Host genotype- and temperature-dependent colonizationof In vitro carnation callus cultures byFusarium oxysporum f.sp.dianthi race 2

Differentin vivo resistance/susceptibility levels of 14 carnation cultivars toFusarium oxysporum f.sp.dianthi race 2, the causal agent of Fusarium wilt disease of carnation, were also expressed in anin vitro system and assayed as the degree of fungal colonization of callus cultures at 20° C. Temperature influenced thein vitro expression of carnation resistance. An incubation temperature of 27° C increased the colonization of calli derived from both the susceptible (‘Corrida’ and ‘Ambra’) and the resistant (‘Pulcino’ and ‘Pallas’) cultivars. At 15°C, the colonization of calli derived from Pulcino and Pallas diminished significantly more than for Ambra and Corrida. Inhibition of fungal growth on resistant calli was correlated to retardation in hyphal development. Both scanning electron microscopy and light microscopy observations showed that hyphae did not penetrate into carnation cells.     

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.     

Passive transport of microconidia of Fusarium oxysporum f. sp. dianthi in carnation after root inoculation

Root inoculation of susceptible carnations withFusarium oxysporum f. sp.dianthi induced characteristic unilateral wilt only if root woundings and use of a microconidial suspension had not been combined at the time of inoculation. The combination, however, induced atypical and sudden stem breaking soon followed by death. In all cases wilt was due to destruction of the xylem. Unilateral wilt appeared to follow sparse natural infection of single roots. Stem breaking was due to destruction of the vascular tissues all around the stem and is ascribed to multilateral infection caused by translocation of microconidia at inoculation through several wounded roots directly into the stem.Microconidia were carried passively 5–10(–10) cm into stems of susceptible and resistant carnations within 24 h both after immersing cut ends of the roots in a conidial suspension and after pouring a suspension on the soil. Passive spore transport is an inoculation artefact which may severely affect interpretation of experimental results; it seems to be unimportant in natural Fusarium wilt development in carnation.Samenvatting Inoculatie van de wortels van vatbare anjers metFusarium oxysporum f. sp.dianthi veroorzaakte de kenmerkende eenzijdige verwelking alleen als wortelbeschadigingen bij de inoculatie niet werden gecombineerd met het gebruik van een suspensie van microconidiën. Die combinatie veroorzaakte namelijk afwijkende symptomen waarbij de planten plotseling omknakten en vervolgens snel afstierven. De verwelking leek in alle gevallen veroorzaakt te worden door afbraak van het xyleem. Eenzijdige verwelking leek te volgen op spaarzame natuurlijke wortelinfecties. Bij omgeknakte planten bleek het vaatweefsel rondom in de stengel aangetast te zijn, hetgeen toegeschreven wordt aan infectie van verschillende kanten van de stengel als gevolg van passief transport van microconidiën bij de inoculatie door verscheidene beschadigde wortels direct de stengel in.Microconidiën werden binnen 24 uur 5–10(–30) cm de stengels van vatbare en resistente anjers ingezogen wanneer de wortels afgesneden en met het uiteinde in een sporensuspensie gehangen werden, maar ook wanneer de suspensie op de grond gegoten werd. Passief transport van sporen is een inoculatie-artefact dat echter belangrijke consequenties kan hebben voor de interpretatie van de resultaten van proeven. Bij de natuurlijke verspreiding vanF. oxysporum in anjers lijkt passief sporentransport van weinig belang.     

