我国河北、浙江、云南及西藏木霉种记述

对从中国河北、浙江、云南及西藏分离的72个木霉菌株进行了鉴定和分类学研究。采纳Gams&Bissett(1998)及Kullnig—Gradinger et al.(2002)的分类观点,鉴定出木霉属的12个种, 其中包括8个已知种:深绿木霉T.atroviride,桔绿木霉T.citrinoviride,哈茨木霉T.harzianum, 康宁木霉T.koningii,长枝木霉T.longibrachiatum,中国木霉T.sinensis,绿木霉T.virens和绿色木霉T.viride;4个我国新记录种是:木霉组(Trichoderma section)的棘孢木霉T.asperellum及粗梗组(Pachybasium section)的淡黄木霉T.cerinum,螺旋木霉T.spirale和茸状木霉T.velutinum。     

辽宁木霉属(Trichoderma)真菌的形态分类研究

对采自辽宁省内14个地方的173份土样和植物组织材料进行分离,获得了54株Trichoderma菌株,采用形态学分类方法鉴定出12种木霉菌,分别是拟康氏木霉(Trichoderma pseudokoningii)、长枝木霉(T.longi-brachiatum)、粘绿木霉(T.virens)、卷曲木霉(T.spirale)、顶孢木霉(T.fertile)、粗壮木霉(T.strigosum)、长孢木霉(T.longipile)、钩状木霉(T.hamatum)、绿色木霉(T.viride)、康氏木霉(T.koningii)、深绿木霉(T.atroviri-de)和哈茨木霉(T.harzianum)。其中长孢木霉为中国新记录种,粗壮木霉和卷曲木霉为东北地区首次报道。文中列有辽宁省木霉属真菌的分种检索表,并附有各木霉菌的生境和分布。     

木霉植酸酶编码基因多样性及作为系统发育标记潜力研究

对现有的148株木霉菌株在含植酸钙的琼脂培养基上进行了产植酸酶能力鉴定,结果表明所有菌株均产生了水解透明图,说明所有测试的木霉菌株都具有植酸酶活性,植酸酶编码基因在木霉群体中具有广泛性.选取14个种类的21株木霉,采用植酸酶保守序列设计简并引物P8205、P500-2扩增获得其中11种17株木霉植酸酶基因片段,进行了序列测定;利用ITS4、ITS5引物扩增17个木霉菌株的ITS序列并测序.分别基于植酸酶基因片段序列以及ITS序列信息,通过邻接法(N-J法)构建系统发育树,结果表明植酸酶基因序列具有多样性的特点,而基于植酸酶基因序列与基于ITS序列的分类结果基本相同,不同的是长枝木霉(Trichoderma longibrachiatum)植酸酶基因序列与哈茨木霉(Trichoderma harzianum)被分到同一分支当中,与ITS序列的进化关系相差较大,表明有可以作为木霉分类的一种新的标记的潜力,并携带部分与ITS序列不同的系统发育相关信息.     

工厂化生产海鲜菇菌包污染霉菌的鉴定及防治

对工厂化生产中海鲜菇菌包污染霉菌进行分离,根据霉菌的形态特征、培养性状及ITS序列分析,鉴定其为哈茨木霉、拟康氏木霉、脉孢霉、长枝木霉、黑曲霉、产红青霉和产黄青霉;在此基础上探讨了常用抑菌剂对霉菌的防治效果及对海鲜菇菌丝生长的影响。结果表明,质量浓度100 mg/L克霉灵对哈茨木霉、黑曲霉、产红青霉、产黄青霉、拟康氏木霉有强抑制作用,质量浓度100 mg/L多菌灵对长枝木霉、产红青霉、产黄青霉、拟康氏木霉有强抑制作用,二者对海鲜菇菌丝生长的抑制都比较弱。可为海鲜菇工厂化生产中污染霉菌的综合防治提供参考。     

