Effects of cyazofamid against Plasmodiophora brassicae Woronin on Chinese cabbage

Cyazofamid (4-chloro-2-cyano-N,N-dimethyl-5-p-tolylimidazole-1-sulfonamide) is a novel fungicide with high levels of activity against Oomycetes fungi and Plasmodiophora brassicae Woronin. The effects of cyazofamid were investigated against P. brassicae, the causal agent of clubroot disease in Chinese cabbage. Cyazofamid at 0.3 mg litre(-1) inhibited resting spore germination of this pathogen by about 80%. Cyazofamid at 3-10 mg litre(-1) exhibited fungicidal activity to resting spores of P. brassicae 1-10 days after treatment. When cyazofamid was applied to infested soil, both root-hair infections and club formation caused by P. brassicae were strongly inhibited at 1-3 mg kg(-1) dry soil. These results suggest that cyazofamid directly inhibits resting spore germination, thereby leading to the inhibition of root-hair infection and club formation. Cyazofamid at 3 mg kg(-1) dry soil also exhibited complete control of clubroot disease. The effect of broadcast soil application using a dust formulation at 2 kg AI ha(-1) (equivalent to 1.3 mg AI kg(-1) dry soil), and plug seedling tray application by a suspension concentrate formulation at 200 and 400 mg AI tray(-1) (30 x 60 x 4 cm3) against P. brassicae was also evaluated. Cyazofamid exhibited good efficacy against the pathogen. The sequential treatment including plug seedling tray application with cyazofamid and pre-plant broadcast soil application with the fungicide fluazinam also exhibited excellent levels of control. These results indicate that cyazofamid has a high potential to be an effective fungicide for the control of clubroot disease.     

Population structure of Phytophthora infestans collected on potato and tomato in Italy

Late blight caused by the oomycete Phytophthora infestans is a disease of potato and tomato of worldwide relevance and is widespread throughout Europe and the Mediterranean region. While pathogen populations in northern Europe have been sampled and characterized for many years, the genetic structure of populations from southern Europe, including Italy, has been less studied. Between 2018 and 2019, we collected 91 samples of P. infestans from potato and tomato crops in Italy, Algeria, and Tunisia on FTA cards and genotyped them using 12-plex microsatellites. These samples were compared to genotypes of P. infestans previously collected within the framework of the EuroBlight network and from published sources. Four clonal lineages were identified: 13_A2 (Blue 13), 2_A1, 23_A1, and 36_A2. Two other isolates collected could not be matched to any currently known clonal lineage. The 13_A2 and 36_A2 lineages were found exclusively in southern Italy and Algeria, while 2_A1 was only found in Algeria. This is the first report of the 36_A2 lineage in Italy. Two isolates from Solanum nigrum were 13_A2, suggesting this weed host could be a reservoir of inoculum. The 23_A1 lineage was found widely on infected tomato crops in Italy and is the same as the lineage US-23 that is widespread in North America. Differences in genotypes across the country suggests that there may be different sources of introduction into Italy, possibly via infected seed tubers from other countries in Europe, tubers for consumption from North Africa, or tomatoes.     

Effectiveness of plant extracts of Paeonia suffruticosa and Hedera helix against diseases caused by Phytophthora infestans in tomato and Pseudoperonospora cubensis in cucumber

Journal of Plant Diseases and Protection - The effects of ethanolic extracts (20 % per fresh weight (Fw) or 6 % per dry weight) of 20 different plant species were tested against late blight...     