Colonization and histopathology of susceptible and resistant carnation cultivars infected with Fusarium oxysporum f. sp. dianthi

Stems of the susceptible Early Sam and resistant Novada carnations were inoculated with a conidial suspension ofFusarium oxysporum f. sp.dianthi. Stem segments of either cultivar were sampled regularly and used for determination of fungal growth and for microscopical investigation.Early Sam showed typicalFusarium wilt symptoms and its stems were colonized intensively. The observed vascular browning appeared to be caused by discolouration of primary walls of infected vessels and surrounding cells. Vessels were rarely occluded with gel. Cell wall degradation led to the formation of stem cavities. Hyperplasia of xylem parenchyma was not seen.In Novada, fungal colonization remained low throughout the experiment. Macroscopic symptoms were absent except for longitudinal bursts in the stem, which appeared to be caused by hyperplasia of xylem parenchyma bordering infection. Vascular gelation occurred in the infected tissues, causing some vascular browning also. Xylem vessel regeneration was observed in the hyperplastic layer. Cavities were not formed, and wall discolouration was rare. Vascular gelation is considered part of theFusarium wilt resistance mechanism. It is followed by xylem vessel regeneration, which expresses a general plant response to vascular dysfunction rather than being part of the resistance mechanism.Although of different origin, vascular browning as such occurs in both susceptible and resistant interactions. In breeding for resistance, care should hence be taken with the current use of browning as an indication of disease.Samenvatting Anjers van de vatbare cultivar Early Sam en de resistente cultivar Novada werden geïnoculeerd met een conidiënsuspensie vanFusarium oxysporum f. sp.dianthi. Van beide cultivars werden regelmatig stengeldelen geoogst om deze microscopisch te onderzoeken en om de schimmelgroei te bepalen.Early Sam vertoonde de voor deze verwelkingsziekte kenmerkende symptomen en werd intensief gekoloniseerd. Aan het vaatweefsel waargenomen bruinkleuring bleek veroorzaakt te worden door verkleuring van de primaire wanden van geïnfecteerde vaten en de hen omringende cellen. Zelden trad er in de vaten gomvorming op. Celwandafbraak veroorzaakte de vorming van holten in de stengel. Hyperplasie van het houtparenchym werd niet waargenomen.In Novada bleef de schimmelgroei gedurende het hele experiment beperkt. Macroscopisch waren er enkel lengtescheuren in de stengel te zien, die veroorzaakt bleken te worden door hyperplasie van aan de infectie grenzend houtparenchym. In het geïnfecteerde vaatweefsel optredende gomvorming veroorzaakte ook enige bruinkleuring. In het hyperplastische weefsel werd regeneratie van houtvaten waargenomen. In de stengel werden geen holten gevormd, en verkleuring van de celwanden kwam weinig voor. De vorming van gommen in de houtvaten maakt waarschijnlijk deel uit van het resistentiemechanisme. De daarop volgende houtvatregeneratie is eerder een algemene reactie van de plant op vaatverstopping dan een deel van het resistentiemechanisme.Vaatverbruining, zij het van verschillende oorsprong, komt voor in zowel vatbare als resistente interacties. Om die reden moet men in de resistentieveredeling bij de anjer voorzichtig zijn met het gebruik van bruinkleuring als ziekteïndicatie.     

Histology of roots of resistant and susceptible carnation cultivars from soil infested with fusarium oxysporum f.sp. dianthi

