Response of carnation cultivars to Fusarium oxysporum f.sp. dianthi in the field

The response of carnation (Dianthus caryophyllus L.) cultivars toFusarium oxysporum f.sp.dianthi (F.o.d.) was evaluated in an artificially infested field from 1988/9 to 1991/2. Disease incidence was highly correlated with disease severity, indicating that disease incidence may be used to estimate the impact ofF.o.d. on the host. Based on the results, the following stepwise procedure was developed for characterizing the response of carnation cultivars toF.o.d. First, the general response of the tested cultivar was classified as resistant or susceptible on the basis of disease incidence values recorded 180 days after planting. Empirical analysis of the data revealed that a disease incidence level of 75% may be taken as a reliable cut-off point for separation of cultivars into the two groups. Within each group, cultivars were then subjected to a more explicit classification: in the resistant group the records of actual disease incidence were used for classification, while in the susceptible group the linear rearrangement of the disease progress curve was calculated according to the Gompertz function, and the value of the intercept was used for classification.Contribution No. 3539-E series, from the Agricultural Research Organization.     

Induced resistance in tomato plants against Fusarium wilt invoked by Fusarium oxysporum f.sp. dianthi

Tomato plants, susceptible toFusarium oxysporum f. sp.lycopersici, were inoculated by immersing the roots in a conidial suspension ofF. oxysporum f. sp.lycopersici race 1,F. oxysporum f. sp.dianthi race 2 or a mixture of both fungi. Plants inoculated withF. oxysporum f. sp.lycopersici showed disease symptoms after 2 weeks, whereas plants inoculated withF. oxysporum f. sp.dianthi or a mixture of both fungi remained symptomless for over 7 weeks, the duration of the experiment. In another experiment root systems of plants were split and each half was separately inoculated. One half was firstly inoculated withF. oxysporum f. sp.dianthi or treated with water, followed after a week by a second inoculation of the other half withF. oxysporum f. sp.lycopersici or by a water treatment. The disease symptoms in the half firstly inoculated withF. oxysporum f. sp.dianthi were significantly delayed, compared to plants of which that half had been treated with water. BecauseF. oxysporum f. sp.dianthi reduced disease symptoms caused byF. oxysporum f. sp.lycopersici without any direct interaction with this pathogen, it is concluded thatF. oxysporum f. sp.dianthi is able to induce resistance againstF. oxysporum f. sp.lycopersici in tomato plants.     

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.     

Genetic diversity in Fusarium oxysporum f.sp. dianthi and Fusarium redolens f.sp. dianthi

Pathogenic isolates were selected representing all known vegetative compatibility groups (VCGs) and races of Fusarium oxysporum sensu lato from Dianthus spp. On basis of differences in the internal transcribed spacer region of the ribosomal DNA, six VCGs were classified as F. oxysporum f.sp. dianthi and four as F. redolens f.sp. dianthi. All VCGs of F. oxysporum f.sp. dianthi were characterized by unique restriction fragment length polymorphisms (RFLPs), unique overall esterase profiles, and unique virulence spectra, supporting a clonal lineage concept. Two VCGs of F. oxysporum f.sp. dianthi nevertheless comprised more than one race, but races within the same VCG shared the same distinct overall virulence spectrum. VCGs belonging to F. redolens f.sp. dianthi also had unique RFLPs and unique virulence spectra, but had grossly identical esterase profiles. Three new races (9, 10 and 11) are described for F. oxysporum f.sp. dianthi, and four for F. redolens f.sp. dianthi. Two races previously considered lost were recovered; race 7 was identified as a member of VCG 0021 of F. oxysporum f.sp. dianthi while race 3 was identified as a distinct VCG and race of F. redolens f.sp. dianthi. A summary of races and VCGs in F. oxysporum f.sp. dianthi and F. redolens f.sp. dianthi is presented.     

