枇杷花腐病病原物的初步鉴定

经调查发现,在重庆发生的枇杷花腐病症状有2种类型:干腐型和湿腐型。通过病原菌形态特征、培养性状和致病性测定,初步鉴定这两种症状分别由拟盘多毛孢菌[Pestalotiopsis eriobotrifolia(Guba)Chen et Cao]和灰葡萄孢菌(Botrytis cinerea Pers.)引起。枇杷花腐病害的发生与环境湿度有关,且湿度越大,病害发生越严重。     

洛阳市牡丹灰霉病病原菌的鉴定

为明确洛阳市牡丹灰霉病的病原菌种类,于2015年从该地区的4个牡丹Paeonia suffruticosa Andr.种植区采集灰霉病样品,采用组织分离法对病原菌进行分离和纯化,结合形态学和基因序列分析对分离物进行鉴定,采用针刺接种法测定不同分离物对牡丹离体叶片的致病性。结果表明,从灰霉病样品中分离获得40株分离物,这些分离物分别属于Ⅰ、Ⅱ、Ⅲ型菌,I型菌不易产孢但产菌核;Ⅱ型菌易产孢,后期产生少量菌核;Ⅲ型菌易产孢,且产生大量菌核。产孢分离物均形成直立状分生孢子梗,孢子梗分枝顶端聚生葡萄穗状分生孢子,分生孢子卵圆形或长卵圆形。供试菌株的ITS序列和G3PDH基因序列与灰葡萄孢Botrytis cinerea Pers.的同源性达到99%;致病性测定结果表明,各菌群代表菌株对牡丹均有致病性,但不同菌群间致病力有差异。研究表明,引起洛阳市牡丹灰霉病的病原菌为灰葡萄孢,且菌群类型多样。     

挥发性化合物2-甲基苯并噻唑对核盘菌和灰葡萄孢的抑制效应

为明确挥发性化合物2-甲基苯并噻唑（2-methylbenzothiazole,2-MBTH）对向日葵菌核病病原菌核盘菌Sclerotinia sclerotiorum和番茄灰霉病病原菌灰葡萄孢Botrytis cinerea的防治潜力,采用密封盘菌丝生长速率法测定其对2种病原菌的抑制作用,利用离体叶片接种试验分析其对2种病原菌致病力的影响。结果显示,2-MBTH对核盘菌和灰葡萄孢具有明显的抑制作用,2-MBTH对核盘菌菌丝的EC₅₀为0.87 μL/L,对灰葡萄孢菌丝的EC₅₀为4.49 μL/L。2-MBTH能够显著降低灰葡萄孢的致病力,受抑制程度与2-MBTH浓度呈正相关;扫描电镜观察发现,2-MBTH能够使灰葡萄孢菌丝形态异常,菌丝干瘪,分枝增多,表面塌陷;使核盘菌菌丝破损,表面塌陷。另外,随着2-MBTH熏蒸浓度的增大,核盘菌的菌核数量和质量都有所减少,且健康菌核在2-MBTH的熏蒸作用下失去了萌发能力。表明2-MBTH对核盘菌和灰葡萄孢均有较高的抑制作用,表现为菌丝生长、菌核萌发受到抑制,利用其熏蒸作用可以为向日葵菌核病和番茄灰霉病的有效防控提供新思路。     

盾叶薯蓣灰霉病(Botrytis cinerea)的研究

对一种引起盾叶薯蓣叶、茎呈暗绿色至黑褐色水渍状软腐的病害进行了调查 ,并作了鉴定。结果明确其为灰葡萄孢菌 (Botrytis cinerea Pers.ex Fr )引起的一种新病害 ,根据症状和病原鉴定将该病称为盾叶薯蓣灰霉病。该病原菌生长温度 0～30℃ ,最适生长温度 10～ 2 5℃。病害在每年春季、夏初和秋末 ,雨水多的年份发生严重。     