Fusarium wilt-resistant Novada and wilt-susceptible Early Sam carnations were planted in soil infested withFusarium oxysporum f.sp.dianthi, and their roots studied after five and ten weeks. Both in Novada and Early Sam, the extravascular tissue of undamaged young root parts were scarsely colonized. In roots of Novada, infected xylem vessels were usually occluded with gums and surrounded by phellem tissue. In mature parts of roots, the phellem surrounding occluded vessels often merged with the external phellem surrounding the vascular cylinder, after which the occluded vessels were shed from the roots. The phellem at the root surface appeared to be a strong barrier to fungal invasion. In roots of Early Sam carnations, as well as in a few roots of Novada carnations, the defence responses did not result in compartmentation of the fungus, and colonization and degradation of the vascular tissues followed. Diseased roots finally healthy. Shoots of Early Sam carnations, and eventually a few shoots of Novada carnations, were colonized and developed wilt symptoms.Samenvatting Resistente Novada en vatbare Early Sam anjers werden geplant in grond besmet metFusarium oxysporum f.sp.dianthi. Na vijf en tien weken werden de wortels van de planten bestudeerd. De extravasculaire delen van onbeschadigde jonge wortelgedeelten waren in beide cultivars nauwelijks gekoloniseerd. In de wortels van Novada waren geïnfecteerde houtvaten meestal verstopt door gommen en omgeven door kurkweefsel. In oudere wortelgedeelten sloot het kurkweefsel rond verstopte vaten vaak aan op het kurkweefsel aan de buitenkant van het vaatweefsel, waarna de aangetaste vaten werden uitgestoten uit de wortel. Het kurkweefsel aan het worteloppervlak leek een belangrijke barrière te vormen tegen het binnendringen van de schimmel. In de wortels van Early Sam en in enkele wortels van Novada mislukte het afweerproces, en werd het vaatweefsel door de schimmel gekoloniseerd en vervolgens afgebroken. Aangetaste wortels werden na afloop van tijd hol. De bovengrondse delen van de meeste Novada anjers werden niet gekoloniseerd en bleven gezond. De bovengrondse delen van de Early Sam anjers, en op den duur die van enkele Novada anjers, werden gekoloniseerd en verwelkten.     

Inheritance of resistance in carnation against Fusarium oxysporum f.sp. dianthi races 1 and 2, in relation to resistance components

Four carnation cultivars, Novada (resistant to races 1 and 2 ofFusarium oxysporum f.sp.dianthi), Elsy (susceptible to race 1), Lena (susceptible to race 2) and Sam's Pride (susceptible to both races), were selfed and crossed. When three months old, the seedlings were inoculated via the roots or via the stems, after which wilting was recorded weekly according to a 5-point ordinal scale.Analyses were carried out on the proportions of diseased plants. For race 1 variation between the progenies could be described by means of general combining abilities only; GCA values were not affected by the inoculation method used. Also for race 2 GCAs were most important but the GCA values appeared different for the two inoculation methods. It is concluded that resistance to both races is inherited in an additive way.Indications for independently inherited root-specific resistance components (extravascular resistance) were only found with race 2. With both races, the ability to confine the pathogen at the infection site appeared the most important resistance component. Resistant progenies were also characterized by longer latent periods and lower wilting rates.Both race 1 and race 2 induced the accumulation of the phytoalexins dianthalexin and methoxydianthramide S, but race 2 induced higher amounts than race 1. The accumulation of phytoalexins was positively correlated to the resistance level of the progenies against the respective races. The progenies of the double-resistant cultivar Novada appeared to produce particularly high levels of phytoalexins.     

Reductive detoxification of the acetophenone skeleton of the carnation phytoanticipin by Fusarium oxysporum f.sp. dianthi

The skeleton of the carnation phytoanticipin, acetophenone, was detoxified by Fusarium oxysporum f.sp. dianthi , the main fungal parasite of carnation. This process consisted of the reduction to ethanol of the acetyl group, leading to the formation of phenylethanol, which has lower fungitoxic activity than the parent molecule. The conversion took place through the activity of an adaptive fungal oxidoreductase, which was NADH-dependent and was released by the fungus as two enzymatic forms within the culture substrate in the presence of acetophenone. Reduction was stereospecific and gave rise to only one of the two possible enantiomeric forms.     

