The Effects of Co-Infection by Sunn-Hemp Mosaic Virus (SHMV) and Fusarium oxysporum on the Growth of French Bean

In the seedlings of three cultivars of French bean (Phaseolus vulgaris), ‘Prince,’‘Masterpiece’ and ‘Pinto’, co-infection by Sunn-hemp mosaic virus (SHMV) with either of the vascular wilt pathogens, Fusarium oxysporum f. sp. phaseoh (Fop) or f. sp. tracbeiphilum (Fot) caused greater losses in total fresh weight and in leaf area compared with uninfected plants or plants infected singly with any one of these pathogens. Co-infection of a fourth cultivar (‘Canadian Wonder’) had no greater effects on growth reduction than single infection. The concentration of SHMV in the leaves of ‘Prince’ and to a greater extent in the leaves of ‘Masterpiece’ increased more after double infection than with infection by the virus alone. The nature and possible mechanisms of the pathogenic effects in French bean are discussed.     

Interactions of Cowpea Mosaic Virus with Cowpea Protoplasts1)

The capability of cowpea mosaic virus to attach to and infect protoplasts of immune, hypersensitive, and susceptible cowpea (Vigna unguiculata) lines was examined by inoculating protoplasts with either purified virus or radioiodinated purified virus ¹²⁵I-CPMV. Systems were used in which plants were immune and protoplasts susceptible, plants were immune and protoplasts resistant, and plants and protoplasts were susceptible to CPMV. No differences were observed in the attachment of ¹²⁵I-CPMV to resistant and susceptible protoplasts. Polycations, proteins, or virus particles were added to the inoculation medium to neutralize potential nonspecific interactions between cells and virus particles. The various additives induced quantitative differences in binding of virus particles to protoplasts.     

Host-specific Fusarium oxysporum Causes Wilt of Jojoba

Jojoba [Simmondsia chinensis (Link) Schneider] plantations in Israel originated from vegetative propagation, planted during 1991–92, have shown symptoms of wilting and subsequent death. Verticillium dahliae was only rarely isolated from these plants and artificial inoculation showed only mild disease symptoms. Fusarium oxysporum caused severe chlorosis, desiccation, defoliation and wilt in leaves of jojoba plants, resulting in plant death. Recovery of the fungus from artificially inoculated stem cuttings and seedlings showed for the first time that F. oxysporum was the primary pathogen. Inoculated cuttings exhibited wilt within 3 weeks, while in seedlings wilt occurred 10–24 weeks after inoculation. Seedlings and cuttings of jojoba which were inoculated with other Fusarium isolates originating from different crops (F. oxysporum f. sp. vasinfectum from cotton, F. oxysporum f. sp. dianthi from carnation, F. oxysporum f. sp. lycopersici from tomato and F. oxysporum f. sp. basilicum from basil) did not develop symptoms. Moreover, cotton, tomato, melon and cucumber seedlings inoculated with several virulent F. oxysporum isolates from jojoba did not show any symptoms of wilt or defoliation. These results indicate a high degree of specificity of the Fusarium isolates from jojoba; therefore, it is suggested that this isolate be defined as F. oxysporum f. sp. simmondsia.     

香蕉幼苗三类有机小分子溶质对尖孢镰刀菌侵染的生理响应

为阐明香蕉枯萎病发病机制,研究了尖孢镰刀菌侵染后,香蕉植株中几种对尖孢镰刀菌生长有显著作用的物质(氨基酸、有机羧酸、酚酸)种类和含量的变化。结果表明:(1)病原菌侵染后,伤害逐渐加剧,株高和生物量显著下降。(2)病原菌侵染后,叶片氨基酸总量显著升高,其中丝氨酸、缬氨酸、组氨酸、异亮氨酸和亮氨酸增幅较大,病原菌侵染16 d,其含量分别为侵染前的7.1、6.2、4.4、3.5和2.3倍;而根氨基酸总量开始显著降低,差异逐渐变小。(3)叶片有机羧酸酸含量在病原菌侵染后显著增加,而在根中显著降低。侵染植株叶片中草酸、柠檬酸、苹果酸、琥珀酸和延胡索酸含量分别是未侵染植株叶片的2.6、1.6、1.9、1.8和2.3倍;根中草酸、柠檬酸、苹果酸、琥珀酸和延胡索酸含量分别是未侵染植株的81%、42%、44%、28%和59%。(4)病原菌侵染后,植株叶片和根中酚酸含量都显著升高。叶片中阿魏酸、肉桂酸和水杨酸含量分别是未侵染叶片的2.9、1.7和2.9倍;而根中对羟基苯甲酸和丁香酸含量分别是未侵染根的4.3和1.5倍。研究结果表明,尖孢镰刀菌侵染后,植物与病原菌的相互作用使得植物体内抑菌物质和促菌物质都会相应的增加,植株对病害有一定的抗性,但促菌物质种类和含量较高最终使得感病植株发病。     

