营养物质对两种炭疽菌潜伏侵染的影响

通过不同浓度的几种糖类对潜伏侵染在青香蕉果实中的colletotrichum musae和芒果果实中的colletotichum gloeosporioides的菌体的影响进行了测定,结果表明,高浓度淀粉可极显著地提高两种炭疽菌的孢子萌发率,并有利于附着胞和分生孢子的形成。单糖和二糖在较低浓度时有利于孢子萌发和产孢,不利于附着胞形成。未成熟果实的坚硬结构和高淀粉含量为病菌提供了以附着胞形式潜伏侵染在寄主中的条件。     

营养物质对两种炭疽菌潜伏侵染的影响

通过不同肖的向种糖类对侵染在青香蕉果实中的Ｃｏｌｌｅｔｏｔｒｉｃｈｕｍｍｕｓａｅ和要果实中的Ｃｏｌｌｅｔｏｔｉｃｈｕｍ ｇｌｏｅｓｐｏｒｉｏｉｄｅｓ的菌体的影响进行了测定,结果表明,高浓度淀粉可极显著地提高两种炭疽菌的子萌发率,并有利于附着胞和分生孢子的形成。单糖和二糖在低浓度进有得盱孢子萌发和产孢,不利于附着胞形成。未成熟果实的坚硬结构和高淀粉含量为病菌提供了以附着胞形式潜伏浸染在寄主中的条件。     

浙江无核柿炭疽病菌鉴定及附着胞形成过程中的核相变化

浙江无核柿炭疽病近年来在浙江淳安地区严重发生,根据形态学特征病原菌鉴定为胶孢炭疽菌ColletotrichumgloeosporioidesPenz.,在枝条病斑上的分生孢子盘通常不产生刚毛,分生孢子顶端顶部钝圆,基部平截,分生孢子盘中的孢子包埋在基质中,紧密结合在一起。分生孢子在自然寄主和人工培养条件下形态特征相似。6个柿树炭疽菌菌株的rDNAITS序列联配显示,其序列是相同的。用UPGMA方法分析ITS1-ITS2序列构建的炭疽菌系统发育树把6个柿树炭疽菌菌株和其它寄主上的胶孢炭疽菌或其有性型围小丛壳菌菌系分入同一个组,与根据形态学的鉴定结果一致。在附着胞形成过程中,用DAPI荧光染色观察到核相发生两次有丝分裂变化。第一次有丝分裂发生在分生孢子固着聚苯乙烯塑料培养皿3-4h后,随后,分生孢子中部形成一个隔膜,把它分成两个细胞;6~7h后,分生孢子发生第二次有丝分裂。分裂后,一个核通过芽管移入附着胞中。     

胶孢炭疽菌C2H2型转录因子CgGcp1的生物学功能

【背景】胶孢炭疽菌(Colletotrichum gloeosporioides)可以寄生于多种植物,侵染方式多样,能够引起严重的农业危害。在胶孢炭疽菌中,CgGcp1是一个C2H2型的转录因子,关于其生物学功能的研究未见报道。【目的】明确CgGcp1的生物学功能,为深入解析该病菌的致病机制奠定一定的理论依据。【方法】构建CgGCP1基因的敲除载体,利用同源重组得到敲除突变体。通过表型分析,包括营养生长、胁迫响应、孢子产生、附着胞形成及致病性分析等,明确该基因的生物学功能。【结果】CgGCP1基因敲除突变体生长速率较野生型减慢,对SDS、刚果红、NaCl和甘油更加敏感,孢子产量显著降低,附着胞的形成率降低且侵入能力减弱,在橡胶叶片上的致病力明显下降。【结论】CgGcp1参与调控胶孢炭疽菌营养生长、细胞壁完整性、分生孢子产生、附着胞形成与侵入和致病性。     

胶孢炭疽菌的菟丝子专化型

本文报道对寄生菟丝子的两个炭疽菌(Colletotrichum Corda)分离系(鲁保一号New-76)的鉴定结果。两分离系按其培养特性、分生孢子和附着孢的形态、大小及产孢细胞、分生孢子盘的特点,应属于胶孢炭疽菌[Colletotrichum gloeosporioides(Penz.)Sacc.],但在对寄主的选择性方面不同于该种的其他分离系,特别是对菟丝子(Cussuta chinensis)具有寄生专化性,故确定为胶孢炭疽菌菟丝子专化型[C.gloeosporioides(Penz.)Sacc.f.sp.cuseutae]。     