Regeneration of vascular tissues in relation to Fusarium wilt resistance of carnation

Fusarium wilt-resistant Novada carnations responded both to stem inoculation with a conidial suspension ofFusarium oxysporum f. sp.dianthi orF. oxysporum f. sp.lycopersici and to root inoculation by planting in soil infected withF. oxysporum f.sp.dianthi by means of a localization mechanism comprising gel formation in the xylem vessels and hyperplasia of adjacent parenchyma cells. Dye translocation experiments showed that xylem transport was limited by the presence of vascular gels, although wilting did not occur. Overcapacity of the vascular system apparently allowed for sufficient water transport to compensate for local vascular dysfunction. Also, vascular regeneration in the hyperplastic tissue next to occluded xylem vessels created new pathways for water transport to compensate for those lost by occlusion. Regeneration of xylem vessels was eventually followed by regeneration of xylem fibers, xylem parenchyma, cambium, and phloem cells.Early Sam carnations, susceptible to Fusarium wilt, responded to stem inoculation withF. oxysporum f. sp.lycopersici by similar localization of infection and vascular regeneration. Stem inoculation withF. oxysporum f. sp.dianthi, however, resulted in colonization of the xylem vessels followed by lysis of the vascular tissues. Vascular gelation, hyperplasia of parenchyma cells, and vascular regeneration did generally not occur. However, if some hyperplasia occurred in attempted defence, some differentiation of hyperplastic cells into single xylem vessel elements was observed which only rarely resulted in complete vascular regeneration next to colonized xylem. In the absence of hyperplasia, differentiation of medulla parenchyma cells bordering destroyed vascular tissue into xylem vessel elements was even more exceptional. Apparently, vascular regeneration in carnation is a normal defence reaction to fungal invasion.Samenvatting Novada anjers, resistent tegen Fusarium-verwelkingsziekte, reageerden op stengelinoculatie met een conidiënsuspensie vanFusarium oxysporum f.sp.dianthi of vanF. oxysporum f.sp.lycopersici en op wortelinoculatie door te planten in metF. oxysporumf.sp.dianthi besmette grond met een lokalisatiemechanisme dat onder meer bestond uit vorming van gommen in de houtvaten en hyperplasie van naburige parenchymcellen. Uit proeven over kleurstoftransport bleek dat de sapstroom door de gomvorming beperkt werd, hoewel dit geen verwelkingssymptomen veroorzaakte. Overcapaciteit van het vaatstelsel zorgde kennelijk voor voldoende compensatie aan watertransport om plaatselijke verstoring van de sapstroom op te vangen. Daarnaast werd het verlies aan functionele houtvaten ook opgevangen door vaatweefselregeneratie in het hyperplastische weefsel grenzend aan door gommen verstopte houtvaten. Na verloop van tijd werden behalve houtvaten ook houtvezels, houtparenchymcellen, cambium- en floeemcellen geregenereerd.Early Sam anjers, vatbaar voor Fusarium-verwelkingsziekte, reageerden op stengelinoculatie metF. oxysporum f. sp.lycopersici met eenzelfde lokalisatiemechanisme en ook met vaatweefselregeneratie. Stengelinoculatie metF. oxysporum f.sp.dianthi echter had kolonisatie en vervolgens lysis van het vaatweefsel tot gevolg. Meestal trad er geen gomvorming, hyperplasie van parenchymcellen of vaatweefselregeneratie op. Als echter bij pogingen tot afweer toch enige hyperplasie optrad, bleken sommige hyperplastische cellen wel tot houtvatelementen te differentieren. Dit leidde echter maar zelden tot totale vaatweefselregeneratie parallel aan het gekoloniseerde vaatweefsel. In afwezigheid van hyperplasie differentieerden mergparenchymcellen vlak naast lyserend vaatweefsel slechts bij hoge uitzondering tot houtvatelementen. Vaatweefselregeneratie bij anjer is kennelijk een gewone afweerreactie op besmetting met pathogene schimmels.     

Physiological races and vegetative compatibility groups of butterhead lettuce isolates of Fusarium oxysporum f. sp. lactucae in Japan

Races were identified among butterhead lettuce isolates of Fusarium oxysporum f. sp. lactucae collected from three geographical areas of Hokkaido, Shizuoka, and Fukuoka in Japan by inoculation tests using Fujinagas race differential cultivars of lettuce (i.e., Patriot, Costa Rica No. 4, and Banchu Red Fire). Eighteen isolates from Shizuoka and Fukuoka were designated race 3, with two unknown vegetative compatibility groups (VCGs) that differed from Ogisos VCG 1 and 2. These two new VCGs were obtained from both Shizuoka and Fukuoka. On the other hand, three isolates from Hokkaido were classified as race 1 and identified as VCG 1, which represents a VCG of crisphead isolates from Nagano.     

