黄皮酰胺类生物碱的提取及对7种水果病原真菌的抑菌活性

利用菌丝生长速率法测定黄皮果核甲醇提取物对芒果炭疽病菌(Colletotrichum gloeosporioides Penz)等7种水果病原真菌的抑制活性,结果表明该提取物10 mg/mL浓度下对芒果炭疽病菌和香蕉枯萎病菌（Fusarium oxysporum f.sp. cubense）菌丝生长抑制率分别达到87.36%和84.55%。从黄皮果核甲醇提取物中分离得到生物总碱,测定其对上述两种真菌的EC50值分别为0.78 mg/mL和0.81 mg/mL。生物总碱进一步分离纯化得到新肉桂酰胺类化合物lansiumamide B, 在0.08 mg/mL浓度下,对芒果炭疽病菌和香蕉枯萎病菌的菌丝生长抑制率分别为 83.33%和60.78%。     

海南芒果蒂腐病菌对吡唑醚菌酯的抗药性测定

从海南芒果主产区采集分离蒂腐病菌Botryodiplodia theobromae。首先采用区分剂量法检测供试菌株对吡唑醚菌酯的抗药性,然后再采用生长速率法测定吡唑醚菌酯对蒂腐病菌的EC_(50)。结果表明:95株芒果蒂腐病菌中有56株在含10mg/L吡唑醚菌酯的PDA培养基中菌落直径大于空白对照的40%,抗性频率为58.95%,来自不同产区的芒果蒂腐病菌对吡唑醚菌酯的抗性频率差异不大。吡唑醚菌酯对供试菌株的EC50在3.12~1525.43mg/L之间,呈连续性正态分布,而抗性最高的菌株的EC_(50)是最敏感菌株EC50的488.92倍。敏感菌株、低抗菌株、中抗菌株及高抗菌株的平均EC_(50)分别为:5.39mg/L、45.34mg/L、144.72mg/L和514.46mg/L。海南芒果蒂腐病菌对吡唑醚菌酯杀菌剂已经产生严重的抗药性。     

三株昆虫病原线虫共生菌胞外产物的抑菌活性

采用生长速率法、孢子萌发法、活体组织法和盆栽试验对3株昆虫病原线虫共生菌胞外产物甲醇提取物的抑菌活性进行了系统研究.结果表明,3个菌株甲醇提取物对番茄灰霉病菌、辣椒疫霉病菌、油菜菌核病菌的菌丝生长,对番茄灰霉病菌、玉米大斑病菌、稻瘟病菌、白菜黑斑病菌、杨树溃疡病菌、小麦根腐病菌的孢子萌发均有很强抑制作用.其中嗜线虫致病杆菌YL001菌株对辣椒疫霉病菌菌丝生长的抑制作用最强,EC50为35.11mg/L;伯氏致病杆菌YL002菌株对番茄灰霉病菌菌丝生长的抑制作用最强,EC50为 42.16mg/L;嗜线虫致病杆菌TB菌株对油菜菌核病菌菌丝生长的抑制作用最强,EC50为18.95mg/L.3个菌株甲醇提取物对玉米大斑病菌孢子萌发都有很强的抑制作用,EC50分别为28.03、36.77和26.40mg/L.3个菌株甲醇提取物在1000mg/L浓度下,对离体番茄果实灰霉病的治疗和保护效果均在70%以上;对盆栽番茄灰霉病和辣椒疫霉病的治疗和保护效果均在65%以上.     

中国芒果炭疽病菌复合种对苯醚甲环唑敏感性测定

芒果炭疽病是芒果上最严重的病害之一?本文采用生长速率法测定了来自不同芒果产区的芒果炭疽病菌对苯醚甲环唑的敏感性?结果表明, 苯醚甲环唑对供试的136个菌株的平均EC50为(1.46±0.14)mg/L, 其中, 最小值为0.09 mg/L, 最大值为9.74 mg/L, 其敏感性频率不符合正态分布?苯醚甲环唑对不同采集地区菌株的平均EC50无显著差异, 对贵州的菌株平均EC50最高, 对海南的最低?供试的芒果炭疽病菌包含13个种, 其中, 苯醚甲环唑对Colletotrichum cordylinicola的平均EC50最低, 对C.gloeosporioides 的平均EC50最高?     

