黄檗鞘锈菌担子发育及担孢子萌发的细胞学研究

为了解黄檗鞘锈菌Coleosporium phellodendri担子发育及担孢子萌发的细胞学变化,采用光学显微镜观察了冬孢子萌发、担子发育及担孢子萌发过程,并利用苏木精染色法对冬孢子萌发过程中细胞核的变化进行了研究。结果表明:未成熟的冬孢子含有两个细胞核,成熟时冬孢子伸长,两个细胞核融合成为1个核;冬孢子萌发时,中间形成一隔膜,将冬孢子分成两部分,上部分含金黄色的原生质体,下部呈透明的柄状结构;上部分发育形成4个细胞的内担子,然后发育形成担孢子。担孢子萌发时,产生1个芽管,顶端形成外观上与担孢子形态大小基本一致的次生孢子。次生孢子萌发产生芽管,形成菌丝。有极个别的次生孢子可再次形成次生孢子。冬孢子和担孢子在5–25℃温度范围内均能萌发,其中以15–25℃为适宜萌发的温度范围。     

环境因子对努利虫疠霉产孢和孢子萌发的影响

探讨努利虫疠霉Pandoranouryi(Remaudi埁re&Hennebert)H櫣mber在田间蚜虫种群中发生与流行的规律,研究外界环境因子对感菌虫尸产孢和孢子萌发的影响。结果表明,处于水琼脂培养平板上感染努利虫疠霉的桃蚜虫尸在8~25℃的温度范围内均能产生大量的初生分生孢子,在30℃下,仅弹射极少量孢子。8℃下,孢子弹射可以持续120h,当温度高于15℃,大部分的孢子会在48h内完成弹射。相对湿度小于95%,虫尸停止产孢。20℃下,光照条件不会影响虫尸弹射孢子的总量。在8℃和30℃时,24h后处于水琼脂培养平板上的孢子萌发率分别为45.23%和61.74%,显著低于15~25℃温度范围内的孢子萌发率(95%以上)。处于叶片上的真菌孢子,当相对湿度大于74%时出现萌发,但在盖玻片的表面,当湿度低于100%时未发现孢子萌发。     

光照和温度对尖叶拟船叶藓孢子萌发及原丝体发育的影响

用组织培养法和光学显微镜技术初步研究了光照和温度对尖叶拟船叶藓孢子萌发及原丝体发育的影响。结果表明：（1）光照是影响孢子萌发的主要环境因子,20℃环境下,24h光照4d的孢子萌发率达83.3％;温度下黑暗培养的孢子30d也不能萌发,转光照后4d的萌发率可达84.6％;（2）温度是影响原丝体发育的主要环境因子,连续光照下,20℃的原丝体生长最快、分枝最多、分化最早,第31天可长达651.64μm;25℃次之,只有379.12μm;而自然光照下5-10℃环境下的孢子萌发率（18d为70.2％）和原丝体生长速度（127.44μm）均最慢;（3）原丝体发育到一定阶段,断裂为单个细胞,单个细胞再萌芽出新原丝体。     

三裂叶豚草锈病发生和流行规律的研究

通过田间调查与定点系统观察,结合沈阳地区的气象条件,对三裂叶豚草锈病在沈阳地区的发生和流行规律进行了研究。结果表明：三裂叶豚草锈病属于喜高温、高湿型病害;该病于6月初在沈阳地区开始发病,可持续至9月,以7～8月发病最重。人工接种试验结果表明：在30℃和相对湿度96．9％条件下接种锈菌冬孢子,4d后三裂叶豚草即可发病;冷冻保藏（-20℃）可打破冬孢子休眠,在沈阳地区冬孢子是第二年锈病发生的初侵染菌源。苍耳柄锈菌三裂叶豚草专化型对三裂叶豚草表现出了显著的致病性、致死性及专一性,证明是防治三裂叶豚草的理想生防菌。     

