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1.
Greenhouse trials conducted in 2003 and 2004 investigated the impact of trichothecenes on the severity of seedling blight and root rot in common wheat ( Triticum aestivum ), durum wheat ( Triticum turgidum var. durum ), barley ( Hordeum vulgare ) and triticale (× Triticosecale 6x ) using two trichothecene-producing and two trichothecene-nonproducing Fusarium graminearum strains. In 2003 seedling emergence and survival following soil infestation of the trichothecene-producing strain (Gz3639) were significantly reduced compared with the trichothecene-nonproducing strain (GzT40), while root-rot incidence and severity were increased significantly. In 2004, two trichothecene-producing strains (Gz3639 and GzT106) reduced seedling emergence and survival ( P  ≤ 0·01) in eight of 10 crops/cultivars based on single-degree-of-freedom contrasts. However, when results from all strains were combined no significant differences were observed between two trichothecene-producing and two trichothecene-nonproducing F. graminearum strains. Inoculation with GzT106, a trichothecene-producing 'add-back' strain, resulted in more severe root rot symptoms in eight of 10 cultivars ( P  ≤ 0·01–0·05) and lower seedling emergence and survival in seven of 10 cultivars ( P  ≤ 0·01–0·10), compared with the wild-type parental strain Gz3639. The presence of trichothecenes may play an important role in the aggressiveness of F. graminearum .  相似文献   

2.
Pink rot of potato, most commonly caused by Phytophthora erythroseptica , is a major field and post-harvest problem in southern Idaho, USA, particularly since 1998 when isolates resistant to the phenylamide fungicide metalaxyl-M (mefenoxam) were detected. Isolates of P. erythroseptica were collected from infected tubers in 2001 and 2002 from six Idaho counties and tested for resistance to metalaxyl-M on amended agar. Metalaxyl-M resistant (MR) and metalaxyl-M-sensitive (MS) isolates were identified in six counties; 160 isolates were highly resistant, seven moderately resistant and 57 sensitive to metalaxyl-M with mean EC50 values of 182, 23 and 0·5 mg L−1 ai metalaxyl-M, respectively. Mycelial growth rates and oospore production in agar were assessed for 20 MS and 20 MR isolates at 10, 15, 20, 25 and 30°C. Growth rates of MR isolates were between 2·5 and 3·1 times greater ( P  < 0·05) than those of MS isolates at 10, 15, 20 and 25°C, and oospore production was between 6·8 and 20·5 times greater ( P  < 0·0001) for MR than for MS isolates at the same temperatures. Colony growth in V8 broth at 18°C was greater for MR than MS isolates ( P  < 0·0032). However, zoospore production at 18°C was greater for MS than for MR isolates ( P  < 0·0109), and zoospore production m m −1 of colony circumference was also greater for MS than for MR isolates, 14 191 and 9959, respectively ( P  = 0·0109). Sexual reproduction of MR isolates in nature may be greater than MS isolates, but MS isolates may be more asexually fit based on the fitness parameters studied.  相似文献   

3.
为明确我国不同种、地理来源和毒素化学型小麦茎基腐病菌的致病力分化情况,采用纸塔法对来自全国9个省市80个采样点分离的224株小麦茎基腐病菌进行致病力分析。结果表明,不同种镰刀菌的致病力不同,黄色镰刀菌Fusarium culmorum,禾谷镰刀菌F.graminearum,假禾谷镰刀菌F.pseudograminearum及亚洲镰刀菌F.asiaticum致病力强于其他种。F.culmorum致病力显著高于F.pseudograminearum和F.asiaticum,而F.pseudograminearum,F.graminearum及F.asiaticum三者之间无显著性差异。中华镰刀菌F.sinensis,木贼镰刀菌F.equiseti,锐顶镰刀菌F.acuminatum致病力较弱,三者间苗期致病力无显著性差异;多数省份F.pseudograminearum群体间致病力无显著差异,仅山东F.pseudograminearum群体的致病性显著低于河南群体;此外,产毒类型为3ADON的F.pseudograminearum群体致病力显著高于15ADON群体。  相似文献   

