印度块菌子囊果内部解剖结构的扫描电镜观察

利用扫描电镜对成熟印度块菌子囊果的内部结构进行了观察,以系统揭示其子囊果内部组织特征,为块菌属的分类以及块菌属真菌子囊果的生理研究奠定基础。观察结果进一步证明,成熟印度块菌子囊果横切面上的白色迷宫状脉络是由不育的侧丝构成,而暗脉则是被侧丝缠绕并包裹着的可育的菌丝组织,即产孢组织,这些白色脉络和暗脉就构成了印度块菌子囊果横切面上迷宫状的纹脉;产孢组织中,可观察到正在发育的大大小小的子囊被缠绕在一起的大量产囊丝与侧丝包裹着,形成密密麻麻微小的类似蜂巢状结构;子囊孢子游离于子囊中,成熟子囊孢子表面有刺状纹饰,刺的顶端有小弯钩。单个子囊内含的子囊孢子大小与其内含的子囊孢子数目有关,子囊内所含的子囊孢子越多,子囊孢子就越小。     

昆明地区印度块菌子囊果的解剖结构研究

通过印度块菌子囊果石蜡切片的显微观察,对印度块菌子囊果包被及子实层的显微结构、子囊及子囊孢子的发育过程进行研究.结果表明:(1)印度块菌子囊果是由包被和子实层构成,子囊果的表面密被大量大小不一的疣状突起,包被由外皮层和内皮层构成;子实层由封闭的、大小不一的产孢组织构成,在产孢组织中包含有侧丝、产囊丝、子囊和子囊内的子囊孢子;(2)成熟印度块菌子囊果横切面上有明暗相间的迷宫状纹脉;(3)子囊卵形,由产囊丝顶端的细胞发育而来;(4)成熟子囊孢子红褐色,椭圆形至近球形不等,子囊孢子双层壁,外壁密布有刺状纹饰.子囊孢子在子囊内的发育过程中常出现败育现象,每个成熟子囊中含有1～5个子囊孢子(常见4个).     

内蒙古贺兰山某块菌的形态解剖学与分子生物学研究

利用形态解剖学和分子生物学方法,对采自内蒙古贺兰山地区实验样地青海云杉林下的块菌两菌株（菌株a和b）进行分析鉴定。研究发现：（1）两菌株子囊果均为黄褐色,表面光滑,没有明显的疣状突起和棱角。（2）菌株a产孢组织乳白色、致密团状,菌肉组织褐色;球形、棒状子囊呈蜂窝状排布,内含有1～4个带包被的、表面具有突起状纹饰的球型子囊孢子;子囊孢子双层壁,厚约1.7 μｍ,直径约20 μｍ（含纹饰）。（3）菌株b产孢组织有裂隙,松散,子实层内除了具有上述蜂窝状排布的子囊和内部的球型孢子外,还具有“口袋”状子囊,该子囊内含有大量两端尖、外壁光滑、褐色的椭球型孢子。（4）分子生物学进化分析表明,两菌株聚为一支,但属于块菌属的支持率相对较低;推断两菌株可能为中国猪块菌属Choiromyces新记录种。     

同形鳞毛蕨成精子囊素系统的初步研究

研究了同形鳞毛蕨成精子囊素对该种和水蕨孢子萌发和配子体发育的影响,结果表明：同形鳞毛蕨配子体能产生成精子囊素,该成精子囊素能抑制同种孢子的萌发,抑制作用随配子体成熟度的增加而增强;同形鳞毛蕨成精子囊素还可促进同种孢子发育为雄配子体;光照条件下,同形鳞毛蕨成精子囊素对水蕨孢子萌发和配子体发育影响不大,黑暗条件下,同形鳞毛蕨成精子囊素能显著的促使水蕨孢子提早萌发,但都不影响其孢子最终萌发率和配子体的性别分化,表明同形鳞毛蕨和水蕨的成精子囊素不属于同一系统。     

中国一新记录属——桃孢壳属

对日本桃孢壳(Persiciospora japonica)菌进行了描述。该菌的子囊具8个子囊孢子,呈近双列排列;子囊孢子暗橄榄色至黑褐色,椭圆形至梭形,两端具芽孔,孢子表面具小的点状凹陷,光学显微镜下呈网状。该属真菌为我国新记录。     

