绿僵菌属的一个新种

本文报导从贵州省贵阳罹病梨虎Rhynchites Coreanus Kono幼虫虫尸分离的一种绿僵菌新种的鉴定结果,该菌与绿僵菌属模式种金龟子绿僵菌Metarhizium anisopliae (Metsch) s-orkin 有明显区别,根据菌落颜色、分生孢子团块大小、连接紧密和牢固程度、孢子链连接方式和分生孢子形态等,我们鉴定为新种——翠绿绿僵菌Metarhizium iadini chen、Guo et zhou sp.nov.     

绿僵菌属的三个新种

作者从广东、贵州、北京等地收集的虫生真菌中,发现在我国具有绿僵菌属特征的真菌可分为四个种。根据菌落颜色、分生孢子团块大小、紧密和牢固程度、孢子链连接方式、分生孢子形状等,将我们收集的属于本属的菌株分为四个种。本文报道绿僵菌属三个新种的描述。金龟子绿僵菌Metarhizium anisopliae(Metsch.)Sorokin为最常见的甘蔗金龟子致病菌,是本属的模式种。平沙绿僵菌Metarhizium pingshaense Chen et Guo sp.nov.、柱孢绿僵菌Metarhizium cylindrosporae Chen et Guo sp.nov.和贵州绿僵菌Metarhizium guizhouense Chen et Guo sp.nov.是三个新种。     

响应面设计优化莱氏绿僵菌SZCY固体发酵培养条件及致病力测定

【背景】莱氏绿僵菌(Metarhizium rileyi)对新入侵我国的草地贪夜蛾(Spodoptera frugiperda)具有较强的致病力和田间流行性,因此具备深入开发的价值。【目的】优化莱氏绿僵菌SZCY固态发酵培养条件,测定所产分生孢子对草地贪夜蛾幼虫的毒力,为提高该菌株分生孢子规模化生产奠定基础。【方法】采用单因素试验确定了相对适宜的固态培养基,利用Box-Behnken响应面法优化该菌株的固态培养基和发酵参数,同时评价不同条件下该菌所产分生孢子对草地贪夜蛾幼虫的毒力。【结果】去颖稻谷(rice)为莱氏绿僵菌SZCY菌株固相产孢最佳载体。培养温度、光周期及酵母浸粉含量是影响莱氏绿僵菌SZCY固态发酵产孢量的主要因素。莱氏绿僵菌SZCY固态发酵最佳工艺参数为温度22.83℃、光周期18.68 h L:5.32 h D、酵母浸粉4.98 g/100 g,在此条件下,莱氏绿僵菌在去颖稻谷固态培养基上的产孢量为5.65×10¹⁰孢子/g,用其制备浓度为10⁷孢子/mL的孢子悬浮液,对草地贪夜蛾3龄幼虫的LT₅₀为3....     

壳聚糖酶产生菌KJ0312的鉴定

通过对产壳聚糖酶菌KJ0312群体形态包括菌落大小、颜色、质地等特征的观察,和个体形态包括菌丝、分生孢子梗、分生孢子等特征的研究,以及部分18sDNA序列的鉴定,初步确定产壳聚糖酶菌KJ0312的分类地位属于淡紫拟青霉.     

竹叶锈病重寄生现象及重寄生菌鉴定

竹叶锈病是竹类主要病害之一,影响竹林生长。为了利用重寄生菌对竹叶锈病进行生物防治,研究了竹叶锈病菌重寄生现象及重寄生菌的形态与分类地位。结果表明,该重寄生现象伴随竹叶锈病的发生而从4月下旬至5月初开始出现;重寄生菌的自然重寄生率于5月至8月初呈快速增长趋势,8月中旬达到高峰期,随后其重寄生率呈缓慢下降趋势;重寄生菌寄生在竹叶锈病夏孢子堆上呈白色的点状霉状物,阻碍了竹叶锈病夏孢子的发育和释放。通过对其分生孢子梗、分生孢子及菌丝等形态特征观察,该重寄生菌被鉴定为赭红枝顶孢Acremonium salmoneu     