Pathogenicity of Fusarium oxysporum Isolates from Malaysia and F. oxysporuin f. sp. elaeidis from Africa to Seedlings of Oil Palms (Elaeis guineensis)

Pathogenicity tests with Fusarium oxysporum isolated form Malaysian oil palm were made with oil palms seedlings raised form Malaysian seed as well s with wilt-susceptible seedlings gown from African seed. Oil palm seedlings grown form Malaysian seed were also inoculated with African isolates of F. oxysporum f. sp. elaeidis and F. oxysporum var. redolens. The experiments were made under normal soil moisture conditions and under water stress. F. oxysporum f. sp. elaeidis isolates form Africa were pathogenic to oil palm seedlings from Malaysian seeds but the Malaysian F oxysporum isolates were non-pathogenic to plams grown from Malaysian seed or the wilt-susceptible palms from African seed. Seedlings from Malaysian seed proved to be highly susceptible to the vascular wilt disease caused by F. oxysporum f. sp. elaeidis as 75–90% of the palms were infected. The susceptibility of the palms from Malaysian seed varied with different African isolates tested. The Yaligimba isolate from Zaire which was found to be F. oxysporum var. redolens was the most virulent. Disease was more severe when oil palm seedlings were subjected to a period of water stress. The incidence of death in the seedlings under stress conditions was 45% as compared with only 15% for palms grown under normal conditions.     

The effects of Fusarium oxysporum f.sp. lycopersici (Sacc.) Snyder & Hansen on tomato in diphenamid-treated soil

Pre-emergence soil application of the herbicide diphenamid in concentrations exceeding the normal field rate increased the resistance of tomato plants towards infection by the wilt fungus Fusarium oxysporum f.sp. lycopersici. This was detected as significant increases in the percentage emergence of seedlings although growth parameters of the raised seedlings were reduced. Treated plants exhibited no wilt symptoms, although the pathogen maintained its population at detectable levels in the rhizosphere of tomato plants. However, the growth inhibition caused by diphenamid alone was much less than that reported for the combined application of pathogen and herbicide. Growth activities of F. oxysporum f.sp. lycopersici were inhibited by high concentrations of diphenamid in vitro. It is possible that the biodegradation of this herbicide by species such as Aspergillus candidus (present in substantial counts in treated rhizospheres) was one of the causes of increased tolerence of the pathogen to the herbicide in situ.     

Fungus gnats vector Fusarium oxysporum f.sp. radicislycopersici1

Fungus gnat adults transported Fusarium oxysporum f.sp. radicis-lycopersici from Petri dish culture and infected host plants to the roots and hypocotyls of healthy tomato and bean plants. The source of the fungus did not affect the ability of fungus gnats to transport the fungus to healthy hosts. The presence of fungus gnat larvae in media in which young tomato plants were grown did not increase the incidence of plant infection by the pathogen. Fungus gnat adults appear to aid in the dissemination of F. oxysporum f.sp. radicis-lycopersici.     

Chromatographic Characterization and Phytotoxic Activity of Fusarium oxysporum f. sp. albedinis and Saprophytic Strain Toxins

