黑龙江和云南葡萄产区灰霉病菌对多菌灵和异菌脲的抗性检测

为明确黑龙江省和云南省葡萄灰霉病菌对多菌灵和异菌脲的抗性频率,从两省分别采集、分离到葡萄灰霉病菌Botrytis cinerea 39株和38株,利用菌丝生长速率法测定其对多菌灵和异菌脲的敏感性,并分别检测上述菌株两种药剂靶标基因β-tubulin和Bos1的突变情况.结果表明,黑龙江省葡萄灰霉病菌对多菌灵和异菌脲的抗...     

太原市郊区黄瓜霜霉病菌对甲霜灵的抗药性检测

对太原市郊区黄瓜主要种植区黄瓜霜霉病菌对甲霜灵的抗药性进行了检测,结果表明:来自于8个不同地区的黄瓜霜霉病菌对甲霜灵的抗性频率均为100%。在检测的27个菌株中有2个中抗菌株,12个高抗菌株,13个特高抗菌株,无敏感菌株。     

番茄晚疫病菌对甲霜灵的抗性

摘要为了掌握我国番茄晚疫病菌对甲霜灵抗性的发生和分布状况，1999～2001年，陆续从全国16个省市的主要番茄产区采集晚疫病样本400余个，分离、纯化出番茄晚疫病菌株183个;采取离体和活体生测法对所有菌株进行了抗甲霜灵的测定。结果表明：我国番茄晚疫病菌对甲霜灵的抗性由北向南逐渐增强，北方地区以中等抗性菌株和敏感性菌株为主，南方地区则以抗性菌株和中等抗性菌株为主。其中东北、西北地区无抗性菌株，中抗和敏感菌株分别为33．3％、66．7％和36．1％、63．9％；华北、华东地区抗性、中抗和敏感菌株分别为5．6％、44．4％、50．0％和38．6％、36．8％、4．6％；华中和西南地区抗性菌株分别占55．6％和44．2％，中抗和敏感菌株则各占33．3％、历．8％和11．1％、18．8％；华南地区无敏感菌株，抗性菌株达66．7％，中抗菌株为33．3％。     

葡萄对霜霉病抗性机理初探

经田间自然鉴定、接种鉴定及室内离体接种鉴定结果表明：葡萄对霜霉病抗性与品种叶内的含水量、可滴定酸、还原糖、淀粉、水解氨基酸、游离氨基酸含量无显著相关，与叶背气孔密度、气孔开闭程度也无相关，而外施高浓度蔗糖及邻苯二酚却显著降低霜霉菌的致病力。     

葡萄对霜霉病抗性机理初探

经田间自然鉴定、接种鉴定及室内离体接种鉴定结果表明：葡萄对霜霉病抗性与品种叶内的含水量、可滴定酸、还原糖、淀粉、水解氨基酸、游离氨基酸含量无显著相关，与叶背气孔密度、气孔开闭程度也无相关，而外施高浓度蔗糖及邻苯二酚却显著降低霜霉菌的致病力。     

黄瓜霜霉病菌对甲霜灵的抗药性治理

霜霉病是黄瓜的主要病害之一,目前主要依靠化学防治来减轻其危害.甲霜灵(metalaxyl)是苯基酰胺类杀菌剂的代表品种.我国20世纪80年代引进该药剂不久,霜霉病菌Pseudoperonospora cubensis就对甲霜灵产生了抗性.在过去10余年中,抗性治理多采用药剂替换或混用技术.作者采用不同作用机制的药剂来制定用药流程,进行了黄瓜霜霉病菌对甲霜灵的抗药性治理.     

河北省黄瓜霜霉病菌对甲霜灵和嘧菌酯的抗性动态及七种药剂的田间防效

为明确黄瓜霜霉病菌对甲霜灵和嘧菌酯的抗性动态及相应药剂对黄瓜霜霉病的田间防效,采用叶盘漂浮法监测了2011—2015年河北省黄瓜霜霉病菌对甲霜灵和嘧菌酯的抗性,采用茎叶喷雾法测定了含甲霜灵或精甲霜灵的混剂及嘧菌酯单剂等7种药剂的田间防效。结果表明:采自河北省不同地区的838株黄瓜霜霉病菌对甲霜灵和嘧菌酯普遍产生抗性,抗性频率为100.00%,抗性倍数为482.99和354.97,黄瓜霜霉病菌对甲霜灵和嘧菌酯的抗性随着监测地区和监测年限而波动,唐山、沧州、石家庄、衡水、廊坊和保定市的菌株对甲霜灵和嘧菌酯的抗性指数高于其它地区,黄瓜霜霉病菌对甲霜灵和嘧菌酯的抗性指数在不同年度间高位波动。按照田间推荐剂量喷施,250 g/L嘧菌酯悬浮剂对黄瓜霜霉病的防效为72.15%~74.13%;68%精甲霜灵·代森锰锌水分散粒剂和58%甲霜灵·代森锰锌可湿性粉剂的防效分别为65.30%~70.19%和63.26%~68.35%,与80%代森锰锌可湿性粉剂防效相当;50%烯酰吗啉可湿性粉剂、250 g/L双炔酰菌胺悬浮剂和687.5 g/L氟吡菌胺·霜霉威盐酸盐悬浮剂的防效达82.00%以上,对黄瓜霜霉病具有较好的防效,在黄瓜霜霉病菌对嘧菌酯、甲霜灵和精甲霜灵产生抗性的地区,可作为替代或混配药剂使用。     

