香蕉上的镰孢菌种类及其系统发育关系(英文)

镰孢菌属真菌是香蕉上的重要病原菌,主要引起香蕉枯萎病以及香蕉冠腐病,在我国已明确引起香蕉枯萎病的病原为尖孢镰孢古巴专化型 Fusarium oxysporum f. sp. cubense(FOC)1号和4号生理小种,但是引起香蕉冠腐病的镰孢菌种类还未明确。为了解香蕉上镰孢菌在种间及种内水平上的多样性,2008–2011 年间作者从华南地区不同的水果市场及香蕉果园采集香蕉样品90份,分离得到143株镰孢菌。通过形态学观察及基于 EF-1α基因的系统进化分析鉴定出10种镰孢菌,即F. oxysporum、F. solani、F. camptoceras、F. pallidoroseum、F. stiloides、F. chlamydosporum、F.verticillioides、F. proliferatum、F. concentricum、F. sacchari,以及藤仓赤霉复合种(Gibberella fujikuroi species complex,GFC)中 3 个未定名的类群。轮纹镰孢 F. concentricum 及甘蔗镰孢 F.sacchari 是香蕉果实中最常见种,前菌为我国首次报道,后菌是首次报道与香蕉有关。对从香蕉上分离的藤仓赤霉复合种(GFC)及尖孢镰孢复合种(FOSC)的EF-1α序列进行了系统发育分析,其GFC中的27个菌株组成的单系群可分为7个不同的亚群,分别为 F.verticillioides、F. proliferatum、F. concentricum、F. sacchari 以及3个没有描述过的菌系 Fusarium sp. 1、Fusarium sp.2和 Fusarium sp.3;FOSC中的50个菌株形成2大分枝共12个谱系,分离自我国华南地区的21株尖孢镰孢形成7个谱系,其中 13株已知的香蕉枯萎病病原菌分布在3个谱系中,我国大陆的香蕉枯萎病病原菌菌株与来源于台湾地区及东南亚的菌株亲缘关系较近,FOC1号生理小种的遗传分化大于4号生理小种,FOC 1号生理小种与分离自香蕉果实上的尖孢镰孢菌的亲缘关系比与FOC 4号生理小种的亲缘关系更近。研究结果表明,我国香蕉上存在着丰富的镰孢菌种类,而且种内遗传多样性丰富。     

Fusarium incarnatum is associated with postharvest fruit rot of muskmelon (Cucumis melo)

Postharvest fruit rot was observed on muskmelon (Cucumis melo) on market shelves at a melon farm in the city of Hatyai, Songkhla, Thailand. Diseased muskmelon fruit displayed cotton-like mycelia on wounds and had brown to dark brown internal tissue. Based on morphological characteristics and molecular analysis of internal transcribed spacer (ITS) and translation elongation factor 1-α (TEF1-α) DNA sequences, the fungal pathogen was identified as Fusarium incarnatum. This species belongs to the Fusarium incarnatum-equiseti species complex (FIESC). A pathogenicity test was conducted to verify Koch's postulates, and F. incarnatum was observed to cause fruit rot on muskmelon; symptoms of the disease were similar to those seen in the field. However, only artificially wounded melons became infected, suggesting that F. incarnatum is an obligate wound-infecting pathogen. To our knowledge, this is the first report of F. incarnatum causing fruit rot of muskmelon in Thailand.     

Characterization of Fusarium spp. associated with pineapple fruit rot and leaf spot in Peninsular Malaysia

Pineapple (Ananas comosus) is one the important fruit crops planted in Malaysia, and this study was conducted to determine Fusarium spp. associated with diseases of the fruit crop as Fusarium is prevalent in tropical countries. Our objective was to identify and characterize Fusarium spp. associated with pineapple fruit rot and leaf spot mainly found on the fruits and leaves in Peninsular Malaysia. Fusarium isolates (n = 108) associated with pineapple fruit rot and leaf spot were characterized by morphological, molecular and phylogenetic analyses, a mating study and pathogenicity testing. TEF‐1α sequence analysis identified Fusarium proliferatum, Fusarium verticillioides, Fusarium sacchari and Fusarium sp. Mating was successful only between tester strains of F. proliferatum and F. verticillioides. Sexual crosses with standard tester strains showed that 82 isolates of F. proliferatum produced fertile crosses with mating population D (Gibberella intermedia) and three isolates of F. verticillioides were fertile with the tester strain of mating population A (Gibberella moniliformis). All isolates were pathogenic, causing pineapple fruit rot and leaf spot, thus fulfilling Koch's postulates.     