4.
Resistance to Xylella fastidiosa was evaluated within a population of 20 interspecific hybrids of Pera sweet orange and Murcott tangor under greenhouse conditions. Efficiency of inoculation, multiplication of bacteria within the plants, xylem vessel morphology, and symptom expression were analysed. The rate of infection ranged from 40 to 100% (average 70%) for all genotypes analysed. Xylella fastidiosa populations ranged from log 0·59 to log 2·13 cells mg−1 tissue for the resistant hybrids. These values were significantly different ( P  = 0·05) from those obtained for the tolerant (no symptoms but bacteria recovered) or susceptible (symptoms and bacteria recovered) hybrids (log 3·02 to log 4·06 cells mg−1). Xylella fastidiosa was recovered from all hybrids (log 2·31 to 5·03 CFU mg−1 tissue) except the resistant ones. The first foliar symptoms appeared at least 90 days post-inoculation, the time varying according to genotype. No correlation between xylem vessel morphology and disease expression was observed, indicating that the resistance was the result of a genetic response of the host. According to this hypothesis, a high broad-based heritability index for resistance was obtained (0·96) at 210 days from X. fastidiosa inoculations, using bacterial quantification by real-time PCR, which indicated that the influence of the number of bacteria was the result of genetic rather than environmental variations.  相似文献   

5.
Several PCR methods have recently been developed for Fusarium analysis in pure cultures. To use these new techniques in mycological studies and in industrial quality control, a protocol was set up for the rapid preparation of fungal DNA from cereals. An ultrasonification probe (sonotrode) and a lysis buffer were used for mechanical lysis of mycelia from infected grains. Following ultrasonification, DNA was isolated using a commercially available kit. DNA preparation was completed within 5 min per sample. The method resulted in DNA of sufficient quality and quantity for diagnostic PCR. Group- and species-specific primers were used to detect DNA of Fusarium graminearum and F. culmorum in species-specific assays as well as trichothecene-producing Fusarium spp. in a group-specific system. A minimum of one F. graminearum -infected grain added to an uninfected 40 g wheat sample was detectable with a species-specific PCR. The PCR signals produced with primers specific for the tri5 gene of trichothecene-producing Fusarium spp. and with primers for the detection of F. graminearum (gaoA) were in accordance with corresponding concentrations of deoxynivalenol (DON) found in samples by HPLC analysis. The speed of the protocol developed may promote the use of PCR in routine applications in an agro-industrial context.  相似文献   

6.
Gibberella zeae (anamorph Fusarium graminearum ) causes head blight of cereals and contaminates grains with mycotoxins such as deoxynivalenol (DON). To determine the correlations among aggressiveness traits, fungal colonization and DON production, 50 progeny from a segregating population of G. zeae were inoculated onto a susceptible winter wheat cultivar in three field environments (year–location combinations). Aggressiveness traits were measured as head-blight rating and plot yield relative to noninoculated plots. Fungal colonization, measured as Fusarium exoantigen (ExAg) content, and DON production were analysed with two ELISA formats. Disease severity was moderate to high based on head-blight rating and relative plot yield. Fusarium ExAg content and DON production ranged from 0·26–1·41 units and from 4·18–43·70 mg kg−1, respectively. Significant ( P  = 0·01) genotypic variation was found for all traits. Heritability for Fusarium ExAg content was rather low because of high progeny–environment interaction and error. DON/ Fusarium ExAg ratio did not vary significantly ( P  > 0·1) among progeny. Correlation between DON production and Fusarium ExAg content across environments was high ( r  = 0·8, P  = 0·01), but no covariation existed between aggressiveness traits and DON/ Fusarium ExAg content ratio.  相似文献   

7.
The genetic diversity and pathogenicity of isolates of Fusarium graminearum and F. asiaticum isolated from wheat heads in China were examined and compared with those of isolates of F. graminearum , F. asiaticum and F. meridionale from Europe, USA and Nepal. Genetic diversity was assessed by SSCP (single strand conformation polymorphism) and AFLP (amplified fragment length polymorphism) analysis and by molecular chemotyping. SSCP analysis of the Fg16F/Fg16R PCR amplicon differentiated F. graminearum , F. asiaticum and F. meridionale and revealed three haplotypes among sequence-characterized amplified region (SCAR) type 1 F. graminearum isolates. AFLP analysis showed a high level of genetic diversity and clustered the majority of Chinese isolates in one group along with other isolates of Asian origin. The second cluster contained F. graminearum isolates from China, Europe and the USA. Of the Chinese isolates, 79% were F. asiaticum and 81% of these were of the 3-AcDON chemotype, with only 9·5% of either chemotype 15-AcDON or NIV. All the Chinese and USA isolates of F. graminearum were 15-AcDON, whereas among the isolates from Europe, 21% were NIV and 8% were 3-AcDON chemotype. No evidence was found for possible differences in aggressiveness between F. graminearum and F. asiaticum . Highly aggressive isolates were present in each region and no evidence was found for any association between aggressiveness and geographical origin or chemotype among the isolates examined. No difference was observed in pathogenicity towards wheat seedlings between Chinese isolates and those from Europe, the USA or Nepal.  相似文献   

