来源于佳木斯茄子上的核盘菌菌株多样性的研究

 从我国黑龙江省佳木斯市同一茄子田采集的菌核样品中分离得到21个核盘菌(Sclerotinia sclerotiorum)单菌核分离物。通过比较其菌丝生长速度、菌落形态、菌核产量及菌核在PDA平板上的萌发特性等将其分成3种类型,记为A、B、C,各包含5、7、9个菌株。A型和B型菌株为正常菌株,而C型菌株为异常菌株,生长慢且呈扇形扩展。对代表性菌株转代培养物及菌核后代的培养特性比较结果说明它们的分化特性是稳定的。致病性的测定结果表明参比菌株Cor-6的致病性最强,A型菌株Ep-1PB和B型菌株Ep-1次之,C型菌株Ep-1PD最弱。3类菌株菌核的可溶性蛋白和酯酶同工酶电泳谱带没有明显的差异。     

津黑两地大白菜褐腐病病原菌鉴定及生物学特性的比较

采用形态特征观察、致病性测定及rDNA-ITS序列分析等方法对天津、黑龙江两地大白菜褐腐病的病原菌进行鉴定。结果表明,天津、黑龙江两地大白菜褐腐病的病原菌为立枯丝核菌(Rhizoctonia solani Kühn),分离鉴定出TJ和DB两个菌株,分属立枯丝核菌AG-2融合群和AG-1融合群;DB菌株为一新菌株。两菌株菌丝生长最适温度均为25℃,最适pH为7,最适光照为12h。DB菌株菌丝生长速率快于TJ菌株;但TJ菌株先于DB菌株形成菌核,且形成量多。两菌株在碳源、氮源利用和菌核致死温度上也存在差异。     

小麦纹枯病菌对三唑酮不同敏感性菌株的生物学特性及对不同杀菌剂的敏感性

为预测小麦纹枯病菌对三唑酮抗药性的发展趋势,采用菌丝生长速率法测定了2013年分离自河南省16个地市的98株菌株对三唑酮的敏感性,比较不同敏感性菌株的生物学性状差异,并分析了其中22株菌株对其它10种杀菌剂的敏感性。结果表明:小麦纹枯病菌群体中已出现对三唑酮敏感性下降的菌株,其中有90株菌对杀菌剂的敏感性属于正态分布,以其EC_(50)均值1.91μg/mL作为小麦纹枯病菌对三唑酮的敏感性基线。敏感菌株菌丝生长速度较快,可达8.50~12.86 mm/d,菌核产量较多,为35.80~42.90 mg/皿,生存适合度较高;敏感性下降及抗性菌株的致病性及菌核萌发等都未发生改变,但菌丝生长速率仅为5.36~10.02 mm/d,菌核产量为9.00~36.90 mg/皿,均下降明显,生存适合度降低;低抗菌株与敏感菌株在菌落形态上相同,但菌丝分枝数增多且分枝菌丝变短。三唑酮对小麦纹枯病菌的毒力较小,EC_(50)大于其它10种杀菌剂;戊唑醇、氟环唑、烯唑醇、咯菌腈和噻呋酰胺的毒力均较大,其EC_(50)≤0.10μg/mL;其余5种杀菌剂的EC_(50)在0.13~0.69μg/mL之间;小麦纹枯病菌对三唑酮的敏感性与其它10种杀菌剂间的决定系数为0.003~0.200,存在微弱的正相关性,表明在生产中这些杀菌剂可与三唑酮轮换或交替使用。     

