木霉菌对灰葡萄孢菌的拮抗作用

对采自宁夏14个市县的170份土样及其它材料进行分离,得到96个木霉菌株,采用形态分类方法鉴定出9种木霉菌以及2种未知名菌种．它们分别为哈茨木霉（Trichoderma harzianum）、绿色木霉（T．viride）、长枝木霉（T．longibrachiatum）、康氏木霉（T．koningii）、拟康氏木霉（T．pseudokoningii）、项孢木霉（T．fertile）、黄绿木霉（T．aureoviride）、深绿木霉（T．atroviride）、钩状木霉（T．hamatum）以及T．sp1和T．sp2。拮抗作用和抑菌活性测定结果表明：11种木霉菌对葡萄、番茄和黄瓜灰霉病的致病菌——灰葡萄孢菌（Botrytis cinema Petssp．）均有不同程度的拮抗作用,其中哈茨木霉、绿色木霉、黄绿木霉、钩状木霉和T．sp1对病原菌的拮抗作用及抑菌活性显著,其生长速度比病原菌平均快1．1～3．0倍：在对峙培养中拮抗系数达Ⅰ级或Ⅱ级．其抑菌率高迭98．78％。     

3株茶藨生柱锈重寄生木霉菌的形态鉴定及ITS序列分析

[目的]探讨更为客观的木霉菌鉴定手段。[方法]在形态分类的基础上结合分子鉴定手段(rDNA-ITS序列分析),对3株茶藨生柱锈重寄生木霉菌进行分类鉴定。[结果]依据TR1的培养性状和显微特征描述,初步鉴定该菌株为深绿木霉(T.atroviride);TR2、TR3的培养性状和显微特征在一定程度上相似,依据其形态特征,初步鉴定这2个菌株为绿色木霉(T.viridePers.ex Fr.)。从Genbank中深绿木霉和绿色木霉的6个菌株与TR1、TR2T、R3所作的系统发育树可知,TR1应该归为深绿木霉,TR2和TR3属同种真菌,应该归为绿色木霉,这与形态学观察结果一致。[结论]形态特征结合ITS序列分析可作为木霉菌分类鉴定的依据。     

3株茶蔗生柱锈重寄生木霉菌的形态鉴定及ITS序列分析

[目的]探讨更为客观的木霉菌鉴定手段.[方法]在形态分类的基础上结合分子鉴定手段(rDNA-ITS序列分析),对3株茶藨生柱锈重寄生木霉菌进行分类鉴定.[结果]依据TR1的培养性状和显微特征描述,初步鉴定该菌株为深绿木霉(T.atroviride);TR2、TR3的培养性状和显微特征在一定程度上相似,依据其形态特征,初步鉴定这2个菌株为绿色木霉(T.viride Pers.ex Fr.).从Genbank中深绿木霉和绿色木霉的6个菌株与TR1、TR2、TR3所作的系统发育树可知,TR1应该归为深绿木霉,TR2和TR3属同种真菌,应该归为绿色木霉,这与形态学观察结果一致.[结论]形态特征结合ITS序列分析可作为木霉菌分类鉴定的依据.     

11种木霉菌对葡萄灰霉病菌的拮抗作用

对采自宁夏14个市县的170份土样及其它材料进行分离,得到96株木霉菌株,采用形态分类方法鉴定出9种木霉菌以及2种未知名菌种,它们分别为哈茨木霉(Trichoderma harzianum)、绿色木霉(T.viride)、长枝木霉(T.longibrachiatum)、康氏木霉(T.koningii)、拟康氏木霉(T.pseudokoningii)、顶孢木霉(T.fertile)、黄绿木霉(T.aureoviride)、深绿木霉(T.atroviride)、钩状木霉(T.hamatum)以及T.sp1和T.sp2。竞争及对峙培养结果表明:11种木霉菌种对葡萄灰霉病菌(BotrytiscinereaPers.)均有不同程度的拮抗作用,尤其前四种木霉菌表现出了很强的竞争优势,在对峙培养中对葡萄灰霉菌的拮抗系数为Ⅰ￣Ⅱ,抑菌率达61.8%￣77.6%。在光学显微镜下可观察到木霉菌或缠绕或穿入葡萄灰霉菌菌丝生长,并发生菌丝断裂、消解以及细胞原生质浓缩等现象。     