8.
Experiments were conducted under controlled environment conditions to study the relationship between environmental conditions, development of fusarium head blight (FHB) and mycotoxin production. A single isolate from each of four Fusarium species ( F. avenaceum , F. culmorum , F. graminearum and F. poae ) was used to inoculate wheat ears separately. Combinations of two or three isolates were also used to inoculate ears simultaneously. Inoculated ears were subjected to various combinations of duration of wetness (6–48 h) and temperature (10–30°C). For all inoculations, both incidence of spikelets with FHB symptoms and concentration of mycotoxins generally increased with increasing length of wetness period and temperature. There were significant positive correlations among disease incidence, fungal biomass (quantified as total amount of fungal DNA) and mycotoxins. Mycotoxin production was also greatly enhanced by high temperatures (≥ 20°C) during initial infection periods. In single-isolate inoculations, F. poae was the least aggressive. There was no evidence to support synergetic interactions between fungal isolates in causing visual symptoms; rather the results suggest, in most cases, the presence of competitive interactions. Furthermore, the competition led to large reductions in fungal biomass compared to single-isolate inoculations, often > 90% reduction for the weaker isolate(s). In contrast, mycotoxin productivity increased dramatically in the co-inoculations, by as much as 1000 times, suggesting that competition resulted in greater production of trichothecene mycotoxins. The F. graminearum isolate was most competitive and isolates of the other three species were similar in their competitiveness.  相似文献   

9.
魏琪  廖露露  陈莉  齐永霞 《植物保护》2019,45(5):221-225
为明确安徽省玉米穗腐病主要致病镰孢菌的种类,采用单孢分离、形态学鉴定和分子生物学鉴定的方法,对2017年采集自安徽省6个玉米主产区——合肥市、蚌埠市、淮北市、阜阳市、亳州市和宿州市的玉米穗腐病病样进行了分离鉴定。在获得的455株镰孢菌中,拟轮枝镰孢菌、禾谷镰孢复合种、层出镰孢菌、尖孢镰孢菌和黄色镰孢菌的分离频率分别为59.13%、21.28%、13.68%、5.12%和0.79%。其中拟轮枝镰孢菌在各地的分布最广,属于安徽省的优势致病镰孢菌。  相似文献   

10.
A 2-year field experiment (1997–98, 1998–99) was conducted to study mummification and subsequent sporulation in spring of apple (cvs James Grieve, Golden Delicious) and pear (cv . Conference) fruits infected by Monilinia fructigena . Most mummified fruits were found in James Grieve and Conference, whereas in late-infected Golden Delicious, fruits were still soft when examined in April. In the first year, these late-infected fruits had a significantly higher sporulation intensity per sporulating fruit ( P  = 0·05) compared with Golden Delicious fruits infected 9 and 5 weeks before harvest maturity, which were partly mummified. It was concluded that early- and late-infected fruits contributed to primary inoculum in the next season. In a postinfection regime of 25°C and 65–75% relative humidity under controlled conditions, the number of Conference fruits sporulating decreased rapidly, and after 12 weeks' incubation sporulation had completely ceased. After 8 weeks' incubation, sporulation intensity in the postinfection regime at 10°C was significantly higher than that at 20 and 25°C in a first experiment with inoculated unripe fruit ( P  = 0·05). Results of a second experiment with ripe fruit were less clear. These results are discussed in relation to orchard disease management.  相似文献   

