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1.
Tests on samples of oilseed rape seed ( Brassica napus ) sown in the UK between 1981 and 1984 indicated that on average 25% of samples were infected with Alternaria brassicae and 61% with Leptosphaeria maculans , with maximum incidences of infection of 19 and 4.2% respectively. Much infection by Alternaria spp. occurred on vegetable and forage brassica seed produced in the UK between 1979 and 1983. In B. oleracea types A. brassicicola occurred most frequently, affecting 88% of samples and up to 55% of seeds. A. brassicae was detected in 44% of B. oleracea samples and in up to 13% of seeds. Little Alternaria infection occurred in swede or forage rape samples (B. napus ), but A. brassicae affected up to 8–5% of seeds in turnip samples ( B. campestris ). L. maculans occurred in 44% of samples of vegetable and forage brassica seed produced in the UK, with a maximum of 4–6% infected seeds. A. brassicicola was present in 73% of samples of imported B. oleracea seed, affecting up to 25.5% of seeds. A. brassicae was absent from these samples and little L. maculans was detected. Pathogenicity tests on isolates of L. maculans from infected seeds indicated that virulent pathotypes were present in 16 rape seed samples but in only one sample (swede) of vegetable or forage brassica seed. The high incidence of seed infection by these pathogens emphasizes the importance of applying fungicide treatments to all types of brassica seed.  相似文献   

2.
Potato blackleg is a seedborne disease that can cause significant economic losses for growers. Disease development depends mainly on two drivers, namely seed inoculum and local climatic conditions. To better establish the relationship between these two drivers, blackleg development was monitored in Swiss field trials at multiple locations from 2010 to 2013 involving three sets of naturally infected seed lots planted in each of three locations. The seed lot itself was thereby the most important factor explaining differences in disease development, rather than environmental factors. In a further on-farm project conducted at various locations in Switzerland and southern Germany from 2013 to 2015, the implementation of a seed-testing procedure was investigated. A total of 177 seed lots were tested for natural latent infection with soft rot Pectobacteriaceae and the corresponding blackleg incidence was tracked in 242 fields. The reliability of the relationship between latent infection and field incidence was found to be strongly linked to the bacterial species. Dickeya spp. field infection could be predicted with an acceptable reliability, whereas Pectobacterium carotovorum subsp. brasiliense, even when detected as latent tuber infection, was not consistently expressed as visual blackleg. Moreover, commonly found mixed latent infections with several bacterial species made it even harder to predict which bacteria would cause blackleg symptoms. Finally, variability in the reliability of seed testing may also be explained by differences in local farming practices. These trials over several years with naturally infected potato seed highlight the usefulness and limits of seed testing to manage blackleg.  相似文献   

3.
 由西瓜嗜酸菌(Acidovorax citrulli)引起的细菌性果斑病是一种毁灭性的种传病害,可为害多种葫芦科作物并造成重大经济损失。该病原菌为检疫性有害生物,种子带菌是田间病害发生的最重要初侵染来源,因此,种子健康检测成为病害综合防控过程中的重要环节。Bio-PCR是当前种子携带细菌检测的常用方法,而特异性引物的选择和使用是检测的关键。本研究使用已报道的7对引物对17株西瓜嗜酸菌、10株嗜酸菌属其它种的菌株和6株其它属的植物病原细菌进行了Bio-PCR检测,筛选出对西瓜嗜酸菌特异性最好的引物为SEQID4m/SEQID5。研究表明:使用该引物对西瓜嗜酸菌MH21纯菌菌悬液的检测限度为102 CFU·mL-1;在人工添加菌悬液的模拟带菌西瓜种子中,使用ASCM和EBBA两种半选择性培养基结合引物SEQID4m/SEQID5进行Bio-PCR检测,ASCM对种子中带菌量的检测限度可达到0.01 CFU·g-1,EBBA对种子中带菌量的检测限度为0.1 CFU·g-1。  相似文献   