拟南芥转录因子IDD4不同突变体对灰葡萄孢菌的抗性分析

为明确IDD家族IDD4基因在拟南芥Arabidopsis thaliana抵抗灰葡萄孢菌Botrytis cinerea侵染过程中的作用,通过统计病情指数检测拟南芥野生型（wild type,WT）植株、过表达植株IDD4-OE和缺失突变体idd4植株感染灰葡萄孢菌情况,利用组织染色检测叶片细胞死亡和H₂O₂的积累情况,采用实时荧光定量PCR（real-time quantitative-PT-PCR,qRT-PCR）技术分析灰葡萄孢菌肌动蛋白基因Bc. ACTIN在3种植株叶片中的表达情况,并施加0.1 mmol/L外源水杨酸（salicylic acid,SA）后测定IDD4-OE植株的病情指数。结果显示,不同株系对灰葡萄孢菌的抗性由高到低依次为idd4＞WT＞IDD4-OE,IDD4-OE植株中病原菌感染部位的寄主细胞死亡程度比idd4植株严重。染色结果表明,病原菌侵染拟南芥后4 h,接种部位已有H₂O₂积累。qRT-PCR反应结果显示,Bc. ACTIN在IDD4-OE中比在idd4植株中的表达水平更高,表明灰葡萄孢菌在IDD4-OE植株中的繁殖速率更快。对IDD4-OE植株外源施加SA后,其病情指数、Bc. ACTIN表达量与WT植株间均无显著差异,说明SA能将感病植株的抗性提高至WT植株的水平,表明IDD4作为负调控因子参与了拟南芥对灰葡萄孢菌的抗性调控,SA在其中发挥着重要作用。     

藜麦灰霉病病原菌鉴定及环境因子对其菌丝生长和分生孢子萌发的影响

为明确我国藜麦灰霉病病原菌的归属及环境因子对其菌丝生长和分生孢子萌发的影响,在藜麦灌浆期从山西省忻州市原平市和五台县采集具典型症状的藜麦标本,应用直接挑取法分离病原菌,选取代表性菌株对其形态学特征、系统发育、致病性及生物学特性进行观察和测定。结果表明,综合形态学特征及基于G3PDH、HSP60和RPB2多基因系统发育分析结果,最终确定引起我国藜麦灰霉病的病原菌为灰葡萄孢Botrytis cinerea。致病性测定发现接种6 d时,接种处密生灰色霉层,籽粒霉变,与田间症状基本一致,说明分离菌株确为藜麦灰霉病的病原菌。灰葡萄孢在燕麦琼脂培养基、马铃薯葡萄糖琼脂培养基和察氏酵母培养基上培养后可产生菌核,而在麦芽浸粉琼脂培养基上培养可产生分生孢子;在察氏酵母培养基上培养的菌丝生长速率最快,平均为16.8 mm/d。适宜灰葡萄孢菌丝生长和分生孢子萌发的温度为15～25℃、水活度≥0.96、pH 5～6。     

环境条件对灰葡萄孢菌菌核萌发的影响

<正>灰葡萄孢菌(Botrytis cinerea)引起的灰霉病可导致植物幼苗猝倒或花朵和果实腐烂,多在冬春季节发生,是保护地蔬菜、水果和花卉生产或储藏过程中一类重要的病原菌。灰霉菌的初侵染主要由菌核或病残体在适宜温湿度时萌发产生的菌丝或孢子侵染植物,关于灰葡萄孢菌的菌丝、分生孢     

苹果黑点病症状及病原菌鉴定

近年在甘肃省天水、陇南苹果产区发现苹果果实黑点病 ,经对其病原菌形态、培养性状和致病性等方面的测定表明 ,其主要致病菌为苹果柱盘孢霉 (Cylindrosporium pomi Brooks) ,其次为苹果茎点菌 (Phomapomi Pass)。该病在苹果不同品种果实上造成的症状不同 ,将其归纳为 5种症状类型。该病在我国尚属首次发现     

纳他霉素对灰葡萄孢不同生育阶段菌体的毒力及其生物学性状的影响

以多菌灵为对照药剂,采用生长速率法、悬滴法和浸渍法分别测定了不同温度下纳他霉素对灰葡萄孢Botrytis cinerea菌丝生长、分生孢子萌发和菌核萌发的毒力,研究了常温下两种药剂对菌株产孢和菌核形成的影响。结果表明:纳他霉素及多菌灵对灰葡萄孢不同生育阶段菌体的抑制作用随温度降低而有不同程度增强;分生孢子对药剂最为敏感;温度对药剂对菌丝毒力的影响最显著;多菌灵对菌株不同发育阶段的抑制活性均高于纳他霉素。供试药剂对菌株产孢时间和菌核产生时间无显著影响,但多菌灵可显著刺激菌株产孢和菌核形成。     