晶杯菌科3个新种（英文）

对来自四川、甘肃和青海的晶杯菌科盘菌采集物进行分类研究,发现了3个新种,它们隶属于黄杯菌属和绒被盘菌属。竹黄杯菌的子囊盘直径0.2-0.5mm,子实层表面米色至米灰色;子囊具8个子囊孢子,孔口在Melzer’s试剂中呈蓝色,58-68×4.5-5.5μm;子囊孢子梭形,具一个分隔,8-12×2.3-3μm。单胞黄杯菌的子囊盘盘状,直径1mm,子实层表面污黄色;子囊具8个子囊孢子,孔口在Melzer’s试剂中呈蓝色,82-92×6-8μm;子囊孢子梭形,14-21×2-3.5μm。隔孢绒被盘菌的子囊盘盘状,直径0.5-1mm,子实层表面白色至淡灰色,子层托表面具短棒状细胞延伸物;子囊棒状至近圆柱形,具8个子囊孢子,孔口在Melzer’s试剂中呈蓝色,73-84×6.8-7.5μm;子囊孢子柱梭形,具3个分隔,18-20.5×2.5-3.3μm。     

利用荧光蛋白标记研究稻瘟病菌有性世代的细胞结构

稻瘟病是水稻最重要的病害之一,同时稻瘟病菌(Magnaporthe oryzae)是植物病原真菌研究的模式生物。稻瘟病菌是异宗配合的子囊菌,但其有性世代的细胞学过程、形态结构、分子机制以及对病菌变异的贡献研究都较少,也缺乏必要的研究手段。该文利用荧光蛋白标记结合荧光染色的方法对稻瘟病菌有性世代结构进行了标记和显微观察,以期为稻瘟病菌有性生殖过程和机制研究提供方法与借鉴。将绿色荧光蛋白GFP和红色荧光蛋白m Cherry分别导入两个相对交配型的稻瘟病菌菌株Guy11(MAT1-2)和70-15(MAT1-1),各转化子及野生型按交配型组合对峙培养,观察有性世代结构中的荧光表达情况。结果表明,组蛋白H3启动子、核糖体蛋白RP27启动子和疏水蛋白MPG1启动子均可在稻瘟病菌有性孢子形成过程中高丰度表达。进一步利用这三种启动子和两种荧光蛋白对稻瘟病菌有性孢子的细胞器(细胞核、过氧化物酶体)进行标记和观察,结果表明,融合组蛋白H2B的m Cherry及融合核定位信号(NLS)的GFP均能有效标记子囊孢子细胞的细胞核,荧光集中而明亮;带有过氧化物酶体定位信号1(PTS1)的GFP可以有效地标记子囊孢子中的过氧化物酶体,每个细胞中均有数量不等的过氧化物酶体,表明子囊孢子中需要过氧化物酶体参与生化代谢。该文还利用脂肪染色剂尼罗红、BODIPY和细胞壁染色剂卡氏白结合荧光蛋白标记对子囊和子囊孢子进行组合染色。结果显示,尼罗红、BODIPY、卡氏白染色剂互不干扰,可以与不同颜色的荧光蛋白相互组合,从而更加清晰地标记子囊和子囊孢子的结构、细胞器和储藏物质。     

狭眼凤尾蕨配子体发育及其成精子囊素对水蕨配子体发育的影响

对狭眼凤尾蕨（Pteris biaurita）配子体发育特征及其外源成精子囊素对模式植物水蕨（Ceratopteris thalictroides）在黑暗和光照条件下孢子萌发和配子体发育的影响进行了研究。结果表明：（1）狭眼凤尾蕨孢子深褐色,三裂缝,孢子萌发为书带蕨型,原叶体发育为水蕨型,无毛状体产生;培养发现,其配子体能产生精子器,但不产生颈卵器,当接种密度适中时,可进行无配子生殖。（2）在光照和黑暗条件下狭眼凤尾蕨成精子囊素有促进和抑制水蕨孢子萌发的作用,但效果均不显著。（3）在光照条件下,狭眼凤尾蕨成精子囊素可以延迟水蕨心脏形配子体分生组织缺刻的形成,但对其配子体形态和性别分化无明显影响;而在黑暗条件下狭眼凤尾蕨成精囊素对水蕨长条形配子体的形态发育具有一定影响,与对照组相比其顶端分生组织发达,整体呈长楔形,对性别分化影响不显著。可见,狭眼凤尾蕨和水蕨不具有同种成精子囊素系统。     

西弗射盾子囊霉研究进展

正西弗射盾子囊霉(Stephanoascus ciferrii)属子囊菌类酵母,与孢子丝菌属和念珠菌属同源,但在形态特征上差别较大,除单细胞的酵母菌外,营养体菌丝中有隔膜,可通过有性繁殖在子囊中产生子囊孢子[1-2]。该菌可造成健康个体的浅表感染,在免疫缺陷或免疫低下人群可造成侵袭性感染且结局较为严重[3]。近年随着激素及其他药物的广泛使用,西弗射盾子囊霉等少见真菌引起的临床感染逐年增多[4]。对该菌的鉴定方法有传统的培养表型分析,     