两株椰心叶甲分离物的鉴定及其系统发育分析

采用形态学方法对2株从自然罹病死亡的椰心叶甲虫尸上分离到的致病菌株Dz01和Ma4进行了鉴定,发现2个菌株在菌丝、瓶梗和分生孢子等形态特征上与金龟子绿僵菌小孢变种基本一致,可将2个菌株鉴定为金龟子绿僵菌小孢变种。基于Dz01和Ma4菌株和其它31个代表绿僵菌主要种或变种菌株rDNA上ITS1-5.8S-ITS2区序列构建的最大简约树显示,Dz01和Ma4菌株均聚在金龟子绿僵菌小孢变种所构成的分支中,这为2个菌株形态学鉴定结果提供了分子依据。     

竹林金针虫对绿僵菌及其代谢产物的行为反应

绿僵菌(Metarhizium spp.)是地下害虫重要的生防真菌,为了明确绿僵菌对金针虫的驱避作用,本文以平沙绿僵菌WP08菌株为研究对象,通过Y型管等嗅觉生测方法测定了筛胸梳爪叩甲幼虫对绿僵菌分生孢子、非挥发物和挥发性物的行为反应。结果表明:金针虫的选择行为与土壤中平沙绿僵菌分生孢子浓度密切相关,当土壤中绿僵菌孢子浓度达到5×10~8个·g~(-1)干土时,绿僵菌对金针虫表现出极显著的驱避作用(P0.001),随着浓度的降低,驱避作用减弱,当浓度达到5×10~6个·g~(-1)干土时,驱避作用消失;平沙绿僵菌孢子发酵液(浓度为1×10~8个·mL~(-1))对金针虫驱避作用极显著(P0.001),而挥发物对金针虫驱避作用不明显(P0.05)。本研究结果为揭示绿僵菌驱虫的内在机制提供了重要的基础信息。     

红火蚁Toll受体家族免疫响应绿僵菌表达模式的研究

为探究红火蚁Solenopsis invicta Buren Toll受体家族基因如何免疫响应绿僵菌Metarhizium的侵染。本研究采用浸渍法测定了不同浓度下绿僵菌对红火蚁大中小3种不同大小工蚁的致病力,并用显微镜观察了绿僵菌侵染后红火蚁3种工蚁的表型变化。利用生物信息学筛选Toll受体基因,并对Toll受体家族基因的理化性质、结构域、染色体位置、系统进化进行分析鉴定。运用实时荧光定量PCR（RT-qPCR）检测了红火蚁大中小型工蚁中Toll受体家族的发育历期和绿僵菌侵染后的表达模式。结果表明,绿僵菌在96 h对红火蚁大中小型工蚁的致死中浓度（LC50）分别为5.8×10^7孢子/mL、3.1×10^7孢子/mL、1.5×10^7孢子/mL;绿僵菌侵染红火蚁第3天,显微镜下观察到红火蚁体壁细小的菌丝,虫体僵硬,寄主死亡后菌丝迅速蔓延并逐渐长成橄榄绿色分生孢子;RT-qPCR结果表明Toll受体基因在红火蚁不同发育阶段中的mRNA表达水平存在显著差异,且Toll受体基因在雌性生殖蚁表达水平显著高于雄性生殖蚁;RT-qPCR结果表明红火蚁大中小型工蚁Toll受体免疫响应绿僵菌不一样。在大型工蚁中,绿僵菌侵染后,Toll受体家族基因能显著上调表达,6 h是一个诱导高峰期,Toll2-1和LRR转录水平最高,响应绿僵菌最强;在中型工蚁中,绿僵菌不能刺激Toll2-2基因的表达,却能强烈诱导Toll1,Toll2-1,Toll6,Toll7和LRR基因的上调表达,Toll1和Toll2-1响应绿僵菌最强。在小型工蚁中,绿僵菌能显著诱导Toll受体家族基因基因的上调表达,在24 h时,LRR基因表达量最高,相比于对照,LRR基因表达提高25倍,LRR在6 h和24 h响应绿僵菌最强。以上研究表明Toll受体可以免疫响应入侵的绿僵菌,且不同的Toll对绿僵菌可能具有不同的响应机制。本研究为进一步阐明Toll受体的功能奠定基础,为进一步利用绿僵菌控制红火蚁提供技术指导。     

虫草一新种及其绿僵菌无性型

从贵州省都匀茶场采得一虫草新种。为缅怀戴芳澜教授对我国真菌学的贡献,我们命名这个新种为戴氏虫草(Cordyceps taii Liang et Liu)。在虫草属于囊壳倾斜埋生群(Cre-mastocarpon)的肉质亚群(Subsect．Carnosae)中,戴氏虫草与已知种的主要鉴别特征是具有较长的次生子囊孢子(21—29μm)。通过分离培养和于囊孢子的微循环产孢(microcycle-conidiation)已基本证明,戴氏虫草的无性型是戴氏绿僵菌新种(Metarhizium taii Lianget Liu)。这种绿僵菌的主要鉴别特征是形成孢梗束并偶有双细胞分生孢子。     

