不同分子量壳聚糖抑菌作用的研究

随着壳聚糖研究的不断深入,其不同寻常的抑菌活性引起了学者们的广泛关注。为了获得壳聚糖对桉树青枯病病原菌极其拮抗菌的抑制活性,在研究中考察了不同分子量的壳聚糖对桉树青枯病的生防菌302-多粘类芽孢杆菌、199-蜡状芽孢杆菌及桉树白枯病的病原菌76-茄青枯拉尔氏菌、RS-桉树青枯病原菌的抑制效果。采用琼脂扩散纸片法研究了不同浓度和分子量的壳聚糖对上述4种菌的抑制效果,利用了系列稀释法研究了不同分子量的壳聚糖的最小抑菌浓度。同时研究了分子量为2200的壳聚糖在不同pH条件下的抑菌效果。结果表明,5种不同分子量的壳聚糖对76-茄青枯拉尔氏菌和RS-桉树青枯病病原菌有明显的抑制作用,其中分子量为2200和5万的壳聚糖对青枯病原菌的抑菌效果最好,且随壳聚糖浓度的增大,壳聚糖的抑菌作用逐渐增强,最小抑菌浓度在0.05%以上。而这2种分子量的壳聚糖对302-多粘类芽孢杆菌和199-蜡状芽孢杆菌的抑菌效果较上述2种病原菌要明显弱很多,最小抑菌浓度在0.1%以上。在上述试验的基础上,本试验进一步探讨了分子量为2200的壳聚糖在不同pH条件下对4种受试菌的抑制效果。结果表明,当pH在5.5~6.5时,对受试菌具有最佳的抑制效果。壳聚糖对上述4种菌的作用效果尚未见文献报道。     

人参主要病原菌生防真菌的筛选及鉴定

【目的】筛选对人参主要病原菌具高效、广谱抑菌作用的生防真菌,丰富人参生防菌菌种资源。【方法】采用稀释平板法,从吉林省2处采样地的人参健株根际土壤中分离真菌;采用平板对峙培养法,通过初筛和复筛,筛选对人参链格孢菌(Alternaria panax)、立枯丝核菌(Rhizoctonia solani)、腐皮镰孢菌(Fusarium solani)、毁灭柱孢菌(Cylindrocarpon destructans)、恶疫霉菌(Phytophthora cactorum)、人参核盘菌(Sclerotinia schinseng)和灰葡萄孢菌(Botrytis cinerea)具有高效广谱抑菌活性的生防真菌;利用传统形态学和rDNA-ITS分子分析技术对分离到的生防真菌进行鉴定。【结果】从人参健株根际土壤中共分离得到400株真菌,其中有15株生防真菌对人参7种主要病原菌具有广谱抑菌作用,特别是对立枯丝核菌有强烈的抑制作用(抑菌率均大于50%),且FSR-106的抑菌作用最强(抑菌率87.41%,下同);FSR-121对人参链格孢菌的抑菌作用最强(60.00%);FSR-74对腐皮镰孢菌的抑菌率高达67.48%;FSR-46对毁灭柱孢菌和灰葡萄孢菌的抑菌率最大(66.67%和68.97%);FSR-97对恶疫霉菌及人参核盘菌的抑制作用最强(73.33%和66.67%)。通过形态学和rDNA-ITS序列分析鉴定发现,15株生防真菌中有8株木霉属(Trichoderma)、3株青霉属(Penicillium)、1株曲霉属(Aspergillus)、1株附球菌属(Epicoccum)、1株毛壳菌属(Chaetomium)和1株轮层炭壳属(Daldinia),其中FSR-38为黑附球菌(Epicoccum nigrum),FSR-10、FSR-43和FSR-106为深绿木霉(T.atroviride),FSR-46为长枝木霉(T.longibrachiatum),FSR-55为钩状木霉(T.hamatum),FSR-72为桔绿木霉(T.citrinoviride),FSR-74为球毛壳菌(Chaetomium globosum),FSR-97为多孢木霉(Trichoderma polysporum),FSR-91为蔡氏轮层炭壳菌(Daldinia childiae),FSR-25、FSR-67和FSR-121为青霉属(Penicilliumsp.)。【结论】从人参健株根际土壤中可快速有效地筛选获得大量广谱性生防真菌;本研究所得的15株生防真菌对7种人参主要病害防治具有较高的潜在应用价值。     

短密木霉新菌株BF06对黄瓜土传病害的防治效果与促生作用

连作导致枯萎病等土传病害发生日益严重,已成为我国黄瓜生产的重要限制因素。采用化学防治和农业措施防治土传根部病害,操作都较为困难,迫切需要开发环境友好的生物防治技术。本研究从森林土壤分离鉴定一株短密木霉菌株BF06,通过对峙培养发现该菌株可以附着和缠绕病原菌的菌丝,对引起黄瓜枯萎病、茎基腐病、菌核病、根腐病和疫病的病原菌Fusarium oxysporum f. sp. cucumerinum,Rhizoctonia solani,Sclerotinia sclerotiorum,F. solani f. sp. cucurbitae,and Phytophthora cryptogea具有较强的拮抗作用。温室盆栽试验发现短密木霉菌株BF06可以迅速附着定殖于黄瓜根部表面,对上述5种黄瓜根部病害的防效达60%以上,对黄瓜枯萎病和茎基腐病的防效分别为90.4和88.8%。此外,利用植物组织培养基培养观察发现BF06显著地促进幼苗黄瓜侧根的形成和生长。本研究的结果表明短密木霉菌株BF06是一种可以有效防治黄瓜根部病害的新生防资源。     

A new genus of Cryphonectriaceae isolated from Lagerstroemia speciosa in southern China

