防治牡丹根腐病的有效药剂筛选

采用菌丝生长速率法和孢子萌发法,分别测定了咪鲜胺、醚菌酯、苯醚甲环唑、百菌清、多菌灵、代森锰锌等6种杀菌剂对牡丹根腐病菌的室内毒力。结果表明,6种药剂对病菌菌丝生长的EC50大小顺序为：咪鲜胺<多菌灵<代森锰锌<苯醚甲环唑<百菌清<醚菌酯,其中以咪鲜胺对牡丹根腐病病菌的菌丝生长抑制作用最强, EC50为0.200 mg/L;多菌灵、代森锰锌、苯醚甲环唑、百菌清依次减弱,醚菌酯的抑制菌丝生长的作用最小,EC50为33.98 mg/L。6种药剂对病菌孢子萌发的EC50大小顺序为：百菌清<咪鲜胺<苯醚甲环唑<多菌灵<代森锰锌<醚菌酯;其中以百菌清对牡丹根腐病菌的孢子萌发抑制作用最强,EC50为0.343 mg/L;咪鲜胺、苯醚甲环唑、多菌灵、代森锰锌依次减弱,醚菌酯抑制孢子萌发的作用最小,EC50为38.4 mg/L。     

三类杀菌剂对牡丹腔孢叶斑病菌生长和发育的影响

腔孢叶斑病是牡丹生产上出现的一种新病害,近几年在洛阳及菏泽牡丹种植园危害越来越重。为了筛选防治该病的化学药剂,本文采用菌丝生长速率法测定了3类8种杀菌剂对病菌菌丝生长的抑制活性,采用涂布平板法测定了多菌灵、戊唑醇及嘧菌酯对病菌孢子萌发和芽管伸长的影响,并在含药平板上测定了3种药剂对病菌产孢量的作用。结果表明:三唑类药剂对病菌菌丝生长的毒力最强,EC50为0.020~0.096μg/mL,且戊唑醇对孢子萌发和产孢也有很强的抑制活性,在0.1μg/mL时孢子即不能萌发;甲氧基丙烯酸酯类药剂对菌丝生长的毒力较强,EC50为0.408~0.939μg/mL,嘧菌酯对芽管伸长和产孢量均表现出很强的抑制活性,10μg/mL时,芽管几乎不再伸长,对产孢的抑制率可达到99%以上;而苯并咪唑类药剂对菌丝生长的毒力较弱,EC50为12.167~30.104μg/mL,多菌灵对孢子萌发的抑制作用弱,100μg/mL时孢子萌发率仍达到90%以上,对产孢的抑制作用也较弱,为63.02%。根据药剂对病菌生长和发育各阶段的影响,生产中应采用适宜的施用方法:戊唑醇和嘧菌酯可作为保护剂在病害发生前期喷施,也可作为治疗剂在病害发生流行期应用,但为防止抗药性菌株的出现,两者应当轮换或复配使用。     

三种杀菌剂对牡丹红斑病病菌分生孢子形成和萌发的影响

为筛选防治牡丹红斑病的杀菌剂,采用涂布平板法测定了多菌灵、戊唑醇、嘧菌酯对病菌分生孢子形成和萌发的影响。结果表明:3种杀菌剂对病菌孢子萌发均有很强的抑制作用,浓度越高,抑制作用越强,以嘧菌酯最强,1μg/mL浓度处理其孢子萌发率低于10%,5μg/mL处理孢子完全不能萌发;3种杀菌剂均可显著抑制病菌芽管的伸长生长,1μg/mL处理下孢子萌发24 h后芽管长度均不足10μm,5μg/mL处理对芽管伸长的抑制率达到90%;3种杀菌剂可使分生孢子及芽管发生畸形,多菌灵的致畸作用最强,0.1μg/mL处理即表现出明显的致畸作用,而戊唑醇致畸作用较弱;嘧菌酯和戊唑醇对病菌产孢结构形成和分生孢子产生数量有强烈的抑制作用,0.05μg/mL浓度下即完全不产孢,但多菌灵在0.05μg/mL浓度下则促进病菌产孢结构形成及产孢量增加。表明3种杀菌剂均可作为保护剂在病害发生前期喷施,但在流行期应尽量使用嘧菌酯和戊唑醇,以减少病菌再侵染数量。     

