甘草叶斑病的发生与病原菌鉴定

近年来,在甘肃省兰州市及渭源地区甘草产区发现一种甘草新病害—甘草叶斑病。2006－2008年对其发病情况进行了调查,甘草叶斑病在6月下旬开始发病,9月中旬为发病高峰期,发病率达80%。高燥地发病率低于低洼地,采用轮作可减轻甘草叶斑病的发生。经过对甘草叶斑病病原菌形态、培养性状和致病性等方面的测定表明,病原菌为豆链格孢（Alternaria azukiae）。通过寄主范围测定表明,该病原菌亦可侵染独活、羌活等药用植物。对大黄、苍耳等不具侵染性。     

Combating plant diseases—the Darwin connection

Although Darwin knew of plant diseases, he did not study them as part of his analysis of natural selection. Effective plant disease control has only been developed after his death. This article explores the relevance of Darwin's ideas to three problem areas with respect to diseases caused by fungi: emergence of new diseases, loss of disease resistance bred into plants and development of fungicide resistance. Darwin's concept of change through natural or artificial selection relied on selection of many small changes, but subsequent genetic research has shown that change can also occur through large steps. Appearance of new diseases can involve gene duplication, transfer or recombination, but all evidence points to both host plant resistance and fungicide susceptibility being overcome through point mutations. Because the population size of diseases such as rusts and powdery and downy mildews is so large, all possible point mutations are likely to occur daily, even during moderate epidemics. Overcoming control measures therefore reflects the overall fitness of these mutants, and much resource effort is being directed towards assessment of their fitness, both in the presence and in the absence of selection. While recent developments in comparative genomics have caused some revision of Darwin's ideas, experience in managing plant disease control measures clearly demonstrates the relevance of concepts he introduced 150 years ago. It also reveals the remarkable speed and the practical impact of adaptation in wild microorganism populations to changes in their environment, and the difficulty of stopping or delaying such adaptation. Copyright © 2009 Society of Chemical Industry     

香草兰根(茎)腐病病原菌鉴定及其致病性测定

香草兰根(茎)腐病是香草兰上重要病害之一,目前国内外尚未有基于rDNA-ITS序列的分子鉴定和系统发育树分析的报道。本研究从海南省万宁市的兴隆、长丰和南林等地的病样中分离获得了9个菌株,经过柯赫氏法则验证、形态学鉴定和分子鉴定,病原菌被鉴定为尖孢镰刀菌,表明海南省万宁市地区的香草兰根(茎)腐病病原菌是尖孢镰刀菌(Fusarium. oxysporum)。然后,基于rDNA-ITS序列构建了系统发育树并对菌株的致病性进行了测定,结果发现9个菌株在系统发育树中被聚类为遗传距离明显的2个分枝,而且分枝Ⅱ的致病性要高于分枝Ⅰ。本研究为香草兰根(茎)腐病病原物快速鉴定提供依据,对香草兰根(茎)腐病防治决策有一定的参考意义。     

粤北地区杉木真菌性病害种类

对粤北地区 ( 小坑林场、龙山林场、大坑山林场 ) 杉木（Cunninghamia lanceolate）真菌性病害进行全面调查,共采集 86 份标本,分离病组织获得菌株 162 份,主要归属于炭疽菌属（Colletotrichum）、拟盘多毛孢属（Pestalotiopsis）、腐皮壳属（Diaporthe）、镰刀菌属（Fusarium）、黑孢菌属（Nigrospora）、拟茎点霉属（Phomopsis）、叶点霉属（Phyllosticta）以及散斑壳属（Lophodermium）等。根据病害症状粤北地区主要杉木病害包括：杉木立枯病、杉木炭疽病、杉木溃疡病、杉木顶枯病、杉木梢枯病、杉木赤枯病、杉木球果坏死。各林场杉木林中,杉木炭疽病普遍发生、分布广;种子园中杉木球果病害危害严重,直接影响球果的产量和质量;在大坑山林场 4-5 年生的林分中,杉木顶枯病大面积发生,但在其它地区少有发生;此外,杉木溃疡病、杉木赤枯病和杉木立枯病在个别无性系或者苗圃中零星发生,但危害较轻。     

Direct quantitative real‐time PCR assay for detection of the emerging pathogen Neonectria neomacrospora