胶孢炭疽菌附着胞形成过程中核相的动态变化

通过DAPI荧光染料染色观察胶孢炭疽菌Colletotrichumgloeosporioides附着胞发育过程中的核相动态变化,结果显示,第2次有丝分裂发生的部位在分生孢子产生芽管的一端中;分裂后,最接近芽管的一个子核移入芽管顶端,或通过芽管移入附着胞中。0.10μg/mL的三环唑可完全抑制附着胞中黑色素形成,但不影响核的分裂。三环唑处理12h后,发生2次有丝分裂数量约为73%,而发生3次有丝分裂的数量约为23.9%;绝大多数附着胞中是单核,双核数量小于5%。     

纳他霉素对芒果采后胶孢炭疽菌的抑菌效果及机理

以纳他霉素为抑菌剂, 实验测定了离体条件下不同浓度纳他霉素对胶孢炭疽菌(Colletotrichum gloeosporioides)的孢子萌发及菌丝生长的抑制效果, 以及活体损伤接种炭疽病菌后, 纳他霉素对芒果(Mangifera indica)果实炭疽病的防治效果。通过测定纳他霉素处理后胶孢炭疽菌的细胞膜相对渗透率、可溶性蛋白含量、细胞膜完整性、孢子内活性氧水平和线粒体分布情况, 初步探明其抑菌机理。结果表明, 3 mg?L ^-1纳他霉素可显著抑制胶孢炭疽菌孢子萌发、芽管伸长和菌落生长, 80 mg?L ^-1纳他霉素可有效抑制芒果贮存过程中果实炭疽病斑的扩展。纳他霉素处理后胶孢炭疽菌细胞膜相对渗透率和可溶性蛋白含量增加; 2 mg?L ^-1纳他霉素处理8小时, 处理组胶孢炭疽菌孢子细胞膜损伤染色率为33.6%, 对照组染色率为13.9%; 处理组胞内活性氧产生染色率达46.9%, 比对照组高39.7%; 同时观察到纳他霉素使胞内线粒体分布不均且荧光信号微弱。以上结果表明, 纳他霉素可以破坏胶孢炭疽病菌细胞膜, 诱导活性氧大量积累, 并降低线粒体活性, 从而干扰菌体正常生理活性, 使其代谢活动受影响, 从而达到抑菌目的。     

纳他霉素对芒果采后胶孢炭疽菌的抑菌效果及机理

以纳他霉素为抑菌剂, 实验测定了离体条件下不同浓度纳他霉素对胶孢炭疽菌(Colletotrichum gloeosporioides)的孢子萌发及菌丝生长的抑制效果, 以及活体损伤接种炭疽病菌后, 纳他霉素对芒果(Mangifera indica)果实炭疽病的防治效果。通过测定纳他霉素处理后胶孢炭疽菌的细胞膜相对渗透率、可溶性蛋白含量、细胞膜完整性、孢子内活性氧水平和线粒体分布情况, 初步探明其抑菌机理。结果表明, 3 mg∙L ^-1纳他霉素可显著抑制胶孢炭疽菌孢子萌发、芽管伸长和菌落生长, 80 mg∙L ^-1纳他霉素可有效抑制芒果贮存过程中果实炭疽病斑的扩展。纳他霉素处理后胶孢炭疽菌细胞膜相对渗透率和可溶性蛋白含量增加; 2 mg∙L ^-1纳他霉素处理8小时, 处理组胶孢炭疽菌孢子细胞膜损伤染色率为33.6%, 对照组染色率为13.9%; 处理组胞内活性氧产生染色率达46.9%, 比对照组高39.7%; 同时观察到纳他霉素使胞内线粒体分布不均且荧光信号微弱。以上结果表明, 纳他霉素可以破坏胶孢炭疽病菌细胞膜, 诱导活性氧大量积累, 并降低线粒体活性, 从而干扰菌体正常生理活性, 使其代谢活动受影响, 从而达到抑菌目的。     

Ca^2＋信号途径参与稻瘟病菌分生孢子萌发及附着胞形成的调控

为了确定Ca^2 信号途径是否参与、在哪一时期参与稻瘟病菌分生孢子萌发及附着胞形成过程的调控,用四种可从不同位点阻断 Ca^2 信号途径的抑制剂分别处理分生孢子,观察抑制剂对孢子萌发及附着胞形成过程的抑制作用。结果表明：Ca^2 螯合剂 EGTA、Ca^2 通道抑制剂 Verapamil、抑制磷脂酶 C 活性的抑制剂 U-73122、影响钙调素与钙调素依赖蛋白激酶作用位点的抑制剂 KN-93,随着浓度的增加,对孢子萌发和附着胞形成过程的抑制作用明显增强;同一浓度下,抑制剂对附着胞形成过程的抑制作用大于孢子萌发过程;抑制剂影响孢子萌发和附着胞形成过程在萌发早期(1-4h)最有效;在完全被抑制、不能萌发的孢子内出现了许多颗粒状囊泡;抑制剂可使附着胞形态明显变小甚至不能形成。以上结果表明钙信号途径参与了稻瘟病菌孢子萌发及疏水条件下附着胞形成过程的调控。     