我国农作物根际土壤木霉菌资源分布调查

通过对我国18个省市地区的农作物根际土壤进行采样,使用木霉菌选择性培养基(TSM)和马铃薯葡萄糖培养基(PDA),对采样土壤中的木霉菌(Trichoderma spp.)进行分离和纯化,再与同一地区的多种植物病原菌进行平板对峙测试,筛选和保存了1 300余株有拮抗功能的木霉菌株。经形态学和分子生物学(ITSTef1-α)鉴定,这些菌株分属9个木霉菌种。其中哈茨木霉(T.harzianum)和棘孢木霉(T.asperellum)为优势菌种,分别占29.07%和29.91%(总数:393和382株);其次是绿色木霉(T.viride),占筛选总数的14%(总数:185株);而渐绿木霉(T.viridescens)和康宁木霉(T.koningii)分别占7.53%和7.00%(总数:99和92株);深绿木霉(T.atroviride)和钩状木霉(T.harmatum)分别只占5%以上(总数:69和67株);长枝木霉(T.longibrachiatum)和橘绿木霉(T.citrinoviride)的群落在我国农作物根际土壤中的分布最少,分别只占1.5%和0.5%以上(总数:21和7株)。同时,为了解不同地区的木霉菌株对上海地区的草莓灰霉菌(Botrytis cinerea)的拮抗作用的差异性,选用14个不同地区的木霉菌株,通过对峙培养拮抗测定,筛选出6个木霉菌株,进一步做了抑制草莓灰霉病的室内生测试验。结果显示,来源于华东地区的木霉菌株TR78和TR85,在平板对峙试验中对草莓灰霉菌生长的抑菌率分别为85.49%和82.90%,室内生测对灰霉病的防治效果分别为80.19%和80.02%,显著高于(P0.05)其他菌株对灰霉病的防治效果。     

设施番茄根围土样中木霉菌多样性及功能活性分析

为探索设施栽培条件下蔬菜根围土壤中木霉种群多样性特征及其抗常见植物病原真菌及有机磷农药降解活性,从连续20多年种植番茄的设施大棚根围土样中以及番茄根系内部分离木霉菌,并以tef1-α序列和系统发育分析进行种类鉴定,估算各菌株的相对种群密度;对其生防和农残降解活性进行了测定。结果共分离不同木霉菌株27株,结合形态学特征和tef1-α序列分析,鉴定出分属6个不同的木霉种,即绿色木霉(Trichoderma virens)、深褐木霉(T.atrobrunneum)、西蒙斯木霉(T.simmonsii)、深绿木霉(T.atroviride)、厚木霉(T.crassum)和1个疑似木霉新种(T.sp.nov.);木霉各菌株产孢簇常呈鲜绿色到暗绿色,分布不均匀,常形成同心圆结构。对峙条件下这些木霉对土壤中常见土传病原菌立枯丝核菌(Rhizoctonia solani)、大丽轮枝菌(Verticillium dahliae)、尖孢镰孢菌(Fusarium oxysporum)均有普遍的不同程度的抑菌活性;除深褐木霉两个菌株对毒死蜱的降解效果不明显外,其它种类的木霉均有菌株能有效降解毒死蜱,木霉菌株在根围土壤中的相对种群密度仅为104 cfu/g,且菌株间差异较大。研究结果表明设施农业土壤中木霉菌具有丰富的种质资源多样性,在形态特征及功能活性方面表现明显的菌株差异;木霉对3种常见植物病原真菌有普遍的抑菌作用,少数菌株对常用有机磷农药降解作用明显,可以发挥生防和农化残留降解方面的协同效应。该结果对于研究设施农业土壤中木霉的多样性,指导功能菌株筛选及应用,更好挖掘利用木霉菌资源有着较好的借鉴意义。     