Fusarium oxysporum f.sp. cucurbitacearum n.f. embracing all formae speciales of F. oxysporum attacking Cucurbitaceous crops

Isolates ofFusarium oxysporum from wilted muskmelons, watermelons, cucumbers and from the muskmelon rootstockBenincasa hispida were screened for pathogenicity on seedlings and adult plants of these crops and related species. In seedling tests the isolates were not typically species-specific, contrary to what might be expected as an implication of their characterization as forma specialis. They often attacked species of several genera of plants, but not beyond the family of theCucurbitaceae. In the adult stage, plants were much more exclusively attacked by their corresponding formae speciales, but essential exceptions occurred. Isolates from cucumber were highly pathogenic to muskmelons, in the adulstage even causing more wilt of the latter than of cucumber.Comparing the results of these experiments with data from the literature, it is argued that the proposed f.sp.cucurbitacearum, embracing all formae speciales which specialize on plants within the family of theCucurbitaceae, would best fit in with the present state of knowledge. A proposition is given for equivalence of old and new classifications of isolates.Samenvatting Isolaten vanFusarium oxysporum uit verwelkte meloenen, watermeloenen, komkommers en uit de meloene-onderstamBenincasa hispida werden getoetst op pathogeniteit voor zaailingen en volwassen planten van deze gewassen en verwante soorten. In zaailingtoetsen waren de isolaten weinig soort-specifiek, in tegenstelling tot wat mocht worden afgeleid uit hun karakterisering als forma specialis. Zij tastten vaak soorten uit verschillende geslachten aan, maar niet buiten de grenzen van de familie derCucurbitaceae. in het volwassen stadium waren de interacties veel specifieker en werden planten slechts aangetast door de bijbehorende formae speciales. Enkele essentiële uitzonderingen kwamen echter voor. Isolaten van komkommer waren zeer pathogeen voor meloen. In het volwassen stadium veroorzaakten zij zelfs sterkere verwelking van meloen dan van komkommer.Vergelijking van de resultaten van deze proeven met gegevens uit de literatuur leidt tot de conclusie dat de voorgestelde f.sp.cucurbitacearum, die alle formae speciales met specialisatie opCucurbitaceae omvat, het best overeenkomt met de huidige stand van kennis. Een voorstet wordt gedaan ter vervanging van de oude classificatie van isolaten door corresponderende nieuwe aanduidingen.     

Resistance sources to Fusarium oxysporum f. sp. cubense tropical race 4 in banana wild relatives

Target trait evaluation in crop wild relatives is an important prerequisite for efficiently using the potential useful genes located in this valuable germplasm. Over recent decades, Fusarium oxysporum f. sp. cubense tropical race 4 (Foc‐TR4) has seriously threatened worldwide banana plantations. Breeding new resistant cultivars from wild banana species is expected to provide invaluable additional resources. However, knowledge on resistance to Foc‐TR4 in wild Musa species is very limited. In this study, eight genotypes of wild banana relatives (Musa acuminata subsp. burmannica, M. balbisiana, M. basjoo, M. itinerans, M. nagensium, M. ruiliensis, M. velutina and M. yunnanensis) were characterized for resistance to Foc‐TR4 in both greenhouse and field conditions. Most wild bananas showed higher resistance levels to Foc‐TR4 than the reference cultivars ‘Brazilian’ (AAA, susceptible) and ‘Goldfinger’ (AAAB, moderate resistance). Among the wild species, M. balbisiana showed the highest levels of disease intensity followed by M. acuminata subsp. burmannica. Some individuals of M. yunnanensis, M. nagensium, M. ruiliensis and M. velutina showed low levels of rhizome discolouration in greenhouse conditions, but were resistant in the field. No symptoms were observed on M. basjoo and M. itinerans, suggesting higher levels of resistance to Foc‐TR4. The results revealed different sources of resistance to Foc‐TR4 in banana wild relatives, which constitute a valuable genetic resource for banana breeding programmes aiming to produce cultivars resistant to fusarium wilt.     

