本文引用: 1]     

[25] Klich M, Mendoza C, Mullaney E, Keller N, Bennett JW, 2001.

A new sterigmatocystin-producing Emericella variant from agricultural desert soils

.Systematic and Applied Microbiology, 24(1): 131-138

[本文引用: 1]     

[26] Kong HZ, Qi ZT, 1985.

Some new records and rare taxa of Aspergillus of China

.Bulletin of Botanical Research, 5(2): 145-150 (in Chinese)

[27] Li DM, Wang DL, Li RY, Wang XH, Horie Y, Fukiharu T, 1998.

Emericella spp. in soils of north China

.Mycosystema, 17(2): 130-136 (in Chinese)

[28] Li YY, Shi Y, Lu XH, Cui XL, Ma Y, Zhao Y, Liu J, Zhang H, 2010.

Morphological and molecular identification of ophiobolins producing strain F02Z2172

.Microbiology China, 37(8): 1205-1210 (in Chinese)

[29] Matsuzawa T, Tanaka R, Horie Y, Hui Y, Abliz P, Yaguchi T, 2012.

The correlation among molecular phylogenetics, morphological data, and growth temperature of the genus Emericella, and a new species

.Mycoscience, 53(6): 433-445

[本文引用: 3]     

[30] Micheli PA, 1729.

Nova plantarvm genera ivxta Tovrnefortii methodvm disposita

. Typis Bernardi Paperinii, Florence.

[31] Peterson SW, 2008.

Phylogenetic analysis of Aspergillus species using DNA sequences from four loci

.Mycologia, 100(2): 205-226

[32] Peterson SW, Varga J, Frisvad JC, Samson RA, 2008.

Phylogeny and subgeneric taxonomy of Aspergillus

. In: Varga J, Samson RA (eds.) Aspergillus in the genomic era. Wageningen Academic Publishers, Wageningen. 33-56

[33] Pitt JI, Samson RA, Frisvad JC, 2000.

List of accepted species and their synonyms in the family Trichocomaceae

. In: Samson RA, Pitt JI (eds.) Integration of modern taxonomic methods for Penicillium and Aspergillus classification. Harwood Academic Publishers, Amsterdam. 9-79

[本文引用: 1]     

[34] Posada D, Crandall KA, 1998.

MODELTEST: testing the model of DNA substitution

.Bioinformatics, 14(9): 817-818

[本文引用: 1]     

[35] Qi ZT, 1997. Flora fungorum sinicorum. Vol. 5 Aspergillus et Teleomorphi Cognati. Science Press, Beijing. 1-198 (in Chinese)

[本文引用: 5]     

[36] Rabie CJ, Steyn M, van Schalkwyk GC, 1977.

New species of Aspergillus producing sterigmatocystin

.Applied and Environmental Microbiology, 33(5): 1023-1025

[本文引用: 1]     

[37] Rank C, Nielsen KF, Larsen TO, Varga J, Samson RA, Frisvad JC, 2011.

Distribution of sterigmatocystin in filamentous fungi

.Fungal Biology, 115(4-5): 406-420

[本文引用: 1]     

[38] Raper KB, Fennell DI, 1965.

The genus Aspergillus

. Williams & Wilkins, Baltimore, MD. 1-686

[本文引用: 4]     

[39] Ronquist F, Huelsenbeck JP, 2003.

MrBayes 3.0: Bayesian phylogenetic inference under mixed models

.Bioinformatics, 19(12): 1572-1574

[本文引用: 1]     

[40] Samson RA, Houbraken J, Thrane U, Frisvad JC, Andersen B, 2010.

Food and indoor fungi

. CBS-KNAW Fungal Biodiversity Center, Utrecht. 1-390

[本文引用: 1]     

[41] Samson RA, Peterson SW, Frisvad JC, Varga J, 2011a.

New species in Aspergillus section Terrei

.Studies in Mycology, 69: 39-55

[本文引用: 1]     

[42] Samson RA, Varga J, Meijer M, Frisvad JC, 2011b.

New taxa in Aspergillus section Usti

.Studies in Mycology, 69: 81-97

[43] Samson RA, Visagie CM, Houbraken J, Hong SB, Hubka V, Klaassen CHW, Perrone G, Seifert KA, Susca A, Tanney JB, Varga J, Kocsubé S, Szigeti G, Yaguchi T, Frisvad JC, 2014.

Phylogeny, identification and nomenclature of the genus Aspergillus

.Studies in Mycology, 78: 141-173

[本文引用: 2]     

[44] Stamatakis A, Hoover P, Rougemont J, Renner S, 2008.

A rapid bootstrap algorithm for the RAxML Web Servers

.Systematic Biology, 57(5): 758-771

[45] Sun ZM, 1993.

Two new records of Emericella in China and their nomenclature

.Acta Mycologica Sinica, 12(3): 246-247 (in Chinese)

[46] Sun ZM, Qi ZT, 1994.

New taxa and a new record of Aspergillus and Eurotium

.Acta Mycologica Sinica, 13(2): 81-87 (in Chinese)

[47] Tai FL, 1979. Sylloge Fungorum Sinicorum. Science Press, Beijing. 1-1527 (in Chinese)

[48] Thom C, Raper KB, 1939.