Numerous species in the Cryphonectriaceae have been recorded on the Myrtales and many of these are economically important pathogens of Eucalyptus. Some species have also recently been shown to be endophytes of native Myrtaceae and to have undergone host jumps to infect Eucalyptus species established as exotics in plantations. Recent surveys in the GuangDong and HaiNan Provinces of South China reveal the presence of a species of Cryphonectriaceae associated with cankers on trees of Lagerstroemia speciosa (Lythraceae, Myrtales). Fungal structures were observed on the surface of dead bark covering cankers and on branch stubs. Multigene phylogenetic analyses were conducted based on DNA sequence comparisons of the partial LSU gene, ITS region of the nuclear ribosomal RNA gene and two regions of the β‐tubulin (BT) gene. The results revealed the presence of a previously undescribed genus and species in the Cryphonectriaceae. The fungus is described here as Chrysomorbus lagerstroemiae gen. et sp. nov. Inoculation tests showed that it is an aggressive pathogen on L. speciosa and that it can also infect Eucalyptus.     

Loop‐mediated isothermal amplification (LAMP) assays for rapid detection of Pyrenopeziza brassicae (light leaf spot of brassicas)

Pyrenopeziza brassicae (anamorph Cylindrosporium concentricum) is an ascomycete fungus that causes light leaf spot (LLS) disease of brassicas. It has recently become the most important pathogen of winter oilseed rape (Brassica napus) crops in the UK. The pathogen is spread by both asexual splash‐dispersed conidia and sexual wind‐dispersed ascospores. Such inoculum can be detected with existing qualitative and quantitative PCR diagnostics, but these require time‐consuming laboratory‐based processing. This study describes two loop‐mediated isothermal amplification (LAMP) assays, targeting internal transcribed spacer (ITS) or β‐tubulin DNA sequences, for fast and specific detection of P. brassicae isolates from a broad geographical range (throughout Europe and Oceania) and multiple brassica host species (B. napus, B. oleracea and B. rapa). Neither assay detected closely related Oculimacula or Rhynchosporium isolates, or other commonly occurring oilseed rape fungal pathogens. Both LAMP assays could consistently detect DNA amounts equivalent to 100 P. brassicae conidia per sample within 30 minutes, although the β‐tubulin assay was more rapid. Reproducible standard curves were obtained using a P. brassicae DNA dilution series (100 ng–10 pg), enabling quantitative estimation of amounts of pathogen DNA in environmental samples. In planta application of the β‐tubulin sequence‐based LAMP assay to individual oilseed rape leaves collected from the field found no statistically significant difference in the amount of pathogen DNA present in parts of leaves either with or without visible LLS symptoms. The P. brassicae LAMP assays described here could have multiple applications, including detection of symptomless host infection and automated real‐time monitoring of pathogen inoculum.     

Dutch elm disease on the island of Gotland: monitoring disease vector and combat measures

The island of Gotland (Sweden) possesses the largest (more than one million trees) wild population of elms in northern Europe, which until recently was not affected by Dutch elm disease (DED). In 2005, DED was observed for the first time in the northeastern part of Gotland, and in the following years, it rapidly spread in all directions, generally following the major distribution of elms. Implementation of intensive control measures including ground surveys and geographic information system mapping of DED-diseased trees, their felling and destruction, and stump treatment with the herbicide glyphosate allowed reduction of the source of infection and probably suppression of an otherwise very rapid spread of the disease. Monitoring the flying periods of Scolytus multistriatus vector beetles between the years 2007 and 2011 showed that the abundance of trapped beetles in most cases differed significantly among individual years. Over the entire monitoring period, 55.2% of the beetles were trapped in June, 31.2% in July and 13.5% in August (difference among individual months significant at p?S. multistriatus, suggesting that the removal of trees attacked in the previous vegetation period as a combat measure should be completed before the beginning of the flying season of S. multistriatus, and that the application of the integrated DED management has the potential to arrest the development of the disease in a geographically isolated area such as Gotland.     

土壤原生病原体对森林植被及生态系统的影响

土壤原生病原体是影响植被和生态系统的重要因子,但是在森林生态系统中的地位和作用未能得到充分认识和研究。文中主要根据国外对土壤原生病原体的研究情况,对其类型、传播途径及植被的防御机制进行阐述,并探讨土壤有益微生物和土壤原生病原体的相互作用及原生病原体对生态系统的影响机制。     

小陇山林区林木病原菌物区系及评价

通过对小陇山林区主要林型内林木病原菌物的广泛调查,利用形态学和分子生物学技术鉴定并记述了该林区的病原菌物154种,隶属71属、41科;其中葡萄座腔菌科(Botryosphaeriaceae)、叶点霉属(Phyllosticta)分别为该林区病原菌物的优势科和优势属;在该林区的7个林型中,油松-锐齿栎林和华山松-栓皮栎林的病原菌物种类最多,云杉林的最少,并根据该林区的发病特点提出了防治建议。     

3种食源性致病菌TaqMan多重荧光定量PCR检测方法的建立

旨在建立一种可同时快速检测大肠杆菌O157 ∶ H7、沙门菌和产单核细胞李氏杆菌3种食源性致病菌的TaqMan多重荧光定量PCR(qPCR)方法.针对大肠杆菌O157 ∶ H7 rfbE基因、沙门菌invA基因和产单核细胞李氏杆菌hlyA基因的保守序列分别设计特异性引物和TaqMan探针,建立多重qPCR反应体系,进行...     

新鲜果蔬流通过程中致病微生物种类及其控制

对新鲜果蔬主要致病微生物种类、侵染途径及加工过程中微生物控制技术进行概述,并提出从采前栽培环境与管理开始到消费整个流通过程中控制的系统工程理论,着重阐述了新鲜果蔬加工产品的安全与控制技术。