木霉对辣椒疫霉菌抑制作用的超微结构与细胞化*

哈茨木霉菌株NF9和TC3对辣椒疫霉病菌具有强烈的抑制作用。超微结构与细胞化学研究表明,木霉菌丝能够缠绕并寄生疫霉菌菌丝,且重寄生作用主要发生在培养基内的菌丝上。但在绝大多数情况下,木霉菌可直接水解和破坏疫霉菌菌丝,说明木霉抑菌作用的主要机制是直接向外分泌水解酶分解疫霉的细胞壁及细胞质,而不是重寄生作用。不同培养基对木霉的重寄生作用有重要影响,减少培养基中葡萄糖含量可以增强木霉的重寄生作用和抑菌效果。此外,在菌落生长后期,疫霉的新生菌丝均可侵染自身的老菌丝。     

拮抗北细辛菌核病木霉菌的分离、鉴定及生防效果

[背景] 菌核病是北细辛根部主要病害之一,木霉菌作为目前应用最广泛的生物防治真菌,利用木霉菌防治北细辛菌核病是目前研究的热点。[目的] 通过稀释分离法对健康北细辛植株根际土壤进行菌株分离,以期筛选出有效拮抗北细辛菌核病的生防木霉菌。[方法] 以北细辛菌核病菌为靶标菌,采用平板对峙培养、挥发性与非挥发性物质抑菌的方法对分离得到的木霉菌进行筛选,采用生长速率法对筛选出的木霉菌的发酵液进行抑菌效果测定,并采用硫代巴比妥酸法测定筛选出的木霉对北细辛菌核病菌的丙二醛（Malondialdehyde,MDA）含量、紫外吸收法测定过氧化氢酶（Catalase,CAT）活性、氮蓝四唑法测定超氧化物歧化酶（Superoxide Dismutase,SOD）活性、愈创木酚法测定过氧化物酶（Peroxidase,POD）活性的影响。[结果] 从土壤中分离出木霉菌共14株,通过形态学和ITS-RPB2双基因联合构建系统发育树,鉴定其为哈茨木霉（Trichoderma harzianum）、钩状木霉（Trichoderma hamatum）、拟康氏木霉（Trichoderma koningiopsis）、深绿木霉（Trichoderma atroviride）、短密木霉（Trichoderma brevicompactum）和装絮木霉（Trichoderma tomentosum）。对峙培养试验表明,钩状木霉A26、拟康氏木霉B30、钩状木霉C6、哈茨木霉A17对北细辛菌核病菌抑制率均在90%以上,挥发性物质抑制测定结果显示钩状木霉C6抑制率最高,为53.73%±0.07%,木霉菌的非挥发性物质抑菌作用较强,哈茨木霉A17、钩状木霉A26、钩状木霉C6的非挥发性物质对细辛菌核病菌的抑制率均在75%以上,而拟康氏木霉B30抑制率可达100%。因此,筛选出的哈茨木霉A17、钩状木霉A26、拟康氏木霉B30、钩状木霉C6为拮抗效果较强的生防木霉菌,这4株木霉菌的发酵液对北细辛菌核病菌的抑制率分别为56.33%±0.12%、77.22%±0.06%、82.28%±0.03%、46.20%±0.04%。经这4株木霉菌的非挥发性物质处理7 d后,菌核病菌MDA含量显著增加,钩状木霉A26是对照组的7.7倍,最为显著;菌核病菌抗氧化酶活性均降低,与对照组相比,CAT、SOD、POD活性分别下降了19.67%-75.84%、4.71%-68.71%和3.57%-67.86%。[结论] 从北细辛健康植株根际土壤中分离的木霉菌株哈茨木霉A17、钩状木霉A26、拟康氏木霉B30、钩状木霉C6对北细辛菌核病菌均有较好的抑制效果,可用于北细辛菌核病的生物防治。     

