Mineral Uptake in Fusarium oxysporum f. sp. vasinfectum

A method for growing Fusarium oxysporum, a mycelial fungus, and a technique for its use in mineral uptake studies have been described. Some general characteristics of the uptake process were determined. The fungus, grown for 54 hours, was found to take up as much K as 15 to 20 meq/100 g dry weight in 2 to 4 hours from a solution of 5 meq/l KCl. Approximately 3 to 5 meq of this uptake was readily removed by a CaCl₂ rinse. The uptake was only slightly sensitive to pH over the range of 4 to 9. Below pH 4 uptake dropped rapidly. The age of the culture appeared to be the dominant factor in determining the rate of uptake. In contrast to other fungi, the presence of glucose during uptake was detrimental to K uptake. Conditions unfavorable for metabolic activity as low temperature, anaerobiosis, or the presence of DNP markedly reduced the uptake rate. Although the fungus took up Na from single salt solutions nearly as well as K, the latter ion was much preferred in mixtures of the two ions. The organism showed no significant metabolic uptake of Ca or Cl. During uptake from KCl solutions, the organic acid content increased. The increase, chiefly in succinic acid and to a lesser extent in acetic and citric acids, amounted to about half the K uptake. The remainder of the K taken up was correlated with a roughly equivalent efflux of cellular Mg.     

Lectins Specificity to Pathogenesis of Fusarium oxysporum f. sp. vasinfectum

Lectins of cotton were isolated from either resistant or susceptible seed cultivars. In agar-gel double diffusion tests, positive reactions took place between lectins of cotton cultivars and the antisera of their corresponding wilt Fusaria. The number of precipitin bands correlated with the degree of susceptibility of the tested cultivars. On the other hand, no visible reaction was detected when these antisera were subjected to react with either the resistant host or nonhost lectins.     

Interactions of Concomitant Species of Nematodes and Fusarium oxysporum f. sp. vasinfectum on Cotton

Meloidogyne incognita, Hoplolaintus galeatus, and North Carolina and Georgia populations of Belonolaimus longicaudatus were introduced singly and in various combinations with Fusarium oxysporum f. sp. vasinfectum on wilt-susceptible ''Rowden'' cotton. Of all the nematodes, the combination of the N. C. population of B. longicaudatus with Fusarium promoted greatest wilt development. H. galeatus had no effect on wilt. With Fusarium plus M. incognito or B. longicaudatus, high nematode levels promoted greater wilt than low levels. The combination of either population of B. longicaudatus with M. incognita and Fusarium induced greater wilt development than comparable inoculum densities of either nematode alone or where H. galeatus was substituted for either of these nematodes. Nematode reproduction was inversely related to wilt development. Without Fusarium, however, the high inoculum level resulted in greater reproduction of all nematode species on cotton. Combining M. incognita with B. longicaudatus or H. galeatus gave mutually depressive effects on final nematode populations. The interactions of H. gateatus with B. longicaudatus varied with two populations of the latter.     

The Inhibition of Fungal Growth in Resistant Cotton Plants Infected by Fusarium oxysporum f. sp. vasinfectum

The vascular colonization of cotton plants by Fusarium oxysporum f. sp. vasinfectum was determined by examining growth of the fungus from free-hand cross sections taken from 0 to six days after inoculation at various distances above the points of root inoculation. Fungal spread in both longitudinal and lateral directions in the susceptible cultivar Rowden was evident four days after inoculation, whereas fungal spread in the resistant cultivar Seabrook Sea Island was restricted. The quantity of viable fungus in infected tissues was determined from macerated tissues plated on Czapek- Dox agar. The colony counts declined within six days after inoculation in resistant Seabrook Sea Island, but not in susceptible Rowden, implying that an inhibition of fungal growth in vascular tissues occurred in resistant Seabrook Sea Island. This inhibition could contribute to the restriction of fungal spread and thus be a factor in the resistance of cotton plants to F. oxysporum f. sp. vasinfectum .     