球孢白僵菌定殖提高番茄对灰霉病抗性及其作用机理

为明确球孢白僵菌Beauveria bassiana定殖对番茄植株抗病性的影响及作用机理,采用灌根法将球孢白僵菌芽生孢子悬浮液接种于番茄植株内,并通过人工接种灰霉菌Botrytis cinerea评价球孢白僵菌定殖后番茄对灰霉病的抗性水平;检测灰霉菌胁迫下番茄植株不同位置叶片内球孢白僵菌的相对含量;测定番茄叶片内草酸氧化酶（oxalate oxidase,OXO）、几丁质酶（chitinase,CHI）和ATP合成酶（ATP synthase,atpA）3种抗病相关基因的表达量。结果表明,球孢白僵菌定殖能够提高番茄植株对灰霉病的抗性,接种灰霉菌第5天,番茄植株发病率、病斑直径和病情指数分别下降了61.6%、41.4%和26.4%;在灰霉菌胁迫下番茄植株内球孢白僵菌偏好于在病原菌感染位置定向聚集,并且引起植物抗病基因OXO、CHI和atpA的表达量上调。表明球孢白僵菌能通过内生定殖与植物互作提高植物抗病性,在植物病害生物防治领域有较大应用潜力。     

Botrytis blight of Taiwanese toad lily caused by Botrytis elliptica (Berkeley) Cooke

In June 1998, leaf spots caused by Botrytis elliptica (Berkeley) Cooke were found on Taiwanese toad lily (Tricyrtis formosana Bak.) in Kanagawa Prefecture, Japan. This is the first report on Botrytis blight of T. formosana caused by B. elliptica.     

Comparative Analysis of the Role of Substrate Specificity in Biological Control of Botrytis Elliptica in Lily and B. Cinerea in Cyclamen with Ulocladium Atrum

Biological control of Botrytis spp. by the fungal antagonist Ulocladium atrum is based on their interaction in plant tissue. U. atrum is effective against B. cinerea in commercial cyclamen crops but not effective against B. elliptica in lily crops. Based on the necrotrophic nature of the Botrytis spp. and the saprophytic nature of U. atrum it is hypothesised, and experimentally confirmed, that the interaction between Botrytis spp. and U. atrum, resulting in a biocontrol effect, only takes place in necrotic plant tissue. The role of necrotic tissue in the epidemiology of B. cinerea in cyclamen and B. elliptica in lily was found to be different. Removal of symptomless senescing leaves resulted in a significant reduction of the area under the disease severity progress curve (AUDPC) for B. cinerea in cyclamen but had no effect on the disease severity in lily. U. atrum applications significantly reduced B. cinerea AUDPC values in cyclamen but were less efficient than the removal of senescing leaves. In lily, disease severity was not affected by applications of U. atrum. It is concluded that necrotic cyclamen tissue, not killed by B. cinerea, plays an important role in the onset of disease. Colonisation of this tissue by U. atrum prevents saprophytic colonisation of those leaves by B. cinerea. In contrast, conidia of B. elliptica directly infect healthy lily leaf tissue. U. atrum applications aimed at blocking the infection pathway from a saprophytic base are therefore not effective against B. elliptica. Control options based on competitive interactions in and around B. elliptica lesions resulted in a reduced production of conidia by B. elliptica but proved ineffective against disease development. The potential of U. atrum as a biocontrol agent against Botrytis spp. and possibly against other necrotrophs appears to be determined by the competitive saprophytic ability of the antagonist in mutual substrates of pathogen and antagonist and by the role of these substrates in disease epidemiology.     