4.
This study investigated conidial dispersal in the field, and effects of simulated wind and rain on the dispersal of A. brassicicola on Chinese cabbage ( Brassica pekinensis ). Spores were sampled using a Burkard volumetric spore sampler and rotorod samplers in a Chinese cabbage crop. Disease incidence in the field was well fitted by a Gompertz curve with an adjusted r 2 of >0·99. Conidia of A. brassicicola were trapped in the field throughout the growing season. Peaks of high spore concentrations were usually associated with dry days, shortly after rain, high temperature or high wind speed. Diurnal periodicity of spore dispersal showed a peak of conidia trapped around 10·00 h. The number of conidia trapped at a height of 25 cm above ground level was greater than that at 50, 75 and 100 cm. Conidial dispersal was also studied under simulated conditions in a wind tunnel and a rain simulator. Generalized linear models were used to model these data. The number of conidia caught increased significantly at higher wind speeds and at higher rain intensities. Under simulated wind conditions, the number of conidia dispersed from source plants with wet leaves was only 22% of that for plants with dry leaves. Linear relationships were found between the number of conidia caught and the degree of infection of trap plants.  相似文献   

5.
Verticillium dahliae is a soilborne fungus that causes Verticillium wilt on multiple crops in central coastal California. Although spinach crops grown in this region for fresh and processing commercial production do not display Verticillium wilt symptoms, spinach seeds produced in the United States or Europe are commonly infected with V. dahliae. Planting of the infected seed increases the soil inoculum density and may introduce exotic strains that contribute to Verticillium wilt epidemics on lettuce and other crops grown in rotation with spinach. A sensitive, rapid, and reliable method for quantification of V. dahliae in spinach seed may help identify highly infected lots, curtail their planting, and minimize the spread of exotic strains via spinach seed. In this study, a quantitative real-time polymerase chain reaction (qPCR) assay was optimized and employed for detection and quantification of V. dahliae in spinach germplasm and 15 commercial spinach seed lots. The assay used a previously reported V. dahliae-specific primer pair (VertBt-F and VertBt-R) and an analytical mill for grinding tough spinach seed for DNA extraction. The assay enabled reliable quantification of V. dahliae in spinach seed, with a sensitivity limit of ≈1 infected seed per 100 (1.3% infection in a seed lot). The quantification was highly reproducible between replicate samples of a seed lot and in different real-time PCR instruments. When tested on commercial seed lots, a pathogen DNA content corresponding to a quantification cycle value of ≥31 corresponded with a percent seed infection of ≤1.3%. The assay is useful in qualitatively assessing seed lots for V. dahliae infection levels, and the results of the assay can be helpful to guide decisions on whether to apply seed treatments.  相似文献   

6.
出口萝卜种子中芸薹生链格孢的分离与鉴定   总被引：1，自引：0，他引：1  
本文对酒泉出境萝卜种子上发现的链格孢菌(Alternariasp.)进行了形态学和分子生物学鉴定,根据结果,将这种链格孢鉴定为芸薹生链格孢(Alternariabrassicicola);该种病原菌是危害十字花科蔬菜种子生产的世界性病原真菌,在甘肃酒泉繁种基地过去未见报道。  相似文献   

7.
Fusarium oxysporum f. sp. phaseoli (Fop) is a devastating pathogen that can cause significant economic losses and can be introduced into fields through infested Phaseolus vulgaris (common bean) seeds. Efficient seed health testing methods can aid in preventing long‐distance dissemination of this pathogen by contaminated seeds. In order to improve detection of Fop in seed, a rapid, accurate and sensitive real‐time PCR assay (qPCR) protocol was developed for detection of Fop in common bean seeds. Seed lots of seven cultivars with infection incidence ranging from 0·25 to 20% were prepared by mixing known amounts of Fop‐infected seeds with Fop‐free seeds. Direct comparisons between SYBR Green and TaqMan qPCR methods were performed using primers based on the Fop virulence factor ftf1. The primers developed in this study produced a 63 bp product for highly virulent strains of Fop but did not produce an amplicon for nonpathogenic or weakly pathogenic isolates of F. oxysporum from P. vulgaris or other hosts. Under optimized conditions, both qPCR assays detected Fop infection at low levels (0·25%); however, the results suggest the TaqMan assay was more reliable at quantification than the SYBR Green assay. Linear regression models were fitted to the relationships between results of qPCR assays and infection incidence, but the models differed among cultivars. Fungal biomass per seed differed among cultivars and was related to seed size. The results indicate that the TaqMan assay developed in this study is a useful tool for the detection and quantification of Fop in bean seeds.  相似文献   