内蒙古自治区向日葵小核盘菌的菌丝亲和组类型及其菌株的生物学特性、致病力和交配型

为明确内蒙古自治区阴山北麓地区向日葵小核盘菌Sclerotinia minor的遗传变异,对自内蒙古自治区乌兰察布市、包头市和呼和浩特市向日葵上分离纯化的110株向日葵小核盘菌菌株进行菌丝亲和组(mycelium compatibility group,MCG)划分,并对5个主要MCG组间和组内各菌株的生物学特性、致病力和交配型进行测定。结果表明,供试的110株菌株被划分为14个亲合组,其中MCG1为主要类型,包含32株菌株,占总菌株的29.1%;MCG2包含来自7个地点的25株菌株;5个MCG仅包含1株菌株;在这14个MCG中,MCG1～MCG5包含92株菌株,占总菌株数的83.6%。MCG1～MCG5组间各菌株在菌落直径和草酸分泌量上存在差异,但在菌核形成量、多聚半乳糖醛酸酶活性和致病力上无显著差异;而MCG1～MCG5组内各菌株在菌落直径、草酸分泌量、菌核形成量、多聚半乳糖醛酸酶活性和致病力上均有一定差异。在MCG1～MCG5各菌株的交配型中,除MCG2中菌株的负反转型与正反转型比例接近1∶1外,其它4个MCG中菌株的负反转型与正反转型比例均偏离1∶1,表明内蒙古自治区向日葵小核盘菌具有较高的遗传变异程度。     

云南省马铃薯晚疫病菌交配型及生物学特性研究

 作者对1998~2000年间采自云南省13个县、23个地点的马铃薯晚疫病菌的交配型、菌落形态、燕麦培养基上生长情况、生长速度和产孢量进行了测定。结果显示,采自云南13个县、23个地点的共157个菌株全部为A1交配型,表明云南马铃薯主产区的晚疫病菌以A1交配型为主,同时,被测的代表菌株在生长速度和产孢量上存在显著差异,表明这一地区的晚疫病菌种群内存在丰富的遗传多样性。此外,结果还显示,晚疫病菌菌株在燕麦培养基上的生长情况与其菌落形态和交配型不相关。     

进境美国苜蓿草中苜蓿黄萎病菌的检疫鉴定

从一批进境的美国苜蓿草样品中分离得到了一株与苜蓿黄萎病菌(Verticillium albo-atrum)相似的分离物M1。该分离物原始菌落为白色,边缘规则呈圆形,菌丝较致密,气生菌丝较少,苜蓿组织块不被菌丝覆盖,菌落生长速度为小于2.5 mm/d;M1在PDA上进行纯培养,前6d内,菌落为白色圆形,容易产生分子孢子轮枝状分生孢子梗,7d后菌落中央表面因产生休眠菌丝开始变成黑褐色至黑色,20d后菌落的表面和背面大部分均变黑色,仍不产生微菌核和厚垣孢子。M1的DNA用V.albo-atrum特异引物Vaa1/Vaa2进行检测,PCR扩增后得到预期330 bp的产物片段,产物序列与V.albo-atrum相应序列的相似性为100%。该分离物接种苜蓿草根部,15d后引起苜蓿黄萎病的典型症状。根据分离物的形态特征、PCR检测结果、PCR产物序列分析,以及致病性测定结果,将进境美国苜蓿草样品中的分离物M1鉴定为苜蓿黄萎病菌。     

口红吊兰菌核病病原鉴定及其生物学特性分析

为鉴定1株分离自口红吊兰叶部病斑上的疑似核盘菌菌株,利用柯赫氏法则验证其致病性并通过形态观察和ITS序列分析对该病原菌进行分类鉴定,结合温度、酸碱度等生理指标研究其生物学特性,并利用菌丝生长速率法测定了扑海因、多菌灵、苯醚甲环唑、腐霉利4种药剂对该病原菌的抑制作用。结果显示:在PDA平板上该病原菌菌丝为白色,均匀生长;约7 d后开始产生菌核,直径3~5 mm;菌核萌发后可产生1~3个子囊盘,内含8个大小为8.0~12.0μm×4.0~5.5μm的子囊孢子。该菌株ITS序列系统进化分析结果显示,其与核盘菌的同源性高达99%。综上所述,初步确定该菌株为核盘菌Sclerotinia sclerotiorum。该病原菌在PDA培养基上的最适生长温度为20~25℃、最适生长p H为5~7。室内毒力测定发现供试4种药剂中扑海因对该病原菌菌丝生长有较好的抑制效果,且其EC_(50)最小,仅为0.62 mg/L,证明扑海因对核盘菌毒力最强。     

