Note: Survey of seedborne fungi of sudanese cultivars of onion,with new records

Through incubation of onion (Allium cepa L.) seed samples on potato dextrose agar and moist filter papers at 28±2°C, 32 species and four varieties belonging to 19 genera of fungi were determined as seedborne in this crop. Among them, 23 species are new reports to the mycoflora of onion seeds.Aspergillus (11 species and three varieties, ∼42.1% of the total colony count of fungi) was the most prevalent genus:A. niger (∼19.7%) was found at the highest rate in the seeds, followed byA. parasiticus (∼17.3%,A. oryzae (∼17.3%) andA. flavus (∼14.5%). The genusAspergillus was followed byPenicillium (∼12.4%),Sclerotium (∼7.1%),Fennellia, Rhizopus, Chaetomium, Drechslera andAlternaria (∼12.4%),Fusarium, Emericella andByssochlamys (∼7.1%), whereas the remaining eight genera displayed a low level of infection (∼10%). Twenty-seven species, three varieties and 12 genera represent Hyphomycetes; four genera, three species and one variety — Ascomycotina; one genus and one species — Zygomycotina; two genera and two species — other Deuteromycotina; and one genus — Oomycetes. Some species, which are known to cause devastating pre- and postharvest diseases to onion crops, were recovered from the seeds of this crop, suggesting the high possibility of their transmission by seed. http://www.phytoparasitica.org posting July 14, 2004.     

北京地区主栽西瓜品种种子携带真菌检测及鉴定

用离体平皿法对北京西瓜产区主栽的11个西瓜品种进行种子带菌检测、分离纯化和形态学、ITS序列比对,确定其属或种地位。结果表明:不同品种所带真菌种类有差异,种子表面携带的优势菌群主要为曲霉属Aspergillus spp.、青霉属Penicilliumspp.、链格孢属Alternariaspp.和镰刀菌属Fusariumspp.;种子内部寄藏真菌主要为青霉属、根霉属Rhizopus spp.和曲霉属。种子外部检出的主要病原菌有镰刀菌属的F.verticillioides,F.proliferatum及茎点霉Phomasp.;种子内部检出的病原菌有F.verticillioides和F.oxysporum。不同品种间种子表面携带真菌量与种子内部携带真菌率差异显著,但种子外部带菌量和内部种仁带菌率之间无显著相关性。本研究对开展西瓜种子处理研究有借鉴意义。     

Seedborne fungi detected on stored solanaceous berry seeds and their biological activities

We isolated 629 fungi from 1296 berry seeds of solanaceous plants, including tomato (Lycopersicon esculentum), eggplant (Solanum melongena), bell pepper (Capsicum annuum), and red pepper (Capsicum annuum var. annuum) preserved for long and short terms. The isolates were classified into 22 genera excluding unidentified fungi, and the fungal floras were divided into two types: the tomato–eggplant and pepper groups. The results of cluster analysis with unweighted pair-group method with arithmetic average also supported these groups. Most tomato seeds infested with Geotrichum candidum germinated and grew the same as uninfested seeds. Cladosporium sphaerospermum and Arthrinium sp. isolated from eggplant seeds strongly suppressed germination, and Penicillium variabile suppressed seminal root elongation on eggplant. Alternaria alternata, Botrytis cinerea, and Myrothecium verrucaria detected from red pepper or bell pepper seeds were pathogenic to the fruits and the seedlings after artificial inoculation.     

手掌参内生真菌及根际土壤真菌的群落组成

为筛选促生菌,采用高通量测序技术对手掌参茎、叶、果荚和根4种组织内生真菌及根际土壤真菌的群落组成进行分析。结果表明:从手掌参所有组织样品及根际土壤中共获得4 192个操作分类单元(operational taxonomic unit,OTU),其中茎中OTU数量最多,为1 096个,其次为根际土壤和叶中,分别为1 048个和983个,根中最少,为397个。所有样品中OTU注释后隶属于4门15纲,优势菌群为子囊菌门,相对丰度为47.25%～81.58%。属水平上主要菌群为枝孢霉属Cladosporium、粉褶蕈属Entoloma、背芽突霉属Cadophora和赤霉菌属Gibberella,分别为果荚、土壤、根、叶和茎中优势菌群。多样性指数表明茎中真菌群落的丰富度及多样性最高,而根部最低,不同样品之间真菌群落的多样性和丰富度差异显著。热图分析和主坐标分析均表明茎、叶和果荚中内生真菌的群落组成较相似,但与根部内生真菌和土壤中真菌的群落组成存在一定差异。     