保护地土壤生防木霉菌种群多样性研究

采用传统的形态学特征和分子方法(ITS、TEF序列和UP-PCR)研究了蔬菜保护地土壤中木霉菌种群多样性及其影响因素.结果表明:提交genbank 11个木霉种的序列,分别为长枝木霉(Trichoderma longibrachiatum)、拟康氏木霉(Trichoderma pseudokoningii)、黄绿木霉(Trichoderma aureoviride)、棘孢木霉(Trichoderma asperellum)、深绿木霉(Trichoderma atroviride)、哈茨木霉(Trichoderma harzianum)、非钩木霉(Trichoderma inhamatum)、微孢木霉(Trichoderma minutisporum)、长孢木霉(Trichoderma longipile)和粘绿木霉(Trichoderma virens);木霉菌24个菌株经UP-PCR扩增,引物AS4、AS19、L45扩增出一条500 bp大小的木霉菌种的特征性谱带,其他谱带则为多态性谱带,多态性达93.5％;营养条件、杀菌剂及土壤因素对不同种木霉菌的影响不同,得到2株适应性较强的木霉菌株,有望成为生防菌株.     

产木聚糖酶菌株JF8的鉴定及固态发酵条件研究

从采集的土壤中分离得到一株高产木聚糖酶的菌株JF8,对该菌株的菌落形态、分生孢子梗及瓶梗着生方式和分生孢子大小等形态学特征进行观察.结果发现,该菌株与棘胞木霉(Trichoderma asperellum)的形态特征极为相似,初步鉴定该菌株为棘胞木霉.进一步对该菌株的rDNA-ITS序列进行克隆并测序,利用DNAstar软件中的Chstal V法建立与该菌株ITS序列同源性较高的木霉属不同种间的系统发育树,发现该菌株的ITS序列与Genebank中已报道的棘胞木霉的ITS序列的同源性高达99.6%,结合形态特征观察结果,证实该菌株为棘胞木霉.以蔗渣和麸皮为基质,先对影响该菌株产木聚糖酶的单个因素进行研究,而后采用L,9(34)正交实验对各因素对产酶影响大小进行分析.结果表明,4种因素对产酶影响从大到小依次是温度、湿度、初始pH和麸皮添加比例,最佳发酵产酶组合为:培养温度28℃,水分添加量12.5 mL,初始pH值3,麸皮添加比例30%.在此优化条件下静止培养72 h,菌株产酶活力可达5 021.1U·g-1干曲.     

生防长枝木霉菌培养特性及形态学与系统发育学鉴定

采用宏观观察法和显微镜镜检的方法,在PDA和MEA培养基上对1株自我采集的具有生防价值的木霉菌株T05进行了培养特性和形态学研究。结果表明:该菌株在PDA平板培养基上生长较快,产分生孢子较早且多,也能产生厚垣孢子;在MEA上能产生分生孢子,但不产生厚垣孢子。该菌株产孢簇为松散羊毛状到紧实的疱状结构;分生孢子梗具有较简单的分枝系统,通常呈直角的1~2次分枝,单生或有时成对。瓶梗不规则地在侧面分布,时常单生,安瓿形或柱形,基部常缩缢但不明显;分生孢子单细胞、长方形到椭圆形、绿色、光滑,大小为(2.0~3.0)μm×(2.0~6.0)μm。根据这些形态特征将菌株T05鉴定为长枝木霉(Trichoderma longibrachiatum)。在此基础上,对T05的rDNA转录间区序列ITS和翻译延伸因子基因tef1-α的第5内含子序列进行了扩增、测序。系统发育学的研究表明,该菌株的ITS序列和tef1-α的第5内含子序列与长枝木霉的多个菌株都具有最大的相似性,因此属于长枝木霉。     