绿僵菌对小麦纹枯病菌的抑制作用研究

在实验室条件下,研究了金龟子绿僵菌（Metarhizium anisopliae）对小麦纹枯病菌（Rhizoctonia cerealis）的拮抗作用及其机理。结果表明,金龟子绿僵菌与小麦纹枯病菌对峙培养以及在培养基中加入金龟子绿僵菌孢子悬浮液,对小麦纹枯病菌菌丝生长均有较好的抑制作用。测定了培养不同天数的金龟子绿僵菌Ma55发酵液对小麦纹枯病菌菌丝生长、菌核产生量及菌核萌发率的影响。结果表明,液体振荡培养25 d的金龟子绿僵菌Ma55发酵液对小麦纹枯病菌的菌丝生长、菌核产生量及菌核萌发具有显著的抑制作用,且Ma55发酵液中的抑菌活性物质具有较好的热稳定性。在光学显微镜下,未观察到Ma55对小麦纹枯病菌的重寄生现象,但发现金龟子绿僵菌与小麦纹枯病菌对峙培养处小麦纹枯病菌营养菌丝的细胞质变稀薄、菌丝部分消解或断裂。上述结果显示,金龟子绿僵菌对小麦纹枯病菌的拮抗机制主要是营养竞争、空间竞争及抗生作用等。     

武汉东湖若干种轮虫休眠卵的形态观察

休眠卵是轮虫在恶劣生态条件下形成的滞育结构。本研究用光学显微镜和扫描电子显微镜对东湖14种轮虫休眠卵形态进行了观察,其中关于尾突臂尾轮虫（Brachionuscaudatus,Barrois＆Daday）、方形臂尾轮虫（Bquadridentatus,hermann）、中型晶囊轮虫（Asplanchnaintermedia,Hudson）、顶生三肢轮虫（Filiniaterminalis,Plate）的休眠卵的形态特征尚未见过报道。这些休眠卵一般卵壳较厚,颜色较深,壳表面有纹饰。不同种轮虫的休眠卵形态、壳饰各不相同,具种的特异性,但在某些轮虫,即使同一种中休眠卵形态有时亦存在较大的差异。     

Contribution to the morphology of the productive strains ofPenicillium chrysogenum thom from the Wisconsin family

The morphology of four productive strains ofPenicillium chrysogenum Thom from the Wisconsin family was studied. The strains Q-176, 47–1564, 49–133, 51–20Z, which were naturally or artificially obtained mutants of thePenicillium chrysogenum NRRL 1951 strain were very variable as to the colony structure and the character of conidiophores. The present study is concerned with the evaluation of their taxonomic position. The macrohabitus of the colonies was not remarkably changed. All different types of colonies (U, D, C, B, rarely A) described by Backus and Stauffer, were found on Czapek agar; they were not recognized on malt agar. Deviations from the normal asymmetric conidiophore were found with every type of colonies, most often with the more floccose or lanose ones showing a higher and a sparser overgrowth. These deviations or changes in the microstructure were divided into three degrees according to their quality and occurrence: (1) A strongly divaricate conidiophore where only metulae and phialides were developed; (2) monoverticilate conidiophore or single phialides on the conidiophore filament; (3) degeneration of phialides or metulae to sterile globose cells or an ultimate reduction of conidiophore to dichotomically branched stump-like hypha. The investigated strains can be involved in the taxonPenicillium chrysogenum Thom; it is necessary, however, to include some additional traits into the characteristics of the taxon: Colonies of the naturally or artificially obtained mutants often have lanose overgrowths sporulating sparsely. Formation of the yellow pigment and the exudate was not obligatory. conidiophores of these strains had a tendency to be more simple. They were scarcer, divaricately open, characterized sometimes by the formation of monoverticilate penicilli. A degeneration was frequently found of the ends of conidiophores (phialides and metulae) to globose enlarged sterile cells as well as the formation of giant cells in the mycelium or reduction of conidiophore to dichotomically branched hypha with stump-like ends.     