Production and Fungitoxicity of the Terpenoid Phytoalexins in Cotton Inoculated with Fusarium oxysporum f. sp. vasinfectum

Four terpenoid phytoalexins, desoxyhemigossypol (dHG), hemigossypol (HG), desoxyhemigossypol- 6-methyl ether (dMHG) and hemigossypol-6-methyl ether (MHG), were identified with HPLC analysis of extracts from the moderately Fusarium wilt resistant cotton var. TAMCOT CAMD-E ( Gossypium hirsutum ) inoculated with Fusarium oxysporum f. sp. vasinfectum (F. o. v.). Concentrations of dHG, HG, dMHG and MG in stem steles at 10 days after inoculation were 45.1, 175. 0, 38. 3 and 1. 6 (g g^–1 fresh tissue, respectively. The bioassays demonstrated that all four phytoalexins were toxic to F. o. v. The ED₅₀'s of dHG, dMHG and HG were calculated as 8. 8, 13. 4 and 29. 3 (g ml⁻¹, respectively. The very low solubility of MHG in the standard assay medium prevented the determination of its ED₅₀ value. Only dHG is water soluble at levels that appear necessary to act as an effective phytoalexin. At 30 μg ml⁻¹, dHG kills all conidia and mycelia of F. o. v . Viable propagules of F. o. v. were recovered from the steles of inoculated plants 10 days after inoculation; however, the pathogen was restricted to a zone 15 cm above the hypocotyl inoculation site. Thus, the fungistatic action of dHG appears to contribute to the resistance of cotton to Fusarium wilt by preventing the systemic distribution of F. o. v . propagules.     

Fusaric acid and other metabolite production in Fusarium oxysporum f. sp. vasinfectum

A thin layer chromatography system is described for the analysis of fusaric acid and other secondary metabolites, to assist in the identification of pathogenic races of Fusarium oxysporum f. sp. vasinfectum . There were differences in the metabolite profiles of the races grown on different media.     

Interaction of Population Levels of Fusarium oxysporum f. sp. vasinfectum and Meloidogyne incognita on Cotton

In autoclaved greenhouse soil without Fusarium oxysporum f. sp. vasinfectum, Meloidogyne incognita did not cause leaf or vascular discoloration of 59-day-old cotton plants. Plants had root galls with as few as 50 Meloidogyne larvae per plant. Root galling was directly proportional to the initial nematode population level. Fusarium wilt symptoms occurred without nematodes with 77,000 fungus propagules or more per gram of soil. As few as 50 Meloidogyne larvae accompanying 650 fungus propagules caused Fusarium wilt. With few exceptions, leaf symptoms appeared sooner as numbers of either or both organisms increased. In soils infested with both organisms, the extent of fungal invasion and colonization was well correlated with the extent of nematode galling and other indications of the Fusarium wilt syndrome.     

Differential interactions among cotton genotypes and isolates of Fusarium oxysporum f. sp. vasinfectum

Abstract

The pathogenicity of nine isolates of Fusarium oxysporum f. sp. vasinfectum (Fov) was evaluated on seedlings of 30 cotton (Gossypium barbadense L.) genotypes in 2005 and 2006. Isolate×genotype interaction was a highly significant (P < 0.01) source of variation in wilt incidence, suggesting that physiologic specialization exists within Fov isolates. Cluster analysis of aggressiveness of isolates and susceptibility of genotypes by the unweighted pair-group method based on arithmetic means (UPGMA) placed the isolates and the genotypes in several groups. Isolates were separated into two distinct groups. One group was closely related to race 5 while the other group was closely related to race 1. Cluster analysis also demonstrated that the Egyptian commercial cultivars had unique susceptibility patterns to Fov isolates remotely related to those of the other genotypes. The interaction between experiments of 2005 and 2006 was mainly due to a differential effect of years on the disease incidence for cotton cultivars.     

The Influence of Phosphate Uptake on Cation Uptake in Fusarium oxysporum f. sp. vasinfectum

Fusarium oxysporum grown in a low phosphate medium was found to take up several times as much K from KH₂PO₄ as from KCI solutions. Large amounts of phosphate also were taken up from KH₂PO₄. Similar large uptakes of Na and phosphate took place from solutions of NaH₂PO₄. Substantial quanties of phosphate were taken up from solutions of Ca(H₂PO₄)₂ in the absence of any appreciable Ca uptake. When the fungus was grown in a medium containing high phosphate, little or no uptake of phosphate from KH₂PO₄ solutions occured and the K Uptake was at the same level as from KCI solutions. During large phosphate uptake sizable reductions in the organic acid content of the fungal cells were observed. Much, but not all, of the data could be explained on the basis of maintenance of charge balance within the cells. – The respiratory rate of fungus, grown in a low P medium, was markedly increased in KH₂PO₄ solution. Fungus, grown in a medium with high phosphate, had a higher respiratory rate which showed only a slight response to KH₂PO₄ solution. Fungus, grown in a medium with high phosphate, had a higher respiratory rate which showed only a slight response to KH₂PO₄.     