宁夏葡萄霜霉病菌致病型鉴定及葡萄品种抗性评价

为合理利用葡萄抗性品种以及田间病害综合治理,采用离体叶盘接种法分别对2018年和2019年自宁夏回族自治区银川市、石嘴山市和吴忠市采集的43株葡萄霜霉病菌Plasmopara viticola菌株进行致病型鉴定及聚类分析,并对17个主栽葡萄品种进行抗性评价及聚类分析。结果表明：2018、2019年供试菌株致病力和供试葡萄品种抗性之间均存在显著差异,菌株平均病情指数与发病率呈显著正相关,皮尔逊相关系数r和回归系数R²分别介于0.45~0.96和0.20~0.92之间;2018、2019年供试菌株的强致病力菌株和弱致病力菌株分别为CS-YL和VI-BYDD、RG-JSJG和CS-YM;2018年和2019年供试菌株致病型被划分为强、中、弱3种类群,2年所占比例分别为10.00%、15.00%、75.00%和34.78%、39.13%、26.09%;供试17个葡萄品种的抗性类型可被划分为感病、抗病、高抗3种类群,2年所占比例均为17.65%、29.41%和52.94%。不同葡萄品种对不同菌株的抗性类型多样化。表明宁夏回族自治区葡萄霜霉病菌菌株致病力分化明显,不同菌株的致病型与其地域来源无关,不同葡萄品种的抗感程度不同。     

葡萄霜霉病菌对甲霜灵抗药性治理及其田间抗药菌株遗传稳定性分析

 为了明确葡萄霜霉病菌对甲霜灵的田间抗药性水平发展态势,于轮换用药前后,采用叶盘漂浮法测定了河北、山西、河南3省葡萄主要种植区11个葡萄园试验地葡萄霜霉病菌对甲霜灵敏感性变化动态。结果表明:田间采集的葡萄霜霉病菌对甲霜灵抗药的菌株其抗药性可以稳定遗传;不同地区轮换用药后,葡萄霜霉病菌对甲霜灵的抗药水平变化态势因用药流程的不同而发生相应的变化。采用不同作用机制的杀菌剂轮换或混合用药进行葡萄霜霉病菌对甲霜灵的抗药性治理时,需制定合理的施药流程,并根据抗药性治理的效果不断完善治理措施。     

鲜食葡萄品种对霜霉病的抗性及抗病机理研究

采用田间自然发病调查，田间接种鉴定和离体葡萄叶圆片室内接种鉴定鲜食葡萄品种对霜霉病的抗性及其抗性机理研究表明，品种间对霜霉病的抗性存在显著差异。可依感病病级分为三类，低感品种有“z工伊豆”、“奥林匹亚”、“山东早红”、“森田尼”、“早玛瑙”。几乎所有的无核品种均高感霜霉病，葡萄对霜霉病的抗性与叶内可滴定酸、还原糖、游离氨基酸含量无显著相关，与叶背气也密度、气孔开度也无相关，而外施高浓度邻苯二酚却显著降低了霜霉病发病程度。     

不同葡萄品种对霜霉病的田间苗期抗性评价

对36个葡萄品种(品系)苗期对霜霉病的田间抗病性进行了评价, 结果表明, 不同葡萄品种(品系) 对葡萄霜霉病的抗性存在明显差异, 绝大多数欧美杂交种的抗性要强于欧亚种葡萄, 供试的10个欧美杂种中, ‘瑞峰无核’对葡萄霜霉病表现为高抗; ‘DEMIR’、‘龙宝’、‘红富士’、‘黑奥林’、‘峰后’5个品种表现为中抗。     

