甘肃大葱贮藏期镰孢菌腐烂病病原鉴定

【目的】大葱在贮藏期频繁发生镰孢菌腐烂病,损失严重。明确该病害病原种类对病害防治具有重要意义。【方法】利用组织分离法对采集自甘肃省兰州市(区)蔬菜市场的16份大葱贮藏期镰孢菌腐烂病病样进行病原物的分离、纯化培养,经单孢分离后根据形态学特征,再结合r DNA-ITS、EF-1a(tef)基因序列分析的方法进行鉴定。【结果】共分离得到80株镰孢菌,经鉴定分属3个种,即层出镰孢菌(Fusarium proliferatum)、尖孢镰孢菌(F.oxysporum)和燕麦镰孢菌(F.avenaceum),其中层出镰孢菌为大葱镰孢菌腐烂病的优势致病菌,分离频率为52.50%。对兰州白葱不同部位进行致病性测定,结果表明层出镰孢菌对大葱鳞茎的致病力最强,而燕麦镰孢菌对大葱鳞茎的致病力最弱。【结论】3种镰孢菌作为该病害的病原,属国内首次报道。     

蚧镰孢菌的生长及产孢研究

蚧镰孢菌孢子萌发及产孢的最适温度是 2 8℃ ,菌生长的适温是 2 4℃～ 2 6℃。孢子萌发受 pH值影响较小 ,菌生长以 pH6.5～ 8.5为最适 ,而 pH8.5时产孢最多。菌的生长也可以调节培养液的pH值 ,使其达到最适生长的 pH范围。光照对该菌的生长及产孢也有一定影响。蚧镰孢菌能利用多种碳源和氮源 ,也能以几丁质为唯一碳源和氮源生长 ,但生长很差。Ca2 、Fe2 等金属离子以及硫胺素、核黄素、叶酸等维生素的加入可使产孢量增加 ,而维生素的作用更明显。该菌的产孢高峰期一般在 2 1天左右 ,而且在相同条件下 ,接种量越大 ,产生最大量孢子所需的时间越短。     

淡紫紫孢菌PLF-1对感染尖刀镰孢菌百合种球的促生作用及其相关防御酶活性变化

为了探究淡紫紫孢菌(Purpureocillium lilacinum)PLF-1对百合种球的促生作用及对百合尖刀镰孢菌的防治效果,采用平板对峙法评估淡紫紫孢菌对尖孢镰刀菌的拮抗效果,以及淡紫紫孢菌对百合抗尖孢镰刀菌的抗性作用。同时监测百合种球中超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）的活性变化情况。研究结果表明：浓度为4.34×10⁴ CFU/mL和4.34×10⁵ CFU/mL的淡紫紫孢菌孢子悬浮液对百合种球表现为促进作用,浓度为4.34×10⁴ CFU/mL时最高茎长达11 cm。平板拮抗实验中该淡紫紫孢菌菌株能有效抑制尖孢镰刀菌生长,抑制率高达72%。接种淡紫紫孢菌和病原菌的百合种球茎长会增长37.6%,根长会增长33%。该菌株能提高感染尖刀镰孢菌百合种球中的超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性,有效抑制尖刀镰孢菌的毒害作用,促进植株健康生长。     

短柄链蠕孢分生孢子发育的研究

短柄链蠕孢(Dendryphion nanum)为暗色丝孢菌,其分生孢子棒状,5～20个隔膜,暗褐色,串生。试验通过玻片培养观察研究了该菌分生孢子的产生方式及发育类型。结果表明:该菌的分生孢子是由产孢细胞的顶端向外突起并生长膨大而形成,因而属于全壁芽生式产孢方式。     