13种海南入侵植物提取物对降香黄檀炭疽病菌的抑制作用

本文研究了13种海南入侵植物不同溶剂提取物对降香黄檀炭疽病菌的生物活性,以筛选出具有较好抑菌活性的植物提取物。结果表明:10种植物对炭疽病菌有一定的抑菌效果,其中效果最好的提取物为垂序商陆乙酸乙酯提取物,EC50为3.413mg/mL,其次是胜红蓟乙酸乙酯提取物以及飞机草乙醇提取物,EC50分别为5.849、21.984mg/mL。3种植物提取物都可以导致炭疽病菌菌丝畸形,高浓度垂序商陆与胜红蓟提取物甚至可以导致细胞壁破裂。垂序商陆提取物的MIC为5.0mg/mL,MBC为10mg/mL;胜红蓟提取物的MIC为7.5mg/mL,而MBC大于12.5mg/mL。     

植物提取物对油茶炭疽菌的抑菌活性筛选

为开发新型植物源农药,探究油茶炭疽病的生物防治方法,以油茶炭疽病菌(Colletotrichum fructicola)为供试病原菌,采用生长速率法测定了49种植物的70种提取物对油茶炭疽病菌的抑制作用,筛选出活性较好的提取物,并通过浓度梯度试验测定其抑菌毒力。结果表明,在提取物浓度为1 mg/mL时,臭灵丹石油醚萃取物和益母草乙酸乙酯萃取物对油茶炭疽菌抑菌效果最好,抑制率分别为54.48%和50.75%,经毒力测定其有效中浓度(EC₅₀)分别为0.7013和0.6246 mg/mL。本研究为进一步利用臭灵丹、益母草开发植物源抑菌剂提供了一定的理论依据。     

苦瓜活性组分物质抑菌活性测定

以苦瓜(Momordica charantia L.)种仁中提取的凝集素备提物和苦瓜籽粗蛋白对8种植物病原真菌,5种植物病原细菌进行了抑菌活性测定。结果表明:两种提取物对小麦赤霉病菌、棉花黄萎病菌和苹果炭疽病菌表现出较好的抑菌活性。凝集素备提物(0.67 mg/mL)对小麦赤霉病菌和棉花黄萎病菌的抑菌率分别为69.12%和64.86%;苦瓜籽粗蛋白(6.67 mg/mL)对苹果炭疽病菌的抑菌率达68.61%。两种提取物对大白菜软腐病菌、花生青枯病菌、水稻白叶枯病菌和根癌农杆病菌等植物病原细菌均无抑菌活性,苦瓜籽粗蛋白(0.1 g/mL)对水稻细条病菌有一定抑制作用。     

红树内生细菌AmS2菌株对芒果炭疽病菌的抑制作用

为了解红树内生细菌AmS2菌株对芒果炭疽菌的抑菌机制及其分类学地位,采用生长速率法测定了菌株抗菌活性对芒果炭疽菌菌丝生长及孢子萌发的有效中浓度(EC50)。结果表明:经形态与培养特征观察、生理生化测定及16S rDNA序列分析,将AmS2菌株鉴定为解淀粉芽孢杆菌Bacillus amyloliquefaciens。抗菌活性物质分析发现,菌株可分泌产生易溶于水和甲醇等强极性溶剂、对热稳定、pH4~7条件下较稳定的非蛋白类抗菌活性物质。毒力测定表明,菌株抗菌物质粗提物对芒果炭疽病菌菌丝生长和分生孢子萌发的EC50分别为0.9772 mg/mL和1.9027mg/mL,当提取物浓度3.0mg/mL时,可致使病菌分生孢子壁消解。     