与成纤维细胞共培养的肺泡II型上皮细胞的生物学特性

建立胎鼠肺泡II型上皮细胞(AECII)与肺成纤维细胞(LF)共培养模型,观察与LF共培养下AECII的生物学特性。倒置相差显微镜观察AECII形态和基本生长情况;RT-PCR和流式细胞术分别检测肺泡表面活性蛋白-C(SP-C)、水通道蛋白5(AQP5)mRNA及蛋白质表达;流式细胞术检测细胞周期及Ki67表达。结果显示,与LF共培养时,AECII能较好地保留其细胞形态,SP-CmRNA及其蛋白质表达明显增加,而AQP5mRNA及其蛋白质表达则明显减少;LF促进AECII增殖,使G2/M、S期细胞及表达Ki67 细胞的比率明显增多。结果提示,AECII与LF共培养时,能更好地保留其细胞形态、分化及增殖特性。     

Consecutive monitoring of lifelong production of conidia by individual conidiophores of Blumeria graminis f. sp. hordei on barley leaves by digital microscopic techniques with electrostatic micromanipulation

Conidial formation and secession by living conidiophores of Blumeria graminis f. sp. hordei on barley leaves were consecutively monitored using a high-fidelity digital microscopic technique combined with electrostatic micromanipulation to trap the released conidia. Conidial chains formed on conidiophores through a series of septum-mediated division and growth of generative cells. Apical conidial cells on the conidiophores were abstricted after the conidial chains developed ten conidial cells. The conidia were electrically conductive, and a positive charge was induced in the cells by a negatively polarized insulator probe (ebonite). The electrostatic force between the conidia and the insulator was used to attract the abstricted conidia from the conidiophores on leaves. This conidium movement from the targeted conidiophore to the rod was directly viewed under the digital microscope, and the length of the interval between conidial septation and secession, the total number of the conidia produced by a single conidiophore, and the modes of conidiogenesis were clarified. During the stage of conidial secession, the generative cells pushed new conidial cells upwards by repeated division and growth. The successive release of two apical conidia was synchronized with the successive septation and growth of a generative cell. The release ceased after 4-5 conidia were released without division and growth of the generative cell. Thus, the life of an individual conidiophore (from the erection of the conidiophore to the release of the final conidium) was shown to be 107 h and to produce an average of 33 conidia. To our knowledge, this is the first report on the direct estimation of life-long conidial production by a powdery mildew on host leaves.     

我国首例肺微小根毛霉病及其致病菌的分离和培养

本文报告1例肺结核和糖尿病并发由微小根毛霉(Rhizomuror pusillus)所致的肺微小根毛霉病。首次在我国从病人的肺组织活检标本分离出该种真菌。此菌可在几种琼脂培养基20°—45℃条件下生长,最适温度为37℃。菌落在初期白色,然后变成褐色厚毡状。在光学显微镜下可看到孢囊梗假轴状分枝,初期无色,然后变为褐色。孢子囊直径50—90μm,呈灰色,后变为褐色。孢子囊成熟后囊壁消解。囊轴卵形或梨形,直径45—48μm。在有性期,接合孢子球形,直径43—63μm,初呈褐色,后变为黑色,表面粗糙或凹凸不平。在透射电子显微镜下,孢囊孢子呈不规则卵形,直径2—5μm;在扫描电子显微镜下,孢囊孢子形态与上述相同。本菌实验感染家兔、豚鼠和小白鼠显示毒力很强,动物于接种后3—10天内全部死亡。从感染动物的脏器分离出本菌。采用中、西医结合治疗,病人痊愈。     

A Study of Pollen Morphology of Sparganium from China

Pollen morphology of 8 species in genus Sparganium from China has been studied. All of them were examined under the light microscope and SEM. It may draw the following conclusions: The characteristics of pollen grains in genus Sparganium are very similar. They are spheroidal, subspheroidal or sometimes slightly angular in shape, 21.8–38.3μm in diameter, monoporate. Porus is slightly sunken, its margin distinct or indistinct, and some granules on the porus membrane. The thickness of exine is 1.7–2.6μ m. The sexine is about as thick as nexine, sometimes thinner than nexine. It is difficult to distinguish one another under the light microscope, but may be seperated under SEM on the basis of the shape or size of lumina and width or height of muff. In some species small excrescenses can be observed under SEM. Some taxonomists (Rendle, 1953) considered that Sparganium is closely related to Pandanus, but others (Hutchison 1934, Takhtajan 1969) to Typha. According to the data of pollen morphology, Sparganium is more close to Typhaceae than to Pandanaceae, thus we agree to put it into family Typhaceae.     

