甘肃省南瓜白粉病菌对己唑醇抗性诱导及抗性菌株生物学性状研究

为了解甘肃省裸仁美洲南瓜白粉菌对己唑醇敏感性及抗性产生情况,采用小株喷雾法对采自兰州、武威、景泰等地的20个南瓜白粉菌菌株对己唑醇的敏感性进行了测定,利用紫外诱导、药剂驯化、先紫外后药剂驯化三种方法对敏感菌株进行了抗药性诱导,并对抗性突变菌株的生物学性状及其对丙环唑、氟硅唑、腈菌唑和戊唑醇的交互抗性进行了研究。结果表明20个南瓜白粉菌菌株对己唑醇的敏感性存在一定差异,EC50值在17.77~285.54μg/mL范围内;与其他两种诱导方法相比药剂驯化方法获得的抗性突变菌株的抗性倍数最高,为8.94倍。敏感菌株和抗性突变菌株孢子萌发的最佳时间分别为36h和48h;抗性突变菌株产孢量和致病力明显高于敏感菌株;适合度测定表明抗性突变菌株与敏感菌株之间存在竞争力,孢子萌发率、芽管个数之间差异显著,菌丝分支数、产孢量之间差异不显著;抗性突变菌株在抗性突变菌株和敏感菌株混合比例为80∶20的群体中,存在频率比较稳定,连续培养7代后仍占85.69%;己唑醇抗性突变菌株对丙环唑、氟硅唑、腈菌唑和戊唑醇4种药剂未表现出交互抗性。研究结果可为甘肃省防治南瓜白粉病提供理论依据。     

黄瓜白粉病菌对己唑醇的敏感基线及实验室抗性获得

为了评价黄瓜白粉病菌对己唑醇的抗性风险,采用子叶法测定了山东地区50个黄瓜白粉病菌菌株对己唑醇的敏感性。结果表明,各菌株敏感性存在较大差异,EC50值最低为0.0017μg/mL,最高达0.16μg/mL,平均值为0.028μg/mL,由于所有菌株均采自未使用过脱甲基抑制剂类杀菌剂的黄瓜日光温室,敏感性频率呈近似正态分布,可作为黄瓜白粉病菌对己唑醇的相对敏感基线。以野生菌株为材料,以紫外线诱导和药剂驯化相结合获得了2个黄瓜白粉病菌对己唑醇的抗性菌株,比较了抗性菌株和敏感菌株的适合度和分生孢子的竞争能力。结果表明,无药情况下2个抗性菌株与野生菌株适合度无显著差异。抗性菌株和敏感菌株分生孢子以80∶20混合培养7代后,抗性菌株的频率变化不明显;50∶50混合培养7代,中抗菌株的抗性频率明显降低,而高抗菌株的抗性频率降低不明显;20∶80混合培养7代,抗性菌株的频率变低甚至消失。     

吉林省西瓜蔓枯病菌对吡唑醚菌酯的抗性监测及抗性风险评估

为明确吉林省西瓜蔓枯病菌Stagonosporopsis citrulli对吡唑醚菌酯的敏感基线和抗性风险,采用菌丝生长速率法测定吉林省151株西瓜蔓枯病菌菌株对吡唑醚菌酯的敏感性及吡唑醚菌酯对西瓜蔓枯病菌的最低抑制浓度,并监测375株西瓜蔓枯病菌菌株对吡唑醚菌酯的抗性水平。结果表明,供试的151株菌株对吡唑醚菌酯的敏感性频率分布呈连续的单峰曲线,接近正态分布,敏感基线为1.800 μg/mL;吡唑醚菌酯的最低抑制浓度MIC为80.000 μg/mL。抗性水平监测结果显示,375株菌株对吡唑醚菌酯的抗性频率为0。室内药剂驯化获得的抗性突变菌株T-LH3-1和T-XZC5-1在连续转接培养8代后的EC₅₀分别为16.139 μg/mL和31.782 μg/mL,抗性倍数分别为15和51,均为中抗水平,抗药性可以稳定遗传。抗性突变体及其亲本菌株生物学性状结果显示,抗性突变体生物适合度与其亲本菌株差异不显著。交互抗性结果表明,吡唑醚菌酯与醚菌酯之间存在正交互抗性,与戊唑醇、异菌脲、多菌灵和肟菌酯之间不存在交互抗性,可以交替使用以达到延缓抗药性的产生。     