The epidemic outbreak in northern Europe of Neonectria neomacrospora, the causal agent of dieback in Abies spp., led the European and Mediterranean Plant Protection Organization (EPPO) to include the pathogen on its alert list in 2017. Effective monitoring of this pathogen calls for a rapid and sensitive method of identification and quantification. A probe‐based real‐time PCR (qPCR) assay based on the β‐tubulin gene was developed for the detection and quantification of N. neomacrospora in infected wood samples, and directly for ascospores. This study presents the first published species–specific molecular detection assay for N. neomacrospora. The analytical specificity was validated on taxonomically closely related fungal species as well as on 18 fungal species associated with the host (Abies sp.). The analytical sensitivity was tested on naturally infected wood, on purified pathogen DNA in a matrix of host DNA and on N. neomacrospora ascospores for detection of airborne inoculum. The latter was tested both with a DNA extraction step prior to qPCR and without DNA extraction by direct qPCR on collected ascospores. The assay was specific to N. neomacrospora, with a sensitivity of 130 fg purified DNA, or 10 ascospores by direct qPCR. Omitting DNA extraction and amplifying directly on unpurified ascospores improved assay sensitivity significantly.     

Fungal colonization patterns in necrotic rootstocks and stem bases of dieback‐affected Fraxinus excelsior L.

European ash (Fraxinus excelsior) trees currently face the major threat of ash dieback caused by an invasive fungus, Hymenoscyphus fraxineus. Collar rots in F. excelsior have been increasingly associated with infections by this pathogen. However, the aetiology of the collar rots is still unclear and remains heavily debated. In contrast to most studies of this kind, entire rootstocks of four diseased ash trees were dug out to examine necrotic tissues in these rootstocks and stem bases in detail and to sample necrotic wood for fungal isolation. With the aid of morphological and molecular identification techniques, five to twelve fungal taxa were detected per tree. Members of the Nectriaceae family and Botryosphaeria stevensii, the causal agent of stem and branch cankers on many tree species, were frequently isolated from outer xylem. In contrast, H. fraxineus was the dominating species in interior wood layers. Microsatellite genotyping of 77 H. fraxineus isolates helped to identify up to six different genotypes per tree. The role of H. fraxineus and other isolated fungi in the aetiology of ash collar rots are discussed.     

Host range of Erwinia psidii and genetic resistance of Eucalyptus and Corymbia species to this pathogen

Dieback caused by Erwinia psidii is currently one of the most important emerging diseases in eucalypt plantations in Brazil. However, little is known in terms of the host range of this pathogen or the potential sources of resistance against the disease it causes. In this study, we inoculated plants of species from nine families to gain insight into the host range of E. psidii. Plants of all inoculated species of Myrtaceae except Acca sellowiana exhibited disease symptoms and therefore represent potential hosts for the pathogen under natural conditions. In addition, the response of four Corymbia species, 29 Eucalyptus species and three interspecific Eucalyptus hybrids to inoculation with E. psidii was evaluated. All Corymbia henryi, Corymbia maculata, Eucalyptus thozetiana, Eucalyptus cloeziana, Eucalyptus viminalis, Eucalyptus dalrympleana and Eucalyptus pilularis plants were highly resistant to the pathogen, whereas differential disease resistance was observed in the other species. This study provides important information on sources of resistance to Erwinia psidii with potential use in the development of clones with enhanced resistance in eucalypt species of economic importance.     

Isolation and characterisation of phages against Pseudomonas syringae pv. actinidiae

Purpose: Pseudomonas syringae pv. actinidiae causes bacterial canker of kiwifruit and is responsible for severe economic losses and emergence of drug-resistant bacteria. Bacteriophages are viruses that infect target bacterial hosts and may be the best strategy to prevent and control kiwifruit canker disease. The objective of this experiment was to monitor the prevalence of Pseudomonas syringae pv. actinidiae and provide insight for the use of phages in biological control.

Materials and methods: In this study, 52 strains of Pseudomonas syringae pv. actinidiae were isolated from 68 stem samples of kiwi plant (cv. Hongyang & Jinkui). Following polymerase chain reaction (PCR) analysis, 15 isolates belonging to biovar 3 were identified, one of which was named XWY0007 and used as the target strain to isolate the phages. Thirty-six phages were isolated and purified from a total of 51 surface water samples collected in Shanghai. All phages were identified by transmission electron microscopy (TEM) and their host ranges were evaluated. Three phages, designated φXWY0013, φXWY0014 and φXWY0026 were selected and further characterised using one-step growth curve and stability at different temperatures and pH.