短短芽胞杆菌对植物病原真菌的抑菌活性和抑菌机理

为探讨短短芽胞杆菌菌株011对植物病原真菌的防控效果,采用平皿打孔法测定菌株011粗提液对14种植物病原真菌的抑制作用,并以番茄早疫病为研究对象,研究了其抑菌机理。结果显示,菌株011粗提液对14种病原真菌均有抑菌活性,其中对番茄早疫病菌的抑菌活性最强,抑菌率达45.9%;菌株011粗提液能导致番茄早疫病菌菌丝生长异常,抑制孢子萌发;对番茄早疫病菌菌丝生长的EC50为71.4 mg/m L,对孢子萌发的EC50和MIC分别为0.12 mg/m L和0.8 mg/m L;该粗提液能引起番茄早疫病菌菌丝的脂质过氧化程度和细胞内过氧化氢含量上升,菌丝细胞膜的蛋白质合成量下降,当粗提液浓度高于50 mg/m L时,菌丝细胞膜的麦角甾醇合成量下降;该粗提液还能降低番茄早疫病菌胞外果胶酶和纤维素酶的活性。研究表明,菌株011粗提液可作为生防制剂应用于植物病原真菌的防治。     

7种龙舌兰属植物提取物对两种炭疽病病菌抑菌活性初探

采用菌丝生长速率法测定了7种龙舌兰属植物提取物对胶孢炭疽病病菌(Colletotrichum gloeosporioides)和香蕉炭疽病病菌(Calletotrichum musae)的抑菌活性。结果表明,在质量浓度10、5 mg/m L下处理72 h,7种龙舌兰属植物提取物对两种炭疽病病菌均有很强的抑菌活性。其中,H.11648、广西76416、有刺番麻和番麻对两种炭疽病病菌抑菌率均为100%,普通剑麻、粤西114和灰叶剑麻对两种炭疽病病菌抑菌率为77.78%~100%。毒力测定表明,有刺番麻对胶孢和香蕉炭疽病病菌抑菌效果最显著,有效中浓度(EC_(50))分别为0.027 7 mg/m L和0.048 2 mg/m L,其次是广西76416,有效中浓度(EC_(50))分别为0.093 4 mg/m L和0.240 9 mg/m L。     

葡萄胶孢炭疽菌对3种麦角甾醇脱甲基抑制剂类杀菌剂的敏感性

系统比较了从云南省主要葡萄产区采集的57株葡萄胶胞炭疽菌Colletotrichum gloeosporioides对三唑类脱甲基抑制剂烯唑醇、腈菌唑及咪唑类脱甲基抑制剂咪鲜胺的敏感性、交互敏感性及敏感性与生存适合度的相关性等指标。结果表明:供试菌株对咪鲜胺的EC50值在0.01~1.58 mg/L之间,高于其对烯唑醇(EC50值为0.05~25.45 mg/L)和腈菌唑(EC50值为0.49~192.93 mg/L)的敏感性;部分菌株对烯唑醇和腈菌唑的敏感性显著降低,而对咪鲜胺仍保持较高的敏感性;供试菌株对烯唑醇和腈菌唑的敏感性具有显著的相关性,而对咪鲜胺的敏感性与对烯唑醇和腈菌唑的敏感性之间则无显著相关性。低敏感性菌株的致病力和菌落生长速率与敏感菌株无显著差异,具有较高的生存适合度。     

Thiophanate-methyl sensitivity and fitness in Lasiodiplodia theobromae populations from papaya in Brazil