青海马铃薯晚疫病菌线粒体DNA单倍型鉴定及分析

 为查明青海地区晚疫病菌单倍型分布情况,本研究对青海不同地区采自2006～2007年的70个晚疫病菌材料的线粒体单倍型进行了分析,结果表明：94.3%的菌株为Ⅱa型,5.7%的菌株为Ⅱb型,其中Ⅱb型均分离自番茄。这说明青海地区马铃薯晚疫病菌群体类型比较单一,主要为Ⅱa型,线粒体单倍型不存在多态性。     

马铃薯晚疫病菌对烯酰吗啉的敏感性基线及其室内抗药突变体的研究

 2003-2004年在内蒙古和黑龙江省共分离获得100株马铃薯晚疫病菌,并测定了这些菌株对烯酰吗啉和甲霜灵的敏感性。结果表明,不同地区、不同年份的马铃薯晚疫病菌对烯酰吗啉的敏感性无显著差异,其敏感性呈连续的单峰曲线分布,EC₅₀在0.0631-0.2206μg/mL之间,最不敏感菌株是最敏感菌株的3.1倍,平均为(0.1153±0.0308)μg/mL,未出现敏感性下降的抗药性亚群体,因此可作为马铃薯晚疫病菌对烯酰吗啉的敏感性基线;64%的供试菌株对甲霜灵的敏感性下降,甲霜灵与烯酰吗啉之间不存在交互抗药性。采用紫外线诱变菌丝的方法仅从3个菌株中的1个菌株获得了3%突变频率的抗烯酰吗啉突变体,抗性倍数为1.5-1.9,没有获得高抗药水平的突变体。这些突变体与氟吗啉之间具有正交互抗药性,而与霜脲氰或嘧菌酯之间无交互抗药性;与野生亲本菌株相比,抗药突变体的生长速率和产孢子囊能力显著下降,不利于抗药群体的发展。但由于马铃薯晚疫病菌被杀菌剂抗性行动委员会认为是高抗药性风险病原菌,建议生产上应将烯酰吗啉与其它无交互抗性的药剂交替使用或混合使用,以避免或延缓抗药性的产生。     

年际间干旱对晚疫病菌生理小种复杂性的影响

由致病疫霉Phytophthora infestans引起的晚疫病是马铃薯生产中最具毁灭性的病害,生理小种是晚疫病菌最重要的生物学特性之一,其组成直接关系到晚疫病的发生与流行。本研究以2007年黑龙江省干旱为契机,对2004-2010年采自黑龙江省的晚疫病菌菌株进行生理小种鉴定,结果表明,382个晚疫病菌菌株中共鉴定出91个生理小种类型,2004-2006年的优势小种为3.4.6.7.8.9.10.11,2007-2009年的优势小种为3.4.7.11,2010年的优势小种为1.3.4.7.8.10.11。2007年生理小种的复杂程度为历年最低,并影响了2008年与2009年的生理小种组成,2010年的生理小种复杂程度则出现明显上升。     

大豆疫霉菌部分生物学特性及其药剂筛选研究

采用大豆疫霉菌在不同条件下菌丝生长速度法研究了大豆疫霉菌的部分生物学特性,并应用杀菌剂室内生测、盆栽药剂防效对药效作了评价。研究结果表明,大豆疫霉菌营养生长的最适温度为25～30℃;最适pH为6;光暗交替有利于该菌营养体的生长,在Rye或CA培养基上生长最快。室内药效测定结果表明,烯酰吗啉EC₅₀为0.165 4μg/mL,抑菌效果最好,甲霜灵、甲霜灵.锰锌和氟吗啉.锰锌的EC_{50分别为0.261 00、.451 0和0.984 2μg/mL,效果次之。盆栽试验结果表明,几种药剂在活体条件下对大豆疫病的防治效果较好,并有较长持效期。}     