Results and conclusions: The isolated phages are promising for use as antimicrobials against bacterial canker in kiwi. This report is regarding Pseudomonas syringae pv. actinidiae and its phages from major areas of kiwifruit cultivation.     

黄淮海夏玉米主产区茎腐病主要病原菌及优势种分析

【目的】 明确黄淮海夏玉米主产区玉米茎腐病的主要病原菌组成及优势种,为病原菌致病机制、抗性育种及茎腐病防治提供依据。【方法】 于2014—2017年采集黄淮海夏玉米主产区3个省（河北、河南、山东）的玉米茎腐病样本850份。通过形态学、通用引物和特异引物对病组织分离物进行鉴定,并采用上述引物直接检测病组织。整合分析分离物鉴定法和组织分子检测法的结果以确定玉米茎腐病的主要病原菌及优势种;分析主要病原菌在各省间以及同一省份不同年度的检出率,揭示主要病原菌的种群动态变化;分析单个样本中病原菌的检出率,探讨多种病原菌的共存模式。【结果】 有667份样本检测出真菌或卵菌,占总样本的78.47%;年度间样本检出率存在差异,2014年的检出率不足50%,而2015—2017年的检出率相近,均高于90%。在所有样本中共检出20属46种真菌或卵菌,其中镰孢菌（Fusarium spp.）的检出率最高,为89.96%,包括禾谷镰孢复合种（F. graminearum species complex）、层出镰孢（F. proliferatum）、拟轮枝镰孢（F. verticillioides）、厚垣镰孢（F. chlamydosporum）、亚粘团镰孢（F. subglutinans）、尖镰孢（F. oxysporum）、黄色镰孢（F. culmorum）、藤仓镰孢（F. fujikuroi）、变红镰孢（F. incarnatum）、木贼镰孢（F. equiseti）和茄镰孢（F. solani）11个种;腐霉菌（Pythium spp.）的检出率为34.18%,包括芒孢腐霉（P. aristosporum）、禾生腐霉（P. graminicola）、棘腐霉（P. acanthicum）、孤雌腐霉（P. amasculinum）和寡雄腐霉（P. oligandrum）5个种。拟轮枝镰孢、禾谷镰孢复合种、芒孢腐霉和层出镰孢为4种主要病原菌,检出率依次为62.07%、46.93%、29.09%和28.04%,其中拟轮枝镰孢为优势种。各省之间,上述4种主要病原菌的检出率存在一定差异。拟轮枝镰孢在河北省的检出率为73.98%,明显高于该菌在河南省和山东省的检出率;禾谷镰孢复合种在3个省的检出率相近;层出镰孢和芒孢腐霉在山东省的检出率分别为35.78%和34.31%,均高于在其他两省的检出率。同一省份不同年度上述4种主要病原菌的检出率呈动态变化,其中任何一种病原菌均有可能上升为优势种。进一步分析表明,单个样本中可以检测出一种或多种病原菌,检测出1种菌的样本占38.38%,检出2种和3种病原菌的样本分别占29.24%和19.04%;2种及以上病原菌共存以镰孢菌和腐霉菌、镰孢菌和镰孢菌模式为主。【结论】 黄淮海夏玉米主产区茎腐病主要病原菌为拟轮枝镰孢、禾谷镰孢复合种、芒孢腐霉和层出镰孢,其中拟轮枝镰孢为优势种;拟轮枝镰孢在河北省的检出率最高,层出镰孢和芒孢腐霉在山东省的检出率最高,禾谷镰孢复合种在3个省的检出率相近;单个样本中存在多种病原菌共存的模式。     

杀菌剂对沙打旺种子萌发和幼苗生长的短期影响

病害是导致沙打旺(Astragalus adsurgens)草地产量下降的主要原因之一。本研究用杀菌剂对沙打旺种子进行预处理,在培养皿和盆栽两种培养条件下模拟沙打旺种子在自然环境中受到不同病原真菌侵染后的萌发和生长状况,探讨杀菌剂预处理的作用和效果。结果表明,人工接种病原菌后,经杀菌剂处理的种子,其发芽率,植株的株高、根长和干重较无菌水处理种子显著提高(P0.05),而发病率、死亡率和平均病情指数则相应地显著降低(P0.05)。不同杀菌剂的防治效果有所不同,在多菌灵、代森锰锌和甲基托布津3种杀菌剂中,代森锰锌预处理防治效果最好。