浙江省稻田藤仓镰孢霉对咪鲜胺的敏感性及抗药性菌株的适合度与交互抗性

为明确浙江省水稻恶苗病主要致病菌藤仓镰孢霉Fusarium fujikuroi对咪鲜胺的抗性现状,以1μg/mL为区分剂量,对2017和2018两年采自浙江省绍兴、嘉兴和金华3个地区稻田的共400株藤仓镰孢霉对咪鲜胺的抗性进行了检测,并测定了抗药性菌株的适合度及其对其他常用甾醇脱甲基抑制剂类(DMIs)杀菌剂的交互抗性。结果表明:两年的抗药性菌株频率分别为53.2%和46.7%;多数田间敏感菌株在产孢能力和孢子萌发能力上强于田间抗药性菌株,在菌丝生长速率和致病性方面则两者差异不大;就稻田藤仓镰孢霉而言,咪鲜胺与丙硫菌唑之间存在交互抗性,但咪鲜胺与戊唑醇、苯醚甲环唑、丙环唑、叶菌唑、己唑醇和三唑酮之间均不存在交互抗性。研究表明,浙江省稻田藤仓镰孢霉对咪鲜胺已经产生了严重的抗药性,需要轮换使用不同作用机制的杀菌剂进行防治。     

福建省草莓炭疽病菌对吡唑醚菌酯的敏感性及与其他药剂的交互抗性

为明确吡唑醚菌酯对福建省草莓炭疽病菌Colletotrichum gloeosporioides的防治潜力和应用前景,采用室内生测法测定草莓炭疽病菌对该菌剂的敏感性及其与6种不同杀菌剂之间的交互抗性。结果显示,吡唑醚菌酯对福建省5个地区分离的62株草莓炭疽菌菌株的EC₅₀介于0.086～0.879μg/mL之间,总变异系数为10.24,且符合正态分布,以其平均值0.363μg/mL为敏感基线的菌株抗性比值均低于5;此外,吡唑醚菌酯EC₅₀与多菌灵、戊唑醇、苯醚甲环唑、咪酰胺、咯菌腈、甲基硫菌灵的EC₅₀间不存在显著相关性。表明福建省不同地区草莓炭疽病菌群体对吡唑醚菌酯的敏感性虽存在一定变异,但仍均较敏感;吡唑醚菌酯与其他6种杀菌剂间不存在交互抗性,可用于防控草莓炭疽病。     

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。     

山西省番茄早疫病菌对苯醚甲环唑的敏感性及抗性突变体的适合度

为了解山西省番茄早疫病菌Alternaria solani对苯醚甲环唑的抗性,采用菌丝生长速率法检测了该病菌对苯醚甲环唑的敏感性,分析了抗性水平和抗性指数;同时通过室内诱导获得了苯醚甲环唑抗性突变体,检测了突变体的抗性遗传稳定性、交互抗性和适合度。结果表明:分离自大同、忻州、长治、晋中和运城5市的101株菌株的EC50范围为0.005～3.315 mg/L,均值为0.428 mg/L,其中敏感菌株占97.03%,低抗菌株占2.97%;抗性指数范围为0.295～0.447,均值为0.343。10株中抗突变体抗性均不能稳定遗传、生长速率减慢、产孢量降低、孢子萌发能力减弱、致病力下降;但在药剂压力下,突变体的致病力强于敏感菌株。苯醚甲环唑与代森锰锌、多抗霉素、啶酰菌胺无交互抗性,而与啶菌噁唑存在交互抗性。因此,今后需加强番茄早疫病菌对苯醚甲环唑的田间抗性监测,在生产过程中轮用或混用不同作用机制的药剂以延缓该病菌对苯醚甲环唑抗性的发展。     

葡萄白腐病菌对抑霉唑的敏感基线及其与不同杀菌剂的交互抗性

为明确山东省胶东地区葡萄白腐病菌Coniella diplodiella对抑霉唑的敏感性及抑霉唑与常规杀菌剂之间的交互抗性,采用菌丝生长速率法测定葡萄白腐病菌对抑霉唑等杀菌剂的敏感性,并通过分析抑霉唑与戊唑醇、吡唑醚菌酯、福美双、多菌灵、代森锰锌毒力的相关性,判断抑霉唑与各药剂之间是否存在交互抗性。结果表明,供试69株葡萄白腐病菌菌株对抑霉唑的EC₅₀在0.13~55.53 μg/mL之间,最高值与最低值相差427.15倍;其频数分布图呈多峰曲线,第一主峰内EC₅₀平均值为6.34 μg/mL,可作为胶东地区葡萄白腐病菌对抑霉唑的敏感基线。与敏感基线相比,田间已出现抑霉唑低抗菌株,占检测总株数的8.70%,未检测到中、高抗菌株。选择3个抗性菌株连续继代培养10代后,其对抑霉唑的敏感性明显提高,说明其抗药性不能稳定遗传。抑霉唑EC₅₀对数值与戊唑醇、吡唑醚菌酯、福美双、多菌灵、代森锰锌的EC₅₀对数值之间的相关系数分别为0.799、-0.143、-0.089、-0.268和0.159,说明抑霉唑与戊唑醇存在一定的交互抗性,但与其他4种药剂之间不存在交互抗性。表明抑霉唑可用于胶东地区田间葡萄白腐病的有效防控。     

