辽宁省不同地区番茄灰霉病菌对咯菌腈的敏感性

为明确辽宁省不同地区间番茄灰霉病菌对咯菌腈的敏感性是否存在差异,以及咯菌腈与其他杀菌剂之间是否存在交互抗性,采用菌丝生长速率法测定番茄灰霉病菌对咯菌腈、腐霉利、多菌灵的敏感性。结果表明,从辽宁省6个不同地区分离的番茄灰霉病菌菌株对咯菌腈的平均EC50差异不显著,番茄灰霉病菌菌株对咯菌腈的敏感性频率呈正态分布,咯菌腈对不同地区共117株番茄灰霉病菌的EC50平均值为(0.007 3±0.004 5)μg/mL,且咯菌腈与腐霉利和多菌灵之间均不存在交互抗药性。     

福建省番茄灰霉病菌对氟啶胺敏感基线建立及与不同杀菌剂的交互抗性

为明确中国福建省番茄灰霉病菌对氟啶胺的敏感性及其与不同杀菌剂的交互抗性,采用菌丝生长速率法测定了106株采自福建省主要番茄产区的番茄灰霉病菌对氟啶胺的敏感性。结果表明,氟啶胺对福建省番茄灰霉病菌菌丝生长的EC50值在0.0037～0.0452 μg/mL之间,平均值为（0.0221 ±0.0098）μg/mL,其敏感性频率分布呈连续单峰曲线,符合正态分布,因此可将该EC50平均值（0.0221±0.0098）μg/mL作为福建省番茄灰霉病菌对氟啶胺的敏感基线,用于其田间抗药性监测。从106株菌株中选择15株对氟啶胺敏感性不同的菌株,测定了其对嘧霉胺、异菌脲、腐霉利和啶氧菌酯的敏感性。结果表明,供试5种杀菌剂对15株番茄灰霉病菌菌丝生长的平均抑制活性依次为氟啶胺 >异菌脲 >腐霉利 >啶氧菌酯 >嘧霉胺,氟啶胺与异菌脲、腐霉利、啶氧菌酯和嘧霉胺之间均不存在交互抗性。     

河南省番茄灰霉病菌对3种杀菌剂的抗药性检测

为了明确河南省番茄灰霉病菌对苯并咪唑类杀菌剂多菌灵、二甲酰亚胺类杀菌剂腐霉利和氨基甲酸酯类杀菌剂乙霉威的抗药性状况,2013年从河南省不同保护地中采集番茄灰霉病病果或病叶,经单孢分离共获得番茄灰霉病菌菌株139株。采用最低抑制浓度法（MIC）测定了其对多菌灵、腐霉利和乙霉威的抗药性。结果显示：番茄灰霉病菌对多菌灵、腐霉利和乙霉威产生抗性菌株的频率分别为81. 29%、80.58%和93.53%; 所测菌株对3类杀菌剂的抗性类型有BenRDicSNPCS、BenRDicRNPCS、BenRDicSNPCR、BenSDicRNPCR、BenSDicSNPCR和BenRDicRNPCR 6种,所占比例分别为2. 88%、3.60%、3.60%、5.04%、13.67%和71. 22%。表明河南省番茄灰霉病菌已对多菌灵、腐霉利和乙霉威产生抗药性,迫切需要筛选新的杀菌剂防治番茄灰霉病。     

辽宁省番茄灰霉病菌对氟吡菌酰胺的敏感性

从辽宁省番茄产区未使用过琥珀酸脱氢酶抑制剂的温室采集番茄灰霉病样本,经分离、鉴定并纯化,得到病原菌(Botrytis cinerea Pers.)单孢菌株206株,采用菌丝生长速率法测定氟吡菌酰胺的有效抑制中浓度EC50,建立了敏感基线,并调查了氟吡菌酰胺与咯菌腈、啶酰菌胺、嘧霉胺、腐霉利、异菌脲之间的交互抗性情况.结...     