Development of sequence tagged site markers to identify races of Fusarium oxysporum f. sp. lactucae

By random amplified polymorphic DNA (RAPD) analysis of the representative isolates of each race of Fusarium oxysporum f. sp. lactucae, RAPD fragments of 0.6, 1.6, and 2.9kb were obtained. The 0.6-kb RAPD fragment was common to the representative isolates of all three races. Amplification of the 1.6- and 2.9-kb fragments were unique to the isolates of races 1 and 2, respectively. Sequence tagged site (STS) marker FLA0001, FLA0101, and FLA0201 were generated from the 0.6-, 1.6-, and 2.9-kb RAPD fragments, respectively. Polymerase chain reaction (PCR) analysis showed that FLA0001 was common to all 49 isolates of F. oxysporum f. sp. lactucae. FLA0101 was specifically generated from all 23 isolates of race 1 but not from races 2 or 3. FLA0201 was specifically amplified from all 12 isolates of race 2 but not from races 1 or 3. In two isolates of F. oxysporum f. sp. lactucum, PCR amplified FLA0001 and FLA0101 but not FLA0201. On the other hand, these STS markers were not detected from isolates of five other formae speciales. Because these STS markers were not generated from isolates of other plant pathogenic fungi, bacteria, or plant materials examined in this study, PCR analysis combined with the three STS markers should be a useful means for rapid identification of races of F. oxysporum f. sp. lactucae.     

Physiologic races ofFusarium oxysporum f.sp.melonis in the southeastern anatolia region of turkey and varietal reactions to races of the pathogen

Thirty-four isolates ofFusarium oxysporum f.sp.melonis (F.o.m.) obtained from 205 fields in melon-producing areas in the southeastern Anatolia Region of Turkey were identified on the basis of colony morphology and pathogenicity by the root dip method. In this region the mean prevalence of wilt disease was 88.1% and the mean incidence of disease was 47.5%. Physiologic races 0, 1, 2, and 1,2 of the pathogen were determined by their reactions on differential melon cultivars ‘Charentais T,’ ‘Isoblon’, ‘Isovac’ and ‘Margot’ in the greenhouse. Race 1,2, representating 58.8% (20/34) of all isolates, was widely distributed. Of the other pathogenic isolates, eight were identified as race 0, five as race 1, and one as race 2. This is the first report of physiologic races ofF.o.m. in Turkey. Of 44 melon cultivars tested in the greenhouse for resistance toF.o.m. races, 36 were found to be moderately resistant to race 0, 17 were susceptible to race 1,2, 34.1% were highly resistant to race 1, and 52.2% had moderate resistance to race 2. http://www.phytoparasitica.org posting July 16, 2002.     

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。     

Comparison of five stem inoculation methods with respect to phytoalexin accumulation and Fusarium wilt development in carnation

Five methods of stem inoculation of carnations with conidial suspensions ofFusarium oxysporum f.sp.dianthi were compared for uptake of the suspension, induction of phytoalexin accumulation and wilt development. Inoculation was performed by incision of the stem across droplets of inoculum placed on leaves, or by injection of droplets into the stem. With both methods, higher inoculum dosages led to higher wilting rates and higher phytoalexin concentrations. Injection was more effective than incision since a lower inoculum dosage was required to obtain the same phytoalexin levels. Injection therefore appears the most promising technique for the development of routine screening methods for resistance based on phytoalexin accumulation.Samenvatting Vijf methoden om anjers via de stengel te inoculeren met een sporensuspensie vanF. oxysporum f.sp.dianthi werden vergeleken wat betreft de opname van de suspensie, de accumulatie van fytoalexinen en het ziekteverloop. Inoculatie vond plaats door de stengel aan te snijden dwars door een druppel inoculum die op een blad was gelegd, of door druppeltjes inoculum te injecteren. Bij beide methoden leidden hogere doses inoculum tot heviger ziektesymptomen en accumulatie van grotere hoeveelheden fytoalexinen. Injectie van inoculum was effectiever dan aansnijden, daar er minder inoculum nodig was voor eenzelfde resultaat. Injectie biedt daarom meer perspectief voor de ontwikkeling van routinematige resistentietoetsen op basis van de accumulatie van fytoalexinen.     

