杀菌剂毒力及其生物测定

杀菌剂毒力是化合物具有开发和应用价值的固有生物学性状,不仅是农药登记部门予以注册登记的前置条件,而且也是反映杀菌剂生物活性及其安全性和有效性的关键技术参数。本文明确了杀菌剂毒力术语的定义,强调了杀菌剂可能的未知作用方式及通过与病原菌、寄主和环境因子相互作用的病害及流行防控效力在杀菌剂毒力评价中的重要性。深入分析了化合物与受体蛋白相互作用的结构特异性和精确性及干扰靶标蛋白功能的毒理学机制,揭示了杀菌剂毒力的选择性原理。综述了杀菌剂毒力生物测定及其结果分析方法,分析了可能影响毒力测定结果评价的常见因素。本文所述内容可为从事农药创制、加工、应用、毒理与抗性及分子互作科研工作者提供参考。     

害螨的生物测定方法

农药的毒力测定是化学防治不可缺少的基础工作之一。在杀螨剂的毒力测定和害螨抗性监测时，简单、准确的生物测定方法也是必不可少的。由于害螨体躯与个体发育过程等与昆虫存在差异，且螨体小，因此，害螨的生物测定方法和害虫的生物测定方法也有所不同。害螨的生测方法有多种，且随着杀螨剂的发展在不断的改进和提高，现就害螨的生物测定方法简介如下：     

防治红枣黑斑病的杀菌剂筛选和复配研究

采用菌丝生长速率法和孢子萌发抑制法, 测定了14种杀菌剂单剂及基于单剂筛选结果的二元复配剂对红枣黑斑病菌Alternaria tenuissima菌丝生长和孢子萌发的抑制作用, 以此评价防治红枣黑斑病的杀菌剂和复配药剂的效果。结果表明, 咯菌腈和嘧菌环胺对靶标病菌菌丝生长的抑制效果最佳, 其EC50分别为0.091 3和0.099 8 μg/mL; 吡唑醚菌酯和啶酰菌胺对病菌孢子萌发抑制效果最佳, 其EC50分别为0.015 3和0.293 4 μg/mL。吡唑醚菌酯与戊唑醇按照8∶2和3∶7(w/w)的比例进行复配, 对病菌菌丝生长的抑制表现出相加作用, 其SR值分别为1.124 5和0.916 9; 两者以5∶5和3∶7(w/w)的比例进行复配, 对孢子萌发的抑制表现出相加作用, 其SR值分别为1.164 6和0.901 0。吡唑醚菌酯、啶酰菌胺、咯菌腈、嘧菌环胺和异菌脲等对病菌菌丝生长和孢子萌发均有抑制效果; 吡唑醚菌酯与戊唑醇3∶7(w/w)复配对病菌菌丝生长及孢子萌发的抑制效果均表现出相加作用。上述结果为红枣黑斑病防治药剂的开发和后续田间应用提供了理论支持。     

烟草疫霉的产孢和接种方法研究

烟草疫霉是重要的病原真菌,但在人工培养条件下难以培养、产孢以及释放游动孢子.作者对烟草疫霉培养、产孢、接种方法进行了研究.芝麻培养基、黑麦培养基、燕麦培养基可用来培养烟草疫霉,烟草疫霉在三种培养基上平均每天生长量为1.10、0.78、0.86cm;芝麻培养基、黑麦培养基为烟草疫霉产孢子囊较好的培养基,产孢能力强,分别为2.52×104、2.07×104个孢子囊/cm2.而且孢子囊释放率高,分别为43.0%、32.1%.游动孢子囊悬浮液灌根接种和注射接种都能使烟草发病,但不同的接种方法和接种浓度下,病情指数不同.     

杀菌剂对烤烟棒孢霉叶斑病菌的作用测定

采用菌丝生长速率法测定12种杀菌剂对烤烟棒孢霉叶斑病菌[Corynespora cassiicola(Berk.Curst.)Wei]的抑菌效果,并对抑菌率高的药剂进行室内毒力测定。结果表明,12种杀菌剂的抑菌效果存在一定的差异,其中咪鲜胺和甲基硫菌灵的抑菌效果最好,抑菌率均为100%,EC50分别为2.00 mg/L和2.10 mg/L,霜霉威盐酸盐和多菌灵对该病原菌有负的抑制作用。     

