日本菟丝子寄生菌致病力的专化性研究

日本菟丝子(Cuscuta japonica)吸取不同寄主植物的营养,经过营养代谢其机体内的过氧化物酶同工酶的活性或酶带数发生变化,从而改变了它对胶孢炭疽菌菟丝子专化型[Col-letotrichum gloeosporioides(Penz.)Sacc.f.sp.Cuscutael]同一菌株的抗性,以此为依据,采用寄生在不同植物上的日本菟丝子做为鉴定寄主,可以鉴别出胶孢炭疽菌菟丝子专化型中不同菌株的致病力差异。 初步认为:龙眼、桂花、柚子和冬青上的紫色菟丝子及泡桐、桂花和冬青上的黄色菟丝子可以作为鉴定寄主。     

酚类物质对两种潜伏炭疽菌生长和繁殖的影响

通过测定丹宁、间苯二酚、邻苯二酚、绿原酸和咖啡因,作者对潜伏侵染在香蕉果实中的芭蕉炭疽菌Colletotrichum musae(Berk & Curt)Arx和芒果果实中的胶孢炭疽菌C.gloeosporioides Penz.的分生孢子萌发,附着胞和分生孢子的形成,以及菌丝体生长的影响进行了研究。结果表明,这几种酚类物质,在一定浓度下,可抑制两种炭疽菌的生长和发育,其中邻苯二酚对芭蕉炭疽菌的作用浓度最低,而间苯二酚对胶孢炭疽菌的作用浓度最低,丹宁则对两种菌的作用浓度最高,在一定浓度下影响着附着胞的形成。     

黄毛草莓F-box蛋白基因FnFBOX1及其启动子的克隆和表达分析

探索黄毛草莓FnFBOX1参与草莓胶孢炭疽菌(Colletotrichum gloeosporioides)侵染过程中的抗病反应和pFnFBOX1启动子的转录活性,为研究FnFBOX1抗炭疽病功能奠定基础.以黄毛草莓(Fragaria nilgerrensis Schidl.)为研究对象,通过RT-PCR技术克隆FnF...     

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。     

生防细菌T132的鉴定及其对采后柑橘炭疽病的抑制效果

【目的】柑橘(Citri)是世界上重要的果树。由胶孢炭疽菌[Colletotrichum gloeosporioides(Penz.)]引起的柑橘炭疽病是柑橘生产的主要病害之一。为探索对采后柑橘炭疽病有效的生防措施,分离鉴定柑橘根围土壤中一株细菌T132,并研究其特性及生防效果。【方法】根据菌株T132的形态特征、生理生化特性以及16S rDNA序列对其进行鉴定;通过连续8次在人工培养基上传代培养,测定该菌株的遗传稳定性;采用柑橘果实刺伤挑战接种和拮抗菌液直接浸泡健康果实两种方法研究该菌株对柑橘炭疽病的抑菌防病效果;利用洋葱伯克霍尔德氏菌致病因子的特异性引物检测菌株T132是否为潜在的人类致病菌。【结果】菌株T132鉴定为越南伯克霍尔德氏菌(Burkholderia vietnamiensis)。连续8次在人工培养基上传代培养,菌株T132抑制胶孢炭疽病菌生长的能力没有发生明显改变。菌株T132对胶孢炭疽菌C.gloeosporioides引起的柑橘炭疽病有明显的防治作用,刺伤接种的防效为88.2%,自然发病的防效为54.9%。未检测到该拮抗菌株有人体致病相关的洋葱伯克霍尔德氏菌致病因子(BCESM)毒力基因。【结论】首次报道对柑橘采后炭疽病具有生防效果、对人类相对安全的越南伯克霍尔德氏菌生防菌株。     

罗汉果果实病害观察初报

经初步研究,罗汉果果实病害主要有炭疽病(Colletotrichum gloeosporioides(Penz) Sacc)、黑斑病(Alternara tenuis Ness)和腐烂病(Fusarium heterosporum Ness)3种,它们多见于成熟期前后。其中炭疽病为害期长,危害严重,在果实成长过程中,可用70％甲基托布津1000—1500倍液来防治。     