吡虫啉胁迫对褐飞虱共生解脂假丝酵母抗性及敏感菌株生长的影响

为明确吡虫啉对褐飞虱共生解脂假丝酵母Candida lipolytica抗性及敏感菌株生长的影响,通过菌株培养、菌落观察和菌丝镜检等方法,比较了褐飞虱共生解脂假丝酵母不同菌株在含不同质量浓度吡虫啉的固体培养基上的菌落数,以及在液体培养基中的生长量差异。结果发现:不同质量浓度吡虫啉对褐飞虱共生解脂假丝酵母敏感和抗性菌株菌落生长均有抑制作用,且吡虫啉浓度越高,抑制作用越强。经500、1 000和2 000 mg/L吡虫啉处理4 d后,敏感菌株的菌落数量分别为对照的46.61%、27.58%和6.25%,均与对照差异显著;500和1 000 mg/L处理组抗性菌株菌落数量与对照无显著差异,而2 000 mg/L处理组与对照差异显著。经吡虫啉处理后,敏感菌株假菌丝形态变得不规则,部分假菌丝不舒展、萎缩或弯曲、顶端出现膨大,酵母出现空泡等,且吡虫啉浓度越高,不规则程度越明显;抗性菌株的假菌丝形态也有类似变化,但与敏感菌株相比,其菌丝体不规则形态的比例明显下降。500 mg/L吡虫啉处理对敏感和抗性菌株的生长量及菌丝干重均无明显影响,1 000和2 000 mg/L吡虫啉对不同菌株前期生长的抑制作用明显,但对后期生长影响不明显。研究表明,吡虫啉对褐飞虱共生解脂假丝酵母抗性菌株生长的影响显著小于对敏感菌株的影响。     

稻瘟菌群体中dsRNA的多样性及稻瘟菌菌株QSP5中病毒对寄主生物学性状影响的研究

 真菌病毒在真菌中广泛存在,一些真菌病毒可以影响病原真菌营养生长、产孢和色素形成等性状。本文研究了真菌病毒在水稻稻瘟菌中的多样性以及其对寄主生物学性状的影响。采用单孢分离的方法从发病水稻叶片上分离获得了120个稻瘟菌菌株,其中约52%携带dsRNA片段。菌株QSP5被2个基因组为dsRNA的病毒即产黄青霉病毒1-C(Magnaporthe oryzae chrysovirus 1-C, MoCV1-C)和稻瘟菌病毒3(M. oryzae virus 3, MoV3)所侵染。对菌株QSP5进行单个的分生孢子纯化,获得了仅携带MoV3的菌株QSP5-9。菌株QSP5与QSP5-9的产孢量、菌丝形态以及菌落形态存在明显差异,经研究推测MoCV1-C与菌株QSP5 的产孢能力衰退、菌丝生长和产色素异常相关。同时对MoV3的稳定性进行了初步的研究。从稻瘟菌菌株QSP5获得了100个单孢分离物,通过对随机挑选的40个单孢分离物进行检测,结果表明这些分离物100%携带MoV3,只有部分单孢分离物携带MoCV1-C。另外,从稻瘟菌菌株QSP5-9获得了94个单孢分离物,对随机挑选的45个单孢分离物进行检测,结果表明这些分离物中均含有与菌株QSP5-9大小相同的dsRNA片段。根据以上结果推定MoV3在稻瘟菌菌株QSP5和QSP5-9及其无性后代中具有一定的稳定性。     

海南省香蕉果实潜伏炭疽菌生物学特性研究

 从海南13个地区分离获得的17个Colletotrichum musae分离菌系经测定,无论是菌丝生长还是孢子萌发均以30℃为适温,pH值6~7为最佳条件;菌丝生长的最佳碳源是葡萄糖,而孢子萌发最佳碳源则是蔗糖,5种碳源都可提高孢子萌发率;自然光照(12h光暗交替)最有利于菌丝生长,30min紫外照射可使分生孢子致死。17个菌系菌落形态特征、菌丝体生长速率和致病力均有很大差异,其中以屯昌3、文昌4、琼山1和万宁1等菌系的菌丝生长快,而临高1、昌江3和昌江6则最为缓慢。     

Interactions between Coniothyrium minitans and Sclerotinia minor affect biocontrol efficacy of C. minitans