中国4省份葡萄霜霉病菌群体对烯酰吗啉的抗性及适合度

为明确我国山西、山东、河北和云南省葡萄霜霉病菌对烯酰吗啉的抗性以及田间病菌群体的适合度,本研究采用Taqman-MGB技术检测了2020年-2021年采自上述4省份292株葡萄霜霉病菌对烯酰吗啉的抗性频率;采用叶盘法测定了田间抗性菌株与敏感菌株的适合度。结果表明,上述4个地区的平均烯酰吗啉抗性频率及抗性等位基因频率分别为64.0%和74.7%,其中山东省蓬莱区最高(91.2%和96.3%),河北省廊坊市(73.8%和85.0%)和云南省宾川县(75.0%和82.6%)次之,山西省清徐县(16.7%和34.7%)最低。适合度分析发现,田间抗性菌株的产孢能力明显高于敏感菌株,但二者的复合适合度指数无显著差异。上述结果表明,4省份霜霉病菌群体均对烯酰吗啉产生了不同程度的抗性,田间抗性菌株与敏感菌株相比无明显适合度变化。     

部分鲜食葡萄品种霜霉病抗性田间调查

以54个鲜食葡萄品种为试材,调查统计不同葡萄品种的霜霉病发病程度、发病速率、落叶率和新梢成熟度。结果表明,不同葡萄品种对霜霉病的抗性存在明显差异,欧美杂种品种群的葡萄对霜霉病的抗性显著高于欧亚品种群;病情指数与落叶率、新梢成熟度呈极显著的相关性,与发病速率之间关系不显著。     

Sensitivity of European Plasmopara viticola populations to cymoxanil

Between 1995 and 1997, 278 grape downy mildew (Plasmopara viticola) populations originating from European vineyards were characterised for their sensitivity to cymoxanil in a leaf-disc assay. The sensitivity profile revealed a wide distribution, with minimum inhibitory concentrations (MIC) ranging from 10 to more than 800 mg litre⁻¹. EC₅₀ values ranged from 1 to more than 800 mg litre⁻¹ with an average of 125 mg litre⁻¹. The sensitivity distribution was stable between 1995 and 1997. Surprisingly, populations from Portugal appeared significantly more sensitive than those from France or Italy, which could not be linked to differential cymoxanil usage in these countries. P viticola populations collected outside Europe and never exposed to cymoxanil appeared significantly more sensitive than exposed European populations, with an average EC₅₀ value of 10 mg litre⁻¹. The level of sensitivity of European P viticola populations was relatively unaffected by the number of cymoxanil applications made during a season or by the number of years of cymoxanil use. No link was found between the level of sensitivity in the leaf-disc assay and the level of performance of the cymoxanil mixtures used in the fields where the populations originated. Specific field trials conducted in Italy and Portugal have shown that the performance of cymoxanil-based mixtures remained good even on populations of the grape downy mildew fungus characterised as less sensitive in the leaf-disc assay. While there are no baseline sensitivity data for pre-commercialisation P viticola populations, the results of our study suggest that a shift in sensitivity (12.5-fold) may have occurred in some areas since introduction of cymoxanil on grapes nearly 20 years ago. Because cymoxanil is never used alone, it is difficult to determine whether or not practical resistance is occurring in European vineyards. © 1999 Society of Chemical Industry     

葡萄霜霉病菌PCR检测方法的建立与应用

通过对已测序和已报道的葡萄霜霉病菌［Plasmopara viticola (Berk et Curtis) Ber.et de Toni］细胞色素c氧化酶亚型2（cytochrome c oxidase Ⅱ,cox2）的基因序列进行同源性比对分析,设计合成了一对用于P.viticola的检测引物F Pv/R Pv。利用该引物对包括P.viticola在内的30种常见植物病原真菌（包括15种Plasmopara属真菌、8种葡萄常见致病菌）的基因组DNA进行了PCR扩增,验证引物F Pv/R Pv的特异性,结果表明：该引物特异性强,仅P.viticola基因组DNA作为模板的PCR扩增产物呈现一条600 bp左右的特异性条带,其他参照菌株及阴性对照均无任何条带;灵敏度验证结果表明,该检测法可以检测出3.3 pg/μL 水平的P.viticola基因组DNA;应用该方法对来自全国不同葡萄产区的不同品种的78份葡萄霜霉病菌进行了检测,结果表明,该检测法适用范围广,且检测准确率达100%。     