桑椹肥大性菌核病病原菌生物学特性及流行性

【目的】对桑椹灾害性真菌病害——桑椹肥大性菌核病病原菌,即桑实杯盘菌(Ciboria shiraiana)的生物学特性进行研究,分析其流行性。【方法】采用人工接种、调查等研究方法,对C.shiraiana在无性生长阶段中菌丝侵染能力,菌核的休眠期,有性生长阶段中子囊孢子的结构、释放、数目以及萌发等进行研究,并对菌核萌发的物候期进行调查。【结果】C.shiraiana菌丝对桑雌花没有侵染能力;C.shiraiana菌核具有休眠期,低温处理6周以上的菌核才能萌发形成子囊盘;1个菌核可萌发1–15个子囊盘,直径为1.5 cm的子囊盘能产生高达(5.6–6.3)×10~7个子囊孢子;C.shiraiana子囊孢子在酸性环境中的萌发率明显高于在中性和碱性环境中的萌发率;C.shiraiana菌核萌发形成子囊盘产生子囊孢子的物候期,从1月下旬开始到4月中旬结束,其中在3月中旬萌发子囊盘的数目达到最高值。【结论】桑椹肥大性菌核病属于典型的流行性侵染病,在果桑栽培上容易造成毁灭性危害,生产上必须高度重视该病的防控。     

Programmed ascospore death in the homothallic ascomycete Coniochaeta tetraspora

Immature asci of Coniochaeta tetraspora originally contain eight uninucleate ascospores. Two ascospore pairs in each ascus survive and mature, and two die and degenerate. Arrangement of the two ascospore types in individual linear asci is what would be expected if death is controlled by a chromosomal gene segregating at the second meiotic division in about 50% of asci. Cultures originating from single homokaryotic ascospores or from single uninucleate conidia are self-fertile, again producing eight-spored asci in which four spores disintegrate, generation after generation. These observations indicate that differentiation of two nuclear types occurs de novo in each sexual generation, that it involves alteration of a specific chromosome locus, and that the change occurs early in the sexual phase. One, and only one, of the two haploid nuclei entering each functional zygote must carry the altered element, which is segregated into two of the four meiotic products and is eliminated when ascospores that contain it disintegrate. Fusion of nuclei cannot be random-a recognition mechanism must exist. More study will be needed to determine whether the change that is responsible for ascospore death is genetic or epigenetic, whether it occurs just before the formation of each ascus or originates only once in the ascogonium prior to proliferation of ascogenous hyphae, and whether it reflects developmentally triggered alteration at a locus other than mating type or the activation of a silent mating-type gene that has pleiotropic effects. Similar considerations apply to species such as Sclerotinia trifoliorum and Chromocrea spinulosa, in which all ascospores survive but half the spores in each ascus are small and self-sterile. Unlike C. tetraspora, another four-spored species, Coniochaetidium savoryi, is pseudohomothallic, with ascus development resembling that of Podospora anserina.     

Fungal cannons: explosive spore discharge in the Ascomycota

The ascomycetous fungi produce prodigious amounts of spores through both asexual and sexual reproduction. Their sexual spores (ascospores) develop within tubular sacs called asci that act as small water cannons and expel the spores into the air. Dispersal of spores by forcible discharge is important for dissemination of many fungal plant diseases and for the dispersal of many saprophytic fungi. The mechanism has long been thought to be driven by turgor pressure within the extending ascus; however, relatively little genetic and physiological work has been carried out on the mechanism. Recent studies have measured the pressures within the ascus and quantified the components of the ascus epiplasmic fluid that contribute to the osmotic potential. Few species have been examined in detail, but the results indicate diversity in ascus function that reflects ascus size, fruiting body type, and the niche of the particular species.     

New information on the mechanism of forcible ascospore discharge from Ascobolus immersus

Many ascomycete fungi spurt their spores from asci pressurized by osmosis. This paper explores the details of this process in the coprophilous species Ascobolus immersus, through a combination of biomechanical and biochemical experiments, and mathematical modeling. A. immersus forms large asci that expel 8 spores as a single, mucilage-embedded projectile. Measurements of ascus turgor using a microprobe attached to a strain gauge showed a pressure of 0.3 MPa or 3 atm. Analysis of ascus sap using GC/MS identified glycerol as a major osmolyte, accounting for 0.1 MPa of the osmotic pressure within the ascus sap. A mathematical model indicated that a pressure of 0.2 MPa would be sufficient to propel the cluster of ascospores over the distance measured in previous studies. The difference between the measured and predicted pressures is ascribed to loss of pressure as the spores are forced through the tip of the open ascus.     

Ultrastructure of asci,ascospores, and spore release inLophodermella sulcigena (Rostr.) v. Hohn.