Effects of antagonistic fungi,plant growth-promoting rhizobacteria,and arbuscular mycorrhizal fungi alone and in combination on the reproduction of <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> and growth of tomato

Antagonistic fungi (Aspergillus niger CA and Penicillium chrysogenum CA1), plant growth-promoting rhizobacteria (PGPR) (Burkholderia cepacia 4684 and Bacillus subtilis 7612) and AM fungi (Glomus intraradices KA and Gigaspora margarita AA) were assessed alone and in combination for their effects on the growth of tomato and on the reproduction of Meloidogyne incognita in glasshouse experiments. Application of antagonistic fungus, PGPR, or AM fungus alone or in combination significantly increased the length and shoot dry mass of plants both with and without nematodes. The increase in shoot dry mass caused by Gl. intraradices KA in plants without nematodes was greater than that caused by PGPR or antagonistic fungi. Similarly, the increase in shoot dry mass caused by Bu. cepacia 4684 in plants with nematodes was greater than that caused by P. chrysogenum CA1. Application of Bu. cepacia 4684 caused a 36.1% increase in shoot dry mass of nematode-inoculated plants followed by Ba. subtilis 7612 (32.4%), A. niger CA (31.7%), Gl. intraradices KA (30.9%), Gi. margarita AA (29.9%) and P. chrysogenum CA1 (28.8%). Use of Bu. cepacia 4684 with A. niger CA caused a highest (65.7%) increase in shoot dry mass in nematode-inoculated plants followed by Ba. subtilis 7612 plus A. niger CA (60.9%). Burkholderia cepacia 4684 greatly reduced (39%) galling and nematode multiplication, and the reduction was even greater (73%) when applied with A. niger CA. Antagonistic fungi had no significant effect on root colonization caused by AM fungi. Applying Bu. cepacia 4684 with A. niger CA may be useful in the biocontrol of M. incognita on tomato.     

Determination of seedborne fungi in Onion and their transmission to Onion sets

Samples of onion (Allium cepa L.) seeds were obtained from seven regions in Turkey. The seed coat, embryo and endosperm were cultured, the seedborne fungi were determined and their transmission to onion sets was investigated in both sterile and field soils. Among the fungi determined,Aspergillus alutaceus Berk, and Curt.,Beauveria bassiana (Bals.) Vuill.,Cladosporium cladosporioides (Fres.) de Vries,Geotrichum sp.,Humicola fuscoatra Traaen,Trichoderma harzianum Rifai andT. pseudokoningii Rifai in onion seeds, andFusarium culmorum (W.G.Sm.) Sacc,F. graminearum Schwabe andF. sambucinum Fuckel in onion sets, were recorded for the first time.Aspergillus niger v. Tieghem was found at the highest rate in seed samples (especially in the seed coat), and in bulbs and roots of onion sets that developed from these seeds, whether in sterile or field soil.Fusarium oxysporum Schlecht was isolated at a higher rate from onion sets grown in sterile or field soil, than from seeds.F. acuminatum Ellis and Everhart,F. sambucinum, F. equiseti (Corda) Sacc. andF. graminearum were isolated only from onion sets grown in sterile soil. In dual culture tests, theseFusarium isolates were inhibited byA. niger and thus, exceptfor F. oxysporum, could not develop in agar plate. TheFusarium spp. appeared in onion-sets grown in sterile soil and were inhibited by other fungi in field soil. It was concluded that all fungi were seedborne and thatA. niger andFusarium spp., but not the other fungi, were transmitted from the seeds to onion sets.A. niger andF. oxysporum were also transmitted through the soil.     