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.     

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     

Use of random amplified polymorphic DNA (RAPD) to identify races 1, 2, 4 and 8 of Fusarium oxysporum f. sp. dianthi in Italy

The RAPD fingerprinting procedure was used in combination with pathogenicity assays on differential cultivars to characterize a representative collection of 72 Fusarium spp. isolates of different geographic origin collected from diseased carnation. In F. oxysporum f. sp. dianthi, isolates were grouped according to the physiologic race: group 1 included isolates of race 4; group 2 was formed by isolates of race 2 and single representatives of races 5 and 6; group 3 included isolates of races 1 and 8. No correlation was found between RAPD data and geographic origin of the isolates tested: representatives of race 2 isolated in Italy, Israel and Japan had the same amplification profile. Three isolates which showed a low level of pathogenicity on all carnation cultivars tested shared an identical amplification pattern and are probably saprophytic F. oxysporum. Finally, two F. redolens isolates from Japan and seven non-pathogenic isolates of F. proliferatum collected from diseased carnation in Italy, Israel and The Netherlands were clearly distinguishable according to their RAPD fingerprint. The results are discussed in relation to previous studies on the genetic diversity of F. oxysporum f. sp. dianthi and to the development of forma specialis- and pathotype-specific diagnostic tools.     

Characterisation ofFusarium oxysporum isolated from carnation in Australia based on pathogenicity,vegetative compatibility and random amplified polymorphic DNA (RAPD) assay

Isolates ofF. oxysporum collected from symptomless carnation cuttings from Australian carnation growers properties, together with isolates from national collections, were screened for pathogenicity and grouped according to vegetative compatibility and random amplified polymorphic DNA (RAPD) patterns. The collection of 82 Australian isolates sorted into 23 different vegetative compatibility groups (VCGs). Of 69 isolates tested for pathogenicity, 24 were pathogenic to carnations, while the remaining 45 were non-pathogenic. All pathogenic isolates were within two VCGs, one of which was also compatible with an isolate obtained from an international culture collection, and which is known to represent VCG 0021 and race 2. Race status of the two pathogenic VCGs remains unknown. The RAPD assay revealed distinct DNA banding patterns which could distinguish pathogenic from non-pathogenic isolates as well as differentiate between isolates from the two pathogenic VCGs.     

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。     

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。     

甘蓝枯萎病菌REMI转化体系的建立

建立高效、稳定的甘蓝枯萎病菌REMI转化体系,为进一步获得特定表型突变体及基因功能研究建立技术储备.利用REMI(restriction enzyme mediate intergration)转化方法,将线性化的含有潮霉素抗性基因的pUCAT-PH质粒转化甘蓝枯萎病菌A6菌株的原生质体,摸索获得转化子最适的潮霉素筛选浓度以及不同限制性内切酶和酶量对转化效率的影响;利用PCR(polymerase chain reaction)技术对潮霉素抗性转化子进行验证.结果表明转化子的最适潮霉素筛选浓度为50 μg/mL;转化效率较高的限制性内切酶为HindⅢ,并且转化效率最高时的酶量为20 U.利用该转化体系构建了含1 050个转化子的甘蓝枯萎病菌转化子库,对转化子进行Southern验证,证明该转化体系是可行的.