三种杀菌剂对牡丹黑斑病菌菌丝生长及分生孢子萌发的影响

为筛选防治牡丹黑斑病的化学药剂,采用菌丝生长速率法和涂布平板法分别测定了多菌灵、苯醚甲环唑和嘧菌酯对牡丹黑斑病菌菌丝生长的抑制活性及对分生孢子萌发和形成的影响。结果表明：苯醚甲环唑对菌丝生长的抑制作用最强,有效中浓度EC₅₀仅为0.30 μg/mL,其次为嘧菌酯6.34 μg/mL和多菌灵92.18 μg/mL;多菌灵可显著降低分生孢子的萌发速率,而苯醚甲环唑和嘧菌酯对孢子萌发的抑制作用较弱,可使分生孢子及芽管发生畸形,并改变分生孢子的萌发方式;3种药剂均可显著抑制芽管的伸长生长,推迟病菌分生孢子梗的形成时间,但对分生孢子的产生数量无显著影响。多菌灵可作为保护剂在病害发生前期使用,苯醚甲环唑和嘧菌酯可在病害发生期轮换使用。     

9种杀菌剂对西瓜炭疽病菌的室内毒力测定及配比试验

为筛选防治西瓜炭疽病的高效低毒杀菌剂, 缓解和治理生产中病菌对药剂的抗性, 在室内离体条件下采用生长速率抑制法及孢子萌发抑制法测定了9种杀菌剂对西瓜炭疽病菌( Colletotrichum orbiculare )的毒力。结果表明, 嘧菌酯、咪鲜胺和甲基硫菌灵对病原菌菌丝生长的EC50在0.093 3~0.118 2 mg/L之间, 均小于1 mg/L, 表明西瓜炭疽病菌对上述杀菌剂比较敏感; 百菌清、烯肟菌酯和戊菌唑的EC50在2.310 1~5.925 9 mg/L, 病菌对药剂的敏感程度相对较低; 代森锰锌、恶霉灵和多菌灵的EC50分别为36.876 3、74.466 6和99.898 5 mg/L, 抑菌活性较差。孢子萌发试验中, 嘧菌酯、咪鲜胺和甲基硫菌灵对病菌孢子萌发的抑制活性最高, EC50在0.069 4~0.167 2 mg/L之间; 百菌清、烯肟菌酯、代森锰锌的抑制活性次之, EC50在1.853 0~9.503 9 mg/L之间; 多菌灵的抑制活性相对最低, EC50为99.335 3 mg/L。将两种不同作用机理的杀菌剂嘧菌酯与咪鲜胺按照2∶1的比例混配, 联合毒力测定和评价结果表明两者混配对抑制西瓜炭疽病菌具有增效作用。     

铁皮石斛黑斑病菌室内药效试验

通过供试的18种药剂对铁皮石斛黑斑病菌体外抑制作用测试结果表明,除克菌特外,所有药剂对其孢子萌发都有显著的抑制作用,其中代森锰锌、易保、代森锌和炭疽福美可完全抑制孢子萌发,霉克特、扑海英、使百克、新太生、甲基托布津和世高对孢子萌发有较强烈的抑制作用。与对照比较,克菌特促进孢子萌发率高于对照23.34%,克菌特和庄园乐都能促进芽管生长。18种药剂对石斛黑斑病菌菌丝体的抑制作用测定结果显示,所有药剂对菌丝生长都有显著的抑制作用,其中使百克和世高对菌丝体抑制效果最好,抑菌率都达91.28%,其次是施佳乐、福星和扑海英。     

红树植物提取物对3种植物病原真菌孢子的抑制活性

用孢子萌发法测定了16种红树植物甲醇提取物对甘蔗凤梨病菌、西瓜枯萎病菌和柑橘疮痂病菌孢子萌发的抑制活性。结果表明,苦郎树和苦槛蓝对3种病菌孢子的萌发有明显的抑制作用,提取物浓度为10 g/L时,抑制率达77.29%以上。桐花树、海漆、榄李和银叶树对甘蔗凤梨病菌孢子萌发有明显的抑制作用,但对其余2种病菌孢子的抑制活性低。其余植物提取物对3种病菌孢子的抑制活性低。榄李和桐花树对甘蔗凤梨病菌孢子的毒力高,EC₅₀分别为0.3852 g/L和0.5714 g/L。     