Coniothyrium minitans, marketed as Contans, has become a standard management tool against Sclerotinia sclerotiorum in a variety of crops, including winter lettuce. However, it has been ineffective against lettuce drop caused by S. minor. The interactions between C. minitans and S minor were investigated to determine the most susceptible stage in culture to attack by C. minitans, and to determine its consistency on S minor isolates belonging to four major mycelial compatibility groups (MCGs). Four isolates of S. minor MCG 1 and 5 each from MCGs 2 and 3 and one from MCG 4 were treated in culture at purely mycelial, a few immature sclerotial, and fully mature sclerotial phases with a conidial suspension of C. minitans. Sclerotia from all treatments were harvested after 4 weeks, air dried, weighed, and plated on potato dextrose agar for recovery of C. minitans. S. minor formed the fewest sclerotia in plates that received C. minitans at the mycelial stage; C. minitans was recovered from nearly all sclerotia from this treatment and sclerotial mortality was total. However, the response of MCGs was inconsistent and variable. Field experiments to determine the efficacy of C. minitans relative to the registered fungicide, Endura, on lettuce drop incidence and soil inoculum dynamics were conducted from 2006 to 2009. All Contans treatments had significantly lower numbers of sclerotia than Endura and unsprayed control treatments, and drop incidence was as low as in Endura-treated plots (P > 0.05). Although the lower levels of lettuce drop in Contans treatments were correlated with significantly lower levels of sclerotia, the lower levels of lettuce drop, despite the presence of higher inoculum in the Endura treatment, was attributable to the prevention of infection by S. minor. A useful approach to sustained lettuce drop management is to employ Contans to lower the number of sclerotia in soil and to apply Endura to prevent S. minor infection within a cropping season.     

Low genetic variability in Sclerotinia sclerotiorum populations from common bean fields in Minas Gerais State,Brazil, at regional,local and micro‐scales

Sclerotinia sclerotiorum, causal agent of white mould, is the most destructive and widely distributed soilborne pathogen of common bean during the autumn–winter season in Brazil. Nevertheless, little is known about the genetic structure of the pathogen population. Microsatellite (SSR) markers and mycelial compatibility groups (MCGs) were used to characterize 118 isolates collected from 20 bean fields located in the most important growing regions of Minas Gerais State (MG). Additionally, the genetic variability among 10 isolates obtained from a single sclerotium was investigated in 10 different sclerotia. Seventy SSR haplotypes and 14 MCGs were identified among the 118 isolates. The genetic differences within bean growing areas accounted for most of the genetic variation (72%). Despite the relatively high genotypic diversity, the SSR loci were at linkage disequilibrium. Moreover, 70% of the isolates were assigned to only two MCGs, and haplotypes of a given MCG were closely related. The discriminant analysis of principal components revealed five groups. There was strong genetic differentiation between isolates collected in one municipality in southern MG when compared to other regions. Common bean resistance to white mould should be assessed with representative isolates of the five genetic groups and, if possible, of the different MCGs detected in the present study. One to five haplotypes were detected among the 10 isolates obtained from a single sclerotium. Therefore, in order to ensure genetic identity of an isolate, hyphal tip or monoascosporic isolates should be used.     

内蒙古和黑龙江的核盘菌菌丝融合群分化及致病性研究

为明确核盘菌的遗传多样性,对采自内蒙古和黑龙江不同地区的44株核盘菌进行了菌丝融合群确定,并比较了不同菌丝融合群间菌丝生长速度、致病力、草酸和总酸产量的差异。结果表明:供试44个菌株分为25个融合群,其中有14个融合群仅由单一菌株组成,所占比例为56.0%。菌丝融合群内和菌丝融合群间菌丝生长速度、致病力、草酸和总酸产量都表现出显著差异(P0.001),并与菌株的地理来源无关。相关分析表明核盘菌菌株的致病力与菌株草酸产量呈正相关(r=0.484,P≤0.01),与pH呈负相关(r=-0.580,P≤0.01),与菌株的生长速度无关;草酸产量与pH高低(表示总酸的分泌量)负相关(r=-0.392,P≤0.01),进一步表明核盘菌菌株产生的总酸中草酸量占了很大的比例。     

