香蕉镰刀菌枯萎病拮抗放线菌的分离筛选及其抑制效果的初步评价

香蕉镰刀菌枯萎病是一种由尖孢镰刀菌古巴专化型Fusarium oxysporum f. sp. cubense侵染引起的维管束系统性病害。本试验对从海南省东方、八所、黄流、三亚和广东省湛江、徐闻、海安等香蕉种植地采集的根际土样进行拮抗放线菌的分离纯化,得到放线菌菌株139个。通过纸片扩散法,筛选出对香蕉枯萎病4号生理小种具有拮抗作用的菌株8个。进一步试验表明,其中4个菌株不仅对香蕉枯萎病生理小种4号的菌丝生长有良好稳定的抑制作用,且对另外14个不同专化型病原菌也有一定的抑制作用。另外,分别将这8株放线菌发酵上清液与香蕉枯萎病病原菌孢子悬浮液混合12h后,有6株放线菌发酵上清液对病原菌孢子萌发的抑制率超过85%。盆栽试验结果表明,2株放线菌对香蕉枯萎病防效达86%以上,极显著地高于恶霉灵药剂处理。     

香蕉假茎细胞对枯萎病菌不同小种及其粗毒素的病理反应

 以香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)1号小种和4号小种及其粗毒素分别接种香牙蕉和粉蕉的组培苗及离体假茎后,用组织切片法观察香蕉假茎细胞的病理反应,以探明香蕉枯萎病菌不同小种及其粗毒素的致病作用。结果表明,枯萎病菌不同小种人工接种仅能感染相应的香蕉种类,但不同香蕉种类的离体假茎细胞用不同小种接种及其粗毒素处理,均产生褐变等病理反应,且病变程度不存在小种间的差异。表明枯萎病菌不同小种对香蕉不同种类的致病力差异可能与存在其它致病因子或专化性识别的因子有关。同时证实了病菌不同小种的毒素对蕉类不存在着选择毒性     

香蕉枯萎病拮抗菌的筛选及其作用机制研究

通过分离和筛选,从香蕉园或者其他果园的土壤中分离获得13株对香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)具有抑制作用的拮抗菌,并对部分拮抗菌抑制病菌菌丝生长和孢子萌发进行了试验。结果表明,拮抗菌株d4、d5、B3和p发酵液对香蕉枯萎病菌生长具有显著的抑制作用,在平板上产生的抑菌圈直径为21.75～34.75 mm,抑菌效果具有持续稳定性,对孢子萌发的抑制率为90.49%～97.18%;拮抗菌对病菌的作用表现为对菌丝的消融、菌丝细胞的泡囊化、抑制病菌分生孢子的萌发、孢子芽管的扭曲。     

Sensitivity of Fusarium oxysporum f. sp. cubense to phosphonate

Phosphonate (0.1 mM) significantly reduced growth of Fusarium oxysporum f. sp. cubense (Foc) race 4 grown at an optimal phosphate concentration of 0.3 mM in vitro. At higher phosphate concentrations, closer to physiological conditions within the plant, the sensitivity of Foc race 4 to phosphonate was greatly reduced, with 25 mM phosphonate required to reduce growth by 50% at 1 mM phosphate. Two isolates of Fusarium oxysporum f. sp. dianthi and another race of Foc, race 1, were shown to be similar to Foc race 4 in their sensitivity to phosphonate, while another species of Fusarium, F. avenaceum , was more sensitive to phosphonate in vitro.     

In planta and soil quantification of Fusarium oxysporum f. sp. ciceris and evaluation of Fusarium wilt resistance in chickpea with a newly developed quantitative polymerase chain reaction assay

Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris can be managed by risk assessment and use of resistant cultivars. A reliable method for the detection and quantification of F. oxysporum f. sp. ciceris in soil and chickpea tissues would contribute much to implementation of those disease management strategies. In this study, we developed a real-time quantitative polymerase chain reaction (q-PCR) protocol that allows quantifying F. oxysporum f. sp. ciceris DNA down to 1 pg in soil, as well as in the plant root and stem. Use of the q-PCR protocol allowed quantifying as low as 45 colony forming units of F. oxysporum f. sp. ciceris per gram of dry soil from a field plot infested with several races of the pathogen. Moreover, the q-PCR protocol clearly differentiated susceptible from resistant chickpea reactions to the pathogen at 15 days after sowing in artificially infested soil, as well as the degree of virulence between two F. oxysporum f. sp. ciceris races. Also, the protocol detected early asymptomatic root infections and distinguished significant differences in the level of resistance of 12 chickpea cultivars that grew in that same field plot infested with several races of the pathogen. Use of this protocol for fast, reliable, and cost-effective quantification of F. oxysporum f. sp. ciceris in asymptomatic chickpea tissues at early stages of the infection process can be of great value for chickpea breeders and for epidemiological studies in growth chambers, greenhouses and field-scale plots.     