马铃薯晚疫病菌对甲霜灵抗性机制的初步研究

 选取对甲霜灵敏感、中抗和高抗的马铃薯晚疫病菌(Phytophthora infestans)菌株各2株,采用电导率法和同位素掺入法,研究了甲霜灵对马铃薯晚疫病菌不同抗药性水平菌株细胞膜通透性和RNA聚合酶活性的影响,借助液相色谱分析方法检测了不同抗药性水平菌株对甲霜灵的降解作用。结果表明:甲霜灵对马铃薯晚疫病菌不同抗药性水平菌株的细胞膜通透性没有明显影响;随着甲霜灵浓度提高,敏感菌株的RNA聚合酶活性受到显著抑制,10μg/mL甲霜灵对敏感菌株RNA聚合酶活性的抑制率达到40%以上,而10和100μg/mL甲霜灵对高抗菌株的RNA聚合酶活性只表现轻微的抑制作用,抑制作用分别小于8%和21%,甲霜灵对中抗菌株RNA聚合酶活性的抑制作用介于对敏感菌株和高抗菌株的抑制作用之间。抗性菌株对甲霜灵有一定的降解作用,培养30d后高抗菌株HL105和SC1的降解率分别达到13.46%和16.14%。     

Development of a quantitative real‐time PCR assay for Phytophthora infestans and its applicability to leaf,tuber and soil samples

A sensitive real‐time polymerase chain reaction (PCR) assay was developed for the quantification of Phytophthora infestans, the cause of foliar and tuber late blight in potato. A primer pair (PinfTQF/PinfTQR) and a fluorogenic probe (PinfTQPR) were designed to perform a quantitative assay for the detection of P. infestans in leaves, tubers and soils. The assay was shown to be specific to P. infestans and the very closely taxonomically related non‐potato pathogen species P. mirabilis, P. phaseoli and P. ipomoea, but did not detect the potato pathogens P. erythroseptica and P. nicotianae. The assay was able to reliably detect P. infestans DNA at 100 fg per reaction and was effective in quantifying P. infestans in infected leaf tissue from 24 h after inoculation and also in infected symptomless tubers and diseased tubers. Attempts to detect oospores of P. infestans in naturally and artificially infested soil samples are described and compared with baiting tests and previous literature. It was not possible to detect oospores in soil samples due to problems with DNA extraction from the oospores themselves. However, the assay was shown to detect even very low levels of asexual inoculum (sporangia and mycelium) in soil. This work assembles all the necessary features of a quantitative P. infestans assay, which have previously been somewhat disparate: the sensitivity, specificity and quantitation are fully validated, the assay is shown to work in common applications in leaf and tuber tissue and the problems with P. infestans oospore detection are explored and tested experimentally.     

马铃薯晚疫病菌拮抗粘细菌YR–35的分离鉴定及其代谢产物稳定性

从乌拉山镇的农田土壤中分离获得致病疫霉的高效拮抗菌株YR-35,通过形态特征、生理生化特征以及16S rDNA序列分析将其鉴定为变绿色粘球菌Myxococcus virescens。菌株YR-35对马铃薯晚疫病致病菌菌丝生长抑制率高达96.67%。抗菌活性物质在30~80℃处理1 h后菌丝生长抑制率可达21.70%,高于80℃时活性逐渐下降,90℃处理后菌丝生长抑制率仍然可达15.38%。活性物质在pH 5.0~9.0条件下菌丝生长抑制率保持在25.27%以上,当pH5.0或pH9.0时,抗菌活性显著降低。活性物质不能被蛋白酶降解,不受紫外线、自然光照射的影响。对马铃薯离体叶片的生防效果检测表明,菌株YR-35的发酵无菌液处理组叶片相对病斑面积仅为0.57%,对照组的相对病斑面积高达87.72%。菌株YR-35的发酵无菌液可以有效抑制致病疫霉侵染马铃薯叶片。本研究首次发现变绿色粘球菌对致病疫霉有显著拮抗活性,为研发有效防治马铃薯晚疫病的生物农药奠定了基础。     