四川猕猴桃灰霉病菌对4种杀菌剂的抗药性检测

为明确猕猴桃灰霉病菌对常见杀菌剂的抗药性状况, 本试验于2016年-2018年调查统计了四川猕猴桃生产中防控灰霉病所用的药剂种类和施药频次, 并从四川8个猕猴桃主产区采集病叶?病花和病果, 经单孢分离获得122个灰葡萄孢 Botrytis cinerea 菌株, 采用最低抑制浓度法(MIC)测定其对4种化学药剂嘧霉胺?咯菌腈?腐霉利和异菌脲的抗药性?结果表明:用于四川猕猴桃灰霉病防控的药剂主要有8种, 其中嘧霉胺?异菌脲和腐霉利3种药剂施用频次最高; 猕猴桃灰霉病菌对嘧霉胺?咯菌腈?腐霉利和异菌脲的抗性频率分别达95.08%?86.07%?80.33%和5.74%, 且不同产区的灰霉病菌对药剂的抗药性频率不同; 所测菌株对4种杀菌剂的敏感性类型共有8种, 其中以Pyr RFlu RPcm RIpr S(对嘧霉胺?咯菌腈?腐霉利表现抗性, 对异菌脲表现敏感)类型为主, 该类型的菌株占65.57%, 对4种杀菌剂均表现抗性的菌株为7株?表明四川省猕猴桃灰霉病菌对嘧霉胺?咯菌腈?腐霉利和异菌脲已经产生了抗药性, 对异菌脲抗药性较低, 迫切需要筛选新的杀菌剂防治猕猴桃灰霉病?     

湖南省草莓灰霉病菌对4种杀菌剂的抗药性检测

为明确湖南省草莓灰霉病菌菌株的抗药性状况,2013-2015年从湖南省不同地区草莓灰霉病病果或病叶上经单孢分离获得草莓灰霉病菌454株,采用最小抑制浓度法(MIC)检测不同地区草莓灰霉病菌株对多菌灵、腐霉利、异菌脲、嘧霉胺的抗药性。结果表明:草莓灰霉病菌对多菌灵、腐霉利、异菌脲、嘧霉胺的抗性频率分别为55.73%,77.31%、3.52%和77.31%;所测菌株对4种杀菌剂的敏感性类型有Cbz~RPcm~RIpd~RPmt~R、CbzSPcm~RIpd~RPmt~R、Cbz~RPcm~RIpdSPmt~R、Cbz~SPcm~RIpdSPmt~S、Cbz~RPcm~SIpd~SPmt~R、Cbz~SPcm~RIpd~SPmt ~R等6种,其所占比例分别为33.26%、5.07%、41.41%、4.63%、3.96%、11.67%,未发现对4种杀菌剂均敏感的类型,表明湖南省草莓灰霉病菌已对多菌灵、腐霉利和嘧霉胺产生抗药性,对异菌脲的抗药性较低。     

莘县地区灰霉病菌对腐霉利的抗药性研究

本研究检测了聊城莘县地区的燕塔街道、张鲁镇、古城镇、燕店镇、董杜庄镇、王奉镇、大王寨乡、俎店乡八个乡镇的芸豆、樱桃西红柿、番茄、茄子和辣椒五种作物的56株灰霉病菌对腐霉利抗药性。以1μg/m L为标准[7],灰霉病菌对腐霉利的抗性频率高达98. 21%,其中8. 93%的菌株为高抗菌株,51. 78%的菌株为低抗菌株。不同乡镇、不同作物的灰霉病菌对腐霉利的抗药性也存在着明显的差异,燕塔街道和大王寨乡出现了高抗菌株,张鲁镇和俎店乡也出现了较大比例的中抗菌株,董杜庄镇和王奉镇的灰霉菌株仍处于较低的抗药性水平。     