Assessment of QoI resistance in Plasmopara viticola oospores

QoI fungicides, inhibitors of mitochondrial respiration at the Qo site of cytochrome b in the mitochondrial bc(1) enzyme complex, are commonly applied in vineyards against Plasmopara viticola (Berk. & MA Curtis) Berl. & De Toni. Numerous treatments per year with QoI fungicides can lead to the selection of resistant strains in the pathogen population owing to the very specific and efficient mode of action. In order to evaluate the resistance risk and its development, two different methods, biological and molecular, were applied to measure the sensitivity of oospores differentiated in vineyards, both treated and untreated with azoxystrobin, from 2000 to 2004. Assays using oospores have the advantage of analysing the sensitivity of bulked samples randomly collected in vineyards, describing accurately the status of resistance at the end of the grapevine growing season. Both methods correlated well in describing the resistance situation in vineyards. QoI resistance was not observed in one vineyard never treated with QoI fungicides. In the vineyard where azoxystrobin had been used in mixture with folpet, the selection of QoI-resistant strains was lower, compared with using solely QoI. In vineyards where QoI treatments have been stopped, a decrease in resistance was generally observed.     

Rapid method for detecting resistance to a QoI fungicide in Plasmopara viticola populations

BACKGROUND: The increasing occurrence of Qo inhibitor (QoI)‐fungicide‐resistant Plasmopara viticola (Berk. & MA Curtis) Berl. & DeToni populations is becoming a serious problem in the control of grapevine downy mildew worldwide. RESULTS: The authors have developed a rapid method for detecting resistance to a QoI fungicide, azoxystrobin, in P. viticola populations using the nested PCR‐RFLP method. With this method, a glycine‐to‐alanine substitution was discovered at codon 143 in the cytochrome b gene of P. viticola populations found in Japan. CONCLUSION: It is proposed that the nested PCR‐RFLP method is a high‐speed, sensitive and reliable tool for detecting azoxystrobin‐resistant P. viticola populations. Copyright © 2009 Society of Chemical Industry     

云南省葡萄霜霉病菌群体遗传结构的SSR分析

为明确云南特色葡萄产区葡萄霜霉病菌的群体遗传结构,本研究采用7对SSR分子标记对采自云南3个地区的155株葡萄霜霉病菌进行了群体遗传多样性研究,并分析了病原菌群体遗传结构与地域之间的关系。7对引物共检测出41个等位基因,114种基因型,病原菌群体的Shannon’s信息指数和Nei’s无偏基因多样性指数分别为0.942和0.600。分子方差分析和群体连锁不平衡分析显示,不同群体间存在相当大的遗传变异。遗传分化系数、基因流以及Structure分析表明,元谋群体与宾川、寻甸两群体之间均具有较高的基因交流,宾川和寻甸两群体间则存在一定的遗传分化。上述结果表明,云南葡萄霜霉病菌群体存在较高水平的遗传多样性,且群体遗传结构与地理距离之间存在一定相关性。     

Monitoring of QoI fungicide resistance in Plasmopara viticola populations in Japan

BACKGROUND: The increasing occurrence of QoI fungicide resistance in Plasmopara viticola (Berk. & MA Curtis) Berl. & DeToni populations is becoming a serious problem in the control of grapevine downy mildew. In Japan, the existence of QoI‐fungicide‐resistant P. viticola was reported in 2009. RESULTS: The QoI fungicide resistance in P. viticola samples collected from vineyards in Japan in 2008 and 2009 was monitored. Resistant P. viticola were detected in the regions where QoI fungicides have been introduced in accordance with the pest management programme, whereas in Hokkaido vineyards, where QoI fungicides have not yet been introduced, QoI‐fungicide‐resistant P. viticola were not found. CONCLUSION: Japan comprises thousands of islands and is physically isolated from other countries by the sea. Monitoring the emergence, incidence and distribution of QoI fungicide resistance in P. viticola populations in Japan is necessary to improve pest management strategies for downy mildew disease in Japanese vineyards. Copyright © 2010 Society of Chemical Industry     

山西省清徐县葡萄霜霉菌对烯酰吗啉的抗药性分析

葡萄霜霉病是葡萄最重要的病害之一,目前防治霜霉病主要依赖化学药剂,其中羧酸酰胺类杀菌剂是一类重要药剂,然而病原菌抗药性对其防效有重要影响。2015年本实验室首次在中国检测到抗烯酰吗啉的葡萄霜霉菌,并开发了Tetra-Primer ARMS PCR快速检测法来检测抗药性。本研究利用此技术对采自山西省清徐县60株葡萄霜霉菌进行了烯酰吗啉抗性检测,结果显示其中1株具有抗药性,其他59株均为敏感菌株,说明该地区霜霉菌已经开始产生烯酰吗啉抗性,但尚未大范围扩展,需要进行持续监测以指导杀菌剂的使用,这也是首次报道山西省葡萄霜霉菌存在烯酰吗啉抗性菌株。