The bayoud disease, vascular fusariosis of date palm tree (Phoenix dactylifera L.), is caused by the pathogenic fungus Fusarium oxysporum f. sp. albedinis. The characteristic symptoms of the bayoud disease were elicited on detached leaves of F. oxysporum f. sp. albedinis‐susceptible cultivars of date palm trees, which were treated either with the F_II (F. oxysporum f. sp. albedinis) fraction purified from the organic extracts of a F. oxysporum f. sp. albedinis liquid culture, or with a solution of fusaric acid. Enniatins, which are secreted by several Fusarium species, were tested at different concentrations and were not capable of inducing symptoms on such detached leaves. The F_II (F. oxysporum f. sp. albedinis) fraction was unable to induce necrosis of potato slices, which indicates that it does not contain significant amounts of enniatins. The high‐performance liquid chromatography (HPLC) profiles of the F_II (F. oxysporum f. sp. albedinis) fraction showed toxic peaks different from fusaric acid. A fraction, named F_II (AZ₄), was obtained from culture filtrates of a saprophytic Fusarium strain maintained in the same cultural conditions as for the F. oxysporum f. sp. albedinis. The HPLC profile of the F_II (AZ₄) fraction did not show the characteristic phytotoxic peaks present in the F_II (F. oxysporum f. sp. albedinis) fraction. This finding well agrees with the fact that the F_II (AZ₄) fraction is not toxic to detached date palm leaves. Moreover, the HPLC profiles of F_II fractions obtained from other special forms of F. oxysporum are different the F_II (F. oxysporum f. sp. albedinis) profile. The phytotoxic compounds purified from the F_II (F. oxysporum f. sp. albedinis) fraction are probably new molecules that may help in understanding the pathogenesis of bayoud disease.     

Insect vectors of cowpea mosaic virus in Nigeria

A yellow strain of cowpea mosaic virus (CPMV) was transmitted in cowpea by two thrips, Sericothrips occipitalis and Taeniothrips sjostedti; two chrysomelid beetles, Ootheca mutabilis and Paraluperodes quaternus; a curculionid beetle, Nematocerus acerbus; and two acridid grasshoppers, Catantops spissus spissus and Zonocerus variegatus. Summarizing trials with single insects, the efficiency of transmission of CPMV averaged 18—21% for N. acerbus and the two grasshoppers, 55% for P. quaternus, and 71% for O. mutabilis. Twenty-two and 40% of the plants exposed to large populations of S. occipitalis and T. sjostedti, respectively, were infected. In three trials with an aphid, Aphis craccivora, 4 of 49 plants were infected with CPMV, but these infections were considered spurious because no infections occurred in any of 63 plants exposed to this insect in four other trials. A coreid bug, Riptortus dentipes, did not transmit CPMV. Mosaic symptoms in infected plants appeared 5—39 days after they were exposed to vectors. Infective virus was recovered from fresh faecal pellets of each grasshopper vector.     

Host perception of jasmonates promotes infection by Fusarium oxysporum formae speciales that produce isoleucine‐ and leucine‐conjugated jasmonates

Three pathogenic forms, or formae speciales (f. spp.), of Fusarium oxysporum infect the roots of Arabidopsis thaliana below ground, instigating symptoms of wilt disease in leaves above ground. In previous reports, Arabidopsis mutants that are deficient in the biosynthesis of abscisic acid or salicylic acid or insensitive to ethylene or jasmonates exhibited either more or less wilt disease, than the wild‐type, implicating the involvement of hormones in the normal host response to F. oxysporum. Our analysis of hormone‐related mutants finds no evidence that endogenous hormones contribute to infection in roots. Mutants that are deficient in abscisic acid and insensitive to ethylene show no less infection than the wild‐type, although they exhibit less disease. Whether a mutant that is insensitive to jasmonates affects infection depends on which forma specialis (f. sp.) is infecting the roots. Insensitivity to jasmonates suppresses infection by F. oxysporum f. sp. conglutinans and F. oxysporum f. sp. matthioli, which produce isoleucine‐ and leucine‐conjugated jasmonate (JA‐Ile/Leu), respectively, in culture filtrates, whereas insensitivity to jasmonates has no effect on infection by F. oxysporum f. sp. raphani, which produces no detectable JA‐Ile/Leu. Furthermore, insensitivity to jasmonates has no effect on wilt disease of tomato, and the tomato pathogen F. oxysporum f. sp. lycopersici produces no detectable jasmonates. Thus, some, but not all, F. oxysporum pathogens appear to utilize jasmonates as effectors, promoting infection in roots and/or the development of symptoms in shoots. Only when the infection of roots is promoted by jasmonates is wilt disease enhanced in a mutant deficient in salicylic acid biosynthesis.     