Seed transmission ofPyrenophora tritici-repentis,causal fungus of tan spot of wheat

Seed transmission ofPyrenophora tritici-repentis, a common foliar pathogen of wheat, was investigated in soft white winter wheat cv. Frankenmuth and found to be non-systemic; the emerging coleoptile was infected externally by hyphal growth from the infected pericarp. Hyphae from the infected coleoptile then infected the first and second seedling leaves as they emerged. Coleoptile symptoms ranged from tiny brown streaks or spots to large, brown necrotic areas accompanied by cracking and distortion of the coleoptile. Small brown spots sometimes occurred on the first and rarely on the second seedling leaves, often accompanied by leaf distortion. Pseudothecial initials of the fungus were present within or on the seed remnants. Seed transmission efficiency was as high as 92%in vitro and 60% in potting soil outdoors. Seed infection did not affect germinationin vitro, but slightly reduced emergence in potting soil. Seedling weight and height were reduced significantly. In potting soil, seed infection also resulted in delayed plant growth and increased tan spot severity at later stages of plant development. Under controlled conditions, seed transmission efficiency and incidence of pseudothecia on seed were negatively correlated with seed germination temperature in the range of 9 to 21 °C, whereas frequency of recovery of the fungus from symptomatic coleoptiles and leaves was positively correlated with seed germination temperature.These results suggest that infected seed may serve as a source of inoculum for tan spot epidemics and for dispersal of strains of the fungus to new areas.     

浙江省铁皮石斛根腐病病原真菌的鉴定

为明确铁皮石斛根腐病病原真菌,于其主产地浙江省金华市武义县收集铁皮石斛根腐病病株,采用平板分离方法对病原真菌进行分离,使用镰刀菌种特异性引物并结合ITS和TEF序列分析及形态学鉴定确定该镰刀菌的分类地位。结果表明,共分离纯获得真菌117株,其中有105株镰刀菌;经分子生物学分析及形态学鉴定结果显示,分离出的镰刀菌为层出镰刀菌Fusarium prolife‐mum、茄病镰刀菌F. solani、尖孢镰刀菌F. oxysporum和厚垣镰刀菌F. chlamydosporum四个种,其中层出镰刀菌在数量上具有优势地位,占总镰刀菌数的44.8%;茄病镰刀菌、尖孢镰刀菌、厚垣镰刀菌分别占总镰刀菌数的21.0%、15.2%和19.0%。在致病性测定中发现层出镰刀菌和茄病镰刀菌并不具备致病性,尖孢镰刀菌的致病性明显弱于厚垣镰刀菌,表明厚垣镰刀菌为浙江省金华市武义县铁皮石斛根腐病的主要致病菌。     

A selective medium for the detection ofAlternaria dauci andAlternaria radicina

A selective medium to detectAlternaria dauci andA. radicina on seed, plant debris and other substrates was developed. Growth and sporulation by most undesired organisms was reduced, but adequate mycelial growth and sporulation byA. dauci andA. radicina was maintained so they could be identified by their unusual mycelial growth or characteristic spores. The medium is based upon carrot leaf extract, which promotes profuse sporulation by both pathogens. Glucose, sodium polypectate and mineral salts further enhanced spore and mycelial production. Streptomycin sulfate and metalaxyl (or mefenoxam), combined with either benomyl or thiophanate-methyl, reduced growth and sporulation of unwanted organisms. There were strong interactions among the fungicides, bactericide, and most other medium components. There were also significant effects on the type of mycelial growth produced by some medium components. If the unique, dark mycelia produced by both pathogens is the desired detection method, the medium components must be optimized for eitherA. dauci orA. radicina because the two fungi responded differently in this regard. All medium configurations allowed both fungi to be identified by their characteristic spores. The sensitivity of theAlternaria dauci-radicina selective medium (ADRSM) to detectA. radicina on carrot seeds was similar to other methods presently in use, but ADRSM was more sensitive than the other methods for detectingA. dauci on infested carrot seeds.     