樱桃番茄叶斑病病原菌鉴定及杀菌剂筛选

采用PDA培养基和PD培养基,对樱桃番茄病株病原菌进行分离、培养,经致病性测定,通过形态学和rDNAITS序列分析,确认其致病菌为多主棒孢(Corynespora cassiicola)。采用菌丝生长速率法分别测定6种杀菌剂对多主棒孢的室内毒力和EC_(50)。结果表明,咪鲜胺锰盐对樱桃番茄多主棒孢菌菌丝抑制效果最好,EC_(50)为0.24 mg/L;氟菌.肟菌酯和苯醚甲环唑有较强的抑制作用,EC_(50)分别为4.18 mg/L和5.46 mg/L;春雷霉素抑制效果居中,EC_(50)为29.01mg/L;嘧菌酯和多菌灵的EC_(50)分别为161.35 mg/L和649.05 mg/L,其抑制效果较差。     

辣椒炭疽病病原鉴定及其杀菌剂毒力测定

从贵阳市花溪区磊庄村辣椒基地采集疑似炭疽病的典型的自然发病辣椒果实,采用组织分离、培养、形态学观察、致病性测定以及多基因分子系统学,确定病原菌的种类;采用菌丝生长速率法研究了8种杀菌剂对该病菌菌丝生长的抑制作用。结果表明,引起贵阳市花溪区磊庄村辣椒炭疽病的病原菌为尖孢炭疽菌Capsicum acutatum。室内毒力测定发现8种杀菌剂对尖孢炭疽菌均有一定的抑制作用,其中25%咪鲜胺EC和30%吡唑醚菌酯SC抑制效果最好,EC_(50)分别为0.253 5 mg/L和0.720 3 mg/L;其次为22.5%啶氧菌酯SC和22.7%二氰蒽醌SC,EC_(50)分别为7.249 5 mg/L和21.664 5 mg/L。将30%吡唑醚菌酯SC和22.7%二氰蒽醌SC按照9∶1、6∶4、3∶7、4∶6的比例混配,联合毒力测定和评价结果显示两者混配对该病菌具有协同增效作用,且25%咪鲜胺EC和22.5%啶氧菌酯SC以6∶4、4∶6、8∶2的比例进行混配时也表现出明显的增效作用。引起贵阳市花溪区磊庄村辣椒基地辣椒炭疽病的致病菌为尖孢炭疽菌。咪鲜胺、吡唑醚菌酯、啶氧菌酯和二氰蒽醌对该菌具有较好的抑制作用,可为辣椒炭疽病的田间防治提供参考依据。     

5种杀菌剂对木霉菌及尖孢镰刀菌的毒力测定

在室内条件下用5种常用土壤杀菌剂对生防木霉菌及尖孢镰刀菌进行了毒力测定。结果表明:木霉菌及镰刀菌对5种杀菌剂敏感性不同,两菌株对多菌灵反应敏感,其EC50最小;5种杀菌剂的毒力在平板上和在土壤中稍有差异,在土壤中福美双和口恶霉灵对木霉菌的抑制作用极小,可与木霉菌结合使用。     

几种杀菌剂防治水稻旱育秧田死苗研究

六种药剂对引起水稻旱育秧死苗的立枯菌和镰刀菌,进行毒力测定和田间小区试验,表明１２．５％烯唑醇和禾康灵Ⅱ不仅抑菌效果较好,相对防效达７０％,并且能提高秧苗素质。     

向日葵菌核病接种方法及品种抗病性鉴定

为建立有效的向日葵菌核病田间接种鉴定方法,以核盘菌菌丝体悬浮液和孢子悬浮液作为接种物,分别对抗、感向日葵品种在现蕾期、始花期和盛花期进行人工接种,并对接种后保湿材料和保湿时间进行比较。试验结果表明:两种接种物均可使向日葵抗、感品种产生盘腐症状。用菌丝体悬浮液和孢子悬浮液接种时,浓度分别为10.0~15.0g/L和200~500个/mL,始花期接种,牛皮纸袋保湿2~4d,即可区分出向日葵品种间抗感性差异。用此方法鉴定出13个对盘腐型菌核病表现抗病的向日葵品种。     