Biodiversity of Fusarium species in Mexico associated with ear rot in maize, and their identification using a phylogenetic approach

Fusarium proliferatum, F. subglutinans, and F. verticillioides are known causes of ear and kernel rot in maize worldwide. In Mexico, only F. verticillioides and F. subglutinans, have been reported previously as causal agents of this disease. However, Fusarium isolates with different morphological characteristics to the species that are known to cause this disease were obtained in the Highland-Valley region of this country from symptomatic and symptomless ears of native and commercial maize genotypes. Moreover, while the morphological studies were not sufficient to identify the correct taxonomic position at the species level, analyses based in the Internal Transcribed Spacer region and the Nuclear Large Subunit Ribosomal partial sequences allowed for the identification of F. subglutinans, F. solani, and F. verticillioides, as well as four species (F. chlamydosporum, F. napiforme, F. poae, and F. pseudonygamai) that had not previously been reported to be associated with ear rot. In addition, F. napiforme and F. solani were absent from symptomless kernels. Phylogenetic analysis showed genetic changes in F. napiforme, and F. pseudonygamai isolates because they were not true clones, and probably constitute separate sibling species. The results of this study suggest that the biodiversity of Fusarium species involved in ear rot in Mexico is greater than that reported previously in other places in the world. This new knowledge will permit a better understanding of the relationship between all the species involved in ear rot disease and their relationship with maize.     

Fusarium fujikuroi associated with stem rot of red‐fleshed dragon fruit (Hylocereus polyrhizus) in Malaysia

Stem rot was recorded as one of serious diseases of red‐fleshed dragon fruit, (Hylocereus polyrhizus), in Malaysia. Fusarium fujikuroi was recovered from stem rot lesion of H. polyrhizus and the species was identified using TEF1‐α sequence and mating study. From maximum likelihood phylogenetic tree using combined TEF1‐α and β‐tubulin sequences, the F. fujikuroi isolates from stem rot were grouped according to three geographical locations, namely Peninsular Malaysia, Sabah and Sarawak. Phylogenetic analysis indicated that F. fujikuroi isolates from stem rot of H. polyrhizus were clustered separately from F. fujikuroi isolates from rice because of intraspecific variation. From amplification of MAT allele‐specific primers, 20% of the isolates carried MAT‐1 allele while 80% carried MAT‐2 allele. From isolates that carried MAT‐1 allele, 65% crossed‐fertile with MP‐C (mating population of F. fujikuroi) tester strain while for MAT‐2 allele, 56% crossed‐fertile with MP‐C. None of the isolates were identified as MP‐D (mating population of F. proliferatum). Pathogenicity test conducted on 40 representative isolates showed that the stem rot symptoms were similar with the symptoms observed in the field, and can be categorized as low, moderate and high aggressiveness, which indicated variation in pathogenicity and virulence among the isolates. This study provides novel findings regarding Fusarium species associated with stem rot of H. polyrhizus and indicated that F. fujikuroi as a new causal pathogen of the disease.     

Fusarium spp. associated with rice Bakanae: ecology,genetic diversity,pathogenicity and toxigenicity

African and Asian populations of Fusarium spp. (Gibberella fujikuroi species complex) associated with Bakanae of rice (Oryzae sativa L.) were isolated from seeds and characterized with respect to ecology, phylogenetics, pathogenicity and mycotoxin production. Independent of the origin, Fusarium spp. were detected in the different rice seed samples with infection rate ranges that varied from 0.25% to 9%. Four Fusaria (F. andiyazi, F. fujikuroi, F. proliferatum and F. verticillioides) were found associated with Bakanae of rice. While three of the Fusaria were found in both African and Asian seed samples, F. fujikuroi was only detected in seed samples from Asia. Phylogenetic studies showed a broad genetic variation among the strains that were distributed into four different genetic clades. Pathogenicity tests showed that all strains reduced seed germination and possessed varying ability to cause symptoms of Bakanae on rice, some species (i.e. F. fujikuroi) being more pathogenic than others. The ability to produce fumonisins (FB₁ and FB₂) and gibberellin A3 in vitro also differed according to the Fusarium species. While fumonisins were produced by most of the strains of F. verticillioides and F. proliferatum, gibberellin A3 was only produced by F. fujikuroi. Neither fumonisin nor gibberellin was synthesized by most of the strains of F. andiyazi. These findings provide new information on the variation within the G. fujikuroi species complex associated with rice seed and Bakanae disease.     

