西藏青稞籽粒燕麦镰刀菌生物学特性研究

以西藏青稞籽粒中分离到的燕麦镰孢(Fusarium avenaceum)Hb39分离株为对象,研究了温度、湿度、pH、光照、营养对其菌丝生长和孢子萌发的影响,以便为进一步研究由燕麦镰孢引起的青稞病害发生规律、防治方法奠定基础,并为研究由此引致的人类疾病提供理论依据.该菌株在供试的10种不同固体、液体培养基上均能生长,但菌丝生长速度和生长量均有很大差异,PDBA、PDA、PSA培养基有利于该菌的生长,而HA、CMA和WA不利于该菌的生长.比较不同碳、氮源对菌丝生长的影响,发现该菌能利用多种单糖、多糖及可溶性淀粉作碳源,其中葡萄糖、蔗糖、麦芽糖较好;能利用蛋白胨等有机氮和硝酸钠等无机氮作氮源,其中蛋白胨最好.该菌最适生长温度为25℃,生长的温度范围为5～30℃.菌丝生长最适pH为6,也能在pH 3～12的范围内生长.连续光照不利于菌丝的生长,而光暗交替或黑暗条件下菌落生长无显著性差异.大型分生孢子萌发的温湿度范围分别为10～30℃、RH 85%～100%,pH值3～12,最适萌发条件为25℃、RH100%和pH 6.该菌株的致死温度为45℃10 min.     

Refinement of PCR-detection of Fusarium avenaceum and evidence from DNA marker studies for phenetic relatedness to Fusarium tricinctum

Existing PCR-based assays for the detection of the cereal pathogen Fusarium avenaceum were found to cross-react with F. tricinctum . An investigation into the phenetic relationship between these two species using two different marker systems revealed a close relationship between them despite their being from separate taxonomic sections. Whilst RFLP differences in the ITS regions surrounding the nuclear 5.8 S rDNA were insufficient to discriminate between isolates of the two species, they were clearly differentiated by RAPD profiling. RAPD fragments from F. avenaceum isolates were screened for hybridization to F. tricinctum DNA on Southern blots. One of 12 selected RAPD fragments showed no hybridization to genomic DNA from F. tricinctum . This fragment was cloned and sequenced, and the sequence obtained was used to design PCR primers. The primers were found to be specific for F. avenaceum, with no cross-reactions obtained with F. tricinctum or any other wheat pathogen assayed. The primers were able to differentiate between the two species in infected plant material, in contrast to the earlier assays.     

Scab Response and Moniliformin Accumulation in Kernels of Oat Genotypes Inoculated with Fusarium avenaceum in Poland

Inoculation experiments with 14 genotypes of oats (10 cultivars and 4 lines) were performed during 1996, 1997 and 1998 in Sitaniec, South-Eastern Poland. Panicles of oats were inoculated with a conidial suspension of Fusarium avenaceum, which caused a reduction in yield by 33% and in 1000 kernel weight (TKW) by 21%. During the period between inoculation and harvest, F. avenaceum was able to accumulate moniliformin (MON) in kernels at an average level of 0.13mgkg^–1 (gg^–1). The highest reduction of yield components caused by the F. avenaceum inoculation was found for cv. Santor, followed by lines CHD 1171, STH 2795 and cvs: Kwant and Farys, while cvs Slawko, Dukat, Borys and Komes exhibited the highest resistance to the disease in terms of TKW and yield reductions after inoculation.     

Germination of Fusarium oxysporum in root exudates from tomato plants challenged with different Fusarium oxysporum strains

The response of microconidia from pathogenic and non-pathogenic Fusarium oxysporum to root exudates from tomato plants inoculated with different pathogenic and non-pathogenic F. oxysporum strains was studied. Root exudates from non-inoculated tomatoes highly stimulated the microconidial germination of the two tomato pathogens, F. oxysporum f.sp. lycopersici strain Fol 007 and F. oxysporum f.sp. radicis-lycopersici strain Forl 101587. In root exudates from tomato plants challenged with the pathogen Fol 007 the microconidial germination of Fol 007 was increased, whereas in root exudates from plants challenged with Forl 101587 the microconidial germination of Fol 007 was reduced. Root exudates of tomato plants challenged with the non-pathogenic unspecific F. oxysporum strain Fo 135 and the biocontrol strain Fo 47 clearly reduced microconidial germination of the pathogenic strain Forl 101587. Moreover, the microconidial germination rate of the biocontrol strain Fo 47 was increased in the presence of root exudates of tomato plants challenged with the tomato wilt pathogen Fol 007. These results indicate that pathogenic and non-pathogenic F. oxysporum strains alter the root exudation of tomato plants differently and consequently the fungal propagation of pathogenic and non-pathogenic F. oxysporum strains in the rhizosphere is affected differently.     