知母提取物对马铃薯晚疫病菌的抑制作用及防病效果

 本研究测试了知母提取物对马铃薯晚疫病菌菌丝生长、游动孢子释放、休止孢萌发和孢子囊直接萌发的抑制作用。结果表明,知母的水提取物对菌丝生长、游动孢子释放和休止孢萌发的抑制中浓度分别为1.91mg/mL、1.23mg/mL和1.04mg/mL;对孢子囊直接萌发也具一定的抑制作用,在10.00mg/mL浓度下,抑制率达72.89%。离体叶片测试发现,浓度高于0.5%的知母提取物对马铃薯晚疫病具有显著的防治效果。盆栽植株测试结果表明,1%的知母提取物对马铃薯晚疫病的保护效果达70%以上,防效优于离体叶片试验结果,且持效期可达5d。2%和1%的知母提取物对马铃薯晚疫病的治疗效果不明显。     

寄主对马铃薯晚疫病菌群体结构的影响研究

对云南省13个寄主品种的马铃薯晚疫病菌群体结构进行研究。结果表明,云南省马铃薯晚疫病菌群体结构复杂,病菌复杂基因型群体增多,生理小种和交配型都趋于复杂化;A1交配型仍然是云南省马铃薯晚疫病菌的主要类型。鉴定生理小种的186个菌株中,共鉴定出100个生理小种类型,优势小种为1.2.3.4.5.6.7.8.9.10.11,占22.1%。品种F44-2、F45-1、PB36、昆引18、F18-1、昆引10和F29-2上都存在优势小种1.2.3.4.5.6.7.8.9.10.11,在各品种中分别占42.1%、31.6%、25.0%、21.1%、61.5%、27.3%和20.0%。滇薯6号、大理7号和克97G10-4的小种类型各不相同,甚至同一品种同一植株上分离菌的小种类型也不相同。在交配型测定过程中,还发现品种F29-2上有2个A2交配型菌株,占这个品种的25.0%,占所测定菌株的1.9%,其小种类型分别为1.2.3.5.6.7.8.9.10.11和2.3.4.6.7.9.10。     

致病疫霉对缬菌胺抗性突变体生物学特性及抗性风险量化评估

 通过研究致病疫霉对缬菌胺抗性突变体的生物学特性,评估致病疫霉对缬菌胺产生抗性的风险。采用菌丝生长速率法检测田间菌株对缬菌胺的敏感性;通过比较抗性突变体在无药条件下继代培养前后对缬菌胺的敏感性来确定其抗性稳定性;采用离体叶片法测定抗性突变体及其亲本敏感菌株的适合度和竞争力;通过量化7个基本抗性因素的风险值评估致病疫霉对缬菌胺的基本抗性风险。结果表明:在2018年从内蒙、河北、贵州、四川、黑龙江等五省(自治区)采集分离的153个菌株和2019年从河北、内蒙采集分离的40个菌株中低抗菌株分别占31.4%和35.0%,敏感菌株分别占68.6%和65.0%,抗性指数均为0.34,抗性倍数分别为1.5和2.1,未发现中抗和高抗菌株;8个抗性突变体在无药条件下继代培养10代后,少数抗性突变体的抗药性不能稳定遗传;6个抗性突变体的适合度均显著低于其亲本敏感菌株;4个抗性突变体与其亲本菌株的孢子囊不同比率混合物分别在离体叶片上继代培养1、3、7代的抗性频率显著低于初始抗性频率或与初始抗性频率无显著差异;致病疫霉对缬菌胺7个基本抗性因素的风险值之和为9,推测致病疫霉对缬菌胺的基本抗性风险为低至中等。建议加强致病疫霉菌群体对缬菌胺的抗性监测,将缬菌胺与不同作用机理的杀菌剂交替或混合使用,以延缓抗药性产生和发展。     

基于无毒基因的晚疫病菌指纹类型与致病型关系研究

由致病疫霉Phytophthora infestans(Mont)de Bary引起的晚疫病是马铃薯生产上可导致绝收的世界性病害,在我国各地均有发生和流行[1].传统的致病型鉴别只局限于表型上的分析,无法对复杂多变的晚疫病菌致病型变化进行快速准确的预测,因此有必要探索新的方法来弥补常规手段的不足,以便更加准确有效地监测其变化趋势.晚疫病菌与寄主马铃薯这一病理体系中的单一抗性基因和相应的无毒基因符合典型"基因对基因"学说,因此基于抗病基因与无毒基因的关系对晚疫病菌分类将更为科学和实用.到目前为止,已知的晚疫病菌无毒基因序列有Avr1、Avr2、Avr3a和Avr4[2,3].本研究试图分析晚疫病菌基于无毒基因的指纹类型与马铃薯鉴别品种划分的致病型之间是否存在对应关系,以期进一步了解病原菌群体结构特征及遗传变异本质,从而更好地指导抗晚疫病育种和品种的合理布局.     