黄瓜霜霉病菌对双炔酰菌胺的敏感基线及其抗性突变体生物学性状研究

为建立黄瓜霜霉病菌对双炔酰菌胺的敏感基线及评估其抗性风险,采用叶盘漂浮法测定了采自河北、山东未使用过羧酸酰胺类(CAAs)药剂地区的69株黄瓜霜霉病菌(Pseudoperonospora cubensis)对双炔酰菌胺的敏感性,并对黄瓜霜霉病菌抗双炔酰菌胺突变体的获得方法及突变体的生物学性状进行了研究。结果表明:69株黄瓜霜霉病菌对双炔酰菌胺的平均EC50值为(0.358±0.144)μg/mL,不同敏感性菌株的频率呈连续单峰曲线分布,未发现敏感性下降的亚群体,因此可将其作为黄瓜霜霉病菌对双炔酰菌胺的敏感基线;通过药剂驯化的方法未获得黄瓜霜霉病菌抗双炔酰菌胺的突变体;而通过紫外诱导的方法获得了6个抗双炔酰菌胺的突变体,其抗性水平介于5.74~22.96倍之间,突变频率为1.09×10-7,适合度显著低于其亲本菌株,且抗药性不能稳定遗传;双炔酰菌胺与甲霜灵、嘧菌酯、霜脲氰、氟吡菌胺之间无交互抗性关系,与烯酰吗啉之间有交互抗性关系。据此推测黄瓜霜霉病菌对双炔酰菌胺的抗性风险为低到中等。     

禾谷丝核菌对井冈霉素的抗性风险预测

采用菌丝生长速率法,测定了4个麦区的5个禾谷丝核菌Rhizoctonia cerealis菌株对井冈霉素的抗药性;在含井冈霉素的PDA平板培养基上对禾谷丝核菌进行继代培养,以诱导抗药性菌系;测定了抗性菌系对其他杀菌剂的交互抗性,并比较了抗性和敏感菌系对渗透压的敏感性及药剂处理后两种菌系培养液内还原糖和可溶性蛋白的含量差异。结果表明,田间禾谷丝核菌菌株对井冈霉素分别产生了7.26、7.75、10.46、14.92和23.31倍的抗性。室内继代培养36代,禾谷丝核菌对井冈霉素的抗性达49.24倍,形成了抗井冈霉素菌系。抗井冈霉素菌系对 口 恶 醚唑、福美双、三唑酮、丙环唑、戊唑醇和咯菌腈分别产生了48.58、21.78、17.62、10.95、2.55和1.78倍的交互抗性。抗性菌系在较低和较高渗透压下其生长抑制率均大于敏感菌系,用井冈霉素处理抗性和敏感菌系后,其体内的电解质都严重外渗,且抗性菌系在10 h 内的外渗量明显大于敏感菌系。     

黄瓜白粉病菌拮抗真菌鉴定及生防效果评价

本文对黄瓜白粉病菌Sphaerotheca fuliginea拮抗真菌进行了筛选和鉴定及防治效果的评价。结果表较高的拮抗活性。菌株JK-13发酵液处理白粉菌孢子48 h后,其对孢子萌发抑制率为89.93%;发酵液处理发病的离体叶片和盆栽幼苗7 d后,其防治效果分别为88.02%和76.34%。利用形态学和分子生物学技术鉴定该菌为腐皮镰刀菌Fusarium solani。     

黄瓜白粉病菌在不同抗性黄瓜材料上的侵染过程

白粉病是黄瓜生产中发生普遍,危害严重的主要病害之一。pm5.1和PM5.2是黄瓜上的2个白粉病抗性位点,本文对7份不同抗病基因型的黄瓜自交系进行了黄瓜白粉病抗性鉴定,并开展了黄瓜白粉病菌侵染过程的研究,对侵染后12、24、72 h的萌发率、菌丝形成率及菌落形成率等进行了分析。结果表明,当基因型为PM5.1PM5.1 pm5.2pm5.2时,黄瓜病情指数最高,表现为高感白粉病;当基因型为PM5.1PM5.1PM5.2 PM5.2和pm5.1pm5.1pm5.2pm5.2时,表现为中感白粉病;当基因型为pm5.1pm5.1PM5.2PM5.2时黄瓜自交系病情指数最低,表现为抗白粉病。分生孢子在抗、感黄瓜自交系植株叶片上均能萌发,但只能在感病黄瓜材料上完成整个无性生长周期,产生分生孢子。此外,黄瓜白粉病菌分生孢子在感病材料上的萌发率、菌丝形成率及菌落形成率均高于抗病材料。     