Effectiveness of antagonistic bacteria,commercial fungicides,and fourth generation quaternary ammonium salts,against Fusarium oxysporum f. sp. cubense race “1 or 2”

European Journal of Plant Pathology - There are three races of Fusarium oxysporum f.sp. cubense (Foc) that significantly affect commercial banana plantations. Most banana cultivars are very...     

福建省香蕉枯萎病菌硝酸盐营养突变体的营养体亲和性测定

 Fourteen strains of Fusarium oxysporum f. sp. cubense were induced to produce 146 nitrate-nonutilizing(nit) mutants on a chlorate-containing medium. Among them, there were 117 nit1 mutants(80.14%), 17 nit3 mutants(11.64%) and 12 nitM mutants(8.22%). These strains were divided into two vegetative compatibility groups(VCGs) by the vegetative compatibility tests. Twelve strains of F. oxysporum f. sp. cubense from Musa AAA belonged to VCG1, two trains from Musa ABB belonged to VCG2.     

Interactions of Pratylenchus thornei and Fusarium oxysporum f. sp. ciceris on Chickpea

ABSTRACT Fusarium oxysporum f. sp. ciceris and the root-lesion nematode Pratylenchus thornei coinfect chickpeas in southern Spain. The influence of root infection by P. thornei on the reaction of Fusarium wilt-susceptible (CPS 1 and PV 61) and wilt-resistant (UC 27) chickpea cultivars to F. oxysporum f. sp. ciceris race 5 was investigated under controlled and field conditions. Severity of Fusarium wilt was not modified by coinfection of chickpeas by P. thornei and F. oxysporum f. sp. ciceris, in simultaneous or sequential inoculations with the pathogens. Root infection with five nematodes per cm(3) of soil and 5,000 chlamydospores per g of soil of the fungus resulted in significantly higher numbers of propagules of F. oxysporum f. sp. ciceris with the wilt-susceptible cultivar CPS 1, but not with the wilt-resistant one. However, infection with 10 nematodes per cm(3) of soil significantly increased root infection by F. oxysporum f. sp. ciceris in both cultivars, irrespective of fungal inoculum densities (250 to 2,000 chlamydospores per g of soil). Plant growth was significantly reduced by P. thornei infection on wilt-susceptible and wilt-resistant chickpeas in controlled and field conditions, except when shorter periods of incubation (45 days after inoculation) were used under controlled conditions. Severity of root necrosis was greater in wilt-susceptible and wilt-resistant cultivars when nematodes were present in the root, irrespective of length of incubation time (45 to 90 days), densities of nematodes (5 and 10 nematodes per cm(3) of soil), fungal inocula, and experimental conditions. Nematode reproduction on the wilt-susceptible cultivars, but not on the wilt-resistant one, was significantly increased by F. oxysporum f. sp. ciceris infections under controlled and field conditions.     

Evaluating the potential of soil management to reduce the effect of Fusarium oxysporum f. sp. cubense in banana (Musa AAA)

European Journal of Plant Pathology - Fusarium oxysporum f. sp. cubense (Foc) causes Fusarium wilt in banana (Musa AAA). Foc Race 1 devastated the subgroup Gros Michel during the first half of the...     

香蕉镰刀菌枯萎病拮抗放线菌的筛选及菌株Da03047的鉴定

通过分离和筛选,从海南原始森林尖峰岭采集的土样中获得10株对香蕉镰刀菌枯萎病病源菌具有抑制作用的拮抗放线菌,其中菌株Da03047活性最强且遗传稳定,通过形态特征、生理生化特征、16S rDNA序列测定及其系统发育分析研究,鉴定为灰肉色链霉菌。     

海洋青霉T03对4种热带植物病原真菌的拮抗作用

T03是从多个海洋青霉菌株中筛选出的对植物病原真菌有拮抗作用的一个菌株。采用对峙培养和无菌发酵液抑制菌丝生长方法比较了T03对4种热带植物病原菌香蕉炭疽菌、橡胶炭疽菌、粉蕉镰刀菌、凤梨镰刀菌的拮抗作用。结果显示青霉T03对香蕉炭疽菌、橡胶炭疽菌、粉蕉镰刀菌、凤梨镰刀菌均有一定的拮抗作用,其中对香蕉炭疽菌菌丝生长的抑制作用最好,EC50为17.1%;其次是橡胶炭疽菌,EC50为49.6%。对粉蕉镰刀菌、凤梨镰刀菌菌丝生长的抑制效果不理想。     