茶树轮斑病的发生及病原菌分生孢子萌发特性

对豫南茶园茶树轮斑病的发生与病原菌分生孢子萌发特性进行了调查和研究。茶树轮斑病在豫南茶区发生较普遍,一般夏秋发病较重,冬春发病较轻;轮斑病的病叶率和病情指数与温度、湿度、光照等生态因子关系密切;病原菌孢子在离体25℃条件下,4 h开始萌发。在茶树叶面上,病原菌分生孢子萌发明显比非叶面条件下好,病原菌菌丝生长较快,说明此病原菌与茶树叶片有高度的亲和力和较强的适应茶树叶面微环境的能力;茶树轮斑病病原菌分生孢子在pH5~7范围内萌发及芽管伸长较好,pH过小或过大均不利于病原菌的分生孢子萌发和菌丝生长。     

中华缩叶藓孢子萌发与原丝体发育特征研究

通过室内人工培养中华缩叶藓的孢子,在光学显微镜下详细观察了其孢子萌发、原丝体发育及配子体发生的全过程.结果表明:中华缩叶藓的孢子在壁内萌发,随后分裂产生块状原丝体;块状原丝体上可产生两种丝状体,一种是具疣的棒状原丝体,另一种是由长圆柱状细胞组成的轴丝体;配子体原始细胞只产生于块状原丝体上.根据中华缩叶藓的孢子萌发和原丝体发育特征,并参照Nishida对藓类植物孢子萌发类型的划分,确定中华缩叶藓的萌发孢子型应属于缩叶藓型(Ptychomitrium-type).     

环境因子对蕨类植物孢子萌发的影响

蕨类植物通过孢子萌发形成独立生活的配子体,配子体能够形成精子器和颈卵器,进而通过受精作用形成新的孢子体。孢子萌发是蕨类植物生活史过程中配子体世代向孢子体世代转变的关键步骤。同时,此过程不仅受到多种环境因子的影响,也是研究细胞核极性移动、细胞不对称分裂、假根极性生长等独特的细胞学事件的良好模型。迄今为止,人们已经研究发现多种环境因子对约200余种蕨类植物孢子萌发有影响。总结了环境因子对蕨类植物孢子萌发影响的规律如下:(1)孢子萌发除了受到光照强度影响外,主要受光质的影响,光质的影响主要表现为4种方式:①孢子萌发受红光刺激与远红光抑制像开关一样调控;②孢子萌发不受远红光抑制;③孢子萌发受蓝光抑制;④孢子只能在黑暗条件下萌发。(2)重力作用会影响孢子细胞核移动,进而影响孢子细胞发育的极性。(3)赤霉素(GA)能增加孢子萌发率或帮助孢子打破休眠。成精子囊素与GA作用相似,启动或促进孢子萌发。而脱落酸(ABA)、茉莉酸(JA)和乙烯等其它激素对孢子萌发的影响相对较小。(4)不同植物孢子有着各自最适的萌发培养基条件,如不同种类孢子对MS培养基中无机盐含量、蔗糖含量、pH值的要求不同。孢子外被中的Ca2+、Mn2+和Mg2+,培养基中的Cd2+和La3+,以及孢子接种密度、萌发空间CO2含量也会对孢子萌发造成影响。(5)多数蕨类植物孢子在15-30℃可以萌发,最适萌发温度为25℃。(6)4℃和液氮储藏可以延长孢子寿命并保持较高萌发率。     