橡胶树白粉病菌分子检测技术的建立

根据橡胶树白粉菌(Oidium heveae Steinm.)基因组中的特有保守序列OHS,设计两对特异性引物OHF1/OHR1和OHF2/OHR2。以不同地区收集的6份O.heveae(OH1~OH6)和橡胶树胶孢炭疽病菌(Colletotrichum gloeosporioides Penz.)等5种非靶标病原菌及健康橡胶树叶片基因组DNA为模板,建立了橡胶树白粉菌PCR及nested-PCR分子检测技术,并验证了检测体系的特异性和灵敏度。结果表明,引物OHF1/OHR1和OHF2/OHR2对橡胶树白粉菌均具有较高的特异性和灵敏度。其中引物OHF2/OHR2能检测到10pg/μL的橡胶树白粉菌DNA,而以OHF1/OHR1和OHF2/OHR2组合进行nested-PCR,其最低DNA检测浓度达到0.01fg/μL。人工接种试验中,当孢子接种量为2×10~3个/叶时,PCR和nested-PCR检测体系可分别在接种4d和24h后检测到目的条带。表明nested-PCR对在叶片组织中处于潜育期的橡胶树白粉菌的检测更有效,可为橡胶树白粉菌的检测提供一种简便而准确的检测方法。     

海南橡胶树白粉菌的群体遗传结构研究

白粉病是海南橡胶树上发生最严重的叶部病害,为了明确海南橡胶树白粉菌群体的遗传结构,本研究对海南7个市(县)的橡胶树白粉菌的ITS序列进行分析。结果表明,95个病菌样品可推导出5种单倍型,其中单倍型H1包含样品数为88个,且在7个市(县)均有分布;三亚种群的单倍型多样性和核苷酸多样性最高。总体和各地理群体的中性检验结果显示,群体扩张遵循中性进化,群体大小保持相对稳定。遗传分化指数(Fst)表明三亚种群与白沙、儋州、海口和琼中等四市(县)种群的遗传分化较大。AMOVA分析显示,遗传变异主要发生在种群内,占总变异的89.66%。因此海南橡胶树白粉菌菌株间虽存在一定的遗传分化,但整体遗传多样性偏低。     

橡胶树白粉病菌分生孢子在不同介质的萌发过程及其内容物的变化

 本研究采用不同的染色方法和显微技术观察橡胶树白粉病菌(Oidium heveae)分生孢子的超微结构,分生孢子在不同介质表面及不同介质溶液中的萌发情况,以及孢子萌发过程中孢内主要物质的变化,以明确O. heveae分生孢子在不同介质萌发的形态变化及萌发所需能量来源。结果表明：O. heveae 分生孢子表面具有较浅的花纹结构,呈椭圆形或者卵圆形,大小为26.1~45.1μm×13.5~21.9μm。O. heveae 分生孢子在亲水和疏水介质上均能萌发产生芽管和附着胞,在葡萄糖水中的萌发率略高于清水中,但萌发率无显著性差异 (P>0.05),芽管和附着胞形成与形态无明显差别。分生孢子萌发过程中,几个液泡会融合形成一个,最终消散而呈不明显可见的泡囊结构。孢子内的糖原、脂质及分裂后的核仁等可通过芽管向附着胞输送,表明O. heveae 分生孢子萌发所需能量物质主要来源于自身能量贮备。     

杀菌剂生物测定技术的发展

概述了杀菌剂生物测定的发展历史,探讨了离体、活体和组织筛选的常规生物测定技术,以及高通量筛选方法,同时从多个角度阐述了杀菌剂生物测定技术的发展趋势。     

杀菌剂生物测定技术的发展

概述了杀菌剂生物测定的发展历史,探讨了离体、活体和组织筛选的常规生物测定技术,以及高通量筛选方法,同时从多个角度阐述了杀菌剂生物测定技术的发展趋势。     

Survival of Oidium anacardii on cashew (Anacardium occidentale) in southern Tanzania

During March and April of 1993 and 1994, surveys on the incidence and severity of cashew powdery mildew ( Oidium anacardii ) were conducted in the Newala, Mtwara, Nachingwea and Tunduru areas of southern Tanzania to determine the variation in perennation between localities. Only immature cashew shoots, panicles and fruit can be infected by O. anacardii conidia. Cashew trees at sites in each district were assessed for shoot and panicle production and cashew powdery mildew. Survival of O. anacardii between seasons, in any area, was determined by the degree of production of shoots that were within the canopy and by the incidence of infection. Immature shoots produced from the main branches within the tree canopy were the main source of active powdery mildew in all districts; trees in the Newala district had the highest numbers of infected immature shoots in comparison with survey sites in the other areas. During the 1994 cashew-growing season (June–August), powdery mildew developed more rapidly and affected more shoots on the inside of the tree canopy than on the outside. Germination of conidia was reduced after aqueous suspension for 3 h. Germination on cashew leaves submerged under 2 mm of water was not affected. Appressorial and hyphal formation by germinating conidia on leaves decreased with increasing duration under water. Germination of conidia on glass slides at 100% r.h. was higher at 25 and 30°C than at 15°C and there was no germination at 35°C.     