蔬菜灰霉病菌对腐霉利抗药性变异及其治理

蔬菜灰霉病菌（Botrytis cinerea）分生孢子经紫外光灯照射，可获得10^-4～10^-5频率的腐霉利抗药性突变体。获得的13个抗性突变体的抗性水平平均EC50值为18.45ug/ml,EC90值为305.17ug/ml。抗性突变体的可溶性蛋白谱带比其亲本敏感菌株多2～4条，其中Rf=0.38的蛋白谱带是抗性菌株所拥有而敏感菌株所共同缺乏的。抗性菌株对腐霉利抗药性稳定遗传，具有与亲本菌株相同的致病性。苗期抗、感测定结果表明，灰霉克杀菌剂对腐霉利突变体具有明显的克抗作用。田间抗性菌治理使用灰霉克400～800倍对大棚番茄灰霉病平均防效为95.93%～75.8%,优于腐霉剂600倍的平均防效(61.32%)。     

白术黑斑病菌对甲基硫菌灵和腐霉利的抗性检测及对咪鲜胺敏感性基线的建立

黑斑病是影响白术种植的主要病害之一,2015—2017年从浙江省磐安、天台、新昌和嵊州4地采集、分离得到137株白术黑斑病菌Alternaria alternata,采用菌丝生长速率法检测其对甲基硫菌灵和腐霉利的抗性。结果表明:供试的黑斑病菌群体 (n = 137) 对甲基硫菌灵的高水平抗性频率为77.4%,中等水平抗性频率为22.6%,未检测到敏感菌株;对腐霉利的抗性频率为20.4%,含21株低水平抗性菌株和7株中等水平抗性菌株。7种杀菌剂的室内毒力测定结果表明:咪鲜胺对白术黑斑病菌的活性最高,EC₅₀值为0.15 mg/L。供试的137株白术黑斑病菌对咪鲜胺的EC₅₀值分布在0.05~0.87 mg/L之间,平均EC₅₀值为 (0.29 ± 0.11) mg/L,可作为敏感性基线。     

辽宁省蔬菜灰霉病菌对嘧霉胺的抗药性

2006年从辽宁省不同地区采集蔬菜灰霉病果或病叶,经分离共获得灰霉病菌株86株。采用菌丝生长速率法测定了其对嘧霉胺的敏感性。试验结果表明,灰霉病菌对嘧霉胺产生中等水平抗药性,抗性频率为17.41%。经紫外、药剂联合诱导获得了高抗菌株,抗性倍数最高达62.78倍。嘧霉胺与多菌灵和腐霉利间不存在交互抗药性,苯胺基嘧啶类杀菌剂之间存在交互抗性。野生抗性菌株具有较好的遗传稳定性,连续转接9次后抗药性无明显下降。不同菌株的菌丝生长速度、鲜重和产孢量存在明显差异,但该差异与灰霉病菌对嘧霉胺的敏感性无相关性。     

Distribution and fitness of isolates of Botrytis cinerea with multiple fungicide resistance in Spanish greenhouses

Forty-nine greenhouses of vegetable crops were surveyed in southeast Spain at the beginning of the disease season in December 1992 to estimate frequencies of resistance to benzimidazoles, dicarboximides and N -phenylcarbamates (NPC) in B. cinerea . Out of 261 isolates collected, 28% were sensitive to both benzimidazoles and dicarboximides, 15% were benzimidazole-resistant and dicarboximide-sensitive, 8% were benzimidazole-sensitive and dicarboximide-resistant and 46% were benzimidazole- and dicarboximide-resistant. Resistance to benzimidazole, dicarboximide and N -phenylcarbamate was determined by measuring the ability of each isolate to grow in the presence of carbendazim, procymidone and diethofencarb fungicides respectively. Carbendazim- or procymidone- resistant isolates were found in all surveyed greenhouses. Three isolates were found with resistance to carbendazim, procymidone and diethofencarb collected in two adjacent greenhouses that were sprayed with the carbendazim and diethofencarb mixture. All other isolates were sensitive to the mixture because they were either sensitive to carbendazim and resistant to diethofencarb or vice versa. Fitness of 31 isolates of B. cinerea was determined in vivo by measuring their sporulation and lesion growth rate on leaf disks. No fitness costs were associated with resistance to iprodione (dicarboximide) and benomyl (benzimidazole). Isolates with EC₅₀ values higher than 101 mg/L for benomyl and 1.6 mg/L for iprodione were considered to be field resistant (they caused visible lesions on cucumber leaf disks treated with each fungicide).     