BTH处理对甜瓜叶片活性氧代谢的诱导作用

为了明确活性氧（reactive oxygen species,ROS）代谢在甜瓜抗病性诱导中的作用,以抗白粉病甜瓜品种Tam Dew和感病品种卡拉克赛幼苗为材料,通过盆栽试验研究了苯丙噻二唑（BTH）喷雾或白粉菌接种后甜瓜叶片超氧阴离子（O2.-）产生速率、过氧化氢（H2O2）含量及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、苯丙氨酸解氨酶（PAL）活性的变化。BTH处理或白粉菌接种均可诱导甜瓜叶片SOD、PAL活性升高,抑制CAT活性,导致叶组织O2.-产生速率和H2O2含量增加,BTH喷雾＋白粉菌接种比二者单独处理效果更好。结果表明,BTH处理后叶片O2.-产生速率提高和H2O2积累是甜瓜抗白粉病能力提高的重要机制,BTH通过诱导ROS代谢酶活性调节H2O2含量,且BTH诱导的甜瓜抗病性与品种的基础抗性有关。     

Control of cucumber powdery mildew by the hyperparasite Ampelomyces quisqualis and fungicides

Application of conidial suspensions of the hyperparasite Ampelomyces quisqualis to greenhouse cucumbers inoculated with Sphaerotheca fuliginea increased cucumber yield compared with an untreated control. Similar yield increases were obtained with triforine and also when a reduced rate of triforine and the hyperparasite were combined. A. quisqualis extensively parasitized S. fuliginea and also, in experiments with commercial cucumber crops, Erysiphe cichoracearum.     

寡聚糖诱导黄瓜对白粉病抗病反应的超微结构研究

 中科6号(2%氨基寡糖素)处理黄瓜植株叶片,5 d后接种白粉菌Sphaerotheca fuliginea(Schlecht.) Poll.,可诱导黄瓜Cucumis sativus产生对白粉病的抗病性,寄主细胞对病原菌的侵入产生了防卫反应结构和物质以及过敏性坏死反应。表现为寄主细胞壁加厚,染色加深,寄主细胞壁下产生多层次结构的乳突,在寄主细胞壁与质膜之间有黑色物质沉积;吸器外质膜皱褶,染色加深,吸器外基质中出现染色加深的颗粒状电子沉积物;寄主细胞质紊乱,细胞器解体,整个寄主细胞解体、坏死。     

Nature of resistance of seven melon lines to Sphaerotheca fuliginea

The nature of resistance of Sphaerotheca fuliginea Poll. race 1 was investigated in leaves of susceptible, moderately resistant and resistant genotypes of Cucumis melo . For this purpose three different tests were carried out. The first one was designed to assess whether resistance was caused by pre- or post-infectional mechanisms, and the other two tests were focused to determine if any differences were present in the progression ratio of the disease, measured as the delaying time in conidiophore and conidia formation, among the different cultivars which were tested. The results showed that in the resistant controls ('PMR5' and 'PI 124111'), the resistant reaction was probably due to pre-infectional mechanisms, or at least very near to the moment of infection, while in the moderately resistant and resistant Spanish local cultivars it was clearly due to post-infectional mechanisms characterized by an important delay in the growth and development of the fungus when compared with the susceptible cultivars. Suggestions are made for the development of more successful breeding programmes.     

甲氧基丙烯酸酯类杀菌剂ZJ 0712对黄瓜 白粉病和霜霉病的毒力及药效评价

ZJ 0712,2- -3-甲氧基丙烯酸甲酯,是我国自行创制的高效广谱杀菌剂。为评价该药剂10%SC对黄瓜白粉病和霜霉病的使用效果,进行了室内和田间药效评价。经室内检测,该药剂对白粉病菌Sphaerotheca fuliginea的EC50和EC90值分别为1.79和 5.47 μg/mL,对霜霉病菌Pseudoperonospora cubensis的EC50和EC90值分别为0.34和 45.15 μg/mL。 田间试验结果表明,该药剂对白粉病在10 ~20 g/hm2、对霜霉病在37.5 ~150 g/hm2剂量下防效分别为85.79% ~91.33%和86.70% ~90.18%。该药剂在本试验剂量范围内对作物安全。     