Variation in severity of cashew powdery mildew (Oidium anacardii Noack) disease in Tanzania: Implications for research and extension

Abstract

Cashew trees in farmers’ fields in two contrasting areas in southern Tanzania were surveyed for growth, powdery mildew development, amounts and applications of sulphur, pest damage and nut yield by village‐based extension workers in 1993. There was a wide range of powdery mildew levels on cashew flowers, apples and nuts at both the Tunduru (inland) and Newala (western Makonde plateau). In both Newala and Tunduru application of sulphur dust was associated with reduced severity of blossom powdery mildew. Sulphur usage commenced later in Newala than Tunduru though this was associated with earlier onset of blossoming in the latter district. Farmers in both areas applied similar numbers of rounds of sulphur, however in Newala more sulphur was applied per tree reflecting the bigger trees found in this district. There was a highly significant (P < 0.001) negative association between use of sulphur and severity of blossom powdery mildew irrespective of nut yield. There was a strong positive association (P < 0.01) between use of sulphur and more than one nut per panicle from harvests in October, no significant association in November and a significant negative association (P < 0.05) between use of sulphur and more than one nut per panicle in December. The survey highlighted areas in Newala where there were higher powdery mildew levels, through either lack of sulphur application or poor control by late application of sulphur, with relatively high numbers of nuts per panicle. Conversely, areas were identified where despite control of cashew powdery mildew severity by application of sulphur nut yield remained relatively low. The results suggest that final yield results from an interaction of factors of which cashew powdery mildew is only one. There was a highly significant (P < 0.01) association between the number of sulphur rounds and severity of cashew powdery mildew (Newala data only) but no significant association between number of rounds of sulphur and yield taken during October. Pest damage estimates from cashew nuts showed no consistent pattern in either region. Survey results suggest that there is considerable potential in the optimization of sulphur usage as a strategy for control of powdery mildew.     

Consecutive monitoring for conidiogenesis by Oidium neolycopersici on tomato leaves with a high-fidelity digital microscope

Conidiogenesis by Oidium neolycopersici KTP-01 on tomato leaves was vitally monitored with a high-fidelity digital microscope. Conidiophores were initially formed 3 days after inoculation and then elongated to a maximum length within at least 12h. The apical part was split into two cells after two successive septations, accompanied by apical expansion. These cells subsequently developed into primary and secondary conidia. An additional septation at the stem portion of the conidiophores produced a generative and a foot cell. Subsequent conidiation occurred during repeated cycles of splitting of the generative cell, maturation of the apical cell into a conidium, and abstriction of the conidium. To our knowledge, this report is the first on the developmental process of conidiogenesis by powdery mildew on host leaves as revealed with the digital microscope.     

Host Range of Oidium lycopersici Occurring in the Netherlands

Nine accessions of three cucurbit species, ten of eight legume species, three of lettuce (Lactuca sativa) and 34 of 14 Solanaceae species were inoculated with a Dutch isolate of the tomato powdery mildew fungus (Oidium lycopersici) to determine its host range. Macroscopically, no fungal growth was visible on sweet pepper (Capsicum annuum), lettuce, petunia (Petunia spp.) and most legume species (Lupinus albus, L. luteus, L. mutabilis, Phaseolus vulgaris, Vicia faba, Vigna radiata, V. unguiculata). Trace infection was occasionally observed on melon (Cucumis melo), cucumber (Cucumis sativus), courgette (Cucurbita pepo), pea (Pisum sativum) and Solanum dulcamara. Eggplant (Solanum melongena), the cultivated potato (Solanum tuberosum) and three wild potato species (Solanum albicans, S. acaule and S. mochiquense) were more heavily infected in comparison with melon, cucumber, courgette, pea and S. dulcamara, but the fungus could not be maintained on these hosts. All seven tobacco (Nicotiana tabacum) accessions were as susceptible to O. lycopersici as tomato (Lycopersicon esculentum cv Moneymaker), suggesting that tobacco is an alternative host. This host range of the tomato powdery mildew differs from that reported in some other countries, which also varied among each other, suggesting that the causal agent of tomato powdery mildew in the Netherlands differ from that in those countries. Histological observations on 36 accessions showed that the defense to O. lycopersici was associated with a posthaustorial hypersensitive response.