Mycelial compatibility groups in Sclerotium cepivorum

Isolates of Sclerotium cepivorum from the Holland–Bradford Marsh, Ontario, Canada ( n  = 146) and other locations ( n  =23) were evaluated for mycelial compatibility in agar culture and by transmitted light microscopy. In compatible interactions, hyphae of paired colonies anastomosed without cell deterioration or death at the point of fusion. In some incompatible interactions, hyphae of paired colonies were initially similar to those of compatible interactions but were later followed by deterioration or death of the fused cells. Incompatible interactions were associated with a red–brown colour that, in mass, produced a dark line at the junction of the two colonies. In other incompatible interactions, hyphae were observed to bypass or repel each other without fusing. Compatible and incompatible reactions were used to define nine mycelial compatibility groups (MCGs) among the isolates examined. Among isolates from Ontario, no incompatibility responses were observed among isolates from individual fields (e.g. no intrafield variability) but two MCGs were found among isolates from different fields (e.g. interfield variability). Among isolates from other regions, seven additional MCGs were detected that consisted of one to eight isolates. Two isolates were variable in reaction and were not assigned to any MCG. Three isolates from Switzerland were compatible with MCG-1 isolates from Ontario; and one isolate from Switzerland, two from England, two from New Zealand, and two from Australia were all intercompatible. The results establish that mycelial compatibility can be used to reflect genetic heterogeneity among isolates of S. cepivorum .     

Molecular,morphological and pathogenic diversity of Sclerotinia sclerotiorum isolates from common bean (Phaseolus vulgaris) fields in Argentina

White mould, caused by Sclerotinia sclerotiorum, is one of the most threatening fungal diseases occurring across major bean production regions worldwide. In Argentina, under favourable weather conditions, up to 100% seed yield losses occur on susceptible common bean cultivars. The aim of this study was to characterize the diversity of S. sclerotiorum isolates from six dry bean fields in the main production area of Argentina by means of molecular, morphological (mycelium colour, number and pattern of sclerotia distribution) and pathogenic approaches. Among 116 isolates analysed, high genotypic and morphological variability was observed. A total of 52 mycelial compatibility groups (MCGs) and 59 URPs (universal rice primers) molecular haplotypes were found. All the MCGs were location specific, while only 12% of the URP haplotypes were shared among locations. The molecular analysis of variance revealed a significant differentiation among populations, with higher genetic variability within the populations analysed than among them. The aggressiveness of the isolates towards bean seedlings was assessed in the greenhouse. Most of the isolates were highly aggressive, while no variation among locations was observed. The information generated in the present study provides, for the first time, information on the variability of S. sclerotiorum associated with white mould in the main common bean production area in Argentina. In addition, the findings suggest the occurrence of both clonal and sexual reproduction in the populations analysed. This work contributes to the development of sustainable management strategies in bean production aimed to minimize yield losses due to white mould.     

Genetic Variability Within and Among Mycelial Compatibility Groups of Sclerotium rolfsii in South Africa

ABSTRACT Isolates of Sclerotium rolfsii, the causal organism of stem rot or southern blight of groundnut, can be placed in mycelial compatibility groups (MCGs) based on hyphal interactions between isolates. The aim of this study was to determine whether amplified fragment length polymorphism (AFLP) analysis was a suitable technique to assess genetic variability between isolates and MCGs of S. rolfsii. For preliminary genetic analysis, 10 isolates were selected from each of two MCGs and compared with each other using the restriction enzymes EcoRI and MseI and 4 primer pairs. The number of polymorphisms ranged from 10 to 36 per primer combination, with an average of 22.5. AFLP analysis clearly showed genotypic differences (22%) among MCGs B and C, with a maximum variation of 6.41% between any two isolates per group using four primer pairs. Certain isolates could not be distinguished from each other. A more in-depth study of 10 isolates from MCG B, using 8 additional primer pairs, showed small genetic differences (maximum of 4.2% and minimum of 0.2%) between isolates. These results suggested that DNA could be pooled for comparison of MCGs. Pooled DNA from isolates within groups using 20 primer pairs confirmed differences between 9 MCGs. This technique effectively differentiated MCGs of S. rolfsii from each other and also detected differences between isolates within a single MCG.     