Further Investigation of the Specificity of Interactions Between Wild Lactuca spp. and Bremia lactucae Isolates from Lactuca serriola

Callus cultures derived from isogenic lines of the tomato cultivars Moneymaker and Craigella, resistant or susceptible to F. oxysporum f. sp. lycopersici, were inoculated with Fusarium oxysporum f. sp. lycopersici race 1. Fungal growth was restricted on callus derived from resistant plants, after inoculation with a conidial suspension, whereas callus derived from susceptible plants was totally overgrown by the fungus within 7 days. The concentration of the phytoalexin rishitin was significantly higher in the callus culture derived from a resistant tomato line compared with the callus culture from a susceptible line, 2 and 3 days after inoculation with mycelium. The results of the experiments were compared with experiments with whole plants. Rishitin production as well as growth of the fungus was comparable with responses in plant-fungus interaction. Therefore callus culture may be useful in studying the interaction between tomato plants and race 1 of F. oxysporum f. sp. lycopersici.     

Management Fusarium wilt on melon and watermelon by Penicillium oxalicum

The potential of the biological control fungus Penicillium oxalicum to suppress wilt caused by Fusarium oxysporum f. sp. melonis and F. oxysporum f. sp. niveum on melon and watermelon, respectively, was tested under different growth conditions. The area under disease progress curve of F. oxysporum f. sp. melonis infected melon plants was significantly reduced in growth chamber and field experiments. In glasshouse experiments, it was necessary to apply P. oxalicum and dazomet in order to reduce Fusarium wilt severity in melons caused by F. oxysporum f. sp. melonis. For watermelons, we found that P. oxalicum alone reduced the area under the disease progress curve by 58% in the growth chamber experiments and 54% in the glasshouse experiments. From these results, we suggested that P. oxalicum may be effective for the management of Fusarium wilt in melon and watermelon plants.     

Fusaric acid accelerates the senescence of leaf in banana when infected by Fusarium

Fusarium oxysporum f.sp. cubense (FOC) is a causal agent of vascular wilt and leaf chlorosis of banana plants. Chloroses resulting from FOC occur first in the lowest leaves of banana seedlings and gradually progress upward. To investigate the responses of different leaf positions to FOC infection, hydroponic experiments with FOC inoculation were conducted in a greenhouse. Fusarium-infected seedlings exhibited a decrease in net photosynthesis rate, stomatal conductance, and transpiration rate of all leaves. The wilting process in Fusarium-infected seedlings varied with leaf position. Measurements of the maximum photochemical efficiency of photosystem II (F _V/F _max) and visualization with transmission electron microscopy showed a positive correlation between chloroplast impairment and severity of disease symptoms. Furthermore, results of malondialdehyde content and relative membrane conductivity measurements demonstrated that the membrane system was damaged in infected leaves. Additionally, the activities of phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase were increased and total soluble phenolic compounds were significantly accumulated in the leaves of infected plants. The structural and biochemical changes of infected plants was consistent with plant senescence. As the FOC was not detected in infected leaves, we proposed that the chloroplast and membrane could be damaged by fusaric acid produced by Fusarium. During the infection, fusaric acid was first accumulated in the lower leaves and water-soluble substances in the lower leaves could dramatically enhance fusaric acid production. Taken together, the senescence of infected banana plants was induced by Fusarium infection with fusaric acid production and the composition of different leaf positions largely contribute to the particular senescence process.     

First Record of Fusarium Wilt of Tobacco in Greece Imported as Seedborne Inoculum

Leaf yellowing and brown discoloration was observed in tobacco plants cv. Burley TN97 in tobacco fields of central Greece in 2002. Fusarium oxysporum f. sp. nicotianae was isolated from symptomatic plants and Koch's postulates were fulfilled. The pathogenicity of the isolated fungus was examined on five tobacco cultivars (Burley TN97, BurleyB21, VirginiaBE9, Virginia Niki and Anatolika KE26/2). The pathogen was present in tobacco seed batches imported in 2000 and 2001, which indicates that the infected seed is most probably the primary source of the disease in Greece. As Fusarium oxysporum f. sp. vasinfectum can also cause vascular wilt in tobacco, the hypothesis that the isolated F. oxysporum strain belongs to f. sp. vasinfectum was excluded by a pathogenicity test to cotton cv. Acala SJ‐2. This is the first report of F. oxysporum f. sp. nicotianae in Greece and the second in the European Union, although the seedborne nature of the pathogen has not been previously reported in Europe.     

