真菌固态载体预处理对不灭菌玉米秸秆降解的影响

采用云芝变色栓菌(Trametes versicolor)、色精木霉菌(Trichoderma chromospermum)、深绿木霉菌(Trichoderma atroviride)和云芝变色栓菌分别与色精木霉菌、深绿木霉菌的混合菌对干黄玉米秸秆进行真菌预处理。结果显示,真菌预处理不灭菌秸秆30 d后,木质纤维素的降解率几乎可以忽略,在不灭菌的环境中,接入的真菌对秸秆本身附着的微生物群落不能形成竞争优势,造成较低的木质素降解率。而云芝变色栓菌、色精木霉菌、深绿木霉菌以及云芝变色栓菌分别与色精木霉菌、深绿木霉菌的混合菌对灭菌秸秆预处理30 d后,木质素的降解率分别为34.0%、38.1%、38.1%、39.1%及40.3%。为降低秸秆的灭菌成本,将真菌预处理30 d后的灭菌秸秆作为真菌固态载体与不灭菌秸秆按照1∶9、1∶4及1∶1的干质量比混合培养30 d,结果显示,采用云芝变色栓菌分别与色精木霉菌和深绿木霉菌的混合菌固态载体预处理不灭菌秸秆,木质素降解率可达到30%~40%。因而,真菌固态载体可以较好地适应非灭菌环境并对秸秆中的木质素进行有效降解,是一种有效降低过程成本的预处理手段。     

Suppression of the fungal pathogen Magnaporthe grisea by Stenotrophomonas maltophilia,a seed-borne rice (Oryza sativa L.) endophytic bacterium

The present work was carried out to study the potential of bacteria isolated from the seeds of rice plant for the biocontrol of five rice pathogenic fungi. Eleven endophytic bacteria isolated from rice seeds were evaluated for their antagonistic potential. Of five pathogens studied, only the growth of Magnaporthe grisea was inhibited by one bacterial isolate in an in vitro dual culture assay. Based on 16S rDNA sequence analysis, and biochemical and morphological characteristics, this strain was closely related to Stenotrophomonas maltophilia. We named this new isolate to be S. maltophilia SEN₁ (seed endophyte). This isolate was further tested for the production of volatile and diffusible antibiotics against M. grisea, for plant growth-promoting (PGP) traits and colonization of some rice cultivars. In addition, S. maltophilia SEN₁ was tested for its ability to promote plant growth and reduce the incidence of rice blast disease under greenhouse conditions. When applied to the soil, this isolate increased seedling growth and suppressed blast disease in plants of three studied cultivars. This study also showed this isolate could colonize the root interior of other rice cultivars. This study indicates that the S. maltophilia isolate studied has an excellent potential to be used as biocontrol agents of M. grisea or biofertilizer under in vitro and in vivo conditions.     

Physiological disorders,bitter rot and other fungal decay of ‘Aroma’ apple fruit stored in controlled atmosphere

Effects of controlled atmosphere (CA) conditions on physiological disorders and fungal fruit decay on apple ‘Aroma’ were investigated. Fruit from three growing seasons were stored at 1% or 2% O₂ (both at 2% CO₂) at either 1°C or 3°C in small research units; controls were kept in the same ventilated rooms at the two temperatures (ambient air). The fruit were removed from storage after four or six months and assessed for fruit decay immediately afterwards and after two weeks at 20°C. Fruit quality parameters were recorded at the end of storage. On a three-year average, fruit stored in CA was less ripe at the end of storage. After both four and six months storage, CA reduced total decay (physiological disorders and fungal decay) by on average 70% and 45%, respectively, compared to storage in ambient air. Senescent breakdown was lower after CA storage for four months, but not after six months and not after simulated shelf life. Soft scald was lower when stored in CA both after cold storage at 1°C and simulated shelf life. After storage at 3°C there was lower incidence of soft scald when stored in CA after four months, but not after six months. For fungal fruit decay in general, there was no effect of low oxygen, however, 2% O₂ gave slightly less bitter rot (Colletotrichum acutatum) than 1% O₂ and significantly less than ambient air after simulated shelf life. Averaged over all oxygen levels, 1°C gave significantly less bitter rot than 3°C. It may be concluded that use of CA for storage of ‘Aroma’ is a good way of reducing development of physiological disorders. However, development of bitter rot seemed to be more influenced by temperature and storage time than by low O₂.     