木霉对辣椒疫霉菌抑制作用的超微结构与细胞化学

哈茨木霉菌株NF9和TC3对辣椒疫霉病菌具有强烈的抑制作用。超微结构与细胞化学研究表明,木霉菌丝能够缠绕并寄生疫霉菌菌丝,且重寄生作用主要发生在培养基内的菌丝上。但在绝大多数情况下,木霉菌可直接水解和破坏疫霉菌菌丝,说明木霉抑菌作用的主要机制是直接向外分泌水解酶分解疫霉的细胞壁及细胞质,而不是重寄生作用。不同培养基对木霉的重寄生作用有重要影响,减少培养基中葡萄糖含量可以增强木霉的重寄生作用和抑菌效果。此外,在菌落生长后期,疫霉的新生菌丝均可侵染自身的老菌丝。     

4种木霉菌对棉花黄萎病菌抑制作用的测定

以绿色木霉(Trichoderma viride)、康氏木霉(Trichoderma koningii)和二株未知木霉菌株(Trichoderma spp.)为供试木霉菌株,采用对峙培养法测定了不同温度处理下对棉花黄萎病菌(Verticilliam dahliae)的拮抗作用。结果表明,木霉在不同温度下对棉花黄萎病菌的抑制作用不一,其中以30℃和25℃黑暗条件下木霉对棉花黄萎病菌菌丝生长的抑制作用最强,对峙培养5d后,绿色木霉、康氏木霉和二株未知木霉菌在30℃条件下对棉花黄萎病菌的抑制率分别达到67.3%、65.9%、56.8%、65.9%,25℃条件下的抑制率分别为65.9%、72.9%、65.9%、78.8%;20℃下木霉菌株对棉花黄萎病菌的抑制作用次之,10℃、15℃和35℃条件下木霉菌丝扩散速度较慢,且孢子产生量少,不能有效地抑制棉花黄萎病菌菌丝的扩展,表明环境温度过高或过低对木霉菌丝的生长及分生孢子的产生均有较大影响。该研究为筛选棉花黄萎病菌更为有效的生防木霉菌株提供理论依据。     

A new cultivation independent approach to detect and monitor common Trichoderma species in soils

A set of primers was developed for the detection, identification and quantification of common Trichoderma species in soil samples. Based on a broad range master alignment primers were derived to amplify an approximate 540 bp fragment comprising the internal transcribed spacer region 1 (ITS 1), 5.8S rDNA and internal transcribed spacer region 2 (ITS 2) from all taxonomic Clades of the genus Trichoderma. The primer set was applied to test strains as well as community DNA isolated from arable and forest soil. For all tested isolates the corresponding internal transcribed spacer regions of Trichoderma spp. strains were amplified, but none of non-Trichoderma origin. PCR with community DNA from soil yielded products of the expected size. Analysis of a clone library established for an arable site showed that all amplified sequences originated exclusively from Trichoderma species mainly being representatives of the Clades Hamatum, Harzianum and Pachybasioides and comprising most of the species known for biocontrol ability. In a realtime PCR approach the primer set uTf/uTr also proved to be a suitable system to quantify DNA of Trichoderma spp. in soils.     

Endochitinase gene-based phylogenetic analysis of Trichoderma

DNA sequences of the single-copy gene coding for the 42 kDa endochitinase enzyme (EC 3.2.1.14) were used for phylogenetic analysis in Trichoderma. A set of 12 primers was developed and the entire gene was sequenced for 16 strains, and nucleotide and deduced amino acid sequences were compared to data from GenBank for additional Trichoderma strains. Analysis of the sequences revealed parsimony informative variation from 2.4 to 43.6% depending on the part of the gene (exons/introns) and the taxonomic level considered. Results are discussed in comparison to previous data from ITS-1 and ITS-2 rDNA sequencing and suggest the 42 kDa endochitinase gene as a potential molecular marker for reconstructing phylogenetic relationships in the genus Trichoderma at species level.     