Fusarium ananatum sp. nov. in the Gibberella fujikuroi species complex from pineapples with fruit rot in South Africa

Pineapple (Ananas comosus) is native to South America and widely planted as a fruit crop in the tropics and sub-tropics. This plant is susceptible to a number of fungal diseases of which the most severe is fusariosis. The disease is caused by Fusarium guttiforme and occurs only in South and Central America. The occurrence of a similar disease on pineapples in South Africa has prompted a re-evaluation of the Fusarium sp. associated with pineapple fruit rot. Phylogenetic relationships of isolates from pineapples collected in Brazil and South Africa were assessed based on sequence data for the translation elongation factor-1-α, histone H3 and β-tubulin gene regions. Analyses showed that the South African isolates represent a species distinct from Brazilian isolates. The South African isolates are characterised by a concentration of aerial mycelium at the centres of the colonies, different to the Brazilian isolates that have an even distribution of aerial mycelium. Both phylogenetic and morphological data show that the disease on pineapple in South Africa is caused by a new Fusarium species described here as F. ananatum sp. nov.     

Colletotrichum species associated with sugarcane red rot in Brazil

Sugarcane is a widely cultivated crop in Brazil and in many parts of the world. However, the red rot causes huge losses due to the reduction of sucrose and deterioration of the juice. The aim of this study was to identify Colletotrichum species associated with the red rot through polyphasic approaches; which included phylogenetic, morpho-cultural analyzes and pathogenicity tests. Nine isolates from the states of Alagoas and two from São Paulo, Brazil, were preliminary analyzed with the glyceraldehyde-3 phosphate dehydrogenase gene (GAPDH), as an initial measure for species diversity. Later on, the representative isolates of each species were sequenced with the β-tubulin (TUB2) gene, calmodulin (CAL), DNA lyase (APN2/MAT IGS) and the ITS-rDNA region. Morphocultural characterization was performed by evaluating the mycelial growth rate (MGR), colony appearance and the shape and size of 50 conidia and appressoria. For the pathogenicity test asymptomatic leaves and stalks of sugarcane were tested with and without injuries. Phylogenetic analysis associated with morphocultural characteristics and the pathogenicity test of the eleven isolates revealed three Colletotrichum species: Colletotrichum falcatum (8 isolates), Colletotrichum siamense (1 isolate) and Colletotrichum plurivorum (2 isolates) causing the red rot disease in sugar cane. All species were pathogenic in wounded leaves and stalks, being C. falcatum the one causing the largest lesions (1.12 cm) in leaves and C. plurivorum in stalks (0.67 cm). Therefore, this study confirms the association of C. falcatum as a sugarcane pathogen and records for the first time worldwide the occurrence of C. siamense and C. plurivorum associated with this host.     

Fusarium sporotrichioides Sherb. and trichothecenes associated with Fusarium-ear rot of corn before harvest

Fusarium sporotrichioides was found to be the predominant fungus in approximately 2 % of corn ears damaged byFusarium species, before harvest during 1984 and 1985 in Poland. Concentrations of up to 1,714.9 mg/kg of total type-A trichothecenes (T-2 Toxin, HT-2 Toxin, Neosolaniol, T-2 Triol, and T-2 Tetraol) were found in hand-selected, heavily damaged kernels obtained fromF. sporotrichioides-molded ears.     

Iridoid glycosides from fruits reduce the growth of fungi associated with fruit rot

Aims Ripe, fleshy fruits generally function as rewards to attract mutualistic seed dispersers, but many fruits also contain high concentrations of toxic secondary metabolites. These compounds may serve a variety of adaptive roles in seed dispersal or as a defense against non-dispersing seed predators or pathogens. We tested the effects of iridoid glycosides from fruits of a hybrid bush honeysuckle, Lonicera × bella, on the growth of two pathogenic fungal strains associated with fruit rot, Alternaria tenuissima and Aspergillus tubingensis.Methods Fungi were isolated from field-collected L. × bella fruits and identified using molecular techniques. Their growth rates were assessed in vitro in the presence of varying concentrations of pure loganin, one of the most abundant iridoid glycosides in fruits, as well as fruit extracts containing a mix of at least seven different iridoid glycosides.Important findings Loganin had strong dose-dependent negative effects on the growth of both fungi. Extracts from fruits had no effect on Aspergillus but a strong antifungal effect on Alternaria that increased with fruit ripening. Total iridoid glycoside concentrations in extracts were not good predictors of variation in fungal growth, but several individual compounds had significant negative effects. Although iridoid glycosides have primarily been studied as antiherbivore defenses in leaves, these results indicate that they can also function to reduce the growth of fungi associated with fruit rot.     