Cowpea mosaic virus chimeric particles bearing the ectodomain of matrix protein 2 (M2E) of the influenza a virus: Production and characterization

The epitope presentation system for the ectodomain of the M2 protein (M2e) of the influenza A virus was constructed on the basis of the cowpea mosaic virus (CPMV) for expression in the plant Vigna unguiculata. CPMV is widely used as a vector to produce immunogenic chimeric virus particles (CVPs) bearing epitopes of various infectious human and animal pathogens. To produce chimeric CPMV particles in plants, two binary vectors were constructed to bear a modified gene coding for the CPMV S-coat protein with insertions of M2e epitopes of human influenza and bird influenza viruses. Antigenic and immunogenic properties of CVPs were investigated in mice immunization experiments. CVPs were shown to induce anti-M2e IgG production and to partly protect mice against a challenge with low doses of the influenza virus. However, low infectivity and immunogenicity of chimeric CPMV particles indicate that the plant virus-based systems for M2e epitope presentation requires further optimization in order to use plants as a possible source of flu vaccines.     

Appearance of a Host Protein in Cucumber Plants Infected with Viruses, Bacteria and Fungi

Electrophoretic analyses of extracts of cucumber leaves infectedwith Colleiotrichum lagenarium, Fusarium oxysporum f. sp. cucumerinum,Pseudomonas lachrymans, Erwinia tracheiphila, tobacco necrosisvirus or cucumber mosaic virus revealed the presence of a proteinband with an R_F value of 0.55–0.60 (based on mobilityof bromophenol blue) on 10% polyacrylamide gel. This band wasnot evident in extracts of healthy or mechanically wounded leaves.The protein was not detected in uninfected leaves of infectedplants, but it was detected in similar amounts in infected leavesand in secondarily challenged leaves of infected plants eventhough symptoms were not apparent on the latter. The proteinhad a molecular weight of approximately 16 000 d, was adsorbedon DEAE-cellulose, did not react with Schiff's reagent, anddid not have ribonuclease activity. When injected into cucumberleaves, it did not inhibit germination of conidia of C. lagenariumor induce resistance against disease caused by the fungus.     

Sciarid and shore flies as aerial vectors of Fusarium oxysporum f. sp. cucumerinum in greenhouse cucumbers

The options for managing Fusarium wilt in greenhouse cucumbers are limited by our poor understanding of the modes of survival and dissemination of the pathogen. This study uses a specific quantitative real‐time PCR assay for Fusarium oxysporum f. sp. cucumerinum to investigate the significance of flying insects as aerial vectors of the pathogen in a commercial cucumber greenhouse. Shore flies were more frequently detected (35.5%) carrying F. oxysporum f. sp. cucumerinum than sciarids (25%), with both species carrying between 1 × 10² and 1 × 10⁶ pathogen genome copies/individual. Sciarid and shore flies acquired F. oxysporum f. sp. cucumerinum following exposures to agar cultures of the pathogen of up to 94 h. Light microscopy revealed that spores were carried externally on the bodies of the adult flies. The ability of adult sciarid flies to vector the pathogen from peat‐grown diseased cucumber plants and infect healthy cucumber plants was demonstrated in a caged glasshouse trial. An inoculum density trial showed that vascular wilt disease was initiated after inoculation of peat‐grown seedlings with as few as 1000 conidia. We conclude that sciarid and shore flies play significant roles as vectors of F. oxysporum f. sp. cucumerinum in greenhouse cucumbers and need to be recognized in developing integrated crop management strategies.     