Occurrence and Pathogenicity of Fusarium Species in Cereals in Finland1

Several Fusarium species occurred abundantly in spring cereal seed samples studied in 1966, 1968 and 1972 in Finland. A total of 17 species of Fusarium were isolated and identified. The most frequently isolated species were F. avenaceum (Fr.) Sacc., F. culmorum (W. G. Sm.) Sacc., F. poae (Peck) Wr. and F. tricinctum (Cda) Sacc. These species were found in more than 40% of the seed samples from 1972. The same 4 Fusarium species in addition to F. graminearum Schwabe were used in the pathogenicity test. Artificially inoculated spring wheat and barley seeds were grown in the field during the growing season of 1973. Of these 5 species F. culmorum was clearly the most pathogenic. This species significantly lowered the shooting percentage and the yield of both cereals. Foot rot assessment also revealed F. culmorum as the most damaging species. Natural soil-borne infection obviously increased the foot and root rot, and thus the soil-borne F. culmorum strongly affected all treatments. In the harvested seed F. poae dominated in all treatments, but F. culmorum still occurred most abundantly in the seed lot harvested from plots originally inoculated with this species.     

瓜类尖孢镰刀菌的营养体亲和群研究

 应用营养亲和性方法研究了尖孢镰刀菌菌株抗氯酸盐突变体和nit突变体的诱发规律及分布特性,以及菌株营养体亲和群(VCG)的划分。研究表明,不同寄主(黄瓜、甜瓜和西瓜)分离的尖孢镰刀菌菌株形成的抗氯酸盐突变体数目差异不显著,平均为每个接种点产生0.89~0.98个;但寄主不同部位(根部、茎基部和茎中部)分离的菌株间差异显著,形成的数目分别为1.27、0.75及0.76个。菌株产生的nit1突变体比例(75.40%)显著高于nitM突变体比例(13.17%);nit1突变体数目会因菌株的寄主及菌株寄主部位的不同而有差异,寄主为黄瓜、甜瓜和西瓜的菌株产生的比例依次为67.73%、83.71%和77.50%,根部、茎基部及茎中部分离菌株产生的比例依次为81.82%、78.48%和68.64%,而在致病菌株与非致病菌株间无显著差异,分别为74.43%和79.63%;nitM突变体数目受菌株寄主影响较小,所占比例在11.17%~13.92%之间;而在寄主不同部位分离的菌株及致病菌株与非致病菌株间差异显著,分离自茎基部的菌株所占比例最高为15.97%,茎中部菌株所占比例最低为9.87%,致病菌株与非致病菌株所占比例分别为14.08%和9.26%。供试菌株分为7个VCGs,其特点为来源于不同寄主的尖孢镰刀菌菌株互不亲和,同一寄主的致病菌株与非致病菌株均不亲和,同一寄主不同部位分离的菌株可亲和。     

Molecular and Morphological Diversity of Fusarium Species in Finland and North-Western Russia

In 2001 the range of the total Fusarium contamination percentage of infected seeds was between 0% and 44%, while in 2002 the contamination level was 2–25% in naturally infected Finnish samples and 5–14.5% in six samples from northwestern Russia. The most common Fusarium species in barley were F. avenaceum, F. arthrosporioides, F. sporotrichioides and F. culmorum, while in spring wheat the most common Fusarium species were F. avenaceum, F. arthrosporioides, F. culmorum, F. sporotrichioides and F. graminearum. In most cases, molecular identification with species-specific primers corresponded to the morphological analyses and allowed the identification of degenerated and otherwise morphologically difficult cultures. It was even possible to separate most of the F. arthrosporioides isolates from Finland from the closely-related F. avenaceum isolates. In the phylogenetic analysis of combined β-tubulin, IGS and ITS sequences most European F. arthrosporioides formed a separate clade from most isolates of F. avenaceum and from all isolates of F. tricinctum. Most of the species-specific primers also amplified DNA extracted from grain samples. It was, for instance, possible to detect F. avenaceum in all barley samples with contamination levels higher than 1% and in all spring wheat samples with contamination levels higher than 3%. The detection level for F. graminearum was at a contamination level of 3–5% and that for F. culmorum at a contamination level of 1–5%. In addition, the first Finnish F. langsethiae isolate was found by means of species-specific primers.     