8.
Routine laboratory testing of 710 bean seed lots from various origins forPseudomonas syringae pv. phaseolicola (Psp) with immunofluorescence microscopy (IF) showed that 27.5% of the seed lots (five subsamples of 1000 seeds tested per sample) contained two or more IF-positive cells in a total of 500 microscope fields (magnification 500×). Simultaneously performed dilution-platings of IF-positive subsamples on King's medium B confirmed presence of Psp for one-third of these IF-positive seed lots. The grey area of disagreement between both laboratory tests was studied by comparison of test data and by field trials.The number of IF-positive cells per subsample was positively correlated with isolation and identification of Psp (R=0.85). The detection level of IF was ca. 102 Psp cells per ml of undiluted subsample extract. The detection level of Psp by isolation on King's medium B was variable, being inversely related with the saprophyte to Psp ratio. The high sensitivity of IF was in part due to high percentages of dormant or dead IF-positive cells in the sample extract. Field trials over two years with 10 000 seeds per seed lot, showed disease incidence for 9 of the 22 seed lots. Of ten IF-positive lots with five positive subsamples per sample, nine were positive in the field test plot (the negative lot gave primary infection spots of Psp when used for commercial growing). By isolation, seven of these ten IF-positive lots were positive. Of the five IF-positive lots with two or less positive subsamples, isolation and field trial were both negative. Based on data on seed transmission from literature, field incidence was unlikely for these five samples in a 10 000 seeds field trial. All seven IF-negative lots were negative in the field trial. The value of IF and isolation for indexing bean seed lots for Psp is discussed.This study was carried out at the Centre for Plant Breeding and Reproduction Research (CPRO-DLO), Binnenhaven 1, P.O. Box 16, 6700 AA Wageningen the Netherlands, to which address correspondence should be addressed.  相似文献   

9.
Over a 5‐year period (2006–2010), 277 certified, visually healthy potato seed lots, imported from Europe to Israel for commercial use, were tested for Dickeya spp. latent infection by PCR analysis (277 seed lots) and ELISA (154 seed lots). Seeds from these lots were grown in commercial potato fields which were inspected twice a season by Plant Protection and Inspection Services (PPIS). Stem samples were tested for the presence of Dickeya spp. by PCR analysis. PCR and ELISA results from seed lot testing correlated with disease expression in 74 and 83·8% of the cases, respectively. Positive laboratory results with no disease symptoms in the field (‘+lab/?field’ results) comprised 24·7 and 9·7% of the PCR and ELISA analyses, respectively, whereas negative laboratory results with disease symptoms in the field results (‘?lab/+field’) were obtained in 1·3 and 6·5%, of cases respectively. Maximum disease incidence, as well as the number of cultivars expressing disease symptoms, increased over the years of this study, indicating an increase in the prevalence of the disease. Severe disease incidence was observed on cvs Dita, Rodeo, Desiree, Mondial, Tomensa and Jelly. Of the 55 imported seed lots from which disease was recorded in the field, 49 originated from the Netherlands, four from Germany and two from France. None originated from Scotland.  相似文献   

10.
Potato seed tubers are imported annually to Israel from northern Europe. Although the seed is registered as certified, a survey carried out over a 9-year period indicated that most lots were affected by latent or active bacterial and fungal infections. Latent infection byErwinia carotovora subsp.atroseptica, the causal agent of blackleg, at a level of 103 cells/g peel, was present in 30% of the lots in most years. Black scurf caused byRhizoctonia solani was present in 20–70% of the imported lots, with a moderate to high level of infection in all years except 1985. In contrast, although many lots were affected by powdery scab, common scab, and Fusarium dry rot in most years, disease incidence within lots was generally low. The gangrene pathogen (Phoma exigua) was rarely detected. The survey findings are of marked importance, due to the extensive use of soil fumigation in Israeli agriculture.  相似文献   