菜心炭疽病菌拮抗细菌的筛选及鉴定

从土壤中分离筛选获得2株对菜心炭疽病菌——希金斯炭疽菌(Colletotrichum higginsianum)具有强烈抑制作用的芽孢杆菌菌株19E2、13A1。通过形态特征观察、生理生化特性测定, 结合16S rDNA序列同源性分析以及部分基因特异性序列分析, 将2株菌鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。抑菌试验的结果表明: 当19E2和13A1的培养液在PDA培养基中的浓度为10%时, 抑菌率分别达到75.62%和85.35%, 当培养液浓度升高到20%时, 可完全抑制菜心炭疽病菌生长。抗菌谱测定结果显示: 2株菌对辣椒炭疽菌(C. capsici)、胶孢炭疽菌(C. gloeosporioides)、灰霉 (Botrytis cinerea)、苞叶芋柱帚霉(Cylindrocladium spathiphylli)和尖镰孢(Fusarium oxysorum)等重要病原菌具有明显的作用效果。     

生防菌株1404的鉴定及其对采后柑橘炭疽病的防治效果

【目的】柑橘是世界上重要的果树。由胶孢炭疽菌[Colletotrichum gloeosporioides(Penz.)Sacc.]引起的柑橘炭疽病是柑橘生产的主要病害。为开发对采后柑橘炭疽病有效的生防措施,对从辣椒根际土壤中分离的一株生防细菌1404进行了鉴定,并对其特性及生防效果进行了研究。【方法】根据菌株1404的形态特征、生理生化特性以及16SrDNA序列对其进行鉴定;通过连续在人工培养基上转代培养,测定该菌株拮抗活性的稳定性;用果实刺伤接种法对采后柑橘炭疽病的防效进行测定。【结果】菌株1404与来自GenBank的短短芽孢杆菌[Brevibacillus brevis(Migula)Shidaetal.]以100%bootstrap水平类聚一群。该菌株的形态特征及生理生化特性与Brevibacillus brevis相符。连续4次在人工培养基上转代培养,菌株1404对柑橘炭疽病菌生长的抑制力没有发生明显改变。该生防菌对柑橘炭疽病的防治效果明显,处理后第20天防效达到64.9%。【结论】根据16SrDNA序列、形态特征、生理生化特性,将菌株1404鉴定为短短芽孢杆菌。本文首次报道对柑橘采后炭疽病具有较好防效的生防菌Brevibacillus brevis。     

胶孢炭疽菌热休克蛋白基因Cghsp90的RNAi突变体获得与功能分析

为揭示胶孢炭疽菌(Colletotrichum gloeosporioides)热休克蛋白基因Cghsp90在抗逆境胁迫中的功能,利用RNAi技术结合PEG介导的原生质体转化方法,获得了Cghsp90的RNAi突变体菌株,经实时荧光定量PCR (qRT-PCR)对野生型菌株和突变体菌株间Cghsp90基因差异表达分析以及NaCl、H_2O_2、SDS、Congo Red、18℃(Cold)和33℃(Hot)等非生物胁迫条件下野生型菌株和突变体菌株的胁迫耐受性分析。通过特异性引物检测鉴定、qRT-PCR分析,获得Cghsp90的RNAi突变体pSilent-1:Cghsp90-1和pSilent-1:Cghsp90-2;与野生型菌株相比,侵染过程中Cghsp90基因的表达量显著低于野生型菌株,8个致病相关基因显著下调表达;突变体菌株的产孢量下降、分生孢子畸形、萌发率下降;在非生物胁迫条件下,突变体菌株较野生型菌株更为敏感;在胶孢炭疽菌的分生孢子中研究Cghsp90基因的形态建成与萌发,对应变逆境胁迫以及调控致病相关基因等方面发挥重要的作用。     