番茄灰霉病菌对嘧霉胺的抗药性

2001年从辽宁省不同地区的保护地中采集番茄灰霉病果或病叶，经单孢分离共获得番茄灰霉病菌70株。采用菌丝生长速率法测定了其对嘧霉胺的敏感性。结果表明，番茄灰霉病菌已对嘧霉胺产生中等水平抗药性，抗性频率为20．0％。经室内药剂诱导获得了高抗菌株，抗性倍数最高达35．7倍。嘧霉胺与多菌灵及嘧霉胺与速克灵间不存在交互抗药性。野生抗性菌株具有较好的遗传稳定性，连续转接9次后抗药性无明显下降。不同菌株的菌丝生长速度、鲜重和渗透敏感性存在显著差异，但该差异与灰霉病菌对嘧霉胺的敏感性无相关性。抗性菌株与敏感菌株具有同样的致病能力。     

灰葡萄孢对腐霉利的抗性分子机制及快速检测技术

为明确灰葡萄孢Botrytis cinerea对腐霉利的抗性现状,于2017—2018年采用单孢分离法从浙江省5个地区的草莓大棚共分离获得200个菌株.通过区分剂量法测定了其对腐霉利的抗性,对抗药性菌株的分子机制进行了分析,并根据抗药性分子机制,建立了B.cinerea腐霉利高抗基因型的环介导等温扩增(loop-med...     

速克灵抗性灰霉病菌菌株性质的研究

研究结果表明,灰霉菌对二甲酰亚胺类与苯并咪唑类药剂之间不存在交互抗药性,与传统的保护性杀菌剂之间也不存在交互抗药性。杀菌剂福美双、克菌丹、百菌清对二甲酰亚胺类抗性菌抑菌效果较好。高抗二甲酰亚胺类药剂的某些菌株产孢能力大于敏感菌株。抗性菌单孢分离后代仍保留抗性。大棚草莓连续9次喷撒速克灵,每次间隔10—14天,未发现速克灵抗性菌株的产生。盆栽草莓试验表明,无药条件下,抗性菌株的致病力明显低于敏感菌;正常用药条件下,速克灵对抗性菌株的防治效果明显低于敏感菌株。     

两种蚕豆赤斑病菌对常用杀菌剂的抗药性检测及比较

对采自浙江、湖北和安徽3省的蚕豆赤斑病样品进行了病原菌的分离和鉴定,采用菌丝生长速率法检测了引起赤斑病的2种病原菌——蚕豆葡萄孢Botrytis fabae和灰葡萄孢B.cinerea 的抗药性发生情况,并在离体条件下通过抗药性诱导试验比较了二者的抗药性风险。结果共分离得到153个菌株,其中蚕豆葡萄孢122株(占79.7%),灰葡萄孢31株(占20.3%)。共检测到37株多菌灵高水平抗药性菌株(其中蚕豆葡萄孢9株)和42株异菌脲低水平抗药性菌株(其中蚕豆葡萄孢17株);嘧霉胺对153个菌株的EC50值在0.01~5.13 μg/mL之间,平均EC50值为0.72±0.15 μg /mL;表明蚕豆赤斑病菌对常见杀菌剂已表现出一定的抗药性,且灰葡萄孢的抗药性问题比蚕豆葡萄孢要严重得多。抗药性诱导试验进一步证实,灰葡萄孢的抗药性风险明显高于蚕豆葡萄孢。     

Stability and fitness of pyraclostrobin- and boscalid-resistant phenotypes in field isolates of Botrytis cinerea from apple