11.
Potato (Solanum tuberosum) is the largest crop in Israel. Production is based on the import of seed tubers from Europe for the spring crop. Imported tubers are generally free from virus infection. The most important virus infecting potato is Potato virus Y (PVY), which may cause severe damage to marketable yields. In Israel, tubers from the spring harvest are stored over the summer for planting in the autumn. It is important to be able to determine the infection rate of seed tuber lots from the spring harvest prior to storage. Commonly, infection is measured by sprouting tubers and measuring virus titre in the leaves using ELISA (the “Growing-On test”), which takes at least 6 weeks to give results. There is a need for a faster method to produce results, such as Taqman Real Time PCR (qPCR), for direct analysis of viral infection in tubers at harvest. To use qPCR as a diagnostic tool, it is necessary to demonstrate that both techniques give comparable results on batches of field-grown tubers. Such a comparison was performed on potential seed tuber lots of 14 different cultivars over three Israeli spring harvests (2013–2015). The agreement between the results of the two techniques was not of high statistical significance. However, the qPCR technique can distinguish well, by binary classification, between tuber lots with a low PVY infection rate (<5% by Growing On test; suitable for seed) and those unsuitable for seed (≥5% by Growing On test). Therefore, qPCR is an appropriate technique for determination of the PVY infection rate of seed tuber lots in Israel.  相似文献   

12.
Tsror  Leah  Aharon  M.  Erlich  Orly 《Phytoparasitica》1999,27(3):215-226
Potato seed tubers are imported to Israel from northern Europe and planted in spring; tubers harvested early from the spring crop are used as seed for the autumn crop. Although only seed lots registered as certified are imported, a previous survey (1984–1994) indicated that most imported lots were affected by latent or active infections caused byErwinia carotovora,Streptomyces scabies, Rhizoctonia solani, Fusarium spp. andSpongospora subterranae. The survey was extended until 1998, and included additional pathogens:Ralstonia solanacearum,Helminthosporium solani, Colletotrichum coccodes andVerticillium dahliae. Most of these pathogens were also monitored in domestic seed tubers, and are reported for the first time. Brown rot was not observed in any of the imported lots. Blackleg and soft rot caused byErwinia spp. were detected in most of the imported lots; however, less than 7% of the lots were contaminated at high levels, while approximately 65% were contaminated at moderate levels. Common scab was detected in most of the imported lots; 51% of the imported lots were contaminated at moderate or high levels, whereas only 6.5% of the domestic seed lots were contaminated at these levels. Black scurf was detected in most of the imported lots; on average, 47.3%, 44.2% and 1.4% of the lots were contaminated at low, moderate and high levels, respectively, and only 7.1% were disease-free. In contrast, most of the domestic lots were either disease-free (45.4%) or had a low disease incidence (37.3%). Only 16.7% of the lots were moderately infected and 0.2% were highly contaminated. Silver scurf was observed in most of the imported lots during all years of the survey, with no differences among the producing countries; on average, 22.7%, 66.1% and 7.5% of the lots were contaminated at low, moderate and high levels, respectively, and only 3.7% were disease-free. Most of the domestic lots (76%) were disease-free and only 6.6% were infected at moderate or high levels. Black dot was observed in a considerable portion of the shipments from Holland during all years of the survey, particularly in 1998, when 34% of the lots were infected. The shipments from France and Germany were infected at low levels, except in 1998, when 19% and 11% of the lots, respectively, arrived infected. In shipments from Scotland and Ireland low incidences of the disease were observed in 1994 and 1995. In the domestic lots, black dot incidence was low (<2.4%) except in 1996, when 11% of the lots were infected.V. dahliae was monitored only in domestic seed tubers. The incidence of disease-free lots was 56–64%, whereas in 20–30% of the lots the level of infection was <5%, and in 6–16% of the lots the level was >5%. The survey findings demonstrate transmission of seedborne pathogens; most of these pathogens can become established in the soil and eventually cause severe outbreaks of disease in potatoes grown in Israel. http://www.phytoparasitica.org posting May 16, 1999.  相似文献   