蕨类植物孢子萌发及原叶体发育的观察

以腐叶土为培养基质,对21种蕨类植物进行了孢子萌发和原叶体发育的研究,结果表明：①不同时期播种的同种蕨类的孢子,发育出原叶体和幼孢子体所历经的时间长短不同;②孢子萌发和配子体生长发育的适宜温度约为15～24℃;⑨稀有蕨类的孢子萌发率低,而在野外能形成较大种群的蕨类的孢子萌发率高;④用GA3处理孢子可以促进萌发;⑤当原叶体上长出幼孢子体时,原叶体由大变小,由绿变黄,21种蕨类的原叶体都在幼孢子体上长出第3片叶时消失;⑥幼孢子体上长出的第1、2片叶在形态上与以后长出的叶不同;⑦孢子萌发需要光;⑧1片原叶体尽管有多个颈卵器,但仅发育出1株幼孢子体;⑨利用腐叶土进行蕨类孢子繁殖是一种经济实用的繁殖方法。     

乌蕨孢子萌发研究

用随机区组试验和方差分析方法, 研究了培养温度、贮藏温度、GA 处理和光照强度对乌蕨( Sphenomeris chinensis) 孢子萌发的影响。与室温(20～25℃ ) 相比, 培养温度在28℃时, 孢子最大萌发率相近而萌发速率明显较高。贮藏温度(A) 极显著( P < 0 . 01 ) 影响孢子萌发率, - 20℃ 贮藏降低萌发率; GA (B) 对孢子萌发率无显著影响。光照强度(C) 极显著( P < 0.01) 影响孢子萌发率, 充足光照和弱光照无显著差异, 黑暗处理降低萌发率。A×B, A×C, B×C 及A×B×C 交互效应不显著。     

球囊霉属几种AM真菌孢子表面消毒与萌发的研究

通过对球囊霉属几种AM真菌进行孢子表面消毒与萌发研究,结果表明,采用"差别灭菌法"即先用含2%氯胺T、0.02%链霉素和0.01%庆大霉素的溶液孢子表面消毒10min,然后在25～27℃下培养3天,再用含2%氯胺T、0.02%链霉素和0.01%庆大霉素的溶液孢子表面消毒10min,其消毒效果最好,孢子萌发率较高,污染率较低.同时发现土壤、草炭和寄主植物根的分泌物对孢子萌发有促进作用.培养基pH值的变化对孢子萌发也有一定的影响.AM真菌孢子用"改良Sandwich法"进行萌发,其萌发率最高.     

Molecular profiling demonstrates limited diversity amongst geographically separate strains of Ustilago scitaminea

Intraspecies diversity within Ustilago scitaminea isolates from South Africa, Reunion Island, Hawaii and Guadeloupe was assessed by RAPDs, bE mating-type gene detection, rDNA sequence analysis, microscopy and germination and morphological studies. Except for sequence data, the other analyses yielded no differences in the isolates that could be used in a phylogenetic separation. Mycelial DNA of the SA isolate shared 100% sequence identity with that of mycelial DNA cultured from in vitro produced teliospores of the parent cultivar. Overall the ITS1 and ITS2 regions were found to have 96.1% and 96.9% sequence identity with a total of 17 and 21 base changes, respectively, amongst the isolates. The Reunion Island isolate was shown to be most distantly related by 3.6% to the other isolates, indicating a single clonal lineage. The lack of germination in teliospores from Guadeloupe may be attributed to changes in temperature and humidity during transportation.     

Exogenous gibberellins inhibit coffee (Coffea arabica cv. Rubi) seed germination and cause cell death in the embryo

The mechanism of inhibition of coffee (Coffea arabica cv. Rubi) seed germination by exogenous gibberellins (GAs) and the requirement of germination for endogenous GA were studied. Exogenous GA(4+7) inhibited coffee seed germination. The response to GA(4+7) showed two sensitivity thresholds: a lower one between 0 and 1 microM and a higher one between 10 and 100 microM. However, radicle protrusion in coffee seed depended on the de novo synthesis of GAs. Endogenous GAs were required for embryo cell elongation and endosperm cap weakening. Incubation of coffee seed in exogenous GA(4+7) led to loss of embryo viability and dead cells were observed by low temperature scanning microscopy only when the endosperm was surrounding the embryo. The results described here indicate that the inhibition of germination by exogenous GAs is caused by factors that are released from the endosperm during or after its weakening, causing cell death in the embryo and leading to inhibition of radicle protrusion.     