枝孢样枝孢霉的研究概况

引起暗色丝孢霉病的枝孢霉属,属半知菌亚门、丝孢菌纲、暗色孢科。该属50余种。枝孢样枝孢霉为腐生真菌,广泛存在于自然界的土壤、某些动物的粪便、蔬菜、腐木、鸟巢、腐烂水果中。人因皮肤外伤接触后有被感染的机会。它也是真菌过敏症的重要病原菌之一。目前对枝孢样枝孢霉的系统报道较少,现就其生理生化代谢、生态学、病理学及临床和抗真菌研究等方面进行综述。     

胶孢镰刀菌产生串珠镰刀菌素的不稳定性

从陕北克山病病区分离到的两株串珠镰刀菌素产生菌株——胶孢镰刀菌(Fusarium subglutinans Wollenw.et Reinking)陕-6号和2-17号进行单孢分离,分别得到23和19个单孢分离株。这些单孢菌株可分为两种培养型:一种形态上与原始菌株相似,产生串珠镰刀菌素,产色素,具有大、小分生孢子,转管八次产毒量有下降;另一种则不产串珠镰刀菌素和孢子,无色素,后者在二株菌的单孢分离菌中所占比例分别为60.9％和15.8％。由此可见,胶孢镰刀菌产毒稳定性受异核体和该菌单核变异性共同影响。     

一株纤维素降解真菌的筛选、鉴定及酶学性质分析

通过对富含枯枝败叶的土壤样品进行富集培养,利用刚果红纤维素培养基初筛和酶活测定复筛得到产纤维素酶的一株真菌,将其命名为GC2-2,并对该菌株进行鉴定及酶学性质研究。结果表明该菌株是一株耐高温、碱性纤维素酶的真菌GC2-2。通过18S rDNA分子克隆测定,该菌为球孢枝孢菌,其滤纸酶的活力优于CMC酶的活力。该菌所产酶的最适反应条件为温度35°C,最适pH值7.5。     

红色毛癣菌和枝孢样枝孢霉混合感染导致银屑病患者甲真菌病1例

目的报道1例银屑病患者出现红色毛癣菌和枝孢样枝孢霉混合感染导致的甲真菌病。方法报告病例,对甲标本进行真菌镜检和培养,对病原菌进行形态学及分子生物学鉴定。结果该病例经临床、真菌镜检和真菌培养鉴定,确诊为红色毛癣菌和枝孢样枝孢霉导致的甲真菌病。病原菌通过菌落和显微镜下形态特征结合rRNA内转录间隔区序列分析证实。结论通过形态学及分子生物学鉴定,证实为真菌红色毛癣菌和枝孢样枝孢霉混合感染导致的甲真菌病。     

脉孢菌lca-1基因调控无性产孢及类胡萝卜素的合成

类胡萝卜素是很多生物细胞内重要的抗氧化剂,具有保护细胞免受紫外线伤害的功能。粗糙脉孢菌是少数几个类胡萝卜素合成基因比较清楚的真菌之一,为了深入了解该菌类胡萝卜素合成调控机制,通过对粗糙脉孢菌基因突变体库中6,087株突变体进行筛选,新发现6个基因敲除突变体营养生长正常,但类胡萝卜素的合成降低,其中表型较好的1个突变体,其无性产孢量与类胡萝卜素合成量均明显降低。鉴定发现该突变体所缺失的基因编码一种依赖ATP的染色体重建复合体的ATP酶链ISW1,将该基因命名为lca-1。进一步测定发现lca-1基因的突变导     

链霉菌质粒pSET152电转化稀有放线菌小单孢菌的研究

利用链霉菌(Streptomyces)噬菌体ΦC31所构建的整合型载体pSET152作为供体质粒,分别以小单孢菌(Micromonospora)40027菌株的萌发孢子和新鲜菌丝体作为受体菌,在不同的电场强度下进行电转化实验,结果表明:以小单孢菌40027菌株萌发孢子为受体菌,未获得电转化子;以小单孢菌40027菌株新鲜菌丝体为受体菌,获得了电转化子。电场强度为13kV/cm时可获得最高转化效率。Southern杂交结果表明:质粒pSET152可通过菌丝体电转化法导入小单孢菌40027菌株,并整合到小单孢菌40027菌株的染色体上,暗示链霉菌噬菌体ΦC31的整合酶基因和整合位点在异源宿主小单孢菌40027菌株中仍具有相同的功能。质粒稳定性检测实验表明:质粒pSET152可稳定地存在于小单孢菌40027菌株中。     