Physiological and biochemical aspects of the resistance of banana plants to Fusarium wilt potentiated by silicon

Silicon amendments to soil have resulted in a decrease of diseases caused by several soilborne pathogens affecting a wide number of crops. This study evaluated the physiological and biochemical mechanisms that may have increased resistance of banana to Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense, after treatment with silicon (Si) amendment. Plants from the Grand Nain (resistant to F. oxysporum f. sp. cubense) and "Ma??" (susceptible to F. oxysporum f. sp. cubense) were grown in plastic pots amended with Si at 0 or 0.39 g/kg of soil (-Si or +Si, respectively) and inoculated with race 1 of F. oxysporum f. sp. cubense. Relative lesion length (RLL) and asymptomatic fungal colonization in tissue (AFCT) were evaluated at 40 days after inoculation. Root samples were collected at different times after inoculation with F. oxysporum f. sp. cubense to determine the level of lipid peroxidation, expressed as equivalents of malondialdehyde (MDA), hydrogen peroxide (H(2)O(2)), pigments (chlorophyll a, chlorophyll b, total chlorophyll, and carotenoids), total soluble phenolics (TSP), and lignin-thioglycolic acid (LTGA) derivatives; the activities of the enzymes phenylalanine ammonia-lyases glucanases (PALs), peroxidases (POXs), polyphenoloxidases (PPOs), β-1,3-glucanases (GLUs), and chitinases (CHIs); and Si concentration in roots. Root Si concentration was significantly increased by 35.3% for the +Si treatment compared with the -Si treatment. For Grand Nain, the root Si concentration was significantly increased by 12.8% compared with "Ma??." Plants from Grand Nain and "Ma??" in the +Si treatment showed significant reductions of 40.0 and 57.2%, respectively, for RLL compared with the -Si treatment. For the AFCT, there was a significant reduction of 18.5% in the +Si treatment compared with the -Si treatment. The concentration of MDA significantly decreased for plants from Grand Nain and "Ma??" supplied with Si compared with the -Si treatment while the concentrations of H(2)O(2) on roots and pigments on leaves significantly increased. The concentrations of TSP and LTGA derivatives as well as the PALs, PPOs, POXs, GLUs, and CHIs activities significantly increased on roots of plants from Grand Nain and "Ma??" from the +Si treatment compared with the -Si treatment. Results of this study suggest that the symptoms of Fusarium wilt on roots of banana plants supplied with Si decreased due to an increase in the concentrations of H(2)O(2), TSP, and LTGA derivatives and greater activities of PALs, PPOs, POXs, GLUs, and CHIs.     

Genetic Variation Among Vegetative Compatibility Groups of Fusarium oxysporum f. sp. cubense Analyzed by DNA Fingerprinting

ABSTRACT Genetic variation within a worldwide collection of 208 isolates of Fu-sarium oxysporum f. sp. cubense, representing physiological races 1, 2, 3, and 4 and the 20 reported vegetative compatibility groups (VCGs), was analyzed using modified DNA amplification fingerprinting. Also characterized were 133 isolates that did not belong to any of the reported VCGs of F. oxysporum f. sp. cubense including race 3 isolates from a Heliconia species and isolates from a symptomatic wild banana species growing in the jungle in peninsular Malaysia. The DNA fingerprint patterns were generally VCG specific, irrespective of geographic or host origin. A total of 33 different genotypes were identified within F. oxysporum f. sp. cu-bense; 19 genotypes were distinguished among the isolates that belonged to the 20 reported VCGs, and 14 new genotypes were identified among the isolates that did not belong to any of the existing VCGs. DNA fingerprinting analysis also allowed differentiation of nine clonal lineages within F. oxysporum f. sp. cubense. Five of these lineages each contained numerous closely related VCGs and genotypes, and the remaining four lineages each contained a single genotype. The genetic diversity and geographic distribution of several of these lineages of F. oxysporum f. sp. cubense suggests that they have coevolved with edible bananas and their wild diploid progenitors in Asia. DNA fingerprinting analysis of isolates from the wild pathosystem provides further evidence for the coevolution hypothesis. The genetic isolation and limited geographic distribution of four of the lineages of F. oxysporum f. sp. cubense suggests that the pathogen has also arisen independently, both within and outside of the center of origin of the host.     