Stem-end rot, caused by Lasiodiplodia theobromae, is an important postharvest disease of papaya in Brazil. The use of fungicides is one of the main disease management measures. However, there are no data available on the sensitivity of L. theobromae to thiophanate methyl (methyl benzimidazole carbamate), the most common fungicide used in papaya orchards in northeastern Brazil. Thus, the effective concentration that results in 50 % of mycelial growth inhibition (EC₅₀) of 109 isolates, representing five populations of the pathogen was estimated in vitro. Seven components of fitness were measured for the 10 isolates with lower and high values of EC₅₀. Of the 109 isolates, 20.2 % were resistant to the fungicide with EC₅₀ values greater than 300 μg ml^?1, whereas the remaining 79.8 % were sensitive with an average EC₅₀ of 1.87 μg ml^?1. The EC₅₀ values for the resistant isolates were significantly (P?≤?0.05) higher than those for the sensitive isolates. When the fitness components were evaluated, only in relation to the spore production was significant difference among sensitive and resistant isolates, and resistant isolates showed sporulation capacity significantly lower than the S isolates, indicating a fitness cost.     

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。     

2013年湖北省小麦赤霉病菌对多菌灵和戊唑醇的敏感性

2013年从湖北省7个小麦主产区分离获得106株禾谷镰刀菌,测定了其对戊唑醇和多菌灵的敏感性。结果表明:戊唑醇和多菌灵对所有供试菌株的EC50值分别为0.064~0.778和0.090~0.858 mg/L。采用SAS软件的W法对EC50分布进行了正态性检验,表明106株菌株对戊唑醇和多菌灵敏感性的频率分布符合正态分布,其EC50平均值分别为(0.383±0.129)和(0.526±0.151)mg/L。不同地区来源的菌株对两种药剂的敏感性存在显著差异,其中襄阳的菌株对两种药剂的敏感性显著低于其他6个地区的。研究结果显示:湖北省小麦赤霉病菌未出现对戊唑醇和多菌灵抗性菌群,两种药剂用于防治小麦赤霉病仍具有使用价值。     

Multiple resistance of Botrytis cinerea from kiwifruit to SDHIs,QoIs and fungicides of other chemical groups

BACKGROUND: Botrytis cinerea Pers.: Fr. is a high‐risk pathogen for fungicide resistance development that has caused resistance problems on many crops throughout the world. This study investigated the fungicide sensitivity profile of isolates from kiwifruits originating from three Greek locations with different fungicide use histories. Sensitivity was measured by in vitro fungitoxicity tests on artificial nutrient media. RESULTS: Seventy‐six single‐spore isolates were tested for sensitivity to the SDHI fungicide boscalid, the QoI pyraclostrobin, the anilinopyrimidine cyprodinil, the hydroxyanilide fenhexamid, the phenylpyrrole fludioxonil, the dicarboxamide iprodione and the benzimidazole carbendazim. All isolates from Thessaloniki showed resistance to both boscalid and pyraclostrobin, while in the other two locations the fungal population was sensitive to these two fungicides. Sensitive isolates showed EC₅₀ values to boscalid and pyraclostrobin ranging from 0.9 to 5.2 and from 0.04 to 0.14 mg L^?1 respectively, while the resistant isolates showed EC₅₀ values higher than 50 mg L^?1 for boscalid and from 16 to > 50 mg L^?1 for pyraclostrobin. All QoI‐resistant isolates carried the G143A mutation in cytb. Sensitivity determinations to the remaining fungicides revealed in total eight resistance phenotypes. No isolates were resistant to the fungicides fenhexamid and fludioxonil. CONCLUSION: This is the first report of B. cinerea field isolates with resistance to both boscalid and pyraclostrobin, and it strongly suggests that there may be a major problem in controlling this important pathogen on kiwifruit. Copyright © 2010 Society of Chemical Industry     

Resistance development in rice blast disease caused by Magnaporthe grisea to tricyclazole