苍耳柄锈菌三裂叶豚草专化型的超微结构观察(英文)

20 0 3年在沈阳的三裂叶豚草 (AmbrosiatrifidaL .)上发现了苍耳柄锈菌三裂叶豚草专化型 (PucciniaxanthiiSchwein .f.sp .ambrosia trifidaeS .W .T .Batra) ,这是在我国三裂叶豚草上发现的一种新病菌。试验采用扫描电镜和透射电镜对该锈菌的冬孢子和吸器的形态结构进行了观察。     

Sugar cane arginase competes with the same fungal enzyme as a false quorum signal against smut teliospores

Sugar cane cultivars resistant to smut produce a pool of glycoproteins after experimental infection, one of them develops arginine activity. This arginase induces cytoagglutination of smut teliospores but impedes germination. Teliospores also secrete a fungal arginase that accelerates their own germination. This fungal arginase binds to teliospore cell walls. The affinity of this arginase for the ligand shows to be higher than that found for plant arginase. In fact, fungal arginase removes sugar cane arginase previously bound to their ligands in the cell wall whereas the inverse process is practically negligible. The enzymatic activity is required to the binding of the protein to teliospore cell wall but it is not related to the germination process. Thus, it can be concluded that the binding of fungal arginase to smut teliospores activates a signal transduction cascade that enhances germination.     

A Comparison of Polypeptides from Teliospores of Tilletia controversa (Kuhn) and Tilletia tritici (Bjerk) Wint.

Teliospores from 12 races of Tilletia tritici (Bjerk.) Wint. and twelve isolates of Tilletia controversa (Kuhn) were sampled from field-inoculated wheat (Triticum aestivum L.) differential cultivars. Proteins were extracted from the teliospores and analysed by one dimensional electrophoresis. An abundant 116 kD polypeptide detected in extracts from teliospores of all isolates of T. controversa was not detected in T. tritici teliospore extracts. Spores which were mechanically disrupted yielded greater quantities of the protein compared to intact teliospores, and this suggested the polypeptide was derived from within the teliospore. The presence of the polypeptide was correlated with dwarf bunt-causing Tilletia- Isolates of dwarf bunt-causing Tilletia that were intermediate between T. tritici and T. controversa in either morphology or germination characteristics contained the polypeptide while a common bunt-causing race of T. tritici (T18) with intermediate characteristics lacked the protein. The 116 kD polypeptide present in all T. controversa isolates may provide a stable biochemical marker for identification of these teliospores in wheat shipments.     

Differential gene expression during teliospore germination in <Emphasis Type="Italic">Ustilago maydis</Emphasis>

Ustilago maydis is a model fungal pathogen that induces the formation of tumors in maize. The tumor provides an environment for hyphal differentiation, leading to the formation of thick-walled, diploid teliospores. Such spores serve as a dispersal agent for smut and rust fungi, and their germination leads to new rounds of infection. The morphological changes that occur during teliospore germination in U. maydis have been described in detail. However, the specific molecular events that facilitate this process have not been identified. Through the construction and hybridization of microarrays containing a set of 3918 non-redundant cDNAs, we have identified genes that are differentially regulated during teliospore germination. Teliospores induced to germinate for 4 and 11 h were selected for comparison with dormant teliospores. Genes identified as differentially expressed included many that are presumably involved in as yet undescribed molecular events during teliospore germination, as well as characterized genes previously shown to be required for the process. This study represents the first large-scale investigation of changes in gene expression during teliospore germination.Electronic Supplementary Material Supplementary material is available for this article at     

A role for sugarcane glycoproteins in the resistance of sugarcane to Ustilago scitaminea