13.
Diplodia sapinea is one of the major pathogens of pines worldwide. Despite the putative critical importance of seed infection in the epidemiology of the disease, this aspect of the biology of the fungus is poorly known. Here, biological and molecular methods were developed for the detection of the fungus and applied to assess D. sapinea infection in Corsican pine seeds. A buffered medium containing tannic acid and malt extract as a nutrient base was the most efficient and selective for D. sapinea recovery. A molecular method based on DNA extraction with a commercial kit and specific amplification, including an internal amplification control, was developed. A high percentage of infection (57% positive isolations) was observed in seeds obtained from fallen cones in a Corsican pine stand with no apparent symptoms of D. sapinea. Seeds collected from trees in a seed orchard showing severe symptoms of dieback caused by D. sapinea had comparatively lower infection (38%). Moreover, very low infection levels (1–5%) were observed after the standard treatment used for seed extraction, which included heating at 40°C. Diplodia sapinea was not recovered from seedlings grown from infected seed lots submitted to water stress. Overall, results suggest that the risk of disease transmission by commercial seeds is probably low, but could be further reduced by thermotherapy.  相似文献   

14.
An indexing system for detectingTomato mosaic virus (ToMV) in commercial tomato seed lots is described. Factors associated with the procedure were analyzed and the following standard two-step working scheme is proposed: (i) mass screening by ELISA for the presence of the virus; (ii) evaluation of virus infectivity within the infested seed lots. A threshold of 10 ng ml−1 was determined for detection of purified ToMV by either ELISA or plant inoculation.Nicotiana tabacum cv. Xanthi NN was found to be a highly sensitive local lesion assay plant for the detection of ToMV. A positive ELISA threshold (1.3 times above the non-specific background) was set for seed samples taken from commercial seed lots by testing the same samples by both ELISA and a bioassay. http://www.phytoparasitica.org posting July 14, 2004.  相似文献   

15.
The potential of the competitive polymerase chain reaction (PCR) assay for quantification of seedborne infection by Rhynchosporium secalis in barley was examined using a primer set (RS1 and RS3) derived from the internal transcribed spacer (ITS) regions of ribosomal RNA genes of this pathogen. Introduction of a heterologous internal control, which competes for the same primer set in the conventional PCR assay, allowed for detection and quantification of R. secalis fungal biomass. In order to generate a standard calibration curve, DNA prepared from infected seeds with different levels of R. secalis infection was subjected to competitive PCR assay. The resulting PCR product ratio for each PCR reaction ( R. secalis -amplified DNA/internal control template-amplified DNA) increased proportionally with increasing levels of infected seed DNA in the reaction mixture. Naturally infected seed lots collected from 1995 to 1999 were used to demonstrate the potential of the competitive PCR assay as an alternative seed health testing method. The results from this competitive PCR assay were compared with those from conventional visual disease assessment and an agar plate assay. Although relatively good correlation between visual disease assessment and the competitive PCR was found in the case of artificially mixed seed samples, there was poor correlation in the experiments using naturally infected seed samples.  相似文献   

16.
A method for detectingClavibacter michiganensis ssp.michiganensis in tomato seeds was evaluated. The method is based on rapid screening of tomato seed lots using indirect immunofluorescence staining (IF), followed by dilution plating of IF positive seed lots. Different polyclonal antisera, prepared againstC. michiganensis ssp.michiganensis were tested for their specificity using IF. All strains ofC. michiganensis ssp.michiganensis tested reacted with the polyclonal antisera. Two of nine saprophytic isolates from tomato seeds were positive with the antisera as well as with the control normal serum, but cells of these isolates were distinct in shape from cells ofC. michiganensis ssp.michiganensis.For extraction of the pathogen from the seed, seeds were either blended with a stomacher or soaked at 4–6 °C. The stomacher method yielded more fluorescent cells in IF than 24 h soaking of seed samples. However, soaking of seeds for 48 h generally yielded less saprophytes and overall higher numbers ofC. michiganensis ssp.michiganensis colonies in dilution plating when compared to blending by a stomacher. SCM medium was generally more selective than KBT and modified CNS medium. However, the efficacy of the medium was dependent on the seed lot and/or extraction method used. Confirmation of suspected colonies with YDC (yeast-dextrose-carbonate medium), IF and a pathogenicity test on tomato seedlings proved to be highly reliable (P>0.95). For routine testing of seed lots it is recommended to screen tomato seed lost after soaking seeds for 24 h at 4–6 °C with IF, followed by plating of IF-positive seed lots on modified CNS and SCM after soaking seeds for an additional 24 h.  相似文献   