河西走廊制种玉米生育中后期叶部真菌病害诊断与病原初步鉴定

对河西走廊制种玉米生育中后期叶部真菌病害进行田间调查,并将采集到的标本进行组织分离、病原鉴定和致病性测定。结果表明,引起制种玉米生育中后期叶部真菌病害的植物病原有13种,其中由Curvulairia lunata(Wakke)Boed病原引起的玉米弯孢菌叶斑病是玉米叶部的一种新病害,在田间发病率较高,病情指数较大,危害十分严重;由Exserohilum turcicum(Pass.)Leonard et Suggs和Bipolaris maydis(Nishik.et Miyabe)Shoemaker病原引起的大斑病和小斑病是玉米生育中后期叶部发生的主要真菌病害;Physoderma maydis Miyabe和Fusarium subglutinans(Wollenwe.et Reinking)P.E.Nelson et al病原引起的褐斑病和顶腐病有加重发生的趋势;Puccinia sorghi Schw引起的普通锈病在局部区域的一些品种和组合上发病较为严重;其他叶部病害零星发生,对制种玉米正常的生长发育影响较小,一般不会造成流行为害。     

单端孢菌素对几种作物病原菌的室内毒力及田间防效

为明确单端孢菌素对主要作物病原菌的抑菌活性和田间防效,采用孢子萌发抑制测定法、菌丝生长速率法和田间药效常规试验方法,测定了单端孢菌素在不同质量浓度下对作物病原菌的室内毒力及田间药效。结果表明:单端孢菌素对供试16个病原菌的MIC为1.9~15.5mg/L,对辣椒炭疽病菌、番茄早疫病菌、水稻恶苗病菌、水稻稻瘟病菌、番茄灰霉病菌、玉米小斑病菌和马铃薯晚疫病菌等7个病原菌的EC50分别为5.93、7.90、9.27、10.78、13.34、14.95和16.50mg/L,500mg/L质量浓度对番茄灰霉病、番茄早疫病、辣椒炭疽病、马铃薯晚疫病、玉米小斑病和水稻稻瘟病的防治效果分别为78.98%、81.29%、85.28%、72.42%、78.99%和72.74%。单端孢菌素具有较强的抑菌活性和较广的抑菌谱,对几种主要作物真菌和卵菌病害具有较好的田间防治效果,应用前景广。     

产多酚氧化酶茶树内生真菌的筛选及产酶条件优化

以愈创木酚、α-萘酚、没食子酸、L-酪氨酸和单宁酸等5种酚类物质为底物,采用鉴别培养基筛选法从14株茶树内生真菌中初筛获得4株产多酚氧化酶的真菌。根据变色圈的大小、颜色深浅和摇瓶发酵的结果复筛获得产多酚氧化酶能力较强的菌株CSN-13。对菌株CSN-13产酶营养条件进行初步分析,结果表明,在供试的6种碳源物质中,以麸皮对菌株CSN-13产多酚氧化酶的促进作用最为明显;供试的5种氮源物质中,以硝酸铵的促进作用最为明显;在发酵培养基中添加茶水,对产酶有明显的促进作用。采用正交设计对CSN-13产酶发酵培养基进行初步优化,优化后的培养基配方为：麸皮（40βg·L^-1）、硝酸铵（15βg·L^-1）、茶水（4βg·L^-1）、KH₂PO₄（2βg·L^-1）、MgSO₄·7H₂O（0.5βg·L^-1）、无水CaCl₂（0.075βg·L^-1）、CuSO₄·5H₂O（0.01βg·L^-1）。采用优化后的培养基,菌株CSN-13在28℃下培养5βd,酶活力达到241βU·mL^-1·min^-1,比优化前提高8.5倍。茶树内生真菌菌株CSN-13及其发酵产酶培养基的研究为多酚氧化酶的进一步开发打下了基础。     