纳米氧化镁对柑橘炭疽菌的抗菌效应

纳米氧化镁(MgO NP)因其自身卓越的物理化学性质,尤其是较高的比表面积,使其在光学、电学、材料学和生物医学等各个领域都表现出重要的应用前景.MgO NP对细菌表现出良好的抗菌性,而对真菌的抗菌性研究较少.本文研究了MgO NP对柑橘炭疽病菌胶孢炭疽菌(Colletotrichum gloeosporioides)的抗菌作用,测定了其对胶孢炭疽菌的菌丝生长和菌丝形态的影响,以及对孢子萌发的抑制作用,并测定了MgO NP对柑橘炭疽病的防控效果.研究结果表明,100~800μg/mL的MgO NP都能较好抑制胶孢炭疽菌菌丝生长和孢子萌发,其中400μg/mL和800μg/mL剂量效果最明显;同时提前施用400μg/mL的MgO NP表现出较好的柑橘炭疽病防治效果,说明MgO NP具有作为植物保护剂的潜力.     

芒果炭疽病菌β-微管蛋白基因的克隆及其与多菌灵抗药性发生的关系

参照豆科合萌属 (Aeschynomene)作物炭疽病菌的tub1和tub2基因序列设计了 2对引物 ,分别从芒果 (Man gifera)炭疽病菌对多菌灵 (MBC)田间抗药性 (MBCR)和敏感 (MBCS)的菌株中扩增 β_微管蛋白基因。结果只有以tub2为参照设计的引物扩增到了特异片段。进一步对全基因进行了克隆和测序。该基因序列全长 1344bp ,编码4 4 7aa ,其核苷酸和氨基酸序列与豆科合萌属炭疽病菌的tub2基因高度同源。对芒果炭疽病菌抗、感菌株 β_微管蛋白氨基酸序列进行比较分析 ,发现第 181、2 37和 36 3位氨基酸发生了突变 ,而其它位置 (如第 198位或 2 0 0位 )均不变     

Endophytic fungi from Amomum siamense.

Endophytic fungi were isolated from apparently healthy organs of the wild ginger Amomum siamense Criab.. including leaves, pseudostems, and rhizomes, collected from two sites in Doi Suthep-Pui National Park, Thailand. Endophytes were relatively common with an isolate prevalence of 70%-83% at the two sites sampled in the wet and dry seasons. The endophyte assemblages from the two sites were diverse and comprised 7 Ascomycetes and 26 mitosporic fungi. Colletotrichum "gloeosporioides" (Penz.) Penz. & Sacc. in Penz., Glomerella spp., xylariaceous fungi, and Phomopsis spp. were consistently dominant as endophytes on Amomum siamense, but many rare species and mycelia sterilia were also recorded. There was no significant difference between the number of isolates recovered from leaves containing vein tissues and those containing intervein tissues, independent of leaf age. Most taxa showed a preference for either leaf tissue or pseudostems. Two new Ascomycetes species. Gaeumannomyces amomi and Leiosphaerella amomi, were discovered from leaves and rhizomes, respectively, and four species of Pyricularia, including three new species, were isolated from leaves.     

Molecular and phenotypic analyses reveal association of diverse Colletotrichum acutatum groups and a low level of C. gloeosporioides with olive anthracnose

Anthracnose (Colletotrichum spp.) is an important disease causing major yield losses and poor oil quality in olives. The objectives were to determine the diversity and distribution pattern of Colletotrichum spp. populations prevalent in olives and their relatedness to anthracnose pathogens in other hosts, assess their pathogenic variability and host preference, and develop diagnostic tools. A total of 128 Colletotrichum spp. isolates representing all olive-growing areas in Portugal and a few isolates from other countries were characterized by molecular and phenotypic assays and compared with reference isolates. Arbitrarily primed PCR data, internal transcribed spacer of rRNA gene and beta-tubulin 2 nucleotide sequences, colony characteristics, and benomyl sensitivity showed Colletotrichum acutatum to be dominant (>97%) with limited occurrence of Colletotrichum gloeosporioides (<3%). Among C. acutatum populations, five molecular groups, A2 to A6, were identified. A2 was widely prevalent (89%), coinciding with a high incidence of anthracnose and environmental conditions suitable to disease spread. A4 was dominant in a particular region, while other C. acutatum groups and C. gloeosporioides were sporadic in their occurrence, mostly related to marginal areas of olive cultivation. C. gloeosporioides, isolated from olive fruits with symptoms indistinguishable from those of C. acutatum, showed same virulence rating as the most virulent C. acutatum isolate from group A2. C. acutatum and C. gloeosporioides isolates tested in infected strawberry fruits and strawberry and lupin plants revealed their cross-infection potential. Diagnostic tools were developed from beta-tubulin 2 sequences to enable rapid and reliable pathogen detection and differentiation of C. acutatum groups.     