Smut is a major disease of sugarcane caused by Ustilago scitaminea. Germination of fungal teliospores is achieved on the internode surface of plants, and it is followed by the formation of appressoria. A primary response of sugarcane plants to the infection seems to be the production of several glycoproteins, defined as mid-molecular mass (MMMG) or high molecular mass (HMMG) macromolecules. Teliospore germination in the presence of both MMMG and HMMG decreased about 50% following 5 h of teliospore contact with glycoproteins. This may be related to the ability of glycoproteins to produce cytoagglutination. Binding of fluorescein-labelled glycoproteins was studied by fluorescence microscopy, showing that staining of cells was not uniform, but mainly in the contact zone between two individual teliospores when aggregated. HMMG was composed of only one fraction that was completely retained by smut teliospores, whereas three of the five different glycoproteins occurring in the MMMG fraction were retained by teliospore cell walls. Moreover, a unique application of salicylic acid, naturally produced by sugarcane stalks after experimental fungal infection, enhanced the production of both glycoprotein pools. A hypothesis about the role of both HMMG and MMMG as defence glycoproteins is discussed.     

Glycoproteins from sugarcane plants regulate cell polarity of Ustilago scitaminea teliospores

Saccharum officinarum, cv. Mayarí, is a variety of sugarcane resistant to smut disease caused by Ustilago scitaminea. Sugarcane naturally produces glycoproteins that accumulate in the parenchymatous cells of stalks. These glycoproteins contain a heterofructan as polysaccharide moiety. The concentration of these glycoproteins clearly increases after inoculation of sugarcane plants with smut teliospores, although major symptoms of disease are not observed. These glycoproteins induce homotypic adhesion and inhibit teliospore germination. When glycoproteins from healthy, non-inoculated plants are fractionated, they inhibit actin capping, which occurs before teliospore germination. However, inoculation of smut teliospores induce glycoprotein fractions that promote teliospore polarity and are different from those obtained from healthy plants. These fractions exhibit arginase activity, which is strongly enhanced in inoculated plants. Arginase from healthy plants binds to cell wall teliospores and it is completely desorpted by sucrose, but only 50% of arginase activity from inoculated plants is desorpted by the disaccharide. The data presented herein are consistent with a model of excess arginase entry into teliospores. Arginase synthesized by sugarcane plants as a response to the experimental infection would increase the synthesis of putrescine, which impedes polarization at concentration values higher than 0.05 mM. However, smut teliospores seem to be able to change the pattern of glycoprotein production by sugarcane, thereby promoting the synthesis of different glycoproteins that activate polarization after binding to their cell wall ligand.     

关于茭白黑粉菌的正名问题*

茭白黑粉菌在文献中出现Yenia esculenta(P.Henn.)Liou及Ustilago esculenta P.Henn.两个名字。现经形态发生学的研究以明确新名字是否有效。此菌冬孢子没有休眠期,孢子成熟后在适宜的环境中即可萌发。萌发的最适pH值为6;最适温度为25％;不需要外界光的诱导,在黑暗条件下也能正常萌发;培养基中的营养成份对孢子萌发的速率有一定影响。冬孢子在不同的环境条件下,其萌发形态及方式都是稳定的:孢子萌发时产生有隔的担子或先菌丝(promycelium)。该先菌丝起初可以是无隔的,但生长到一定程度,便产生分隔,分隔多为2—4个。初生的先菌丝和小孢子是单核的,但成熟的先菌丝、小孢子及短菌丝细胞内均是双核的。可以认为,该菌的双核期在整个生活史中占有相当长的时间,它只有短暂的单核期。没有发现先菌丝、小孢子彼此之间或相互间融合的现象;未见到双核的融合及减数分裂过程。茭白黑粉菌在人工组合培养基上生成厚壁的孢子,较自然界中的冬孢子为大,但萌发方式则相同。最后作者等认为茭白黑粉菌仍应保留在Ustilago属中,而Yenia esculenta(P.Henn.)Liou的名字是无效的。