一株飞龙斩血内生真菌的分离鉴定及其活性研究

从飞龙斩血内分离得到一株广谱、高活性抑菌内生真菌F-46／2,其对细菌、植物病原真菌和皮肤致病真菌24种病原微生物有不同程度的抑制作用。形态特征表明,该菌株与木霉属Trichoderma中的康氏木霉Trichoderma koningiopsis的特征基本一致。ITS序列分析显示,本菌株与康氏木霉Trichoderma koningiopsis同源性高达98％,因此将菌株F-46／2命名为康氏木霉Trichoderma koningiopsis F-46／2。     

4种木霉对核盘菌的抑制作用

通过对峙培养以及对木霉菌(Trichoderma spp.)次生代谢物对病原菌的抑菌率进行了测定,结果表明,供试哈茨木霉(T.harzianum)、绿色木霉(T.viride)、康氏木霉(T.koningii),以及一株未知种名的木霉菌株TY(Trichoderma sp.)均对核盘菌(Sclerotinia sclerotiorum (Lib) de Bary)有明显的抑制作用.在对峙培养中,木霉菌菌落与核盘菌的菌落相向生长一定时间后,挑取核盘菌菌落内部的菌丝进行镜检,发现有木霉菌孢子存在,且核盘菌发生菌丝变形,如原生质浓缩,菌丝断裂,或菌丝消解等现象.并发现不同种木霉所产生的次生代谢物的活性不同,对核盘菌的抑制作用也存在显著差异.     

保护地蔬菜土壤中木霉菌种群影响因素

采用传统的形态学特征和ITS、TEF序列比对鉴定出11个木霉种,其中长枝木霉Trichoderma longibrachiatum、深绿木霉Trichoderma atroviride和哈茨木霉Trichoderma harzianum是蔬菜保护地土壤中的优势种群.同时研究表明种植年限、蔬菜种类、轮作制度、杀菌剂等均会影响木霉菌数量和分布,木霉菌的数量与种植年限、蔬菜种植结构密切相关,连作时,当种植年限超过10 a,木霉菌数量减少;轮作地比连作地木霉菌数量虽然下降,但在真菌中的比例却呈上升趋势;种植葱蒜类蔬菜的地块比种植番茄、黄瓜的地块木霉菌数量多.从蔬菜田中分离到的木霉菌对杀菌剂的抗性较强.土壤因素中以土壤有机质含量对木霉菌数量的影响最为重要,达到极显著水平,回归直线方程为y=0.13x-1.07.表明提高土壤中有机质含量及轮作是提高木霉菌生物种群多样性的主要因子.     

Trichoderma species collected from Iran

In order to identify Trichoderma species isolated from Iran, Trichoderma selective media and malt extract agar (MEA) were used to isolate Trichoderma species from the soil samples. All the cultures were purified on 2% water agar by hyphal tip method prior to morphological examination.Morphological observations were carried out on the cultures grown on 2 % MEA and oat meal agar at 20℃ under ambient laboratory conditions.     

Trichoderma species from China

Seventeen species of Trichoderma, isolated from soil or tree bark from Ch ina are identified based on morphological and physiological characters, and from their phylogenetic position inferred from parsimony analyses of nucleotide sequ ences of the internal transcribed spacer regions of the rDNA cluster (ITS1 and 2) and partial sequences of translation elongation factor 1-alpha (te f1) . There were T.citrinoviride, T.longibrachiatum, T.sinensis in section Long ibrachiatum, T.…     

Population dynamics of Trichoderma species in the rhizosphere of tobacco and four species form China

To study the effect of tobacco growth on Trichoderma population, we in vestigated the occurrence of Trichoderma species in the rhizosphere of tob acco plant during the period from transplanting (June) to harvesting (October) and measured relative environmental factors. Eleven species of Trichoderma we re isolated, among which T.harzianum, T.viride, T.hamatum, T.atroviride, T.lo ngibrachiatum, T.virens, T.koningii were identified, other four species Ty1, Ty2, Ty3, Ty4 are new s…     