乙烯产生的抑制与氟乐灵诱导棉苗对枯萎病的抗性有关(英文)

氟乐灵处理可诱发棉苗产生对枯萎病的诱导抗性,表现为推迟显症,降低发病率和病情指数。经1．0－4．0μg／g的氟乐灵处理后,棉苗受枯萎病菌侵染时其叶片和根茎部组织中乙烯的产生量明显下降（表1）,而且病菌侵染后氟乐灵处理棉苗中乙烯产生的抑制与其发病率和病情指数的下降呈正相关（图1）。经氟乐灵处理的棉苗受病菌侵染后喷以138．4μmol/L的乙烯利可同时提高其组织中乙烯的产生量以及发病率和病情指数（图2）,而未经氟乐灵处理的棉苗受病菌侵染后喷以乙烯合成抑制剂──2,4-二硝基苯酚和乙烯作用抑制剂──硫代硫酸银则明显降低发病率和病情指数（表2）。由此认为,乙烯产生的抑制可能与氟乐灵诱发棉苗产生对枯萎病的诱导抗性有关。     

棉枯萎病菌异核体及其不同表型分离子在基因转录水平上的差异

采用银染mRNA差异显示技术,研究了棉花枯萎病菌异核体菌株(Ag149)及其两个不同表型分离子(Ag149-Ⅰ和Ag149-Ⅲ)之间差异表达的基因,观察到在300～700bp之间出现了19个差异条带,经反向RNA印迹,证实其中2条差异带为阳性：编号为C6的差异条带在Ag149和Ag149-Ⅰ菌株中呈高表达,而编号为G5的差异条带在菌株Ag149和Ag149-Ⅲ中呈高表达。这两条差异片段经测序后在GenBank中分别进行同源比较,发现由C6片段编码的氨基酸序列与多种生物(包括动物、植物和微生物)NADH脱氢酶的第6个亚基有不同程度的同源性(30％～70％);而编码G5片段的氨基酸序列与龟裂链霉菌(Streptomyces rimosus)的OtrB(tetracycline efflux protein)蛋白有35％的同源性。首次证明在异核体菌株及其不同表型分离子之间存在基因转录水平差异,为深入研究丝状真菌异核体形成的分子遗传机制提供了线索。     

己二酸二异丁酯对辣椒种子萌发及枯萎菌的化感效应

采用模拟的方式,利用生物测定和室内培养的方法,研究了不同浓度的己二酸二异丁酯对辣椒种子萌发、幼苗生长及辣椒枯萎菌的化感效应.结果表明:己二酸二异丁酯对辣椒种子萌发和幼苗生长具有低浓度促进、高浓度抑制的化感效应;己二酸二异丁酯对辣椒和茄子种子的化感效应存在较大差异,在低浓度时对辣椒种子萌发和幼苗生长的促进作用强于茄子种子,而高浓度时对辣椒种子萌发和幼苗生长的抑制强度弱于茄子种子;各浓度己二酸二异丁酯对辣椒枯萎菌菌丝生长有极显著的抑制作用,且作用强度随着浓度增加而增强,当浓度为1 mnol·L~(-1)时抑制作用最强,抑制率为10.75%;在田间抗病性调查期间内各浓度处理均提高了辣椒幼苗的抗病性,其中以1 mmol·L~(-1)处理抗病性最好,病情指数比对照降低了49.88%.

Abstract：

By the methods of laboratory incubation and bioassay, a simulation test was conducted to study the allelopathic effects of different concentration diisobutyl adipate on pepper seed germi-nation and its seedling growth and Fusarium oxysporum f. sp. vasinfectum. For the pepper seed germination and seedling growth, diisobutyl adipate exhibited allelopathic promotion at low con-centration, but allelopathic inhibition at high concentration. There existed greater differences in the allelopathic effects of diisobutyl adipate on the seed germination and seedling growth of pepper and eggplant. The promotion effects of low concentration diisobutyl adipate on pepper were stron-ger than those on eggplant, while the inhibition effects of high concentration diisobutyl adipate on pepper were weaker than those on eggplant. All test concentration diisobutyl adipate had signifi-cant inhibition effects on the mycelium growth of Fusarium oxysporum f. sp. vasinfectum, and the effects increased with increasing diisobutyl adipate concentration, being the strongest (an inhibi-tory rate of 10. 75%) at 1 mnol·L~(-1). Field text indicated that diisobutyl adipate at all test con-centrations enhanced the disease resistance of pepper seedlings. The best effect was observed at 1 mmol·L~(-1) of diisobutyl adipate, with the disease index decreased by 49.88%, compared to the control.     