Physiological Specialization of Fusarium oxysporum f. sp. lactucae, a Causal Organism of Fusarium Root Rot of Crisp Head Lettuce in Japan

In 1995, Fusarium root rot of crisp head lettuce, caused by Fusarium oxysporum f. sp. lactucae, was simultaneously found in the Shiojiri and Kawakami areas of Nagano Prefecture, Japan. The Shiojiri and Kawakami isolates differed in pathogenicity to lettuce cultivars. Because of this distinct physiological specialization, these Shiojiri and Kawakami isolates should be designated as race 1 and race 2, respectively, using lines VP1010 (highly resistant to race 1), VP1013 (highly resistant to race 2) and variety Patriot (highly susceptible to both races) as differential varieties. This is the first report of races of Fusarium oxysporum f. sp. lactucae, Received 21 September 2000/ Accepted in revised form 21 March 2001     

两种葱属植物提取液对香蕉枯萎病菌的毒力测定及GC-MS分析

 以韭菜和葱的乙酸乙酯提取液对香蕉枯萎病菌进行了抑菌试验,并在高温条件下,采用气相色谱-质谱法(GC-MS)对葱的各部位、韭菜根的乙酸乙酯提取液进行分析。结果表明:韭菜和葱各部位乙酸乙酯提取液对香蕉枯萎病菌均有一定的抑制作用,以韭菜茎的抑制效果最好,为29.85%,韭菜根的抑制效果次之;葱的根、茎、叶和韭菜根的乙酸乙酯提取液检出物分别为34、20、12和21种,其中韭菜根提取液中有较多的含硫化合物,脒基硫脲最多,为17.01%。葱提取液中除含硫化合物外,还检出噻吩、吡嗪和喹啉类物质。     

一种快速、有效的香蕉枯萎病病原菌FOC4的分子检测方法

以ITS测序与SCAR分子标记相结合的香蕉病原菌FOC4分子检测方法,利用通用引物ITS1和ITS4对病原菌SF001的rDNA中ITS序列进行PCR扩增,获得560bp左右的特异条带。经Blast分析比对,确定该菌为FOC。再利用FOC4的特异引物PCL/PDL对基因组上的特征序列扩增区域(SCAR)进行PCR扩增,获得677bp的特有序列,鉴定菌株SF001是尖孢镰刀菌古巴专化型4号生理小种。经过致病性测试,菌株SF001表现出典型4号生理小种的病理特征。     

Inoculum Densities of Fusarium oxysporum f. sp. vasinfectum and Meloidogyne incognita in Relation to the Development of Fusarium Wilt and the Phenology of Cotton Plants (Gossypium hirsutum)

ABSTRACT Development of Fusarium wilt in upland cotton (Gossypium hirsutum) usually requires infections of plants by both Meloidogyne incognita and Fusarium oxysporum f. sp. vasinfectum. In this study, the soil densities of M. incognita and F. oxysporum f. sp. vasinfectum and the incidence of Fusarium wilt in three field sites were determined in 1982-1984. Multiple regression analysis of percent incidence of Fusarium wilt symptoms on population densities of M. incognita and F. oxysporum f. sp. vasinfectum yielded a significant fit (R (2) = 0.64) only on F. oxysporum f. sp. vasinfectum. Significant t-values for slope were also obtained for the interaction of M. incognita and F. oxysporum f. sp. vasinfectum, but densities of M. incognita and F. oxysporum f. sp. vasinfectum were also related on a log(10) scale. The physiological time of appearance of first foliar symptoms of Fusarium wilt, based on a degree-days threshold of 11.9 degrees C (53.5 degrees F), was used as a basis for determining disease progress curves and the phenology of cotton plant growth and development. Effects of Fusarium wilt on plant height and boll set were determined in three successive years. Increases in both of these plant characteristics decreased or stopped before foliar symptoms were apparent. Seed cotton yields of plant cohorts that developed foliar wilt symptoms early in the season (before 2,000 F degree-days) were variable but not much different in these years. This contrasted with cohorts of plants that first showed foliar symptoms late in the season (after 2,400 F degree-days) and cohorts of plants that showed no foliar symptoms of wilt. Regression analyses for 1982-1984 indicated moderate to weak correlations (r = 0.16-0.74) of the time of appearance of the first foliar symptoms and seed cotton yields.     