农杆菌介导的西瓜枯萎病菌遗传转化

为获得带GFP标记的西瓜枯萎病菌转化株,用于后期观察病原菌侵染过程,采用农杆菌介导的方法,对西瓜枯萎病菌1号生理小种进行了遗传转化。结果表明:共培养时间为36h,枯萎病菌孢子和农杆菌AGL1比例为1∶1时该菌株的遗传转化效率最高,可以达到117.33个转化子/107个孢子。转化株的孢子、菌丝体及萌发的孢子均能发出稳定而强的绿色荧光。转化株的致病力检测显示其致病力与转化前的野生菌株致病力无明显差异。结果表明本研究获得的带GFP标记的西瓜枯萎病菌转化株可用于观察病菌在西瓜根系的侵染过程。     

Phylogenetic relationships of Fusarium oxysporum f. sp. melonis in Iran

Fusarium wilt of melon caused by Fusarium oxysporum f. sp. melonis is a destructive fungal disease in melon growing regions. Isolates of F. oxysporum obtained from six major melon producing provinces in Iran, from melons and other hosts, were characterized based on pathogenicity to melon, vegetative compatibility groups (VCGs) and nuclear ribosomal DNA intergenic spacer (IGS) sequencing. Thirty-four of 41 isolates from Iran in this study were identified as race 1,2 which belonged to either VCG 0134 or an unassigned VCG, which based on IGS sequencing grouped with the VCG 0135 tester isolate. The seven remaining isolates were identified as nonpathogenic to melon belonging to two undescribed VCGs. Based on sequence analyses of the IGS region of Iranian and foreign isolates, nine lineages were identified, each including one VCG. The separation of VCGs into distinct lineages based on IGS sequences is mostly consistent with Repetitive extragenic palindromic PCR (Rep-PCR) results. Exceptions are VCGs 0130 and 0131, which could be differentiated with IGS sequences, but not with Rep-PCR. Different races from the USA, France and Iran associated with VCG 0134 grouped into one IGS lineage but could be differentiated with Rep-PCR, suggesting that this VCG is more diverse than previously thought. Given the long history of melon cultivation in Iran and the Rep-PCR diversity of isolates belonging to this VCG, it could be speculated that VCG 0134 perhaps evolved in Iran.     

香蕉枯萎病菌生理分化研究摘要本研究

本研究对香蕉枯萎病菌菌株FOCAAA9(来自香蕉)和FOCABB1(来自粉蕉)进行培养试验和接种试验;在含粉蕉和香蕉组织浸提液的培养基上2个菌株的培养性状、菌丝生长速度、孢子形态、大小型孢子比率和产孢量显示出差异;接种结果FOCAAA9能侵染香蕉(MusaAAA)品种巴西蕉、红香蕉和台蕉引起枯萎病,而FOCABB1对3个香蕉品种无致病性。研究结果表明侵染香蕉和粉蕉的古巴尖镰孢[Fusariumoxysporumf.sp.cubense(E.F.Smith)Snyder]存在生理分化现象。     

假单胞菌对香蕉枯萎病菌的抑制作用

从已感染香蕉枯萎病的果园中分离到1株对香蕉枯萎病菌（Fusarium oxysporum f.sp.cubense）抑制作用很好的菌种,命名为G1。采用固体和液体培养法从不同角度证实了G1对香蕉枯萎病菌生长的抑制作用。结果表明,G1的发酵液、无菌滤液、挥发性物质、非挥发性代谢产物的平均抑菌率分别为92.5%、27.4%、73.8%、57.7%,可见抑制作用与活菌体有直接关系,其中产生挥发性物质尤其重要。液体培养的病原菌浓度为1.0×107cfu/mL经G1作用10d后,取样镜检发现G1通过抑制病原菌菌丝正常生长以至不能产孢,从而导致菌丝消融;通过吸附在孢子的周围,融解细胞壁,造成原生质泄露致使孢子死亡。     

Physiological races of Fusarium oxysporum f.sp. melonis in Tunisia

Fusarium wilt of melon, caused by Fusarium oxysporum f.sp. melonis (Fom), is an important disease; races of the pathogen were identified by inoculating differential standard host cultivars. A total of ten isolates that were obtained from 23 fields located in four different geographical regions were identified as pathogenic. Results indicate that all four known Fom races, namely, 0, 1, 2 and 1.2, were found in north and middle Tunisia. Race 1.2 was the most prevalent.     