Virulence variation and DNA polymorphism in Sphaerotheca fuliginea, causal agent of powdery mildew of cucurbits

Strains of Sphaerotheca fuliginea, one of the causal agents of powdery mildew of cucurbits, were examined for differences in virulence, mating type and DNA polymorphism. The 28 strains were chosen to be diverse according to host and geographic origin. Characterization of virulence phenotypes was based on the expression of symptoms on 4 species of cucurbits and 6 cultivars of melon. Two pathotypes, capable of attacking either cucumber cv. Marketer and melon cv. IranH and squash cv. Diamant or cucumber cv. Marketer and melon cv. IranH were observed. Tests on melon cultivars revealed 3 races. In tests of sexual compatibility with reference strains, heterothallism was observed for all isolates. Frequency of the two mating types differed significantly in the population. DNA polymorphism was determined both by restriction fragment length polymorphism (RFLP) of the ribosomal internal transcribed spacers (ITS) and 5.8S DNA amplified by the polymerase chain reaction and by random amplified polymorphic DNA (RAPD). For any one of the 11 restriction enzymes tested all strains presented an identical pattern of ITS RFLP. RAPD analysis, using 22 primers which provided reproducible patterns, revealed a relatively low degree of polymorphism. Furthermore, cluster analysis based on RAPD data (152 markers) did not separate groups within the species S. fuliginea. No association could be found between virulence, mating type, geographical and host origin and RAPD patterns. The lack of association between phenotypic and molecular markers and the close fit to linkage equilibrium for the characters examined suggest that recombination may play a role in populations of S. fuliginea.     

Ultrastructural and Cytochemical Investigations of the Antagonistic Effect of Verticillium lecanii on Cucumber Powdery Mildew

ABSTRACT Chronological events of the intercellular interaction between Verticillium lecanii and cucumber powdery mildew, caused by Sphaerotheca fuliginea, were investigated at different times after inoculation by transmission electron microscopy. V. lecanii hyphae colonized host structures by tight binding, apparently mediated by a thin mucilaginous matrix. As early as 24 h after application of the antagonist, increased vacuolation and disorganization of the cytoplasm of the pathogen hyphae were easily detected. By 36 h after treatment, plasmalemma retraction and local cytoplasm aggregation were typical features of damage. Labeling chitin with the wheat germ agglutinin (WGA)/ovomucoid-gold complex showed that intracellular invasion of S. fuliginea by V. lecanii did not cause extensive host cell wall alterations, except in the area of hyphal penetration. By 48 h after inoculation, further cytoplasm disorganization was observed, as evidenced by the loss of cell turgor and contortion of the cell wall. Such deformation suggests that penetration of the antagonist results from mechanical pressure or localized enzymatic hydrolysis through the action of chitinases, as confirmed by the pattern of labeling obtained with the WGA/ovomucoid-gold complex. By 72 h after contact between the fungi, S. fuliginea cells were markedly collapsed, depleted of their protoplasm due to extensive multiplication of the antagonist, and totally encircled by the antagonist. Based on the current observations, the antagonism of S. fuliginea by V. lecanii appears to involve the following events: (i) attachment of the antagonist to the powdery mildew fungus; (ii) mechanical pressure and production of cell-wall degrading enzymes such as chitinases; (iii) penetration and active growth of the antagonist inside the pathogen hyphae; and (iv) digestion of host tissues and release of the antagonist from dead cells of S. fuliginea. The interaction between V. lecanii and S. fuliginea also affected the morphological and structural features of the haustorial bodies, as shown by increased vacuolation, distortion, and necrotization of the haustorial lobes. These observations provide the first experimental evidence that V. lecanii, primarily known as an entomopathogenic fungus, also has the potential to colonize mycelial structures of S. fuliginea. V. lecanii, therefore, may become a valuable alternative to current management of cucumber powdery mildew in greenhouses.     

硅对瓠瓜酚类物质代谢的影响及与抗白粉病的关系

研究了硅酸盐和诱导接种白粉菌对瓠瓜叶片苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、过氧化物酶(POD)活性的动态变化和木质素含量的影响.结果表明,诱导接种提高了瓠瓜叶片的PAL、PPO、POD活性,加硅后接种处理植株叶片酶活性更明显增加;接种后瓠瓜叶片中木质素含量增加,加硅处理植株叶片木质素含量是不加硅处理的1.43倍.说明酚类物质代谢在植物抗病机制中起着重要作用,硅能提高酚类代谢的酶活性.1.7mmol/L的硅能显著降低瓠瓜白粉病的病情指数,提高其对白粉病的抗病能力.