Recent advances and future prospects in peptaibiotics, hydrophobin, and mycotoxin research, and their importance for chemotaxonomy of Trichoderma and Hypocrea

Fungi of the genus Trichoderma with teleomorphs in Hypocrea are abundant producers of a group of amphiphilic, non-ribosomal peptide antibiotics, which are rich in the non-proteinogenic amino acid Aib (alpha-aminoisobutyric acid). They are referred to as peptaibiotics, or peptaibols, if a 1,2-amino alcohol is present at the C-terminus. Trichoderma/Hypocrea, like other ascomycetous fungi, also produce hydrophobins, a class of small, cysteine-rich proteins. Advanced soft ionization mass spectrometric techniques such as LC-CID-MS, LC-ESI-MS(n), and IC-MALDI-TOF-MS enabled the high-throughput analysis, simultaneous detection and sequence determination of peptaibiotics and hydrophobins from minute quantities of fungal materials. Some Trichoderma species have been recognized to produce peptaibiotics as well as simple mycotoxins of the trichothecene group. The combination of sequence data of both groups of peptides with the pattern of low-molecular-weight secondary metabolites, including trichothecene-type mycotoxins, independently confirmed the results of morphological, molecular, and phylogenetic analyses. This approach established a new lineage in Trichoderma/Hypocrea, the Brevicompactum clade, comprising four new and one redescribed species. Notably, commercial preparations of single or mixed cultures of Trichoderma species, in particular T. harzianum, and T. koningii, are registered as biocontrol agents for soil and plant pathogens. In this context, it is emphasized that the four mycotoxin-producing species of the recently established Brevicompactum clade (T. brevicompactum, T. arundinaceum, T. turrialbense, and T. protrudens) are not closely related to any of the Trichoderma species currently used as biocontrol agents. Furthermore, possible health concerns about release of peptaibiotics in the biosphere are discussed with respect to their bioactivities and their use as drugs in human and veterinary medicine. Finally, future prospects regarding novel bioactivities and further research needs, including interdisciplinary taxonomic approaches, are outlined.     

Extracellular proteases of Trichoderma species. A review

Cellulolytic, xylanolytic, chitinolytic and beta-1,3-glucanolytic enzyme systems of species belonging to the filamentous fungal genus Trichoderma have been investigated in details and are well characterised. The ability of Trichoderma strains to produce extracellular proteases has also been known for a long time, however, the proteolytic enzyme system is relatively unknown in this genus. Fortunately, in the recent years more and more attention is focused on the research in this field. The role of Trichoderma proteases in the biological control of plant pathogenic fungi and nematodes has been demonstrated, and it is also suspected that they may be important for the competitive saprophytic ability of green mould isolates and may represent potential virulence factors of Trichoderma strains as emerging fungal pathogens of clinical importance. The aim of this review is to summarize the information available about the extracellular proteases of Trichoderma. Numerous studies are available about the extracellular proteolytic enzyme profiles of Trichoderma strains and about the effect of abiotic environmental factors on protease activities. A number of protease enzymes have been purified to homogeneity and some protease encoding genes have been cloned and characterized. These results will be reviewed and the role of Trichoderma proteases in biological control as well as their advantages and disadvantages in biotechnology will be discussed.     

Phylogeny of Philippine slender skinks (Scincidae: Brachymeles) reveals underestimated species diversity, complex biogeographical relationships, and cryptic patterns of lineage diversification