Ca2+信号途径参与稻瘟病菌分生孢子萌发及附着胞形成的调控

为了确定Ca2+信号途径是否参与、在哪一时期参与稻瘟病菌分生孢子萌发及附着胞形成过程的调控,用四种可从不同位点阻断Ca2+信号途径的抑制剂分别处理分生孢子,观察抑制剂对孢子萌发及附着胞形成过程的抑制作用.结果表明Ca2+螯合剂EGTA、Ca2+通道抑制剂Verapamil、抑制磷脂酶C活性的抑制剂U-73122、影响钙调素与钙调素依赖蛋白激酶作用位点的抑制剂KN-93,随着浓度的增加,对孢子萌发和附着胞形成过程的抑制作用明显增强;同一浓度下,抑制剂对附着胞形成过程的抑制作用大于孢子萌发过程;抑制剂影响孢子萌发和附着胞形成过程在萌发早期(1～4h)最有效;在完全被抑制、不能萌发的孢子内出现了许多颗粒状囊泡;抑制剂可使附着胞形态明显变小甚至不能形成.以上结果表明钙信号途径参与了稻瘟病菌孢子萌发及疏水条件下附着胞形成过程的调控.     

香蕉炭疽菌菌株亲缘关系的RAPD分析*

应用筛选的8个Operon随机引物对供试菌株的DNA进行扩增,所产生的RAPD图谱揭示了香蕉炭疽菌具有丰富的种内遗传多样性。UPGMA聚类分析的结果表明：26个香蕉炭疽菌被分为两个与地理来源相关的RAPD聚类群,它们分别由广东菌株和海南菌株为主组成。在两个香蕉菌群的外侧,两个胶胞炭疽菌菌株聚为一个小的外群。这些结果表明香蕉炭疽菌存在有与地域相关的种下类群分化。     

香蕉炭疽菌菌株亲缘关系的RAPD分析

应用筛选的8个Operon随机引物对供试菌株的DNA进行扩增,所产生的RAPD图谱揭示了香蕉炭疽菌具有丰富的种内遗传多样性。UPGMA聚类分析的结果表明：26个香蕉炭疽菌被分为两个与地理来源相关的RAPD聚类群,它们分别由广东菌株和海南菌株为主组成。在两个香蕉菌群的外侧,两个胶胞炭疽菌菌株聚为一个小的外群。这些结果表明香蕉炭疽菌存在有与地域相关的种下类群分化。     

A Microscopic Characterization of the Infection of Green and Red Pepper Fruits by an Isolate of Colletotrichum gloeosporioides

Colletotrichum gloeosporioides is a common pathogenic fungus in many plants. To investigate the specificity of the isolate C. gloeosporioides to green fruits, fungal behaviours and anthracnose development on green and red pepper fruits were compared using light and stereo microscopic techniques. When the isolate of C. gloeosporioides was inoculated on both green and red fruits, conidial germination, appressoria, and infection hyphae were observed on both fruits within 24 h after inoculation. The fungal invasion and colonization continued to the epidermal cells of green fruits, but not to those of red ones. Initial anthracnose symptoms were detected only on green fruits at 2 days after inoculation resulting in typical sunken necrosis within 5 days after inoculation. Thus the specificity of the isolate to green fruits may be due to successful invasion and colonization of the infection hyphae from appressoria into the epidermal cells through epicuticular layers of green pepper fnats, but not on red ones.     

Isolation of defense-related genes differentially expressed in the resistance interaction between pepper fruits and the anthracnose fungus Colletotrichum gloeosporioides

Unripe mature green fruits of pepper (Capsicum annuum) are susceptible to Colletotrichum gloeosporioides, whereas ripe red fruits are not. We established this pepper-C. gloeosporioides interaction as a model system to study the fungal resistance that develops during ripening of nonclimacteric fruit. Histochemical examination of transverse sections suggested that fungal invasion 24 h after inoculation (HAI) and colonization 48 HAI are critical events that differentiate between resistant and susceptible interactions. Based on this observation, we used messenger RNA differential display to isolate defense-related genes differentially expressed at 24 and 48 HAI. RNA gel blot analysis showed that six out of eighty cloned cDNAs were differentially expressed after infection of ripe fruit. The proteins encoded by these six clones, ddP1, ddP3, ddP4, ddP6, ddP13, and ddP47, showed significant homology to aldehyde dehydrogenase, P23 protein, NP24 protein, cytochrome P450 protein, esterase, and MADS-box protein, respectively, and may be involved in the resistance of ripe fruit to C. gloeosporioides infection.     