Phenotype stability, fitness, and competitive ability of pyraclostrobin- and boscalid-resistant isolates of Botrytis cinerea from apple were investigated. Stability of resistance was determined after consecutive transfers on potato dextrose agar (PDA) or being cycled on apple fruit. In vitro fitness components mycelial growth, osmotic sensitivity, conidial germination, and sporulation were evaluated on agar media. Pathogenicity, virulence and sporulation on apple fruit were evaluated at both 20 and 0°C. Competition between fungicide-resistant and -sensitive isolates on apple fruit also was evaluated. Resistance to the two fungicides was retained at levels similar to that of the initial generation after 20 and 10 transfers on PDA and five and three disease cycles on apple fruit at 20 and 0°C, respectively. Great variability in individual fitness components tested was observed among isolates within the same phenotype groups either sensitive or resistant to the fungicides but, when compared as phenotype groups, there were no significant differences in the mean values of these fitness components between resistant and sensitive phenotypes except that the phenotype resistant only to boscalid produced fewer conidia in vitro than sensitive isolates. Resistant isolates were as pathogenic and virulent on apple fruit as sensitive isolates. There was no significant correlation between the values of individual fitness components tested and the level of resistance to pyraclostrobin or boscalid, except that virulence at 20°C positively correlated with the level of resistance to the two fungicides. The final frequency of pyraclostrobin-resistant individuals in the populations was significantly decreased compared with the initial generation and no boscalid-resistant individuals were detected after four disease cycles on apple fruit inoculated with a pair mixture of a dual-sensitive isolate and one isolate each of the three phenotypes resistant to pyraclostrobin, boscalid, or both. The results suggest that resistance of B. cinerea to pyraclostrobin and boscalid was stable in the absence of the fungicides and that resistance to the two fungicides did not significantly impair individual fitness components tested. However, both pyraclostrobin- and boscalid-resistant isolates exhibited competitive disadvantage over the dual-sensitive isolate on apple fruit.     

Dicarboximide resistance in Botrytis cinerea in protected lettuce

In surveys of protected lettuce crops in the Humberside area of the UK during 1984-5, 47.3% of Botrytis cinerea isolates were resistant to dicarboximides and 25% to benomyl. Dicarboximide-resistant isolates obtained direct from the field had a moderate degree of resistance with ED₅₀ values for mycelial growth on agar of 1.0-2.4 μg/ml vinclozolin for all except two of 30 isolates.
Isolates which were resistant to vinclozolin were also resistant to iprodione, myclozolin and procymidone. Thirty dicarboximide-resistant isolates obtained direct from the field were capable of infecting cucumber cotyledons sprayed with field-strength Ronilan (250 μg/ml vinclozolin) whereas sensitive strains were not. Strains showing different degrees of resistance to dicarboximides developed after incubation of sensitive and resistant strains on agar supplemented with vinclozolin. Some of these strains had ED₅₀ values for mycelial growth on agar of more than 100 μg/ml vinclozolin. No highly resistant isolates were obtained direct from the field. Resistance to dicarboximides in field isolates was stable.     

北京地区番茄灰霉病菌对嘧霉胺的抗药性检测

为了检测北京地区番茄灰霉病菌对嘧霉胺的抗药性,指导生产上科学用药,从北京12个区、县采集番茄灰霉病样150份,分离纯化获得109个番茄灰霉病菌(Botrytis cinerea)的单孢菌株,采用菌丝生长速率法测定了番茄灰霉病菌对嘧霉胺(pyrimethanil)的抗药性。所测菌株中嘧霉胺抗性菌株出现的比例高达82.57%,且以高抗菌株为主,占78.89%,抗性水平最高的菌株EC50是最敏感菌株的5 096倍;不同区县番茄灰霉病菌对嘧霉胺的抗药性存在差异,门头沟区、密云县、延庆县、怀柔区和平谷区、大兴、通州和顺义8个区县的高抗菌株所占比例达66.67%～100%,海淀、朝阳和房山3个区未检测到高抗菌株。上述结果说明北京地区番茄灰霉病菌对嘧霉胺的抗药性非常普遍,且以高抗菌株为主,生产上应替换新型杀菌剂防治番茄灰霉病。