一株仙人掌植物内生真菌的分离鉴定及活性研究

从元江仙人掌的茎中分离到一株产广谱、高活性抑菌物质的内生真菌,经测定对细菌、植物病原真菌和皮肤致病真菌共21种病原微生物有较为明显的抑制作用。形态特征表明,该菌株与曲霉属（Aspergillus）中的土生曲霉（Aspergillus terreus）的特征基本一致,18SrDNA序列分析显示本菌株与土生曲霉的同源性高于99％,但该菌株的分生孢子梗上有明显的瘤状突起,不同于模式菌株。因此认为该菌株为土生曲霉的一个变种,命名为土生曲霉云南变种（Aspergillus terreus vat．yunnanensis）。并对其活性物质的生产条件进行了初步摸索,确定用查氏培养基为最佳种子培养基,PDA培养基为最佳发酵培养基,4d为最适发酵时间。     

Identification and pathogenic characterization of endophytic <Emphasis Type="Italic">Fusarium</Emphasis> species from cowpea seeds

Isolates of Fusarium were obtained and identified from seeds of cowpea, Vigna unguiculata (L.) Walp., by means of blotter tests and slide cultures. Species were differentiated according to the morphology of the macroconidia, microconidia and their arrangement in chains or false heads, the size and type of conidiophore, and the presence or absence of chlamydospores. The species were identified as F. semitectum, F. equiseti, F. oxysporum, F. solani, F. anthophilum, F. sporotrichioides, F. moniliforme, and Fusarium sp. Among the species, F. semitectum was the most frequently detected. None of these species were pathogenic when inoculated in susceptible cowpea cultivar (BR 17- Gurgueia). But, an isolate of F. oxysporum f. sp. tracheiphilum used as a standard of comparison for pathogenicity (control) induced symptoms of yellowing, vascular wilting, and death of a susceptible cowpea cultivar under the same environmental conditions.     

Ecotypes of the fungus Bipolaris oryzae with various responses of the mycochrome system

Two types of fungi based on the light required for induction of conidiophore formation are known as 'photo-induced' and 'non-photo-induced' sporulators. Conidial development in a 'photo-induced' strain of Bipolaris oryzae (formerly, Helminthosporium oryzae ) is controlled by antagonistic action of blue and near-UV radiation mediated through the 'mycochrome' system at two developmental stages: conidiophore induction and conidiophore maturation. Otherwise, conidial development in a 'non-photo-induced' strain of B. oryzae , unlike that in the 'photo-induced' strain, is controlled by antagonistic action of blue and near-UV radiation at the conidiophore maturation stage alone. In this study, we collected fungi, B. oryzae , from brown lesion spots on rice leaves cultivated in paddy fields, and investigated the possible existence of a new ecotype different from known 'photo-induced' and 'non-photo-induced' sporulators. We also investigated the sensitivity of conidial development of each isolate to blue and near-UV radiation.
The isolates were identified as B. oryzae by morphology of hyphae and conidia and the symptomatic brown lesion spots on rice leaves caused by each isolate. There was no distinction between 'photo-induced' and 'non-photo-induced' strains on rDNA base. Moreover, the existence of a new type of 'non-photo-induced' sporulator different from the one presently known was confirmed. In this newly identified sporulator, conidial development was not affected by light irradiation. Among 153 isolates, one isolate belonged to a previously known 'non-photo-induced' sporulator, five isolates belonged to a new 'non-photo-induced' sporulator ecotype, and over 95% of the isolates were "photo-induced" sporulators. Among 'photo-induced' strains, there were several subtypes having different sensitivities to near-UV radiation required for inducing conidiophore formation.     

柿叶黄酮对BSD-2菌株芽胞数量及抑菌活性的影响

研究PB液体培养基中加入不同浓度的柿叶黄酮对内生枯草芽胞杆菌生长、芽胞形成及抑菌活性的影响,利用光学显微镜观察菌体形态、稀释平板法统计芽胞数量、平板扩散法和病原菌生物测定的方法测定抑菌效果。结果表明,加入柿叶黄酮浓度为0.5%的处理对菌体生长无影响,使芽胞数量提高100倍,同时增强对灰霉、黑斑、黄萎病原菌的抑菌效果。