Characterization of the membrane sensor PenJ for β-lactam antibodies from Bacillus licheniformis by amino acid substitution

Abstract The beta scanner and pattern matching software integrated in the automated microbiology identification system (AMBIS) were used to assess the relatedness of isolates of Colletotrichum acutatum, C. kahawae, C. fragariae, C. gloeosporioides and C. musae based on mitochondrial DNA restriction fragment length polymorphisms. The dendrograms generated reflected the intra-specific variation and inter-specific relationships of these species as determined by other previously reported methods. The adaptability of AMBIS as a rapid method for assessing genetic relatedness of fungal species based on DNA polymorphisms is discussed.     

Biological Activity of Four Antifungal Compounds in Immature Avocado

Anthracnose caused by Colletotrichum gloeosporioides in ripe avocados originates as latent infections in the immature fruits. Concentrated ether extract of the peel of apparently healthy, immature avocados when bioassayed on thin layer chromatographic plates with conidia of either Cladosporium cladosporioides or C. gloeosporioides produced four inhibition areas at Rf 0.30, 0.32, 0.70 and 0.75 (these were denoted as AvIV, AvIII, AvII and AvI respectively). A hot chloroform extract was partitioned on a silica gel column and the four antifungal compounds were separated. Spectroscopic data revealed that one of these compounds, AvII, was similar to cis-l-acetoxy-2-hydroxy-4-oxo-heneicosa-12,15-diene and another (AvIV) was a long chain saturated compound comprising hydroxyl group(s) having molecular weight of 268. Toxicity of AvII to C. gloeosporioides was 2 times that of AvIV and 6.5 and 7.5 times that of AvIII and AvI respectively. The amounts of these four antifungal compounds increased gradually during fruit development and reached their maxima at harvest. The concentration of AvI, AvII, AvIII and AvIV was 1300, 920, 1050 and 780 μg/g fresh peel respectively in the fruit at harvesting maturity. The amount of AvII and AvIV decreased to 53 and 64 μg/g fresh peel respectively in the fruit at the ripe stage at which neither AvI nor AvIII was detected. This took place in coincidence with the onset of progressive lesion development by the fungus.     

Colletotrichum gloeosporioides s.l. associated with Theobroma cacao and other plants in Panama: multilocus phylogenies distinguish host-associated pathogens from asymptomatic endophytes

Colletotrichum interacts with numerous plant species overtly as symptomatic pathogens and cryptically as asymptomatic endophytes. It is not known whether these contrasting ecological modes are optional strategies expressed by individual Colletotrichum species or whether a species' ecology is explicitly pathogenic or endophytic. We explored this question by inferring relationships among 77 C. gloeosporioides s.l. strains isolated from asymptomatic leaves and from anthracnose lesions on leaves and fruits of Theobroma cacao (cacao) and other plants from Panamá. ITS and 5'-tef1 were used to assess diversity and to delineate operational taxonomic units for multilocus phylogenetic analysis. The ITS and 5'-tef1 screens concordantly resolved four strongly supported lineages, clades A-D: Clade A includes the ex type of C. gloeosporioides, clade B includes the ex type ITS sequence of C. boninense, and clades C and D are unidentified. The ITS yielded limited resolution and support within all clades, in particular the C. gloeosporioides clade (A), the focal lineage dealt with in this study. In contrast the 5'-tef1 screen differentiated nine distinctive haplotype subgroups within the C. gloeosporioides clade that were concordant with phylogenetic terminals resolved in a five-locus nuclear phylogeny. Among these were two phylogenetic species associated with symptomatic infections specific to either cacao or mango and five phylogenetic species isolated principally as asymptomatic infections from cacao and other plant hosts. We formally describe two new species, C. tropicale and C. ignotum, that are frequent asymptomatic associates of cacao and other Neotropical plant species, and epitypify C. theobromicola, which is associated with foliar and fruit anthracnose lesions of cacao. Asymptomatic Colletotrichum strains isolated from cacao plants grown in China included six distinct C. gloeosporioides clade taxa, only one of which is known to occur in the Neotropics.