Prototheca associated with banana

Prototheca stagnora was found to be a habitant of older harvested banana (Musa sapientum) and plantain (M. paradisiaca) stumps while P. wickerhamii colonized fresh Musa sp. stumps and flower bract water of Heliconia sp. While Prototheca sp. were known to habituate woody plants, this is the first evidence that herbaceous plants also serve as habitats.     

Evaluation of Fusarium wilt disease in passion fruit species inoculated with Fusarium oxysporum f.sp. passiflorae

One of the main diseases that reduces production of passion fruit crops is Fusarium wilt, caused by the fungus Fusarium oxysporum f.sp. passiflorae (FOP). The use of resistant rootstocks, such as the species Passiflora cincinnata, is one of the management strategies used to control this disease. The objective of this work was to evaluate the pathogenicity of different isolates of FOP on P. edulis and P. cincinnata in order to identify its potential for use in areas with a history of the disease. Thirteen isolates of the fungus were used, and the inoculums were produced at a concentration of 10⁶ CFU/ml. Seedlings were produced in coconut fibre, and the root system was then immersed for five minutes in the conidial suspension before being replanted in the 770-ml pots. Inoculated seedlings of P. edulis and P. cincinnata at the three-leaf stage were daily evaluated from the second day after inoculation (DAI) until day 90. All isolates were pathogenic in both Passiflora species; however, the incidence, severity and mortality were higher in P. edulis. There was a statistically significant difference for the incubation period of the FOP 23 and FOP 57 isolates, being higher in P. edulis. We concluded that P. cincinnata was susceptible to FOP.     

Ethylene production and toxigenicity of methionine and its derivatives with riboflavin in cultures of Verticillium, Fusarium and Colletotrichum species exposed to light

Ethylene was produced by Verticillium dahliae Kleb. grown in liquid Czapek's medium. The rate of ethylene production was enhanced by light but was not affected by shaking or the growth rate of the cultures. L-, D- and DL-methionine, DL-ethionine and a -keto- y -methylthiobutyric acid (KMBA) were good substrates for ethylene production. KMBA may be an intermediate in ethylene production and it appears to be degraded to ethylene either enzymatically by peroxidase or photochemically in the presence of riboflavin. Addition of riboflavin or briefly heating the cultures to 100°C enhanced ethylene production greatly, while the addition of sodium azide, potassium cyanide and catalase were very inhibitory. The SS4 (non-defoliating) pathotype of V. dahliae produced significantly more ethylene (up to 108.4 nl ethylene h₁ from 20 ml-10-day-old cultures) than did the T9 (defoliating) pathotype with all substrates tested. The results suggest that the in vitro rate of ethylene production is not related to the relative virulence of pathotypes of V. dahliae on cotton. A number of Verticillium species, Fusarium oxysporum f. sp. vasinfectum and Colletotrichum dematium var. truncatum were able to produce ethylene in liquid Czapek's medium containing 1 m M L-methionine under continuous light. Riboflavin, although highly stimulatory to ethylene production, caused a fungicidal reaction to all the fungi tested in Czapek's medium containing L-methionine under continuous light. The fungicidal effect of the riboflavin-methionine-light combination occurred at concentrations of riboflavin and methionine less than 1.33 μ M and 0.5 m M , respectively. No fungicidal activity was detected when the cultures were grown in total darkness or when either methionine or riboflavin was omitted from the culture medium.     

Emetic material associated with Fusarium species in cereal grains and artificial media

Substances which cause emesis in pigeons were extracted from corn (Zea mays) artificially inoculated with Fusarium graminearum and from liquid culture medium inoculated with F. moniliforme, F. roseum, F. poae, F. culmorum, and F. nivale. Emetic preparations were obtained also from infected wheat (Triticum aestivum L. em. Thell), (Hordeum vulgare L. em. Lam), and durum (Triticum durum Desf). Partial purification resulted from chromatography with columns of cellulose and DEAE cellulose and with thin layers of silica gel. Two active materials were obtained from liquid culture of F. moniliforme but only one from infected cereals. Emetic preparations from F. moniliforme and infected cereals contained a polypeptide as a minor component. Ultraviolet and infrared spectrums, elemental analyses, refractive indices, and amino acid composition of the emetic from corn and one of the emeties from liquid culture of F. moniliforme were similar but not identical. Attempts to crystalline these emetics and to characterize them were unsuccessful.