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。     

杀菌剂对昆虫病原真菌块耳霉孢子萌发的影响

研究了10种杀真菌剂在田间使用浓度的中间浓度及其10倍稀释液下,对昆虫病原真菌块耳霉孢子萌发的影响。结果表明各杀菌剂对块耳霉孢子萌发有不同程度的抑制,浓度愈高,抑制作用愈大。多菌灵、普力克对孢子的抑制作用最小,与块耳霉表现出良好的相容性,是与块耳霉实施菌药合用的候选药剂。施保功、大生、达科宁3种杀菌剂对块耳霉孢子的杀伤力最大,在两种浓度下孢子萌发率均为0。施佳乐、速克灵和敌力脱在低浓度下,与块耳霉的相容性良好,处理后24～48 h与对照无显著性差异;但在中间浓度水平下,处理后48 h孢子萌发率分别为12.45％,0,0。其余药剂随处理时间和浓度不同,对块耳霉孢子萌发有不同程度的抑制。     

月季黑斑病病原菌鉴定及室内药剂初步筛选

为有效防控月季黑斑病,于2017年6—7月自运城学院花圃采集具有典型黑斑症状的月季病叶,分离纯化获得病原菌,利用形态学特征、分子生物学特征及致病性测定对其进行鉴定;同时室内测定5种常用药剂对该病原菌的防治效果,筛选能有效抑制该病原菌菌丝生长及孢子萌发的药剂。本试验共分离纯化得到12株菌株,其病原菌形态特征一致、rDNA-ITS、GPD、EF-1α基因扩增序列均相同。结合形态学特征观察以及rDNA-ITS、GPD、EF-1α基因联合构建的系统发育树将运城月季黑斑病病原菌鉴定为链格孢Alternaria alternata。在5种药剂中,50%咯菌腈和50%异菌脲各浓度对月季黑斑病病原菌的抑菌效果均较好,对菌丝生长抑制率和孢子萌发抑制率分别达到95.2%和96.6%以上;其次是70%代森锰锌,3个浓度的菌丝生长抑制率在84.1%～87.2%之间,3个浓度的孢子萌发抑制率均高于93.5%,其中600倍液的孢子萌发抑制率最高,达98.1%;80%多菌灵和12.5%腈菌唑对月季黑斑病病原菌的抑制效果最差,菌丝生长抑制率和孢子萌发抑制率分别低于42.4%和46.3%。毒力测定结果显示,50%咯菌腈毒力最强,抑制中浓度EC_(50)为25.8 mg/L;12.5%腈菌唑毒力较弱,EC_(50)为160.3 mg/L,其它3种药剂的EC_(50)介于30.4～60.7 mg/L之间。因此,50%咯菌腈可湿性粉剂和50%异菌脲悬浮剂可作为防治由链格孢引起的月季黑斑病的优选药剂。     

不同杀菌剂对稻粒黑粉病菌冬孢子萌发的影响

稻粒黑粉病(Rice kernel smut)是一种真菌病害。在离体条件下,采用孢子萌发测定法,测定了嘧菌酯、灭菌唑、拌种灵、多菌灵、咪酰胺和二硫氰基甲烷抑制稻粒黑粉病菌冬孢子萌发的最低抑制浓度(MIC);同时研究了药剂浸种处理对稻种的安全性以及浸种处理效果。结果表明,嘧菌酯和二硫氰基甲烷对稻粒黑粉病菌冬孢子萌发的抑制活性较高,MIC为0.4μg/mL,多菌灵的抑制活性次之,MIC为0.8μg/mL,其他3种药剂的抑制活性不高,MIC均大于1.6μg/mL。种子安全性试验表明,嘧菌酯在2.0、4.0、8.0μg/mL和多菌灵在4.0、8.0、16.0μg/mL处理3d和7d后,稻种的萌芽率与对照相比无显著性差异;而二硫氰基甲烷在4.0和8.0μg/mL处理下,所选稻种的萌芽率显著低于对照,说明该药剂在浸种处理后会影响所选稻种的萌芽,但其在低剂量处理下对稻种的萌芽率影响较小。关于稻粒黑粉病浸种处理的合适农药剂型仍需要进一步研究。     

Teliospore Survival and Plant Quarantine Significance of Tilletia Indies (causal agent of Karnal bunt) Particularly in Relation to China