Disease symptoms caused byBotrytis cinerea inRuscus hypoglossum plants and their control

Small spots are a newly-defined manifestation ofBotrytis cinerea infection on phylloclades of ruscus. The dark, water-soaked spots encircled by a faint halo, which are found on young phylloclades, become brown lesions later but do not grow in size. Control of all symptoms caused byB. cinerea on ruscus (Ruscus hypoglossum L.),i.e., phylloclade rot or restricted lesions, upper branch rot, and stem canker, was achieved by sprays of fenetrazole (alone), diethofencarb + carbendazim, vinclozolin (alone) or alternately by sprays of diethofencarb + carbendazim, polyoxin B, and polyoxin D. Delay of disease development was achieved by copper sulfate pentahydrate and by the spray adjuvant Nufilm at a high rate of application. Mixing vinclozolin with copper, fenetrazole, TMTD or Nufilm did not control disease better than vinclozolin or fenetrazole alone. Postharvest control of gray mold (up to 72% reduction) in mature branches stored at 18°C was achieved by some of the above mentioned fungicides and by dichloftuanid, tebuconazole (alone or mixed together), iprodione, polyoxin AL, and vinclozolin + TMTD. Measures designed to improve field control of Botrytis epidemics in ruscus are discussed.     

Physiological response of Bacillus cereus C1L-induced systemic resistance in lily against Botrytis leaf blight

Bacillus cereus C1L has been demonstrated to induce systemic disease resistance against Botrytis elliptica in lily. The objective of this study was to investigate physiological responses of B. cereus C1L-triggered systemic resistance in lily cv. Star Gazer against B. elliptica. By histological and biochemical analyses, leaves inoculated with B. elliptica displayed cell death, H₂O₂ accumulation and lignin deposition. As plants were elicited with B. cereus C1L, cell death, H₂O₂ accumulation and lignin deposition in leaves caused by B. elliptica infection were suppressed, revealing that suppression of oxidative burst might be associated with B. cereus C1L-induced systemic resistance. In reactive oxygen species inhibitors assays, B. elliptica-caused lesion numbers and H₂O₂ accumulation in lily leaves were significantly reduced as leaves were pretreated with catalase or diphenylene iodonium. Furthermore, the expression of LsGRP1 and LsPsbR in leaves elicited with B. cereus C1L and inoculated with B. elliptica was decreased. The same expression pattern was also observed in leaves pretreated with catalase or diphenylene iodonium and inoculated with B. elliptica. These results suggest that B. cereus C1L-induced systemic resistance may be related to suppression or alleviation of oxidative stress and cell death of lily caused by B. elliptica.     

Genetic structure and fungicide sensitivity of Botrytis cinerea populations isolated from grapevine in northern Italy

Grey mould, caused by Botrytis cinerea, is a disease severely affecting grape production in northern Italy. However, little information is available on the variability of B. cinerea populations associated with grapevine. The mode of reproduction, sensitivity to fungicides, and for the first time in Italy, the genetic structure of B. cinerea populations isolated from grapevine in a northern Italian region are reported. Botrytis cinerea isolates (317) were completely genotyped for six microsatellite loci and characterized for the presence of the transposable elements Boty and Flipper, for the mating type and for resistance to cyprodinil, fludioxonil, boscalid and fenhexamid. All the isolates were found to belong to B. cinerea Group II, indicating the absence of B. pseudocinerea in the investigated areas. The populations possess a high genotypic diversity, different frequencies of transposable elements and a mixed mode of reproduction. At a regional level, B. cinerea populations belong to a large and interconnected pathogen population that includes the major grape‐growing districts. The populations were generally sensitive to fungicides, with a low proportion (8%) of isolates resistant to cyprodinil, fludioxonil and boscalid. A small genetic distance was found between B. cinerea populations. However, the populations geographically isolated from the others by a mountain range showed a small but statistically significant genetic differentiation and a different pattern of fungicide resistance. The results show that northern Italian B. cinerea populations possess a high evolutionary potential and adaptive capacity.     