Genetic Diversity of Fungi Occurring in Nests of Three <Emphasis Type="Italic">Acromyrmex</Emphasis> Leaf-cutting Ant Species from Córdoba,Argentina

It is assumed in current literature that the fungus garden cultivated by leaf-cutting ants consists of a single fungus species, the putative mutualistic fungus. However, most studies report a very high rate of fungi contamination (fungi isolated from fungus gardens that are considered not to be the mutualistic fungus). In this article, we report a genetic similarity analysis of all fungi (regardless of their mutualistic condition) isolated from 16 fungus gardens of three Acromyrmex species in Córdoba, Argentina, using intersimple sequence repeat (ISSR) as genetic markers. We isolated 60 clones, of which the three primers employed yielded 53 loci. The patterns revealed a high interclone polymorphism, with a few bands shared by the clones. Of all possible pairwise comparisons, 99% showed a genetic similarity (S) lower than 0.5, the threshold level assumed for fungus Operational Taxonomy Unit (OTU). We found more than one fungus OTU in all studied nests (range 2–11). Cumulative number of OTUs increased linearly with the number of nests sampled. The number of fungus OTUs common to both ant species and sites was very small. We did not find a core group composed of few very common OTUs, as expected if a set of truly mutualistic OTU was present. A simple explanation for the high number of OTUs found is that they are regular components of the fungus garden, which may be used as food source by the ants.     

Sequential batch culture studies for the decolorisation of reactive dye by Coriolus versicolor

The white rot fungus Coriolus versicolor could decolorise reactive dye Remazol Brilliant Violet to almost 90%. The fungal mycelia removed color as well as COD up to 95% and 75%, respectively, in a batch reactor. Decolorising activity was observed during the repeated reuse of the fungus. It was possible to substantially increase the dye decolorising activity of the fungus by carefully selecting the operational conditions such as media composition, age of fungus and nitrogen source. The fungal pellets could be used for eight cycles during the long term operation, where medium and dye was replenished at the end of each cycle and the fungus was recycled. Presence of a nitrogen source and nutrient content of media played an important role in sustaining the decolorisation activity of the fungus. The form of nitrogen source (e.g. peptone vs. urea) was also important to maintain the decolorising activity with peptone showing better decolorisation.     

Symbiotic fungi produce laccases potentially involved in phenol degradation in fungus combs of fungus-growing termites in Thailand

Fungus-growing termites efficiently decompose plant litter through their symbiotic relationship with basidiomycete fungi of the genus Termitomyces. Here, we investigated phenol-oxidizing enzymes in symbiotic fungi and fungus combs (a substrate used to cultivate symbiotic fungi) from termites belonging to the genera Macrotermes, Odontotermes, and Microtermes in Thailand, because these enzymes are potentially involved in the degradation of phenolic compounds during fungus comb aging. Laccase activity was detected in all the fungus combs examined as well as in the culture supernatants of isolated symbiotic fungi. Conversely, no peroxidase activity was detected in any of the fungus combs or the symbiotic fungal cultures. The laccase cDNA fragments were amplified directly from RNA extracted from fungus combs of five termite species and a fungal isolate using degenerate primers targeting conserved copper binding domains of basidiomycete laccases, resulting in a total of 13 putative laccase cDNA sequences being identified. The full-length sequences of the laccase cDNA and the corresponding gene, lcc1-2, were identified from the fungus comb of Macrotermes gilvus and a Termitomyces strain isolated from the same fungus comb, respectively. Partial purification of laccase from the fungus comb showed that the lcc1-2 gene product was a dominant laccase in the fungus comb. These findings indicate that the symbiotic fungus secretes laccase to the fungus comb. In addition to laccase, we report novel genes that showed a significant similarity with fungal laccases, but the gene product lacked laccase activity. Interestingly, these genes were highly expressed in symbiotic fungi of all the termite hosts examined.     