The influence of temperature, light and soil conditions on teliospore germination of Tilletia (Neovossia) Indica was investigated. The teliospores were germinated by incubation in an aqueous suspension in a concavity on a water agar surface and the germination percentage was scored directly under low magnification of the compound microscope. The optimal temperature range for germination was found to be 15 to 22°C. Germination was extremely low at 2°C, but total inhibition was only observed after prolonged exposure to 35®C (in darkness) or to - 18°C under dry conditions. Light was shown to have a stimulating effect on germination (artificial daylight was found to be better that near ultraviolet light). No stimulation was achieved by prolonging the illumination from 12 to 24 h. Teliospores from 5-year-old wheat samples were shown to be viable. The weather conditions of the various wheat- growing areas of China were compared with the results obtained for the survival of teliospores of T. indica. It is concluded that a considerable risk exists for establishment and spread of the Karnal bunt disease if first accidentally introduced into China. Strict quarantine precautions, and policies can minimize or even eliminate this risk.     

进境碱茅上腥黑粉菌新记录的初步研究

腥黑粉菌主要侵染禾本科植物。2004年曾从来自美国的碱茅草种中检出一种腥黑粉菌,是碱茅上腥黑粉菌的新记录,迄今国内外尚无其相关研究的报道,本文从冬孢子形态、萌发生理特性及基因序列比较等几方面对该菌进行了初步研究,讨论了该菌的分类地位,根据研究结果,该菌与Tilletia bromi具有较近的亲缘关系,为腥黑粉菌潜在新种。     

Survival of Tilletia indica teliospores under European soil conditions

As part of developing a European Pest Risk Analysis (PRA) for Tilletia indica , the causal agent of Karnal bunt of wheat, teliospore survival studies were done outside under quarantine containment at three European sites (Norway, UK, Italy). At each site, experiments were set up in three consecutive years (Experiments 1, 2 & 3) to determine teliospore survival over time (1–3 years) at 5, 10 and 20 cm depths. Experiments were sampled annually and survival assessed in relation to teliospore recovery and to germination at recovery (T0) and 3 months after recovery in case of burial-induced dormancy ( T3 ). Teliospores survived at all three sites at all depths over all the time periods studied. At each site, there was no evidence of a marked decline in teliospore recovery between sampling years, except in one set of years in one Norwegian experiment. There was no consistent effect of depth on recovery. In general there was little evidence for a marked decline in teliospore germination between sampling years. There was some evidence of a decrease in germination with increasing depth in the UK, and for some time-depth interactions. After 3 years' incubation (Experiment 1), mean teliospore recovery and mean germination were: UK: 61% recovery and 31% ( 33% ) germination for T0 (and T3 ); Italy: 30% recovery and 36% ( 29% ) germination; and Norway: 12% recovery and 19% ( 49% ) germination. Germination for laboratory controls ranged from 20–59% (UK), 18–41% (Italy) and 28–59% (Norway). There was no evidence for burial-induced dormancy except in Norway. Teliospores of T. indica can survive for at least three years in European soils. This prolonged period of survival could support establishment of the pathogen if it were introduced into areas of European cereal production.     

Pheromone-Related Inhibitors of Ustilago hordei Mating and Tilletia tritici Teliospore Germination

ABSTRACT Ustilago hordei, the causal agent of barley covered smut, produces mating pheromones that break down to smaller peptide compounds that act as potent inhibitors of mating and germination in several fungi. The pheromones are members of the farnesylated family of proteins. Synthetic peptide analogs of the pheromone derivatives, ranging in size from 4 mers to full length pheromones, were farnesylated, methyl esterified, or both and tested for mating or teliospore germination inhibition with U. hordei or Tilletia tritici, respectively. N-Acetyl-S-farnesylcysteine, which inhibits processing of Ras, and other sulfur-containing compounds such as homocysteine or methionine, were likewise modified and tested. The most potent inhibitors were methionine methyl ester and modified 4-mer peptides from both pheromones. Alanine scanning of the inhibitory 4 mers determined that the native amino acid sequence was specific for a high level of activity. The sulfur amino acids appear to be required for inhibition. Glasshouse studies using selected antagonists of mating and teliospore germination as seed treatments inhibited covered smut of barley and common bunt of wheat, although the level of control was inconsistent. The use of pheromone-related antagonists to mating or teliospore germination is a promising, novel strategy for control of smut and bunt diseases.     