BHT对苹果采后灰霉病的防效及防御酶活性和丙二醛含量的影响

为探究2,6-二叔丁基-4-甲基苯酚(2,6-di-tert-butyl-4-methylphenol,BHT)对苹果采后灰霉病的防效和防病机制,采用平板法和刺伤接种法测定了BHT对灰霉病菌Botrytis cinerea的抑制作用及果实内防御酶活性和丙二醛含量的影响。结果表明,0.1 mmol/L BHT对苹果灰霉病的防效最佳,处理苹果后间隔48～96 h接种灰霉病菌,其防效可达68.59%～73.55%,其次为0.2、1.0 mmol/L BHT处理,但防效均低于52.94%。0.1 mmol/L BHT处理对灰霉病菌菌丝生长和分生孢子萌发无显著抑制作用,但可显著增强果实内超氧化物歧化酶、过氧化物酶和过氧化氢酶的活性以及总抗氧化能力,其峰值是对照的1.82～5.28倍,且均显著高于单独接种灰霉病菌的处理。此外,单独接种灰霉病菌的苹果果实内丙二醛含量快速升高,最大增幅达251.49%,而BHT处理后丙二醛含量变幅较小,最大增幅仅为83.95%。表明BHT可通过增强苹果果实内防御酶活性水平和总抗氧化能力来降低丙二醛的积累,从而提高果实对灰霉病的抗性。     

Effect of Film-forming Polymers on Control of Lily Leaf Blight Caused by Botrytis elliptica

The effectiveness of film-forming polyelectrolytes for the control of lily leaf blight caused by Botrytis elliptica was evaluated using laboratory a leaf disk assay, greenhouse tests and field trials. Among the six polyelectrolytes, 400ppm FO4240SH, FO4490SH and FO4550SH reduced the disease severity of lily leaf blight in leaf-disk tests. Both FO4240SH and FO4490SH also suppressed sporulation of the pathogen on leaf disks. In greenhouse tests, the number and size of lesions on leaves of Lilium oriental hybrid cv. Star Gazer were markedly reduced by FO4490SH and FO4550SH. Field trials showed that the effectiveness of FO4490SH was similar or better than that of procymidone on the reduction of lily leaf blight disease severity. The polymers had no harmful effects on the lily plants. The cationic polyelectrolytes FO4240SH, FO4490SH and FO4550SH reduced the percentage of conidial germination, inhibited germ-tube growth, and also suppressed the esterase production by germ tubes of B. elliptica. All the above evidence indicates that the disease control achieved with polyelectrolytes is due, at least in part, to the reduction of esterase secretion by B. elliptica.     

Survival of dicarboximide-resistant strains ofBotrytis cinerea in plant debris during summer in Israel

Survival- ofBotrytis cinerea was monitored during two summer seasons. Mycelium and conidia were found dead on the surface of plant debris within 2 months of incubation, whereas a high level of viability was detected in thallus of the pathogen which was 1–2 mm inside the dry host tissue. Of the 148 samples of infected senescing cucumber female fruits, 8% survived seven warm months; half of these isolates ofB. cinerea were resistant to dicarboximides (5 (μ/ml iprodione). Of the stems of cucumber infected withB. cinerea in winter, 18% yielded the pathogen at the beginning of the following winter; 15% of the surviving isolates were resistant to dicarboximides. Cucumber seedlings artificially infected byB. cinerea did not yield the pathogen longer than 9 weeks after establishment of infection, even when incubated in the shade. Plant debris with symptoms of gray mold were kept in the shade during the summer; at the beginning of winter it was possible to establish infection ofB. cinerea from the dry debris.     

Mechanisms involved in the biological control ofBotrytis cinerea incited diseases

Mechanisms involved in the biological suppression of infection and inoculum potential ofBotrytis cinerea are numerous and variable and the involvement of two or more mechanisms has been demonstrated in several systems. Reported combinations include antibiosis with enzyme degradation ofB. cinerea cell walls; competition for nutrients followed by interference with pathogenicity enzymes of the pathogen or with induced resistance; and alteration of plant surface wettability combined with antibiosis. Since germinatingB. cinerea conidia are dependent on the presence of nutrients, competition for nutrients is regarded as important in systems where biocontrol is involved. Conidial viability and germination capacity are also potentially affected by the presence of antibiotics produced by biocontrol agents and present in the phyllosphere. Slower in action are mechanisms involving induced resistance in the host plant and production of hydrolytic enzymes that degradeB. cinerea cell walls. The latter has been demonstrated much more convincinglyin vitro than in the phyllosphere. Biocontrol in established lesions and reduction of sporulation on necrotic plant tissues is a means to minimize the pathogen inoculum.Abbreviations BCA bio-control agent - Bc Botrytis cinerea - PG polygalacturonase - PL Pectin lyase - PME Pectin methyl esterase - PR pathogenesis related - VPD vapour pressure deficit