Colletotrichum acutatum var. fioriniae (teleomorph: Glomerella acutata var. fioriniae var. nov.) infection of a scale insect

An epizootic has been reported in Fiorinia externa populations in New York, Connecticut, Pennsylvania and NewJersey. Infected insects have profuse sclerotial masses enclosing their bodies. The most commonly isolated microorganism from infected F. externa was Colletotrichum sp. A morphological and molecular characterization of this fungus indicated that it is closely related to phytopathogenic C. acutatum isolates. Isolates of Colletotrichum sp. from F. externa in areas of the epizootic were similar genetically and were named Colletotrichum acutatum var. fioriniae var. nov, based on our findings. In vitro and in planta mating observed between isolates of C. acutatum var. fioriniae could serve as a possible source of genetic variation and might give rise to new biotypes with a propensity to infect insects. Only one other strain, C. gloeosporioides f. sp. ortheziidae, has been reported to show entomopathogenic activity.     

小桐子内生真菌及其抗真菌活性研究

从药用植物小桐子的根和茎中分离到内生真菌57株,经鉴定,它们分属于15个不同的分类单元。其中拟盘多毛孢为优势属,其次为拟茎点霉属和茎点霉属。用杨桃炭疽菌,香蕉疫霉和镰刀菌作指示菌对这些内生真菌的抗真菌活性进行检测,结果表明,有2株菌对杨桃炭疽菌具有抑菌活性。     

Bestimmung pektolytischer Enzyme bei mehreren Schadpilzen von Citrusfrüchten

Characterization of pectolytic enzymes From several fungi causing post-harvest decay of citrus fruits From culture filtrates of Penicillium digitatum Sacc., Trichoderma viride Penz., Phomopsis citri (Faw.) cf. Diaporthe citri (Faw.) Wolf, and Colletotrichum gloeosporioides Penz. endo-polygalacturonase and endo-polymethylgalacturonase have been isolated. Both enzymes were active under acid conditions only. The last named fungus additionally produced pectic acid-transeliminase, the activity was found under alkaline conditions.     

Antagonism of Bacillus spp. Isolated from Marine Biofilms Against Terrestrial Phytopathogenic Fungi

We aimed at determining the antagonistic behavior of bacteria derived from marine biofilms against terrestrial phytopathogenic fungi. Some bacteria closely related to Bacillus mojavensis (three isolates) and Bacillus firmus (one isolate) displayed antagonistic activity against Colletotrichum gloeosporioides ATCC 42374, selected as first screen organism. The four isolates were further quantitatively tested against C. gloeosporioides, Colletotrichum fragariae, and Fusarium oxysporum on two culture media, potato dextrose agar (PDA) and a marine medium-based agar [yeast extract agar (YEA)] at different times of growth of the antagonists (early, co-inoculation with the pathogen and late). Overall antagonistic assays showed differential susceptibility among the pathogens as a function of the type of culture media and time of colonization (P < 0.05). In general, higher suppressive activities were recorded for assays performed on YEA than on PDA; and also when the antagonists were allowed to grow 24 h earlier than the pathogen. F. oxysporum was the most resistant fungus while the most sensitive was C. gloeosporioides ATCC 42374. Significant differences in antagonistic activity (P < 0.05) were found between the different isolates. In general, Bacillus sp. MC3B-22 displayed a greater antagonistic effect than the commercial biocontrol strain Bacillus subtilis G03 (Kodiak). Further incubation studies and scanning electronic microscopy revealed that Bacillus sp. MC3B-22 was able to colonize, multiply, and inhibit C. gloeosporioides ATCC 42374 when tested in a mango leaf assay, showing its potential for fungal biocontrol. Additional studies are required to definitively identify the active isolates and to determine their mode of antifungal action, safety, and biocompatibility.