Summary The croziers were formed from large multinucleate cells at the base of the hysterothecium. The diploid ascus had basal and apical vacuoles and there was prominant endoplasmic reticulum near the extending tip of the ascus. The spore delimiting membranes were continuous with the plasmalemma and possibly arose from it. The spore walls were formed between the two membranes. The ascus had a simple apical ring around a thinner region of the wall which became the pore through which the spores were released. Just before spore release the outer layer of the ascospore wall became vesiculated and eventually mucilagenous. The long clavate ascospores were released one at a time, stretching the neck of the ascus as they emerged.     

Possible involvement of Pseudomonas fluorescens and Bacillaceae in structural modifications of Tuber borchii fruit bodies

Previous studies on Tuber borchii fruit bodies in early maturation stages suggested a role of bacteria in sporocarp structural modifications. In order to verify this hypothesis, in the present study we investigated by means of microbial and ultrastructural approaches, the bacterial population of T. borchii sporocarps from intermediate maturation phases to advanced decomposition stages, paying particular attention to chitinolytic and cellulolytic bacteria and to their relationships with ascii and ascospores. We found that Pseudomonas fluorescens and spore-forming Bacillaceae, both able to degrade cellulose and chitin, are present inside the sporocarps in all maturation stages investigated. Moreover, rod-shaped bacteria seem able to erode ascus walls and colonize the interior of ascii containing mature spores. These results suggest a possible role of these bacteria in the process of ascus opening. Moreover, the presence of P. fluorescens and Bacillaceae on isolated mature spores after decontamination suggests an intimate association between these bacteria and the ascospores.     

A study of copulation,sporulation and meiotic segregation in Candida lipolytica

Summary We have attempted to optimize conjugation and sporulation in Candida lipolytica, by studying the conditions of culture and growth.Copulation between compatible strains is a rare event, particularly in the case of auxotrophic mutants. However, diploids can be selected for on minimal medium provided parents are suitable auxotrophs. These diploids can multiply vegetatively for many generations. They can also be induced to sporulate at a very high frequency.Free ascospores were isolated by means of paraffin oil and segregations of markers could be studied. At first quite irregular, these segregations improved following a number of brother-sister matings. At the same time, the mean number of spores per ascus as well as spore germinability were considerably increased.     

The release of elongated, sheathed ascospores from bottle-shaped asci in Dipodascus geniculatus

Yeasts use different mechanisms to release ascospores of different lengths from bottle-shaped asci. Round to oval-shaped ascospores are enveloped in oxylipin-coated compressible sheaths, enabling ascospores to slide past each other when they reach the narrowing ascus neck. However, more elongated ascospores do not contain sheaths, but are linked by means of oxylipin-coated interlocked hooked ridges on the surfaces of neighboring ascospores, thereby keeping them aligned while they are pushed towards the ascus tip by turgor pressure. In this study, we found elongated, oxylipin-coated sheathed ascospores in Dipodascus geniculatus that are released effectively from bottle-shaped asci without alignment. This is possible because the ascus neck and opening have a diameter that is the same as the length of the ascospore, thus allowing the ascospores to turn sideways without blocking the ascus when they are released. We found that increased concentrations of acetylsalicylic acid inhibit both ascospore release and 3-hydroxy oxylipin production in this yeast, thereby implicating this oxylipin in sexual reproduction.     

Ascospore formation and morphology in two species of the genusErysiphe remaining immature on the living host plant

Erysiphe cumminsiana andE. galeopsidis, which have immature asci in the current season, have been recorded from Japan, but the ascospores of both fungi have not been described. In the present experiments, some observations before and after overwintering were made on the cleistothecia ofE. cumminsiana on three species ofCacalia and two species ofLigularia, andE. galeopsidis onGeranium thunbergii. After overwintering, the former fungus developed six to eight, rarely four spores in an ascus and the latter fungus always four spores in an ascus. Their teleomorphic characteristics including those of ascospores are also described.     

Substructural studies on sporulation of Saccharomycopsis lipolytica

During sporulation of diploids from crosses between different strains of the yeast Saccharomycopsis (Candida) lipolytica irregular numbers of ascospores per ascus have been observed. Using the serial section method it could be shown now by means of electron microscopy that in one-, two-, and three-spored asci unenclosed "naked" nuclei occur additionally to nuclei incorporated in mature spores. It was demonstrated that the production of less than four spores per ascus in this yeast is not the result of a lack of meiotic products but of the nonutilization of nuclei from meiosis. In 2--4 spored asci usually four products of meiosis in form of enclosed and free nuclei could be demonstrated which indicate a normal meiotic division. All ascospores derived from asci with different spore numbers are uninuclear. It is assumed that a defect in spore formation caused by structural changes of chromosomes or aneuploidy should give rise to the occurrence of non incorporated nuclei and spore irregularity. It was concluded that meiosis and spore formation in Saccharomycopsis lipolytica seem to represent parallel and coordinated processes which generally resemble those recorded for Saccharomyces cerevisiae and Hansenula species.