印度梨形孢真菌诱导提高花生抗旱性研究初报

将印度梨形孢真菌(Piriformospora indica)接种于受不同干旱胁迫花生幼苗根部,研究接种该真菌对花生抗旱性的影响.试验设置3个处理,分别是:接菌+干旱胁迫处理、未接菌+干旱胁迫处理、未接菌+未干旱胁迫处理(CK).试验结果表明:接菌+干旱处理后的花生主根长、根鲜重、根干重、茎鲜重、茎干重、叶鲜重、叶干重、叶绿素含量(SPAD值)等农艺性状表现分别为14.96 cm、0.64g、0.08 g、1.48 g、0.20 g、1.61 g、0.29 g、42.63.与未接菌+干旱处理相比均显著增加,增幅分别为26.8％、25.50％、60％、15.63％、17.65％、15.59％、26.09％、7.43％.与对照相比,在主根长、根鲜重、根干重、茎干重、叶干重、叶绿素含量(SPAD值)等性状方面均没有显著性变化.以上结果说明:在干旱胁迫条件下,接种印度梨形孢真菌能够维持花生正常生长,使得花生从植株外部形态上表现出较强的抗旱性.     

Pathogenesis by fungi and by parasitic plants: Similarities and differences

The processes by which fungi and parasitic plants infect their plant host are compared. The steps considered are: location of the host by the pathogen, the concept of the haustorium, attachment of the pathogen to the host, mechanism of penetration into the host, responses of the host to infection by fungi and by parasitic plants, and the suppression of the host response by the pathogen. Although superficially similarities between fungal infection and infection by parasitic plants exist, the underlying mechanisms appear to be different and only in the penetration step are similar strategies used, including the use of lytic enzymes. These differences are important when seeking ways of combating the parasitic plants. The strategies used against the parasitic plants must be different from those used to combat fungal infections and should not be based only on analogies with host resistance to fungal pathogens. http://www.phytoparasitica.org posting Nov. 2, 2005.     

Suppression of rice blast by phylloplane fungi isolated from rice plants

Rice phylloplane fungi were evaluated for their potential as biocontrol agents for rice blast disease caused by Magnaporthe grisea. A total of 1923 fungal isolates were obtained from rice plants in fields at Ishigaki and Iwama and from potted plants placed in a cedar woods in Iwama as bait. Although 82.9% of isolated fungi could not be identified, species of Epicoccum were the most prevalent among identified isolates. Of the 1923 isolates, 967 were randomly selected for screening against rice leaf blast. Nine isolates (MKP5111B, MKP5112, J2JMR3-2, K2J131-2, I5R3-1, NOP541, K1KM134-1, NOP5112, MKP33222) suppressed the disease when a conidial or hyphal suspension of both the phylloplane fungus and pathogen were simultaneously used to inoculate rice plants cultured in pots in a growth chamber. Five of the isolates originated from potted plants in the woods and four from Ishigaki, a subtropical island. Five (MKP5111B, MKP5112, NOP541, NOP5112, MKP33222) of the nine isolates strongly suppressed conidial germination of M. grisea (0.7%) and formed inhibition zones (3–5mm width) in dual cultures with the pathogen. Methanol extracts from the isolates also inhibited mycelial growth of the pathogen. These results suggest that the five isolates produced antibiotic(s). These five isolates are likely identical or closely related fungal species because the sequence of their ITS regions were 100% similar. ITS sequence analysis also suggested that J2JMR3-2 was associated with a species of Fusarium. Under field conditions, J2JMR3-2 reduced both leaf and panicle blast severity, and three other isolates (MKP5111B, K1KM134-1, K2J131-2) suppressed leaf blast in one of the three experiments.     

Identification of Trichoderma SKT-1, a biological control agent against seedborne pathogens of rice

Trichoderma SKT-1 was previously reported as a powerful biological control agent against seedborne pathogens of rice, but the taxonomic disposition of the fungal isolate was not clear. Trichoderma SKT-1 produced irregular pyramidal warts on conidia and had an optimum growth temperature of 30°C. Morphological characteristics and colony growth were identical to those of known species of Trichoderma, including the newly recognized species T. asperellum. The 5.8S rDNA with the internal transcribed spacer (ITS) region (ca. 514 bp) of the fungus was compared with those of known species to determine the phylogenetic placement of the fungus. The length and sequence of the regions from Trichoderma SKT-1 were completely identical to those of an isolate of T. asperellum NRRL 5242 (AJ230669). On the basis of these results, we concluded that Trichoderma SKT-1 was T. asperellum.     