Analyses of Lettuce Drop Incidence and Population Structure of Sclerotinia sclerotiorum and S. minor

ABSTRACT To understand the geographical distribution of lettuce drop incidence and the structure of Sclerotinia minor and S. sclerotiorum populations, commercial lettuce fields were surveyed in the Salinas, San Joaquin, and Santa Maria Valleys in California. Lettuce drop incidence, pathogen species, and mycelial compatibility groups (MCGs) were determined and analyzed using geostatistic and geographical information system tools. Lettuce drop incidence was lowest in the San Joaquin Valley, and not significantly different between the other two valleys. Semivariogram analysis revealed that lettuce drop incidence was not spatially correlated between different fields in the Salinas Valley, suggesting negligible field-to-field spread or influence of inoculum in one field on other fields. Lettuce drop incidence was significantly lower in fields with a surface drip system than in fields with furrow or sprinkler irrigation systems, suggesting that the surface drip system can be a potential management measure for reducing lettuce drop. In the San Joaquin Valley, S. sclerotiorum was the prevalent species, causing drop in 63.5% of the fields, whereas S. minor also was identified in 25.4% of the fields. In contrast, in the Salinas Valley, S. minor was the dominant species (76.1%) whereas S sclerotiorum only observed in only 13.6% fields, in which only a few plants were infected by S. sclerotiorum. In the Santa Maria Valley, both species frequently were identified, with S. minor being slightly more common. Although many MCGs were identified in S. minor, most of them consisted of only one or two isolates. In all, approximately 91.4% of the isolates belonged to four MCGs. Among them, MCG-1 was the most prevalent group in all three valleys, accounting for 49.8% of total isolates. It was distributed all over the surveyed areas, whereas other MCGs were distributed more or less locally. Populations of S. sclerotiorum exhibited greater diversity, with 89 isolates collected from the Salinas and San Joaquin Valleys belonging to 37 different MCGs. Among them, the most recurrent MCG-A contained 16 isolates, and 30 MCGs contained only 1 isolate each. Many MCGs occurred within only one or a part of the two valleys. Potential reasons for this abundant diversity are discussed.     

Clonality in Sclerotinia sclerotiorum on Infected Cabbage in Eastern North Carolina

ABSTRACT Eighty-four isolates of Sclerotinia sclerotiorum from four cabbage production fields in North Carolina and 16 isolates from an experimental cabbage field plot in Louisiana were DNA-fingerprinted and tested for mycelial compatibility. In a comparison with 594 unique DNA fingerprints of S. sclerotiorum from Canadian canola, no fingerprints were shared among Canadian, North Carolina, and Louisiana populations. DNA fingerprints from the North Carolina sample were distinctive from those of the Canadian and Louisiana samples, with significantly more hybridizing fragments in the 7.7- to 18-kilobase range. Forty-one mycelial compatibility groups (MCGs) and 50 unique DNA fingerprints were identified from the North Carolina sample. Three MCGs and three fingerprints were identified from the Louisiana sample. From the North Carolina sample, 32 MCGs were each associated with a unique fingerprint; of these, there were 11 clones (i.e., cases in which two or more isolates belonged to the same MCG and shared the same DNA fingerprint). Six clones sampled from two or more fields represented approximately 29% of the total sample (24 of 84 isolates), with six clones recovered from fields 75 km apart. There were 10 cases in which one MCG was associated with more than one DNA fingerprint and two cases in which one DNA fingerprint was associated with more than one MCG. The small sample from Louisiana was strictly clonal. The North Carolina sample had a clonal component, but deviated from one-to-one association of MCG with DNA fingerprint to an extent consistent with more recombination or transposition than the other two populations sampled.     

Interaction betweenSclerotinia sclerotiorum andConiothyrium minitans strains with different aggressiveness

The effects ofSclerotinia sclerotiorum live and autoclaved sclerotia, and sclerotial exudates, and commercial oxalic acid were testedin vitro on sevenConiothyrium minitans strains differing in aggressiveness towardsS. sclerotiorum. Only sclerotial exudates and autoclaved sclerotia affected the mycelial growth rate of almost all the strains tested, whereas a change in theC. minitans mycelial growth pattern was observed in the presence of autoclaved sclerotia and live sclerotia germinating by the myceliogenic eruptive germination. In addition, sclerotial exudates had a stimulatory effect on spore germination. These findings indicate that the various treatments could influence theC. minitans strains regardless of their aggressiveness.