Sporulation of Fusarium oxysporum f. sp. lycopersici on Stem Surfaces of Tomato Plants and Aerial Dissemination of Inoculum

ABSTRACT Plants exhibiting symptoms of wilt and xylem discoloration typical of Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici were observed in greenhouses of cherry tomatoes at various sites in Israel. However, the lower stems of some of these plants were covered with a pink layer of macroconidia of F. oxysporum. This sign resembles the sporulating layer on stems of tomato plants infected with F. oxysporum f. sp. radicis-lycopersici, which causes the crown and root rot disease. Monoconidial isolates of F. oxysporum from diseased plants were assigned to vegetative compatibility group 0030 of F. oxysporum f. sp. lycopersici and identified as belonging to race 1 of F. oxysporum f. sp. lycopersici. The possibility of coinfection with F. oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis-lycopersici was excluded by testing several macroconidia from each plant. Airborne propagules of F. oxysporum f. sp. lycopersici were trapped on selective medium in greenhouses in which plants with a sporulating layer had been growing. Sporulation on stems was reproduced by inoculating tomato plants with races 1 and 2 of F. oxysporum f. sp. lycopersici. This phenomenon has not been reported previously with F. oxysporum f. sp. lycopersici and might be connected to specific environmental conditions, e.g., high humidity. The sporulation of F. oxysporum f. sp. lycopersici on plant stems and the resultant aerial dissemination of macroconidia may have serious epidemiological consequences. Sanitation of the greenhouse structure, as part of a holistic disease management approach, is necessary to ensure effective disease control.     

Isolation of Nonpathogenic Mutants of Fusarium oxysporum f. sp. melonis for Biological Control of Fusarium Wilt in Cucurbits

ABSTRACT Two nonpathogenic mutant strains 4/4 and 15/15 of Fusarium oxysporum f. sp. melonis (race 1,2) were isolated by a continuous dipinoculation technique following UV mutagenesis of the virulent wild-type isolate FOM1.2. No disease symptoms or detrimental effects were observed following inoculation of muskmelon seedlings by strain 4/4. In contrast, strain 15/15 caused mortality of susceptible cultivars although to a lesser extent than the wild-type isolate. Strain 4/4 colonized a variety of muskmelon and watermelon cultivars. In muskmelon cv. Ein Dor, seedlings were dipped in a conidial suspension of strain 4/4 and planted in medium amended with the mutant to achieve 100% colonization of roots and between 30 to 70% of the lower stem tissues 7 days after planting. Similar percent colonization of watermelon seedlings by strain 4/4 was recorded. In cross-protection experiments with muskmelon cultivars, significant reduction in seedling mortality was observed between 4/4-colonized FOM1.2. challenged plants compared with that of wild-type challenged plants alone. Similarly, strain 4/4 was able to significantly reduce mortality of watermelon seedlings caused by F. oxysporum f. sp. niveum race 2. This novel approach of generating nonpathogenic mutants for biological control in Fusarium spp. and other fungal pathogens from virulent wild-type isolates may be beneficial for control, because the mutant strains, lacking only in pathogenicity, may compete more efficiently than other biocontrol organisms against the pathogen of origin.     

Panama Disease: Cell Wall Reinforcement in Banana Roots in Response to Elicitors from Fusarium oxysporum f. sp. cubense Race Four

ABSTRACT The biochemical basis of tolerance in banana to Fusarium wilt, caused by the pathogen Fusarium oxysporum f. sp. cubense race four, was investigated. Tissue culture banana plants from tolerant cv. Goldfinger and susceptible cv. Williams were maintained in a hydroponic system and inoculated with conidial suspensions to evaluate the degree of tolerance to susceptibility between the two clones and to investigate the effectiveness of this technique as a potential tool for early screening for resistance in breeding programs. Similarly, defense responses were induced by treatment of the plants with an elicitor preparation from the mycelial cell walls of the pathogen. Differences in the induction of lignin and callose deposition, phenolics, and the enzymes involved in cell wall strengthening; phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, peroxidase, and polyphenol oxidase were determined. Root tissue of the tolerant cv. Goldfinger responded to the fungal elicitor through the strong deposition of lignin, preceded by the induction or activation of the enzyme activities involved in the synthesis and polymerization thereof, whereas only slight increases were observed for the susceptible cv. Williams. No increase in callose content was observed for either clone. These results indicate an important role for cell wall strengthening due to the deposition of lignin as an inducible defense mechanism of banana roots against F. oxysporum f. sp. cubense race four.