雷帕霉素对4种植物病原真菌的抑菌活性

采用菌丝生长速率法,测定了雷帕霉素对番茄灰霉病菌、油菜菌核病菌、水稻纹枯病菌和棉花枯萎病菌的抑菌活性,比较了嘧菌酯、丙烷脒及雷帕霉素对番茄灰霉病菌的抑菌效果,并通过电子显微镜观察了雷帕霉素对番茄灰霉病菌菌丝生长的影响。结果表明:雷帕霉素对供试4种植物病原真菌菌丝均表现出了极强的抑制活性,其中对油菜菌核病菌的抑制作用最强,EC50值为2.23×10-4μg/mL,对番茄灰霉病菌、水稻纹枯病菌和棉花枯萎病菌的EC50值分别为1.32×10-3、4.05×10-3及3.82×10-3μg/mL;雷帕霉素对番茄灰霉病菌菌丝的抑制活性显著高于嘧菌酯(EC50值为3.24μg/mL)和丙烷脒(EC50值为3.81μg/mL)。电镜观察发现,经雷帕霉素处理后,番茄灰霉病菌菌丝表现出提前衰老等症状。研究结果可为深入探讨雷帕霉素对植物病原真菌的作用机制奠定基础。     

白僵菌及其伴生菌发酵液对线虫的毒力研究

采用发酵液浸没法,以4种线虫作为靶标,对6株白僵菌、7株伴生菌菌株进行了杀线虫活性筛选,以期获得新的、高效的杀线虫生防菌资源。其中:处理24 h对南方根结线虫Meloidogyne incognita 2龄幼虫及大豆胞囊线虫Heterodera glycines 2龄幼虫校正死亡率均达80%以上的菌株仅有白僵菌Snef2598及伴生菌Snef5;供试菌株对水稻干尖线虫Aphelenchoides besseyi的毒力均较低,其中仅有1株白僵菌校正死亡率达到40%以上。处理48 h对南方根结线虫2龄幼虫校正死亡率达90%以上的菌株有白僵菌4株和伴生菌5株;对大豆胞囊线虫2龄幼虫校正死亡率超过90%的菌株有白僵菌3株和伴生菌1株;供试菌株对水稻干尖线虫毒力均较低,校正死亡率达60%以上的菌株有白僵菌3株和伴生菌2株;几乎所有供试菌株,对小杆线虫Caenorhabditis sp.的毒力均很高,处理24 h及48 h的校正死亡率均达97%以上,且被杀死的小杆线虫体壁以及虫体内部物质均被不同程度地消解。研究表明,球孢白僵菌菌株Snef2598及其伴生菌菌株Snef5对4种靶标线虫的毒力均较好,具有进一步研究的价值。     

Pathogenic fungi involved in root rot of peas in the Netherlands and their physiological specialization

Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.     

两株虫生真菌对草地贪夜蛾的致病力及高毒力菌株与卵寄生蜂的相容性

为有效防控新入侵我国的重大农业害虫草地贪夜蛾 Spodoptera frugiperda,采用浸叶法测定球孢白僵菌 Beauveria bassiana菌株Bb1237、 Bb201017、 Bb20091317及金龟子绿僵菌 Metarhiziumanisopliae菌株Ma189和莱氏绿僵菌 M. rileyi菌株MrCDTLJ1对草地贪夜蛾3龄幼虫的致病力,筛选对草地贪夜蛾3龄幼虫有较强致病力的菌株,并测定该菌株对草地贪夜蛾3龄幼虫的致病力和对其天敌的致死率。结果显示,球孢白僵菌Bb20091317和莱氏绿僵菌MrCDTLJ1对草地贪夜蛾3龄幼虫的累计校正死亡率显著高于其他菌株,分别为95.59%和97.06%,致死中时LT₅₀分别为3.42 d和5.72 d, LC₅₀分别为7.05×10⁶个/mL和2.25×10⁵个/mL。2种菌在高浓度1×10⁸个/mL下对草地贪夜蛾天敌夜蛾黑卵蜂 Telenomus remus和玉米螟赤眼蜂 Trichogramma ostriniae成蜂的致死率也仅介于15%~20%之间。表明球孢白僵菌Bb20091317和莱氏绿僵菌MrCDTLJ1不仅对草地贪夜蛾有高致病力,同时与夜蛾黑卵蜂和玉米螟赤眼蜂有较好的相容性。     