Sensitivity to tricyclazole of 129 single-conidial isolates of rice blast fungus, Magnaporthe grisea, was determined. EC₅₀ values ranged from 0.06 to 1.12 mg/L with an average value of 0.46 ± 0.09 mg/L according to the detached leaf segment tests. No significant difference of sensitivity was observed between isolates from Guangdong and Jiangsu where decreased efficacy was reported, and from two other provinces where tricyclazole provided excellent disease control. In seedling tests, tricyclazole could control the most tolerant isolate GY-6 successfully with a efficacy of 81.5% at the concentration of 40 mg/L. Sensitivities of GY-6 and DY-2, the most sensitive isolate, to tricyclazole were both unstable in sub-cultured single-conidial offspring isolates, with respective mean EC₅₀ values of 5.40 ± 0.97 and 4.50 ± 0.88 mg/L calculated from seedling tests. There was no amino acid difference between them in the coding sequences of 1,3,6,8-tetrahydroxynaphthalene reductase and 1,3,8-trihydroxynaphthalene reductase. These results suggested that the decreased control reported in Guangdong and Jiangsu could not be attributed to the occurrence of resistance. When continuously “inoculated-reisolated-reinoculated” under the selection of tricyclazole in vivo, sensitivity of DY-2 decreased 10-fold after 20 generations, although the sensitivity of GY-6 did not shift significantly.     

番茄早疫病菌对啶菌噁唑敏感基线建立及药剂作用方式初探

从中国山西、内蒙古、山东、海南、江苏、福建和陕西省 (自治区) 不同市、县从未使用过啶菌噁唑的露地或保护地采集并经单孢分离获得 166 株番茄早疫病菌,采用菌丝生长速率法测定了病原菌群体对啶菌噁唑的敏感性。结果表明:啶菌噁唑对番茄早疫病菌的 EC50 值范围在 0.11~2.29 mg/L 之间,敏感性频率呈正态分布,其均值 (0.56 ± 0.39) mg/L 可作为番茄早疫病菌对啶菌噁唑田间抗性监测的敏感基线;不同采集地菌株对啶菌噁唑的敏感性存在差异,其中采自福建省的菌株敏感性最高,而山西省大同、晋中和长治市的菌株敏感性最低。200 mg/L啶菌噁唑对番茄早疫病防治具有优异的保护和治疗作用,且接菌前 4 d 内施药和接菌后 1~2 d内施药防效最佳。研究表明,啶菌噁唑对番茄早疫病防治具有良好的实际应用前景。     

不同杀菌剂对黄瓜靶斑病菌的毒力作用特性比较

为筛选防治黄瓜靶斑病的有效药剂,采用菌丝生长速率法、孢子萌发法和芽管伸长法测定了19种杀菌剂对黄瓜靶斑病菌的毒力,探究了4种毒力较高杀菌剂对病原菌菌丝形态的影响。结果表明:氟啶胺、吡唑萘菌胺、咯菌腈和四霉素对黄瓜靶斑病菌菌丝生长、孢子萌发和芽管伸长各生长发育阶段均有较强的抑制活性,其中,4种药剂抑制菌丝生长的平均EC₅₀值分别为0.516 3、1.538、1.605和0.648 2 mg/L,抑制孢子萌发的平均EC₅₀值分别为0.164 4、1.359、1.127和0.002 5 mg/L,抑制芽管伸长的平均EC₅₀值分别为0.189 6、0.144 7、0.060 4和0.001 5 mg/L;啶酰菌胺、百菌清和肟菌酯3种不同作用机制杀菌剂对黄瓜靶斑病菌孢子萌发和芽管伸长的抑制活性较强,3种药剂抑制孢子萌发的平均EC₅₀值分别为1.564、0.373 0和0.021 3 mg/L,抑制芽管伸长的平均EC₅₀值分别为0.629 5、0.233 4和0.405 0 mg/L,但对菌丝生长的抑制作用较弱,平均EC₅₀大于16 mg/L;氟啶胺、吡唑萘菌胺、咯菌腈和四霉素可引起黄瓜靶斑病菌菌丝不同程度的异常生长,主要表现为菌丝隔膜间距变小、菌丝节间肿大、主菌丝上二次分枝发生频繁、菌丝变形黏连、菌丝新分枝处生长受抑制或分生孢子生长变形。研究表明,氟啶胺、吡唑萘菌胺、咯菌腈和四霉素对黄瓜靶斑病菌具有较高的毒力,在黄瓜靶斑病的田间防治中具有较好的开发应用潜力。     