17.
A real-time PCR assay was designed to quantify seed-borne infection of Pyrenophora graminea in barley (Hordeum vulgare). Conventional tests such as the freezing blotter method cannot distinguish P. graminea from the closely related P. teres. The seed infection threshold for P. graminea is lower than the one for P. teres and is therefore applied for both species although P. graminea may be absent. This results in unnecessary rejections of seed lots. PCR primers and a TaqMan probe were designed to target a P. graminea-specific DNA sequence. The potential of the real-time PCR assay for quantifying seed-borne infection of P. graminea was investigated by examining seed lots harvested from P. graminea-infected fields. The major part (84%) of the variation in the amount of P. graminea DNA measured by real-time PCR could be attributed to variation between seed lots while only about 8% was due to variation within seed lots. DNA quantities of P. graminea were positively correlated with seed infection incidence detected by the freezing blotter method as well as with the infection incidence of plants examined in the greenhouse. Both correlations were highly significant (P < 0.001) but the DNA quantities accounted only for 59% (R 2 = 0.59) and 56% (R 2 = 0.56), respectively, of the variation in the results obtained by the two conventional methods. Seed lots of varieties resistant to P. graminea contained considerable amounts of P. graminea DNA but showed no or only few leaf symptoms in the greenhouse test suggesting that the recommended seed infection thresholds could be raised for resistant varieties.  相似文献   

18.
用白菜、假臭草和红毛草为受体植物,采用生物测定的方法研究了石刁柏新鲜茎叶4种浸提液的化感作用,结果表明：红毛草受石刁柏茎叶浸提液的抑制作用最强;4种溶剂的浸提液对假臭草萌发率的作用效果表现出“低浓度促进,高浓度抑制”的双重效应,而对其幼苗根茎生长具有明显的抑制作用,且浓度越大抑制作用越强;4种溶剂的浸提液对白菜萌发率和根长的作用在总体上也表现出显著的抑制作用,但对其茎长却有明显的促进作用。说明4种浸提液对受体植物种子萌发率、幼苗的根长和茎长均有不同程度的化感作用,这为利用石刁柏的化感作用控制杂草提供了理论基础。  相似文献   

19.
A method for detection and quantitative estimation of tomato seedborne pathogenic bacteria has been developed. It enables detection in a 7 g tomato seed sample of as few as ten colony-forming units per gram tomato seeds of the following seedborne pathogens of tomato:Pseudomonas syringae pv. tomato,Pseudomonas corrugata, Xanthomonas campestris pv.vesicatoria, andClavibacter michiganense subsp.michiganense. With representative seed samples, the method employs dry grinding, weighing, bacterial extraction and quantitative calculation on selective or semi-selective medium. The efficiency of this method was tested by diluting pathogen-free seed lots with naturally or artificially infested tomato seeds. This procedure enables one to determine the minimal threshold of pathogen which can be detected by this method on media, in comparison with the percentage of diseased seedlings developed from the same seed lots in the growth chamber or in the greenhouse.  相似文献   

20.
Pseudothecia containing abundant ascospores of Mycosphaerella brassicicola were produced in vitro on Brussels sprout decoction agar at 15°C under a 16-hour photoperiod of different light regimes. Spermogonia containing spermatia were also produced on the decoction agar. Ascospores were released when cultures were misted with SDW and placed under continuous light. Germination of ascospores was highest between 20°C and 25°C and spores remained viable at relative humidities above 93.5%. Exposure of ascospores to 55% relative humidity for 24 h reduced their germination to 75%. A polyclonal antiserum raised against whole ascospores was used to detect, by immunofluorescence, the ascospore and mycelial wall of M . brassicicola , following reaction with anti-rabbit IgG FITC conjugate. Autofluorescence of spore and mycelial components of other fungal species could be eliminated using the counterstains Evan's blue and eriochrome black at 0.2% and 0.5%, respectively, in phosphate buffered saline (pH 7.2). A procedure was developed to detect, by immunofluorescence, ascospores of M . brassicicola on artificially inoculated Melinex spore tape. Coating of the spore tape with bovine serum albumin provided a suitable support medium and blocking agent for detection of ascospores in the field. The potential use of the system for selective detection of ascospores of M . brassicicola in infected crops of vegetable brassicas in the presence of other ascosporic fungi is discussed. Keywords : ascospores, immunofluorescence, Mycosphaerella brassicicola , spore production, spore trapping .  相似文献   

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