青海海东地区马铃薯晚疫病菌生理小种的组成及分布

 50 isolates of Phytophthora infestans potato plants were collected and tested with 11 monogene R1-R11 of potato late blight differential host in Qinghai province from 2006 to 2007.The results showed that there were 15 races in the Haidong Region of Qinghai province and the dominant race was race 3.4.10 with frequency of 36.0%.They were distributed in all collection areas.Race 3.10 with frequency of 16.0% was distributed in Huzu,Ledu,Huangyuan,Datong and Ping'an.Race 3 with frequency of 10.0% was distributed in Huzhu,Xunhua,Huangyuan and Xining.     

A review of biocontrol agents in controlling late blight of potatoes and tomatoes caused by Phytophthora infestans and the underlying mechanisms

Phytophthora infestans causes late blight on potatoes and tomatoes, which has a significant economic impact on agriculture. The management of late blight has been largely dependent on the application of synthetic fungicides, which is not an ultimate solution for sustainable agriculture and environmental safety. Biocontrol strategies are expected to be alternative methods to the conventional chemicals in controlling plant diseases in the integrated pest management (IPM) programs. Well-studied biocontrol agents against Phytophthora infestans include fungi, oomycetes, bacteria, and compounds produced by these antagonists, in addition to certain bioactive metabolites produced by plants. Laboratory and glasshouse experiments suggest a potential for using biocontrol in practical late blight disease management. However, the transition of biocontrol to field applications is problematic for the moment, due to low and variable efficacies. In this review, we provide a comprehensive summary on these biocontrol strategies and the underlying corresponding mechanisms. To give a more intuitive understanding of the promising biocontrol agents against Phytophthora infestans in agricultural systems, we discuss the utilizations, modes of action and future potentials of these antagonists based on their taxonomic classifications. To achieve a goal of best possible results produced by biocontrol agents, it is suggested to work on field trials, strain modifications, formulations, regulations, and optimizations of application. Combined biocontrol agents having different modes of action or biological adaptation traits may be used to strengthen the biocontrol efficacy. More importantly, biological control agents should be applied in the coordination of other existing and forthcoming methods in the IPM programs. © 2023 Society of Chemical Industry.     

丁吡吗啉对致病疫霉的作用机制初探

丁吡吗啉(pyrimorph)是一种结构新颖的杀菌剂,室内生物测定结果表明,其对致病疫霉Phytophthora infestans的菌丝生长、孢子囊产生、休止孢萌发具有较强的抑制作用,其EC50值分别为0.066、0.059和0.550 μg/mL,但对游动孢子释放的抑制作用较弱,其EC50值为 9.78 μg/mL。显微镜下观察结果表明,经丁吡吗啉处理后的菌丝分枝相对较少,分枝间距拉长,但对 菌丝直径无影响;从致病疫霉对番茄叶片的侵染过程来看,丁吡吗啉浓度为100 μg/mL时,处理24 h后在叶片表面只能看到少量休止孢萌发,96 h后未产生孢子囊。丁吡吗啉在1 μg/mL和10 μg/mL 时对菌丝体细胞膜的电导率无影响,但当浓度为50 μg/mL时其电导率值明显上升;初步测定结果表明,在100 μg/mL浓度下,其对致病疫霉菌丝体的蛋白质合成有一定的抑制作用,但对其菌丝体内的DNA合成没有明显的抑制作用。