Production of 3-acetoxyscirpene-4,15-diol from anguidine (4,15-diacetoxyscirpene-3-ol) by Fusarium oxysporum f.sp. vasinfectum.

Growing cells of Fusarium oxysporum f.sp. vasinfectum (ATCC 7808) formed 3-acetoxyscirpene-4,15-diol from anguidine (4,15-diacetoxyscirpene-3-ol) by way of the intermediates triacetoxyscirpene, 3,4-diacetoxyscirpene-15-ol and 3,15-diacetoxyscirpene-4-ol. The new 3-acetoxy analog was found to be less active than anguidine and the other monoacetoxy derivatives when tested against a series of fungal strains and against HeLa cells in vitro.     

Production of 3-acetoxyscirpene-4,15-diol from anguidine (4,15-diacetoxyscirpene-3-ol) by Fusarium oxysporum f.sp. vasinfectum.

Growing cells of Fusarium oxysporum f.sp. vasinfectum (ATCC 7808) formed 3-acetoxyscirpene-4,15-diol from anguidine (4,15-diacetoxyscirpene-3-ol) by way of the intermediates triacetoxyscirpene, 3,4-diacetoxyscirpene-15-ol and 3,15-diacetoxyscirpene-4-ol. The new 3-acetoxy analog was found to be less active than anguidine and the other monoacetoxy derivatives when tested against a series of fungal strains and against HeLa cells in vitro.     

Cross Protection Between Strains of Fusarium oxysporum f. sp. vasinfectum and its Effect on Vascular Resistance Mechanisms

The severity of symptoms induced in cotton plants by stem inoculation with Fusarium oxysporum f. sp. vasinfectum was reduced by inoculation with a non-pathogenic strain (NPS) of the fungus 3 days previously. Symptom reduction was associated with reduced populations of the pathogen in the stele tissue of the stem. This cross-protective response became effective within 24 h of inoculation with the NPS. Although both vascular occlusion and terpenoid aldehyde synthesis were induced by theNPS, the short time required for cross protection to become effective and the duration of the effect, indicated that vascular occlusion rather than fungitoxicity of the terpenoid aldehyde compounds was the mechanism of protection.     

Gene expression profile changes in cotton root and hypocotyl tissues in response to infection with Fusarium oxysporum f. sp. vasinfectum

Microarray analysis of large-scale temporal and tissue-specific plant gene expression changes occurring during a susceptible plant-pathogen interaction revealed different gene expression profile changes in cotton root and hypocotyl tissues. In hypocotyl tissues infected with Fusarium oxysporum f. sp. vasinfectum, increased expression of defense-related genes was observed, whereas few changes in the expression levels of defense-related genes were found in infected root tissues. In infected roots, more plant genes were repressed than were induced, especially at the earlier stages of infection. Although many known cotton defense responses were identified, including induction of pathogenesis-related genes and gossypol biosynthesis genes, potential new defense responses also were identified, such as the biosynthesis of lignans. Many of the stress-related gene responses were common to both tissues. The repression of drought-responsive proteins such as aquaporins in both roots and hypocotyls represents a previously unreported response of a host to pathogen attack that may be specific to vascular wilt diseases. Gene expression results implicated the phytohormones ethylene and auxin in the disease process. Biochemical analysis of hormone level changes supported this observation.     

The effect of root-knot nematode on vascular resistance to Fusarium oxysporum f. sp. vasinfectum in the stems of cotton plants

The effect of root-knot nematode (Meloidogyne incognita) on external wilt symptoms and on the cotton plant's vascular response to stem-inoculation with Fusarium oxysporum f. sp. vasinfectum was investigated. Wilt symptoms were more severe in all plants inoculated with both organisms than with the fungus alone but relative wilt resistance of the cultivars was maintained. Greater symptom severity was associated with greater fungal proliferation in the stele and this was related to the ability of the nematode to reduce the efficiency of vascular occlusion. The nematode had no effect on the accumulation of infection-induced terpenoid aldehyde compounds.