11.
Susceptibility of eight commercial European wheat cultivars to fusarium head blight (FHB) disease caused by Fusarium graminearum , F. culmorum , F. poae and Microdochium nivale (formerly known as Fusarium nivale ) was compared under controlled environment conditions (16°C). FHB did not differentially affect cultivars in terms of disease symptoms, fungal DNA content of grain or deoxynivalenol (DON) contamination. However, the Hungarian-grown cultivars GK-Othalom and Fatima 2 (of Romanian origin) showed greater type V resistance (yield tolerance) to FHB than did the others. Also, nivalenol was produced by F. poae in these two cultivars and in Italian cultivar Norba, but not in other cultivars. Overall, significant relationships were found between the FHB and seedling blight resistance in vitro of these eight cultivars, but such relationships were generally highly dependent on cultivar, and therefore it is likely that the in vitro test is at best measuring components of FHB resistance and/or genotype-specific resistance components.  相似文献   

12.
Doohan  Parry  & Nicholson 《Plant pathology》1999,48(2):209-217
Quantitative PCR analysis and visual disease assessment (VDA) were used to study Fusarium culmorum and F. poae ear blight of wheat and its fungicidal control in three glasshouse trials (1994–5 and 1996–7). VDA indicated high levels of both diseases in the trials, while PCR analysis showed that the amounts of F. poae detected in infected plants were low relative to the amounts detected for F. culmorum . PCR and VDA analysis both indicated that the demethylase-inhibiting (DMI) fungicides prochloraz and tebuconazole significantly decreased F. culmorum and F. poae ear blight. The PCR results, however, revealed levels of disease control by fungicide treatments that were consistently higher than those suggested by VDA. Overall, both fungicides appeared equally effective in controlling the two pathogens. PCR and VDA analysis indicated that the anilino-pyrimidine fungicide pyrimethanil had no significant effect on F. culmorum ear blight. Correlations between VDA at cereal growth stage 80 and PCR analysis were similar for F. culmorum and F. poae . Yield analysis, as measured by 1000 grain weight, indicated that DNA content more accurately predicted yield loss than did VDA scores. Inoculation with F. culmorum significantly reduced yield and significant relationships were observed between F. culmorum disease (as assessed visually or by PCR) and yield, with yield decreasing as disease increased. In contrast, inoculation with F. poae had no significant effect on yield and no significant relationships were observed between F. poae disease and yield. These results have important implications for field studies of Fusarium ear blight of wheat because they highlight the importance of identification of the causal organisms to the species level.  相似文献   

13.
Experiments in controlled environments were carried out to determine the effects of temperature and leaf wetness duration on infection of oilseed rape leaves by conidia of the light leaf spot pathogen, Pyrenopeziza brassicae . Visible spore pustules developed on leaves of cv. Bristol inoculated with P. brassicae conidia at temperatures from 4 to 20°C, but not at 24°C; spore pustules developed when the leaf wetness duration after inoculation was longer than or equal to approximately 6 h at 12–20°C, 10 h at 8°C, 16 h at 6°C or 24 h at 4°C. On leaves of cvs. Capricorn or Cobra, light leaf spot symptoms developed at 8 and 16°C when the leaf wetness duration after inoculation was greater than 3 or 24 h, respectively. The latent period (the time period from inoculation to first spore pustules) of P. brassicae on cv. Bristol was, on average, approximately 10 days at 16°C when leaf wetness duration was 24 h, and increased to approximately 12 days as temperature increased to 20°C and to 26 days as temperature decreased to 4°C. At 8°C, an increase in leaf wetness duration from 10 to 72 h decreased the latent period from approximately 25 to 16 days; at 6°C, an increase in leaf wetness duration from 16 to 72 h decreased the latent period from approximately 23 to 17 days. The numbers of conidia produced were greatest at 12–16°C, and decreased as temperature decreased to 8°C or increased to 20°C. At temperatures from 8 to 20°C, an increase in leaf wetness duration from 6 to 24 h increased the production of conidia. There were linear relationships between the number of conidia produced on a leaf and the proportion of the leaf area covered by 'lesions' (both log10-transformed) at different temperatures.  相似文献   