Physiological and Biochemical Aspects of Symbiotic Nitrogen Fixation in Cowpea (Vigna unguiculata (L.) Walp. var. Tuy) Plants Infected by Cowpea Mosaic Virus

Three-day-old cowpea seedlings were inoculated with the severestrain of Cowpea Mosaic Virus (CpMV) and 24 h later with Bradyrhizobiumsp. cowpea, strain I-125, when virus translocation to rootsstill had not taken place. Plants were harvested at 22, 30,45 and 59 d after germination. Active virus replication wasassociated with increased protein content, detected in the leavesof 22-d-old plants. CpMV infection reduced the total leaf area,dry weight of shoots and the chlorophyll content of the firsttrifoliolate leaf at all experimental times. Low sugar contentwas recorded in leaves of 22- and 30-d-old CpMV-infected plantsand in nodules and roots of 30- and 45-d-old CpMV-infected plants.Up to 45 d, the nodule mass of CpMV-infected plants was lowerthan in controls, but reached control values at the 4th harvest.In CpMV-infected nodules, massive agglomeration of virus particles,crystalline virus inclusions and the proliferation of the endoplasmicreticulum were observed only in those cells containing bacteroids.In 30- and 45-d-old plants, CpMV infection decreased the contentof ureides in nodules, roots, and petioles. Virus infectiondid not alter the -amino-N content of roots and nodules butinduced a transient 74% reduction in the level of -amino-N inpetioles of 45-d-old plants. At the 1st and 2nd harvests theactivity of uricase (EC 1.7.3.3 [EC] ) in the nodules and of pyruvatekinase (EC 2.7.1.40 [EC] ) in the nodules and leaves were decreasedseverely by virus infection. CpMV did not hinder the allantoinase(EC 3.5.2.5 [EC] ) activity in the leaves but caused a 9% transitorydecrease in the activity of this enzyme in nodules of 45-d-oldplants. Measurements of NAD-malic enzyme (EC 1.1.1.38 [EC] ) in nodulesalso showed the non-effect of CpMV on this enzyme, except fora temporary 16% reduction at the 2nd harvest. As compared tocontrols, the relative abundance of ureides in 30-d-old CpMV-infectedplants indicated a 15%, 10%, and 51% reduction in the nodules,roots, and petioles, respectively. Results indicate that atthe time of the 4th harvest the symbiotic process, measuredin terms of ureide content and enzymatic activities, was functioningat a near normal level despite nodule infection by CpMV. Key words: Cowpea Mosaic Virus, nitrogen fixation, cowpea, enzymes, ultrastructure     

Biological control ofFusarium oxysporum f.sp.cubense in banana by inoculation withPseudomonas fluorescens

Native strains ofPseudomonas fluorescens exhibitedin vitro antibiosis towards isolates of races 1 and 4 ofFusarium oxysporum f.sp.cubense, the Panama wilt pathogen of banana. The seedlings ofMusa balbisiana seedlings treated withP. fluorescens showed less severe wilting and internal discolouration due toF. oxysporum f.sp.cubense infection in greenhouse experiments. In addition to suppressing Panama wilt, bacterized seedlings ofM. balbisiana also showed better root growth and enhanced plant height.     

Genetic Diversity of Fusarium oxysporum Populations Isolated from Tomato Plants in Tunisia

Fusarium crown and root rot of tomato (Lycopersicon esculentum) caused by Fusarium oxysporum f. sp. radicis‐lycopersici is a new devastative disease of tomato greenhouse crops in Tunisia. Nothing is known neither about the population of this pathogen in this region, nor about the population of F. oxysporum f. sp. lycopersici the causal agent of Fusarium wilt of tomato. In order to examine the genetic relatedness among the F. oxysporum isolates by intergenic spacer restriction fragment length polymorphism (IGS‐RFLP) analysis and to elucidate the origin of the formae specialesradicis‐lycopersici in Tunisia by looking for genetic similarity of Tunisians isolates with isolates from a foreign source, the genetic diversity among F. oxysporum f. sp. radicis‐lycopersici and F. oxysporum f. sp. lycopersici populations was investigated. A total of 62 isolates of F. oxysporum, obtained from symptomless tomato plants, were characterized using IGS typing and pathogenicity tests on tomato plants. All Fusarium isolates were highly pathogenic on tomato. Fusarium oxysporum f. sp. radicis‐lycopersici isolates were separated into five IGS types. From the 53 F. oxysporum f. sp. radicis‐lycopersici isolates, 34 isolates have the same IGS types (IGS type 25), and the remaining 19 isolates were distributed into four IGS types. However, the only nine isolates of F. oxysporum f. sp. lycopersici have six different IGS types. This difference of diversity between the two formae speciales suggests that F. oxysporum f. sp. radicis‐lycopersici isolates have a foreign origin and may have been accidentally introduced into Tunisia.