A real-time qPCR assay for the detection of the nifH gene of Methanobrevibacter smithii, a potential indicator of sewage pollution

Aims: To degrade ether‐type polyurethane (ether‐PUR), ether‐PUR–degrading micro‐organism was isolated. Moreover, ether‐PUR–degrading mechanisms were analysed using model compounds of ether‐PUR. Methods and Results: A fungus designated as strain PURDK2, capable of changing the configuration of ether‐PUR, has been isolated. This isolated fungus was identified as Alternaria sp. Using a scanning electron microscope, the grid structure of ether‐PUR was shown to be melted and disrupted by the fungus. The degradation of ether‐PUR by the fungus was analysed, and the ether‐PUR was degraded by the fungus by about 27·5%. To analyse the urethane‐bond degradation by the fungus, a degraded product of ethylphenylcarbamate was analysed using GC/MS. Aniline and ethanol were detected by degradation with the supernatant, indicating that the fungus secreted urethane‐bond–degrading enzyme(s). PURDK2 also degraded urea bonds when diphenylmethane‐4,4′‐dibutylurea was used as a substrate. Conclusions: The enzyme(s) from PURDK2 degraded urethane and urea bonds to convert the high molecular weight structure of ether‐PUR to small molecules; and then the fungus seems to use the small molecules as an energy source. Significance and Impact of the Study: Ether‐PUR–degrading fungus, strain PURDK2, was isolated, and the urethane‐ and urea‐bonds–degrading enzymes from strain PURDK2 could contribute to the material recycling of ether‐PUR.     

Ecological studies on Pythium splendens braun in oil palm (Elaeis guineensis Jacq.) plantation soils

Summary The isolation of Pythium splendens Braun by a root-baiting technique showed that the infective propagules of the fungus were numerically 12 times as great in cultivated soil as in non-cultivated secondary forest soil.In cultivated oil palm nursery soil, the propagules of the fungus were found to be most abundant at a depth of between 15 and 30 cm below ground level. The recovery of the fungus from the soil around the roots of diseased oil palm seedlings was 4 times as great as that from around the roots of healthy seedlings. Farmyard manure increased the recovery of the fungus from naturally infested soil, whilst green manure reduced it.The recovery of the fungus from soil declined sharply with decrease in soil moisture content and also with the length of storage of air-dried soil.The recovery of the fungus from nursery soil was high in July–October and low in November–January and these variations corresponded with seasonal fluctuations in soil moisture content.     

Selecting hyperparasites for biocontrol of Dutch elm disease

Hyperparasites in the form of cytoplasmic RNA elements have been proposed as a biological control agent for Dutch elm disease. We characterized the range of outcomes likely to follow the introduction of such an agent by modelling the resultant population dynamics as an ecological interaction between the wild, ''target'', fungus and the hyperparasitized ''control'' fungus. We used data from the 1970s epidemic of Dutch elm disease in the UK to parameterize the population dynamics of the target fungus, and considered the success of control across a wide range of possibilities for the lethality and transmissibility of the modified control fungus. We decomposed hyperparasite transmissibility into horizontal transmissibility (the ability to colonize previously unparasitized target fungal hosts) and vertical transmissibility (the ability of control fungus to establish new colonies). There is an invasion threshold for both horizontal and vertical transmissibility. As vertical transmission is further increased, there is another threshold at which the target fungus is eradicated because of competitive exclusion by the control fungus. In contrast, eradication by raising horizontal transmission may never succeed because the target fungus needs to be present to support new cases through this route. Between these two thresholds for invasion and exclusion, control and target fungus may coexist. Using a stochastic, spatially extended model, we showed that predictions of success based on high competitive ability of the control fungus (i.e. high vertical transmission) are likely to be more robust than those based on the high degree to which the control fungus can cause target fungus to be hyperparasitized (i.e. high horizontal transmission).     