Fine Structure of the Early Interaction of Lily Roots with Fusarium oxysporum f.sp. lilii

The early interaction of lily roots with the cortical rot pathogen Fusarium oxysporum f.sp. lilii was studied using roots of lily bulblets grown in Hoagland's solution, inoculated with the pathogen, and sampled up to 48h later. Conidia produced germ tubes within 6h, which extended towards and into the mucilage covering the root elongation zone, and along and into the anticlinal grooves and middle lamellae of epidermal cells. By 24–48h, infecting hyphae had reached the periclinal walls and intercellular spaces between the epidermis and the outermost cells of the cortex. Penetration of intercellularly growing hyphae directly across host cell walls was not observed; invasion of the cell lumen only occurred by gradual infringing of hyphae upon successive primary wall layers. Non-cellulosic wall appositions rich in vesicles and covered by a cellulosic protective-like layer were formed in response to approaching hyphae in resistant cv.Connecticut King, but rarely in susceptible cv. Esther which seemed more susceptible to plasmolysis and rot. Finger-like projections of the appositions into the host cell cytoplasm likely represent early stages of transfer cell formation.     

香蕉枯萎病菌生理分化研究

本研究对香蕉枯萎病菌菌株FOCAAA9（来自香蕉）和FOCABB1（来自粉蕉）进行培养试验和接种试验；在含粉蕉和香蕉组织漫提液的培养基上2个菌株的培养性状、菌丝生长速度、孢子形态、大小型孢子比率和产孢量显示出差异；接种结果FOCAAA9能侵染香蕉（Musa AAA）品种巴西蕉、红香蕉和台蕉引起枯萎病，而FOCABB1对3个香蕉品种无致病性。研究结果表明侵染香蕉和粉蕉的古巴尖镰孢[Fusarium oxysporum f．sp．cubeilse（E．F．Smith）Snyder]存在生理分化现象。     

Potential role for salicylic acid in induced resistance of asparagus roots to Fusarium oxysporum f.sp. asparagi

Pre-inoculation of asparagus ( Asparagus officinalis ) roots with selected nonpathogenic isolates of Fusarium oxysporum (np Fo ) has previously been shown to induce systemic resistance against infection by F. oxysporum f.sp. asparagi ( Foa ) through activation of plant-defence mechanisms. To elucidate the putative np Fo -mediated defence pathways, the effect of salicylic acid (SA) was examined in a split-root system of asparagus where one half of the seedling root system was drenched with SA and the activation of defence responses was measured subsequently on the remaining roots. SA-treated plants exhibited enhanced systemic resistance, with a significant reduction in disease severity of the roots inoculated with Foa , compared with untreated plants. SA activated peroxidase and phenylalanine ammonia-lyase, as well as lignification, upon Foa attack, in a manner similar to that observed with np Fo pretreatment. In addition, application of diphenyleneiodonium, an SA biosynthesis inhibitor, led to failure of np Fo to induce lignin deposition and systemic resistance. Treatment of fungal spores with SA did not affect germination and growth of either np Fo or Foa in in vitro antifungal assays. Production of SA at the site of np Fo infection may be involved in the induction of Foa resistance in asparagus roots.     

Phylogeny and origin of Fusarium oxysporum f. sp. vanillae in Indonesia

Vanilla stem rot, caused by Fusarium oxysporum f. sp. vanillae (Fov), is the main constraint to increasing vanilla production in the major vanilla‐producing countries, including Indonesia. The current study investigated the origin of Fov in Indonesia using a multigene phylogenetic approach. Nineteen Fov isolates were selected to represent Indonesia, the Comoros, Mexico and Réunion Island. The translation elongation factor 1 alpha gene and the mitochondrial small subunit ribosomal RNA gene phylogenies resolved the Fov isolates into three distinct clades in both phylogenetic species of the F. oxysporum species complex, indicating a polyphyletic pattern of evolution. In addition, Fov isolates from Indonesia were also polyphyletic. These results suggest that the vanilla stem rot pathogen in Indonesia has a complex origin. The implications for disease management are discussed.     