14.
In a series of field experiments in eastern England over 5 years, severe ear blight developed only in plots of winter wheat that were inoculated by spraying with conidial suspensions of Fusarium culmorum during anthesis, and in which infection was encouraged by rainfall or mist irrigation. In the absence of artificial inoculation of the ears, F. culmorum caused less extensive ear blight, and only where soil-surface inoculum was available after its application on infested plant material (colonized oat grains) up to 3–4 weeks before anthesis; it then developed most where significant rainfall occurred close to the time of anthesis. A warm, dry period following application of inoculum to the ground in late March contributed to increased infection of grain by F. culmorum , although ear blight was not increased. Ear infection therefore depended on adequate viable inoculum on infested plant debris within the crop, and conditions tending to favour brown foot rot development as well as, subsequently, rainfall and moist conditions during anthesis. These conditions did not occur together naturally during this period. Seedling infection by F. culmorum or Microdochium nivale made no significant contribution to ear blight. Inoculation of ears at anthesis with M. nivale or a locally obtained isolate of F. langsethiae did not produce ear blight symptoms. Possibilities for minimizing the availability of inoculum of F. culmorum and the implications for various options for ear-blight control are discussed.  相似文献   

15.
The incidence of pathogenic Fusarium and Microdochium species in stem bases of winter wheat was investigated in a total of nine crops in 3 years, i.e. 1987, 1988 and 1989. Four Fusarium species were isolated: F. nivale ( = Microdochium nivale ), F. avenaceum , F. culmorum and F. graminearum . The predominant species was F. nivale followed by F. avenaceum and F. culmorum . Isolations of F. graminearum were made only from shoots collected during August 1989. The highest incidence of F. nivale occurred during April 1989 in the cultivar Brock when the fungus was isolated from 65% of the shoots sampled. The highest incidence of F. avenaceum was 60% (August 1988, cv. Slejpner) and F. culmorum 37% (August 1989, cv. Mercia). A delay in the isolation of Fusarium spp. during 1987 was attributed to the low January temperatures, and an upsurge of F. culmorum and F. graminearum during 1989 to the warm dry summer. The incidence of F. nivale fluctuated during the 1988 and 1989 seasons, particularly during spring. The effects of fungicide spray programmes and the growth and development of the wheat crop are discussed as possible contributory factors to this.  相似文献   

16.
Effects of weather variables of mould development on sorghum grain were studied over three consecutive seasons in South Africa. Five sorghum hybrids planted at different dates ensured developing seeds were exposed to different weather conditions. Incidence of grain mould fungi was determined at harvest by incubating seeds on 2% malt extract agar. Averages of different weather variables (maximum and minimum temperatures, maximum relative humidity, total precipitation and frequency of precipitation) were determined for all permutations of weekly time intervals for a 2-month postflowering period to identify when these variables and pathogen incidence were significantly correlated. Significant correlations were used to develop models to quantify relationships between variables. Significant positive correlations were observed between the incidence of mould fungi and weather 4–6 weeks after flowering in the shorter season hybrid cv. Buster, and 5–8 weeks after flowering in the remaining hybrids. In most hybrids, correlations between the incidence of grain mould pathogens, including Alternaria alternata , Curvularia spp. ( C. lunata and C. clavata ), Fusarium spp. ( F. proliferatum and F. graminearum ), and Drechslera sorghicola , and average minimum temperature, total rainfall and frequency of rainfall were significant ( P =  0·05). In four hybrids, models showing a linear relationship between the logarithm of pathogen incidence and minimum temperature, and in one hybrid, between pathogen incidence and rainfall frequency, were developed. Depending on the hybrid, models that used minimum temperature as predictor described 60–82% of variation in the incidence of pathogens. Frequency of rainfall explained 93% of the variation in pathogen incidence in one sorghum hybrid genotype. Evaluation of the models using an independent data set yielded average prediction errors near zero, indicating that the models were acceptable.  相似文献   

17.
In controlled environment experiments, sporulation of Pyrenopeziza brassicae was observed on leaves of oilseed rape inoculated with ascospores or conidia at temperatures from 8 to 20°C at all leaf wetness durations from 6 to 72 h, except after 6 h leaf wetness duration at 8°C. The shortest times from inoculation to first observed sporulation ( l 0), for both ascospore and conidial inoculum, were 11–12 days at 16°C after 48 h wetness duration. For both ascospore and conidial inoculum (48 h wetness duration), the number of conidia produced per cm2 leaf area with sporulation was seven to eight times less at 20°C than at 8, 12 or 16°C. Values of Gompertz parameters c (maximum percentage leaf area with sporulation), r (maximum rate of increase in percentage leaf area with sporulation) and l 37 (days from inoculation to 37% of maximum sporulation), estimated by fitting the equation to the observed data, were linearly related to values predicted by inserting temperature and wetness duration treatment values into existing equations. The observed data were fitted better by logistic equations than by Gompertz equations (which overestimated at low temperatures). For both ascospore and conidial inoculum, the latent period derived from the logistic equation (days from inoculation to 50% of maximum sporulation, l 50) of P. brassicae was generally shortest at 16°C, and increased as temperature increased to 20°C or decreased to 8°C. Minimum numbers of spores needed to produce sporulation on leaves were ≈25 ascospores per leaf and ≈700 conidia per leaf, at 16°C after 48 h leaf wetness duration.  相似文献   