土壤湿度对小麦矮腥黑穗病菌(TCK)冬孢子萌发的影响

通过土壤湿度对小麦矮腥黑穗病菌(Tilletiacontronversa Kühn,TCK)萌发率的影响试验研究表明,其冬孢子在土壤质量含水量为1%～28%(相对含水量3.57%～100%)范围内均可萌发,其适宜萌发的土壤质量含水量范围为10%～25%(相对含水量17.85%～89.3%),最佳土壤相对含水量范围在65%～75%之间;不同分离物在相同土壤湿度培养下,多数分离物冬孢子的萌发率之间差异不显著。根据分离菌T_t1和T_t2在不同土壤湿度下培养50d和60d的冬孢子萌发率,建立了TCK冬孢子萌发率与土壤相对含水量的关系模型,此结果为TCK的风险评估提供了基础数据。     

Arguments for a low risk of establishment of Karnal bunt disease of wheat in Europe

Tilletia indica, which causes Karnal bunt of wheat, has been perceived as a pathogen that has a high risk of establishment in the UK and other countries in Europe. This viewpoint is challenged with arguments for the risk of establishment being much lower. The present distribution of the pathogen and its likely widespread dissemination with consignments of germplasm and wheat seed from Mexico over a 20 year period without establishment outside the hot arid and semi-arid zones strongly suggests that T. indica is unsuited to environments other than those similar to where it is now found. A critical density of teliospores needs to remain viable and ungerminated between periods of wheat anthesis and then germinate during a narrow window of opportunity to ensure infection and establishment. Despite the results of outdoor teliospore longevity experiments, it is by no means certain that this can occur under European conditions. Generally cooler conditions and more frequent periods of rain in Europe are likely to trigger teliospore germination at times unsuitable for infection making establishment much more unlikely. A model based on year-round soil conditions, especially moisture content, is seen as appropriate to predict teliospore survival and germination potential. It is only after areas where teliospores can survive and germinate in sufficient numbers to guarantee regular infection cycles have been defined can another model, based on temperature, rainfall and/or humidity prior to anthesis, be applied to determine if environmental conditions are suitable for infection. David Jones is a retired employee of the Central Science Laboratory (CSL). Statements and opinions expressed in this paper are those of the author and are not the views of the Central Science Laboratory.     

从进口草籽中截获绒毛草腥黑穗菌

本文介绍了小麦矮腥黑穗病菌（ＴＣＫ）的近似种－绒毛草腥黑穗菌（ＴＨＳ）的检验鉴定情况,并详细比较了两种黑粉菌,发现两种冬孢子萌发对温度条件要求不同。     

温度对小麦矮腥黑穗病菌冬孢子萌发的影响

温度对小麦矮腥黑穗病菌冬孢子萌发影响的试验结果表明,TCK冬孢子在-2～12℃范围内都可以萌发,5℃为最佳萌发温度。同一分离菌冬孢子在不同温度处理的萌发特性和萌发率有差异;不同分离菌冬孢子在相同温度下的萌发特性和萌发率有很大差异。在5℃下5个分离菌中,冬孢子萌发起始时间最短的为15 d,最长为35 d,培养60 d时分离菌T_t1、T_t2、T_y、T_m和T_u冬孢子的萌发率分别为86.4%、23.9%、23.3%、44.3%和81.0%。根据分离菌T_t2不同温度下培养50 d时的萌发率,建立了萌发率(Y)与温度(X)的模型为Y=0.150 8 exp[-0.02949(X-4.957 6)]2,此结果为TCK的风险评估提供了基础数据。     

Enhancement of teliospore germination in wheat- and wild grass-infecting species of tilletia on activated charcoal medium

ABSTRACT The effect of activated charcoal as an amendment to water agar medium on teliospore germination was analyzed for two species of wheat-infecting bunts, Tilletia controversa and T. tritici, and two related wild-grass infecting species, T. bromi and T. fusca. Final percentages of teliospore germination, area under the germination progress curves (AUGPC), and a standardized AUGPC (SAUGPC) on carbon agar and water agar were compared among strains. Carbon agar (CA) significantly increased the final germination percentage of teliospores, AUGPC, and SAUGPC when compared with water agar (WA) for all taxa under study. Additionally, CA reduced significantly the incubation (i.e., lag) period when compared with WA for teliospores of T. bromi, T. controversa, and T. fusca. Bovine serum albumin and polyvinyl pyrrolidone were used as alternative chemical adsorbent amendments to WA to establish the role of activated charcoal in the medium. Only media amended with bovine serum albumin and activated charcoal improved the final germination percentage of all taxa. Polyvinyl pyrrolidone was not significantly better than water agar.