石斛内生细菌DEB-2对5种辣椒病原真菌的抑制作用

为了探明石斛内生细菌DEB-2菌株对贵州辣椒病原真菌的抑制作用,采用生长速率法测定了菌株DEB-2发酵液对辣椒黑斑病病原菌Alternaria alternate、辣椒疫病病原菌Phytophthora capsici、辣椒黑点炭疽病病原菌Colletotrichum capsici、辣椒红色炭疽病病原菌Colletotrichum gloesporioide和辣椒早疫病病原菌Alternaria solani的毒力。结果显示,内生菌DEB-2菌株能显著抑制5种辣椒病原真菌菌丝生长和孢子萌发,菌丝尖端均有不同程度变粗、膨大、崩解或者菌丝分枝增多的现象;分生孢子及芽管产生畸形。DEB-2菌株发酵液对5种病原菌菌丝生长的抑制中浓度EC₅₀值分别是28.5、76.2、80.7、79.9、72.0 μL/mL;对辣椒疫病病原菌和早疫病病原菌分生孢子萌发的EC₅₀分别为73.5 μL/mL和68.7 μL/mL。研究表明,DEB-2菌株能有效地同时抑制5种辣椒病原真菌菌丝生长和孢子萌发,对该5种辣椒真菌病原菌具有一定的生防潜力,可作为生防菌进行开发和应用。     

First report of ophidiomycosis in Asia caused by Ophidiomyces ophiodiicola in captive snakes in Japan

Ophidiomycosis is an emerging infectious disease caused by the fungus Ophidiomyces ophiodiicola, which has been affecting wild and captive snakes in North America, Europe, and Australia. We report 12 cases of suspected ophidiomycosis in captive colubrid snakes in Japan. Pathological and microbiological examinations were performed, and the results confirmed the diagnosis of ophidiomycosis in two snakes, which indicated that the remaining sympatrically raised snakes also had ophidiomycosis since they exhibited similar lesions. This is the first report of ophidiomycosis in Asia caused by O. ophiodiicola. To prevent the expansion of ophidiomycosis in the natural environment in Japan, there is a need to evaluate the ophidiomycosis carrier status of imported snakes, the pathogenicity of the infection in native snakes, and the prevalence and distribution of O. ophiodiicola in wild and captive snakes. Measures also must be taken to prevent endemicity globally.     

Relative abundance and activity of melanized hyphae in different soil ecosystems

Here we report on the frequency of melanized fungal hyphae in 323 soils, covering different land use types. The proportion of total hyphae that was melanized averaged 61%. Arable fields with loamy sand, heathlands and city parks on sandy soils had the highest percentage of melanized hyphae. In addition to the frequency determinations, a microcosm study was performed on the role of melanized hyphae in two different ecosystems: an ex-arable field and a forest. Melanized hyphae appeared to be part of the active hyphae in the forest soil but not in the ex-arable soil. In conclusion, our results indicate that (1) melanized hyphae represent a large proportion of the total fungal biomass in soils and that (2) their function might differ between ecosystems.     

In vitro susceptibility patterns of Aspergillus and Fusarium species isolated from equine ulcerative keratomycosis cases in the midwestern and southern United States with inclusion of the new antifungal agent voriconazole

Objective  To evaluate and compare the in vitro susceptibility of Aspergillus and Fusarium spp. isolated from horses with ulcerative keratomycosis, address regional differences in equine keratomycosis isolates, and provide susceptibility data to update prior studies.
Animal studied  Fourteen horses with ulcerative keratomycosis.
Procedures  Banked fungal isolates from equine corneal ulcers (eight Aspergillus spp. and six Fusarium spp.) were identified at The University of Texas Health Science Center at San Antonio. In vitro minimum inhibitory concentration and susceptibility to natamycin, fluconazole, itraconazole, voriconazole, ketoconazole, and miconazole were determined for each isolate.
Results  Fungi were significantly more susceptible to voriconazole than to natamycin, itraconazole, fluconazole, and ketoconazole, but miconazole susceptibility did not differ significantly from voriconazole. Aspergillus spp. were most susceptible to voriconazole, miconazole, and itraconazole, which were significantly better to fluconazole and ketoconazole. Fusarium spp. susceptibility was greatest to natamycin and voriconazole and lowest to itraconazole and ketoconazole. Fusarium spp. were significantly less susceptible to itraconazole and ketoconazole compared to natamycin. No significant differences in susceptibility were found when isolates from Florida were compared with isolates from other states.
Conclusions and clinical relevance  Based on in vitro evidence, voriconazole appears to be the most effective antifungal for initial treatment of equine keratomycosis in the midwestern and southern United States. Results are comparable with previous studies in that isolated fungi from equine keratomycosis cases showed consistently poor susceptibility to fluconazole. Organisms isolated in different geographic locations of the midwestern and southern United States appeared to have similar patterns of antifungal susceptibility.