天山野果林塞威士苹果和短距凤仙花叶斑病病原真菌的鉴定

为了解新疆天山野果林中塞威士苹果Malus sieversii与其林下伴生植物短距凤仙花Impatiens brachycentra两种植物叶斑病病原菌的多样性及同源性,采用组织分离法获得病原菌,基于rDNA-ITS序列构建系统发育树,并进行ITS序列BLAST同源性比对,对病原菌进行鉴定分类,并依据科赫氏法则测定致病性。结果显示,从新疆新源县天山野果林中的塞威士苹果及短距凤仙花病叶上共分离得到18株菌落形态各异的病原菌,分属于2属4种,绝大多数属于半知菌亚门。其中,链格孢属Alternaria sp.为塞威士苹果和短距凤仙花叶斑病病原真菌中的优势菌群。致病性测定结果显示,其中11株病原真菌对塞威士苹果具有致病性,7株病原真菌对短距凤仙花具有致病性;塞威士苹果所有的病原真菌对短距凤仙花均有致病性,而且短距凤仙花所有的病原真菌对塞威士苹果也有致病性。推测2种植物叶斑病可能由相同来源的病原真菌引发,短距凤仙花染病加剧了塞威士苹果叶斑病暴发,可能是野果林退化的重要原因。     

Comparison of immunofluorescence microscopy and dilution-plating for the detection of Xanthomonas campestris pv. campestris in crucifer seeds

The correlation between immunofluorescence microscopy (IF) and dilution-plating on nutrient starch cycloheximide agar (NSCA) or NSCA with the addition of nitrofurantoin and vancomycin (NSCAA) was studied for the detection ofXanthomonas campestris pv.campestris (Xcc) in crucifer seeds. When checking 50 l of the seed extract in IF, IF and dilution-plating gave corresponding results (both positive or negative) for 45.4–56.4% of the samples tested. No differences were observed in this respect between tests using a polyclonal antiserum (PCA 94) and replicate tests using monoclonal antibodies (MCA 20H6). When 20 l of the seed extract was checked in IF, 67.3–71.3% of the samples tested were both positive or negative with dilutionplating and IF. IF negative and dilution-plating positive samples were found for 0.0–7.3% of all samples tested. The percentage of IF positive and dilution-plating negative samples ranged from 26.7–29.2 (20 l seed extract checked) to 41.8–47.3% (50 l seed extract checked). Generally, the probability of isolating Xcc increased with increasing numbers of fluorescent cells found in IF. Above 10 000 cells per ml the probability of isolating Xcc ranged from 57.1–81.8%. Increasing the extraction time from 5 min to 2.5 h shaking showed no significant increase of the number of samples found positive in IF and dilution-plating. However, when using both 5 min and 2.5 h shaking as compared to 5 min shaking only, more samples can be found positive in IF (1.0–14.5%) and dilution-plating (3.0–18.5%). Examining 1 l instead of 50 l of the sample smear, would increase the correspondence between IF and dilution-plating results up to minimally 69.1% (MCA 20H6). However, the risk of false-negative results in IF as compared to dilution-plating would also increase.     

几种进口花卉种子寄藏真菌的检测与鉴定

采用分离培养检验法分别对8种进口花卉种子共计14个批次的种子样品进行了种子奇藏真菌的检测、纯化和鉴定。研究结果表明，每种样品均带菌，共检测出进口花卉种子寄藏真菌13属24种，链格孢菌、枝孢菌、镰孢菌、毛壳菌是这些进口花卉种子寄藏真菌的优势菌群。