Search for sources of wide-spectrum resistance to Fusarium oxysporum f. sp. basilici isolates in accessions of Ocimum species

Ocimum (Lamiaceae) is an important plant genus, with many species used for food flavorings and for essential oils. Fusarium wilt (Fusarium oxysporum f. sp. basilici, FOB) is the most important disease of basil (O. basilicum L.). Twenty-five accessions of O. basilicum, five of O. americanum and two of O. campechianum were initially evaluated for resistance to one FOB isolate (named as FOB-1). Eight accessions (identified as resistant to FOB-1) and one susceptible control were reevaluated to FOB-1 and four other FOB isolates of distinct geographic origins. The FOB isolates varied in aggressiveness and interacted differentially with the Ocimum accessions. Two accessions of O. americanum, one of O. campechianum, and one of O. basilicum had high levels of resistance to all five FOB isolates. The Ocimum germplasm identified here could represent useful sources of resistance genes for developing cultivars with wide-spectrum resistance (i.e., effective against a broad range of FOB isolates). In addition, having a set of potential differential accessions might be useful for large-scale analysis of FOB isolates to demonstrate the presence of physiological races in the Ocimum–FOB pathosystem.     

Occurrence of Fusarium oxysporum f. sp. melonis Race 1 in Japan

In 1994, Fusarium wilt of melon cultivars which are resistant to races 0 and 2 of Fusarium oxysporum f. sp. melonis was observed in southern area of the Lake Biwa region, Shiga prefecture. In commercial fields, mature plants of cv. Amus which were grafted onto cv. Enken Daigi 2, and of cv. FR Amus showed yellowing, wilting and finally death before harvesting of fruits. Diseased plants had vascular and root discolorations, and their stem sections yielded typical colonies of F. oxysporum. When the Shiga strains were tested for their pathogenicity to 12 species of cucurbits, they caused wilts only on melon. Using race differential cultivars of melon, the Shiga strains were classified as race 1 of F. oxysporum f. sp. melonis, which has not been reported in Japan. To further characterize their pathogenicity, the strains were used to inoculate 46 additional cultivars of melon, oriental melon and oriental pickling melon. All the race 1 strains were pathogenic to the cultivars tested, and their host range was apparently different from those of strains belonging to other races (races 0, 2 and 1,2y). DNA fingerprinting with a repetitive DNA sequence, FOLR3, differentiated race 1 strains from strains of races 0 and 2, but not from race 1,2y strains. Received 2 July 1999/ Accepted in revised form 30 September 1999     

Variation in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined by inoculation tests and molecular markers

Screening of genotypes of melon ( Cucumis melo ) for resistance to wilt caused by Fusarium oxysporum f.sp. melonis is often characterized by wide variability in their responses to inoculation, even under carefully controlled conditions. The variability at the seedling stage of 17 genotypes susceptible to race 1 was examined in growth-chamber experiments. Disease incidence varied from 0 to 100% in a genotype-dependent manner. Using four combinations of light (60 and 90  µ E m⁻² s⁻¹) and temperatures of (27 and 31°C), only light intensity showed a statistically significant effect. Marker-assisted selection for fusarium resistance breeding using cleaved amplified polymorphic sequence (CAPS) and sequence-characterized amplified region (SCAR) markers were compared using a single set of genotypes that included 24 melon accessions and breeding lines whose genotype regarding the Fom-2 gene was well characterized. The practical value of the markers for discriminating a range of genotypes and clarifying the scoring of phenotypes was also tested using a segregating breeding population which showed codominant SCAR markers to be useful in marker-assisted selection.