牛至精油及其主要成分香芹酚和百里酚对16种植物病原真菌的抑制活性

为评价牛至精油及其2种主要成分香芹酚和百里酚在植物真菌病害防治方面的应用前景，采用菌丝生长速率法和孢子萌发法测定3种物质对16种植物病原真菌菌丝生长和孢子萌发的抑制活性，并采用黄瓜子叶法测定其对黄瓜灰霉病的防治效果。结果表明，牛至精油在500 mg/L浓度处理下可完全抑制番茄早疫病菌Alternaria solani、黄瓜灰霉病菌Botrytis cinerea和小麦根腐病菌Bipolaris sorokiniana等12种植物病原真菌的菌丝生长，对棉花黄萎病菌Verticillium dahliae的菌丝生长抑制率也达到97.66%。牛至精油对这13种病原真菌的EC₅₀在83.09~236.58 mg/L之间，香芹酚和百里酚对这13种病原真菌的毒力均高于牛至精油，其中香芹酚对黄瓜灰霉病菌、油菜菌核病菌Sclerotinia sclerotiorum、小麦全蚀病菌Gaeumannomyces graminis、棉花黄萎病菌和小麦茎基腐病菌Fusarium pseudograminearum菌丝生长的毒力较强，EC₅₀分别为9.09、22.45、24.28、28.40和28.80 mg/L；百里酚对黄瓜灰霉病菌和小麦全蚀病菌菌丝生长的毒力较强，EC50分别为21.32 mg/L和27.08 mg/L。香芹酚和百里酚对棉花黄萎病菌、玉米大斑病菌Setosphaeria turcica、苦瓜枯萎病菌Fusarium oxysporum f.sp.momordicae、黄瓜灰霉病菌和黄瓜炭疽病菌Colletotrichum orbiculare的孢子萌发亦有较强毒力，其EC₅₀为3.78~289.07 mg/L。1 000 mg/L牛至精油对黄瓜灰霉病的预防效果与400 mg/L嘧霉胺悬浮剂的预防效果无显著差异；500 mg/L百里酚对黄瓜灰霉病的预防效果和治疗效果均与400 mg/L嘧霉胺悬浮剂的效果相当，而香芹酚的治疗效果显著优于400 mg/L嘧霉胺悬浮剂。表明牛至精油及其主要成分香芹酚和百里酚的抑菌谱广且活性强，具有作为植物源杀菌剂的开发应用潜力。     

Resistance to metalaxyl in isolates of the sunflower pathogen Plasmopara halstedii

Plasmopara halstedii isolates showing an atypical reaction to metalaxyl were collected in France, in 1995 and 1996, and tested in the laboratory for their level of sensitivity to this fungicide. Primary and secondary infections caused by one of these isolates were not controlled by the metalaxyl concentration registered for seed treatment. The EC₅₀ of this isolate was 12 800 mg a.i. kg^-1 compared with 22 mg a.i. kg^-1 for sensitive isolate, indicating a 582-fold decrease in sensitivity to the compound. There was no reduction in the agressiveness of the resistant isolate. Using other anti-oomycete fungicides, it appeared that propamocarb, contact fungicides (fluazinam, folpet, mancozeb) and the mixed formulations dimethomorph + mancozeb, cymoxanil + mancozeb and ofurace + folpet were effective against primary infections made with metalaxyl resistant and sensitive isolates, but not against secondary infections. Metalaxyl mixed with fluazinam, folpet or mancozeb was more effective against primary infections with the resistant isolate than metalaxyl alone. The EC₅₀ of five other isolates ranged from 5 800 to 32 900 mg a.i. kg^-1, indicating a variability in metalaxyl sensitivity of resistant sunflower downy mildew isolates. This is the first report of physiological resistance to metalaxyl in Plasmopara halstedii.