The spectacular, virtually endemic radiation of Philippine semi-fossorial skinks of the genus Brachymeles represent one of the few radiations of scincid lizards to possess both fully limbed and limbless species. And yet, nothing is known of the phylogenetic relationships of this exceptional group. Morphologically similar body plans have made it difficult to assess species-level diversity, and the genus has long been recognized as one of the more modest radiations of southeast Asian lizards. However, recent large-scale survey efforts have resulted in the discovery of numerous new species, and taxonomic studies indicate that the diversity within the genus Brachymeles is grossly underestimated. In this study we provide the first robust estimate of phylogenetic relationships within the genus Brachymeles using a multi-locus dataset and nearly complete taxonomic sampling. We provide statistical tests of monophyly for all polytypic species and two widespread limb-reduced species and our results indicate wholesale deviations from past summaries and taxonomic evaluations of the genus. With few exceptions, we are able to reject the monophyly of all polytypic and widespread species, thereby validating the need for large-scale taxonomic revisions. Our results reveal that the limbless, monotypic, genus Davewakeum is nested within Brachymeles. Mapping of body form on our preferred phylogenetic tree suggests that limb-reduction and digit loss has occurred on multiple occasions in the history of the genus. A Bayesian reconstruction of ancestral areas indicates strong statistical support for a minimum of five major dispersal events that have given rise to a major component of the observed species diversity on separate Pleistocene aggregate island platforms of the archipelago.     

钝头蛇属Pareas（蛇亚目：钝头蛇科）系统分类研究进展

钝头蛇属（Pareas）蛇类形态高度保守,种间形态差异微弱。以分子系统学方法为中心的整合分类方法的应用,为本类群分类难题的解决作出了重要贡献,近8年有9个新种得以描述,其中,仅在近3年就增加了7个新种,并有6个同物异名被恢复。本文依据最新研究成果,对近期钝头蛇属蛇类系统分类研究成果进行了综述,并斟酌中文种名,整理了物种名录,并编制了分类检索表。截止目前,钝头蛇属共有26种,其中在我国有分布的18种,中国特有种9种。同时,对研究中存在的问题进行了探讨,并对下一步工作提出了建议：本属物种多样性估计过低、标本采集覆盖范围不足、证据使用不甚全面,大范围密集采样以及系统发育基因组学方法的应用有助于本类群系统关系的最终解决。     

木霉属真菌的生物降解及生物转化作用研究进展

木霉（Trichoderma spp．）属于半知菌亚门、丝孢纲、丝孢目,粘孢菌类,是一类具有很大应用生产潜力的真菌,目前国际上已查明并命名的木霉属真菌,共计60种和2个变型,国内正式发表的木霉菌有10种,记录种名20个。木霉属真菌的生物降解、转化功能与分泌纤维素酶、葡聚糖酶、几丁质酶、脂肪酶、木聚糖酶等酶的能力相关,综述了木霉属真菌生物降解和生物转化底物及其他方面的研究进展。     

赤眼蜂属系统进化研究进展

赤眼蜂是一类重要的天敌昆虫,该属包括的种类达180多种,并且广泛地分布于世界各地。通过对形态特征的分析,赤眼蜂属的系统发育研究在不同的研究者中得到不同的分类系统。由于赤眼蜂的个体微小,仅04mm左右,现有的化石记录稀少,并且可用作系统发育研究的形态特征很少,导致该类群的系统发育研究仍有许多不清楚的地方。同时,近年来分子生物学的发展为赤眼蜂系统发育研究提供了很好的途径,也取得了一些共识。但仍然未能提出一套精确的、更可靠的分类系统。作者对近年来的赤眼蜂属系统进化研究进行了综述。     

Autecological studies of the genus Porphyra

Among the algal associations of exposed northern shores in Britain, those formed by species of the genus Porphyra deserve greater recognition than has so far been accorded to them. The present paper discusses some of the taxonomic criteria of this plastic genus and delimits five species which are widespread on northern British shores.     

Evolution in indigenous west African rice

The genus Oryza contains two cultivated species,O. sativa L., the Asiatic rice, andO. glaberrima Steud. which is limited to Africa. Despite much taxonomic work on the relationships of the wild species of the genus, they are still imperfectly known and there has not been unanimous agreement on nomenclature. Most of the species group into complexes the delimitation of which is a matter of opinion. This paper presents results from experimental work, and a theory for the evolution of African cultivated rice is put forward based on taxonomic studies.