Composting of lead-contaminated solid waste with inocula of white-rot fungus

The treatment of the simulated lead-contaminated solid waste by composting with white-rot fungus was studied at laboratory scale. The composting system without the inocula of white-rot fungus was prepared as control, and the composting of the uncontaminated solid waste with the inocula of white-rot fungus was carried out as the other control. The results indicated that the solid waste inoculated with white-rot fungus could be successfully processed. The final compost was mature with 70.5% of lead (Pb) in residual fraction and none in exchangeable fraction. Germination index reached 120%. All the results indicated that the bioavailability of Pb in compost was reduced and the potential harm of Pb in compost was alleviated by composting with the inocula of white-rot fungus.     

Evidence that the fungus cultured by leaf-cutting ants does not metabolize cellulose

Leaf‐cutting ants are a very specialized group of ants that cultivate fungus gardens in their nests, from which they obtain food. The current opinion is that the fungus cultivated by leaf‐cutting ants digests cellulose. Here we reassess the cellulose‐degrading capability of the fungus by using two complementary approaches tested in four Attini species (genera Atta and Acromyrmex): (1) ability of fungus to grow in cellulose; and (2) lignin/cellulose ratio in the refuse material dumped outside the nest, as an indicator of cellulose consumption. We found that (1) the fungus did not grow in cellulose, and (2) the lignin/cellulose ratio was much lower in the ants' refuse than in material digested by cellulose‐digesting organisms, such as brown‐rot fungus, termites, and ruminant mammals. This evidence strongly suggests the inability of the fungus to degrade cellulose. Therefore, the fungus–ant symbiosis and the ecological role of leaf‐cutting ants need to be reconsidered.     

Carbon mineralization by termites in tropical forests,with emphasis on fungus combs

A role of termites in decomposition processes was quantitatively evaluated in a dry evergreen forest (DEF) in Thailand, using respiration rates and biomasses of fungus combs as well as of termites themselves. The termite population and fungus combs mineralized 11.2% of carbon (C) in the annual aboveground litterfall (AAL) by their respiration. Fungus combs were responsible for a major part (7.2% of the AAL) of the C mineralization mediated by termites. For comparison, fractions of AAL mineralized by respiration from termite populations and fungus combs were estimated for tropical forests and savannas where termites have been well studied, assuming that there is the same ratio as for the DEF between biomass of fungus combs and abundance of fungus growers. Termites in dry tropical forests (annual rainfall<2,000 mm) are shown to mineralize about 10% of C in the AAL by respiration from their populations and fungus combs, and their ecological impact in savannahs is comparable in this aspect. A significant negative correlation between fraction of AAL and annual rainfall demonstrates that the importance of termites in decomposition processes is greater in dry tropical forests than in moist tropical forests. Considering that fungus combs contributed significantly to AAL mineralization in most of the tropical forests and savannas, fungus growers are a much more influential group than previously expected in tropical ecosystems.T. Abe deceased on 27 March 2000     

Evolutionary aspects of ant-fungus interactions in leaf-cutting ants

Leaf-cutting ants are highly successful herbivores because they are able to use a wide variety of plants as food The workers harvest and process plant material to be used as substrate for a fungus on which they feed. New hypotheses concerning the evolution of the ant-fungus relationship have now been proposed. Although the relationship between the ants and the fungus is mutualistic, if may appear that the fungus has little control over the ants. However, evidence suggests that the fungus may be exploiting the ants to provide it with substrate and antimicrobial defence. Furthermore, experimental evidence suggests that the fungus can select its substrate by controlling the foraging behaviour of the ants, by means of an ingenious chemical feedback mechanism.