The Aspergillus nidulans group

.Mycologia, 31(6): 653-669

[本文引用: 1]     

[49] Thom C, Raper KB, 1945.

A manual of the Aspergilli

. Williams & Wilkins, Maryland, MD. 1-373

[本文引用: 2]     

[50] Tsang CC, Hui TWS, Lee KC, Chen JHK, Ngan AHY, Tam EWT, Chan JFW, Wu AL, Cheung M, Tse BPH, Wu AKL, Lai CKC, Tsang DNC, Que TL, Lam CW, Yuen KY, Lau SKP, Woo PCY, 2016.

Genetic diversity of Aspergillus species isolated from onychomycosis and Aspergillus hongkongensis sp. nov., with implications to antifungal susceptibility testing

.Diagnostic Microbiology and Infectious Disease, 84(2): 125-134

[本文引用: 1]     

[51] Tzean SS, Chen JL, Liou GY, Chen CC, Hsu WH, 1990

Aspergillus and related teleomorphs from Taiwan. In: Anon. (ed.) Mycological monograph

. Food Industry Research & Development Institute, Hsinchu. 1-113

[52] Udagawa SI, Muroi T, 1979.

Some interesting species of Ascomycetes from imported spices

.Transactions of the Mycological Society of Japan, 20(1): 13-22

[本文引用: 1]     

[53] Udagawa SI, Uchiyama S, 2002.

Neocarpenteles: a new ascomycete genus to accommodate Hemicarpenteles acanthosporus

.Mycoscience, 43(1): 3-6

[本文引用: 1]     

[54] Varga J, Frisvad JC, Samson RA, 2009.

A reappraisal of fungi producing aflatoxins

.World Mycotoxin Journal, 2(3): 263-277

[本文引用: 1]     

[55] Varga J, Frisvad JC, Samson RA, 2010a.

Aspergillus sect. Aeni sect. nov., a new section of the genus for A. karnatakaensis sp. nov. and some allied fungi

.IMA Fungus, 1(2): 197-205

[本文引用: 1]     

[56] Varga J, Frisvad JC, Samson RA, 2010b.

Polyphasic taxonomy of Aspergillus section Sparsi

.IMA Fungus, 1(2): 187-195

[本文引用: 1]     

[57] Varga J, Houbraken J, Van Der Lee HA, Verweij PE, Samson RA, 2008.

Aspergillus calidoustus sp. nov., causative agent of human infections previously assigned to Aspergillus ustus

.Eukaryotic Cell, 7(4): 630-638

[58] Wang L, 2012.

Four new records of Aspergillus sect. Usti from Shandong Province, China

.Mycotaxon, 120: 373-384

[本文引用: 1]     

[59] Wang L, 2013.

Aspergillus keveioides, a new species of Aspergillus sect. Usti from Shandong Province, China

.Mycosystema, 32(S1): 136-144 (in Chinese)

[本文引用: 2]     

[60] Yu Y, Zhang YH, Wang L, 2015.

Aspergillus germanicus, a new Chinese record of Aspergillus section Usti

.Microbiology China, 42(4): 674-682 (in Chinese)

[本文引用: 1]     

[61] Yue Q, Chen L, Zhang XL, Li K, Sun JZ, Liu XZ, An ZQ, Bills GF, 2015.

Evolution of chemical diversity in echinocandin lipopeptide antifungal metabolites

.Eukaryotic Cell, 14(7): 698-718

[本文引用: 1]     

[62] Zalar P, Frisvad JC, Gunde-Cimerman N, Varga J, Samson RA, 2008.

Four new species of Emericella from the Mediterranean region of Europe

.Mycologia, 100(5): 779-795

[本文引用: 2]     

[63] Zhang LC, Chen J, Lin WH, Guo SX, 2013.

A new species of Emericella from Tibet, China

.Mycotaxon, 125: 131-138

[本文引用: 2]     

[64] 戴芳澜,1979. 中国真菌总汇. 北京:科学出版社. 1-1527

[65] 孔华忠,齐祖同,1985.

中国曲霉属的几个新记录和罕见分类群

. 植物研究,5(2): 145-150

[66] 李冬梅,王端礼,李若瑜,王晓红,堀江义一,吹春俊光,1998.

我国北方地区裸胞壳属(Emericella)的种

. 菌物系统,17(2): 130-136

[67] 李业英,石英,路新华,崔晓兰,马瑛,赵颖,刘静,张华,2010.

一株产蛇孢假单壳素真菌F02Z2172的形态和分子鉴定

. 微生物学通报,37(8): 1205-1210

[68] 齐祖同,1997. 中国真菌志·第五卷·曲霉属及其相关有性型. 北京:科学出版社. 1-198

[69] 孙曾美,1993.

中国裸胞壳属的两个新记录及其有关命名问题

. 真菌学报,12(3): 246-247

[70] 孙曾美,齐祖同,1994.

曲霉及其有性型散囊菌的新分类群和新记录种

. 真菌学报,13(2): 81-87

[71] 王龙,2013.

类开费曲霉Aspergillus keveioides–曲霉属焦曲霉组一新种

. 菌物学报,32(增刊): 136-144

[72] 余芸,张永红,王龙,2015.

日耳曼曲霉(Aspergillus germanicus)——曲霉属焦曲霉组一个我国新记录种

. 微生物学通报,42(4): 674-682

/