Molecular and symptom analyses of phytoplasma strains from lettuce reveal a diverse population

ABSTRACT Epidemics of aster yellows in lettuce in Ohio are caused by at least seven distinct phytoplasma strains in the aster yellows (AY) group. Five of the strains are newly reported: AY-BW, AY-WB, AY-BD3, AY-SS, and AY-SG. All seven strains were characterized based on symptoms in aster and lettuce, and by polymerase chain reaction (PCR). Strain AY-BD2 (formerly 'Bolt') causes yellowing and leaf distortion in lettuce and bolting in aster, whereas strain AY-S (formerly 'Severe') causes stunting, leaf clustering, and phyllody. Strain AY-WB causes yellowing and wilting in lettuce and witches'-broom in aster. Strain AY-SG induces horizontal growth in lettuce and aster plants. Strain AY-BW causes chlorosis of emerging leaves and abnormally upright growth of leaf petioles. AY-SS causes symptoms similar to those caused by AY-S but has a different PCR-restriction fragment length polymorphism (RFLP) banding pattern. Strains AY-BD2 and AY-BD-3 cause mild leaf and stem distortion in lettuce but are differentiated by PCR-RFLP. All phytoplasma strains collected from lettuce in Ohio belong to the 16SrI group. AY-WB belongs to the 16SrI-A subgroup and the other six belong to the 16SrI-B subgroup. Five of the seven strains were distinguished from each other by primer typing. The results of phylogenetic analyses of sequences of the 16S rRNA genes were basically consistent with the classification based on PCR-RFLP, in which AY-WB clustered with phytoplasmas of the 16rIA subgroup and the other Ohio lettuce strains clustered with phytoplasmas in the 16SrI-B subgroup.     

High concentrations of antagonistic bacterial strains from diseased sanqi ginseng rhizosphere suppressed Fusarium root rot

European Journal of Plant Pathology - Sanqi ginseng Fusarium root rot caused by Fusarium oxysporum, is a serious soil-borne disease that threatens the sustainable development of the sanqi ginseng...     

Symptoms and yield reduction caused by Dickeya spp. strains isolated from potato and river water in Finland

Biochemical characterisation of Dickeya strains isolated from potato plants and river water samples in Finland showed that the majority of the strains were biovar 3. They thus resembled the strains recently isolated from potato in the Netherlands, Poland and Israel and form a new clade within the Dickeya genus. About half of the Finnish isolates resembling strains within this new clade were virulent and caused wilting, necrotic lesions and rotting of leaves and stems. Similar symptoms were caused by D. dianthicola strains isolated from one potato sample and from several river water samples. Frequently, the rotting caused by the Dickeya strains was visible in the upper parts of the stem, while the stem base was necrotic from the pith but hard and green on the outside, resulting in symptoms quite different from the blackleg caused by Pectobacterium atrosepticum. The presence of Dickeya in the symptomatic plants in the field assay was verified with a conventional PCR and with a real-time PCR test developed for the purpose. The virulent Dickeya strains reduced the yield of individual plants by up to 50% and caused rotting of the daughter tubers in the field and in storage. Management of Dickeya spp. in the potato production chain requires awareness of the symptoms and extensive knowledge about the epidemiology of the disease.     

Diversity and pathogenicity of Fusarium graminearum species complex from maize stalk and ear rot strains in northeast China

The Fusarium graminearum species complex (FGSC) is an important group of pathogens distributed in maize‐producing areas worldwide. This study investigated the genetic diversity and pathogenicity of 40 FGSC isolates obtained from stalk rot and ear rot samples collected from 42 locations in northeastern China during 2013 and 2014. A phylogenetic tree of translation elongation factor (EF‐la) sequences designated the 40 isolates as F. graminearum sensu stricto (67.5%) and F. boothii (32.5%). By using inter‐simple sequence repeat analysis (ISSR), it was shown that the isolates were divided into two clades, which corresponded to the species identity of the isolates. However, the isolates from the two different diseases could not be distinguished in pathogenicity. The disease severity index of seedlings inoculated with stalk isolates was slightly higher than that of seedlings inoculated with isolates from infected ears, whereas the pathogenicity of the stalk and ear isolates were identical.     