18.
大豆根腐病原镰孢菌种群多样性DGGE分析及其致病性研究   总被引:4,自引:0,他引:4  
 应用变性梯度凝胶电泳(DGGE)技术以及主成分分析方法,结合植物病原菌传统分离、鉴定及致病力测定,检测位于黑龙江省海伦市大豆连作定位试验区根腐病原镰孢菌的种群构成及其致病力。采用真菌传统分离和鉴定方法共确定镰孢菌6个种,包括燕麦镰孢Fusarium avenaceum、木贼镰孢F. equiseti、禾谷镰孢F. graminearum、腐皮镰孢F. solani、尖镰孢F. oxysporum和拟轮枝镰孢F. verticillioides。其中,尖镰孢菌的分离频率最高,为56.7%。结合DGGE条带克隆测序及系统发育分析鉴定出8种镰孢菌,较传统方法增加了黄色镰孢F. culmorum和一个尖镰孢近似种。同时,DGGE图谱中尖镰孢所占百分比下降为37.4%。采用主成分分析方法将大豆生育指标相关的多种变量进行降维分析,得到第一主成分贡献值为91.2%。按照贡献值的正负结果,将30株镰孢菌分为致病和非致病类群。第二主成分贡献值为5.8%,根据其正负值可以矫正被错误估计的致病能力。综合所有研究结果,该定位试验区大豆根腐病主要病原镰孢菌为尖镰孢、禾谷镰孢和燕麦镰孢,且以尖镰孢菌为优势病原菌。  相似文献   

19.
Quantitative real-time polymerase chain reaction differentiating 10 Fusarium spp. and Microdochium nivale or M. majus was applied to a total of 396 grain samples of wheat, barley, triticale, oat, and rye sampled across Denmark from 2003 to 2007, along with selected samples of wheat and barley from 1957 to 2000, to determine incidence and abundance of individual Fusarium spp. The mycotoxins deoxynivalenol (DON), nivalenol, zearalenone, T-2, and HT-2 were quantified using liquid chromatography-double mass spectrometry. Major differences in the Fusarium species complex among the five cereals as well as great yearly variation were seen. Fusarium graminearum, F. culmorum, and F. avenaceum were dominant in wheat, with DON as the dominant mycotoxin. F. langsethiae, F. culmorum, and F. avenaceum were dominant in barley and oat, leading to relatively high levels of the mycotoxins T-2 and HT-2. F. graminearum, F. culmorum, and F. avenaceum dominated in triticale and rye. The nontoxigenic M. nivale/majus were present in significant amounts in all cereal species. Wheat and barley samples from 1957 to 1996 exhibited no or very low amounts of F. graminearum, indicating a recent increase of this pathogen. Biomass and mycotoxin data exhibited good correlations between Fusarium spp. and their corresponding mycotoxins under field conditions.  相似文献   

20.
Total DNA from a cycloheximide-resistant mutant of Fusarium graminearum producing zearalenone was incubated with the protoplasts of a non-zearalenone-producing cycloheximide-susceptible isolate of F. culmorum. The regenerated protoplasts were inoculated on a 5-mM cycloheximide medium. The cultures which developed were examined for zearalenone production. Sixty-five cycloheximide-resistant isolates, including two isolates producing zearalenone, were obtained from approximately 3.6 × 104 viable F. culmorum protoplasts. Similar incubations with homologous F. culmorum DNA and non-fungal DNA did not elicit zearalenone production. The results suggest the transformation of F. culmorum by the genetic factors responsible for cycloheximide resistance and zearalenone production from F. graminearum. The epidemiological and taxonomic significance of these observations is discussed.  相似文献   

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