海南五指山野生美花兰根部内生真菌研究

美花兰(Cymbidium insigne)为大花蕙兰(Hybrid cymbidium)杂交育种的主要原生种之一,具有极为重要的保护和开发利用价值。内生真菌在兰科植物整个生活史中起着重要的作用。为了解美花兰内生真菌菌群种类,本研究采用组织分离法首次对海南岛五指山地区的野生美花兰根部的内生真菌进行分离,并根据所分离真菌的形态和ITS序列进行鉴定。共分离到内生真菌132株,确定为14属,隶属于半知菌类的Pestalotiopsis 5株,Aspergillus 16株,Penicillium 27株,Trichoderma 13株,Cylindrocarpon 14株,Curvularia 4株,Nigrospora 3株;Nodulisporium 11株,子囊菌门的Chaetomium 3株,Pezicula 1株,Neonectria 6株,Cryptosporiopsis 6株;担子菌门的Tulasnella 9株;Ascomycete sp.9株,因未见产孢暂不能确定其具体分类地位;5株菌(W2-2 1株、W2-6 2株及W6-5 2株)经诱导未见产孢,且其ITS序列与GeneBank中的序列相似性较低,可能是新的属或种。将所得ITS序列提交GeneBank,获得新的登陆号HQ889705-HQ889725。     

Genetic regulation of conidiation in Trichoderma hamatun

Achieving a balance between vegetative growth and spore production is essential for successful biocontrol by fungi. Low sporulation rates in the field can result in poor establishment and survival,whereas failure of conidia to recognise hosts can lead to persistence without efficacy. Commercial biocontrol products involve bulk preparations of conidia, however considerable variability in conidiation rates exists between biocontrol agents, which can restrict choice of strain for production. The majority of studies on Trichoderma conidiation have focused on the species T. viride and T. atroviride.These species form conidia in response to blue and near-UV light and/or nutrient deprivation and conidiation proceeds in a highly co-ordinated fashion, however relatively little is known on the genetic basis of Trichoderrma conidiation. In addition, whilst photoconidiation appears to be a general response detailed studies in other Trichoderma species are absent. In this study, conidiation in the lesser known biocontrol species T. hamatum is being investigated using a combined morphological and molecular approach. In contrast to T. atroviride, conidiation in response to blue-light was weaker and variable and suggested that additional triggers may be required for the T. hamatum photoresponse. A series of comparative photoconidiation assays are currently being undertaken investigating the effect of inoculum type and abiotic factors on timing and intensity of the response.Results will be discussed in relation to the current knowledge on conidial morphogenesis in Trichoderma. In addition to these morphological assays, a selection of genes implicated in sporulation and the blue-light responses are currently being isolated and characterised from T. hamatum. Two genes, phr1 and cmp1 , which were isolated previously from T. atroviride will be used as early and late markers of gene expression during the photoresponse in T. hamatum in order to define time points for harvesting comparable stage-specific RNA from T. hamatum and T. atroviride. Using degenerate PCR putative sporulation gene orthologues have also been identified in T. hamatum.Work is currently underway to isolate genomic clones of these genes from T. hamatum and T.atroviride. Sequence and expression analysis of orthologues, including expression in response to abiotic factors will be presented and discussed in relation to the current knowledge of the molecular basis of conidiation in Trichoderma and other filamentous fungi.……     

深绿木霉(Trichoderma atroviride)T23降解有机磷农药敌敌畏转运蛋白TaPdr2基因的克隆与功能预测分析

根据前期研究,进行了对比分析后明确了深绿木霉中7个敌敌畏耐受相关ABC转运蛋白,其中TaPdr2基因是在短时间内对敌敌畏胁迫应答最为明显的一个基因。本研究通过简并引物同源克隆,在深绿木霉T23中克隆出TaPdr2基因,并对其全长序列进行了测序;明确了其外显子序列和内含子序列;构建了TaPdr2的系统发生树,发现深绿木霉TaPdr2与木霉属其他种PDR5亚家族ABC转运蛋白有很高的同源性,通过生物信息学分析后预测TaPdr2蛋白的ATPase及部分跨膜结构域序列相对保守且具有11个跨膜结构域;通过构建TaPdr2的敲除载体pC1300qh-F,为后续研究TaPdr2蛋白对麦角甾醇和鞘脂类物质含量影响的研究奠定了基础。