利用伏马毒素合成基因对玉米种子寄藏镰刀菌的分子检测

我国玉米种子普遍受到拟轮生镰刀菌(Fusarium verticillioides)的侵染,该菌产生的伏马毒素对国家种质库中玉米种质的安全保存有着潜在的影响。fum1基因是F.verticillioides等镰刀菌中伏马毒素生物合成的必需基因。选用3对已知引物Fum5F/Fum5R、P1/P2和P3/P4分别对从玉米种子中分离出的9株镰刀菌和1株阴性对照菌链格孢菌进行PCR扩增检测,在F001和F003菌株中扩增出相应的特异性表达片段。测序表明,这些片段的长度分别为846bp、888bp和703bp,与已知fum1基因(AF155773)的同源性达99%以上。3对引物的检测灵敏度达到88pg/mL。结果表明,这3对引物都能够有效地检测产生伏马毒素的镰刀菌菌株,但引物Fum5F/Fum5R所扩增片段完全来自fum1基因内部,具有更强的特异性。在此定性检测基础上,需要进一步研究定量检测方法并进一步研究伏马毒素对农作物种质保存的影响。     

Characterization and fitness of carbendazim-resistant strains of Fusarium graminearum (wheat scab)

BACKGROUND: Carbendazim (MBC) has failed to control wheat scab, caused by Fusarium graminearum Schwabe, on the eastern coast of China in recent years after about 30 years of application. RESULTS: MBC resistance was found to be common in pathogen populations on the eastern coast and along areas of the Yangtze River. EC(50) and minimum inhibitive concentration (MIC) values of MBC inhibiting mycelium growth of wild-type isolates were less than 0.9 and 1.4 microg mL(-1) respectively, while EC(50) values of resistant collections averaged 7.02 +/- 11.86 microg mL(-1). The slope of the MBC dosage-response curve (DRC) for resistant isolates of F. graminearum was flat: 1 < b < 2.8 for resistant isolates and 3.5 < b < 11 for sensitive isolates). Both field resistant and sensitive MBC strains shared similar temperature sensitivity, fitness and virulence on ears. Field resistant strains and UV-induced mutants showed positive cross-resistance to other benzimidazole derivatives and were mainly at intermediate MBC resistance level. Highly resistant field MBC strains rarely appeared, but only some of the highly resistant MBC UV mutants were insensitive to N-phenylaminecarbamates. No mutation in beta-tubulin was found in F. graminearum, in contrast to mutation in this tubulin which has led to MBC resistance in other plant pathogens. CONCLUSION: MBC(R) isolates have high fitness and competition in field, conferred by a novel molecular mechanism.     

小尺度空间果生刺盘孢遗传多样性分析

 本研究将来自16个油茶品种的247个Colletotrichum fructicola的多基因序列进行遗传多样性分析,结果表明,247个C. fructicola多基因序列有21个单倍型,其中Hap2占总数的54.7%,Hap4占总数的34%,为两个主要的单倍型,来自所有16个油茶品种。基本上每一个油茶品种都存在C. fructicola独特的单倍型。遗传分化系数(Fst)表明不同品种来源菌株种群间存在一定的遗传分化。对所有品种种群进行中性检测及核苷酸不配对分析,发现小尺度空间的果生次盘孢菌经历大规模的种群扩张,群体间存在有效基因流。系统发育分析表明,来自不同品种的菌株散乱地分布在系统发育树中。     

Novel primers developed from mitochondrial intergenic spacers are useful for multi-locus sequence typing of Fusarium oxysporum strains

The Fusarium oxysporum species complex causes serious vascular diseases of a wide range of crops. In an effort to locate new highly variable regions for multi-locus sequence typing (MLST) of Fusarium oxysporum populations, mitochondrial intergenic regions were targeted. Five regions were amplified and sequenced from five randomly selected isolates. For these isolates, all regions contained at least as many variable sites as the mitochondrial ribosomal RNA small subunit region commonly used in MLST. Most regions contained short indels; however, 80–120 base indels were present in the nad5-arg2 intergenic region. Only the nad5-arg2 region was found to be more variable than the single copy nuclear translation elongation factor (1α) gene introns. The nad5-arg2 region is likely to be a useful inclusion in MLST of Fusarium oxysporum populations.     

Variability amongst strains of Fusarium poae assessed by vegetative compatibility and RAPD polymorphism

Vegetative compatibility tests and random amplification of polymorphic DNA (RAPD) were used to assess genetic relationships amongst 54 strains of Fusarium poae obtained from various geographical regions. Twenty-seven strains were assigned to eight multiple member vegetative compatibility groups (VCGs), while the other 27 isolates were found to form single-member VCGs. There was a partial correlation between VCG and geographical origin, but the relationship was not always clear. However, no correlation was observed between the VCG and the host plant of origin. RAPD patterns were closely associated with VCGs in all cases. Members of VCGs that were interconnected by bridging isolates formed common branches in the phenogram constructed on the basis of the RAPD patterns, while strains that belonged to single-member VCGs were scattered throughout the phenogram. These data demonstrate that the combination of traditional and molecular methodologies allows reliable intraspecific subdivisions in an asexual fungus, which is a secondary invader of a wide range of host plants, and so has never been subject to the intense selection pressure of a single host species and lacks pathogenic subgroups.