Spread of Phoma exigua var. foveata to healthy potato plants

Contamination of stem-cutting bait plants exposed at various locations in and around a potato crop was greatest within the crop, less outside it and least at a site c. 20 km from commercial crops. Within the crop about half the stem infection and tuber contamination detected had occurred before haulm destruction. The incidence of contamination on tubers of bait plants placed within the crop was similar to that on tubers from the surrounding plants. Phoma exigua var. foveata was recovered from green leaves sampled at intervals between June and early September. In one year contamination by P. exigua var. foveata of tubers sampled from healthy plants surrounding a diseased plot was greatest in samples adjacent to the plot and decreased progressively away from it. This pattern was less apparent in two other years. These results are discussed in relation to the spread of airborne inoculum.     

Differentiation of Phoma foveata from P. exigua using a RAPD Generated PCR-RFLP Marker

Phoma foveata and P. exigua variety exigua both infect potatoes and are morphologically very similar. P. foveata produces a pigment which allows differentiation from P. exigua in culture. Discrimination of the two species based on the production of a secondary metabolite, which is dependent on the growth conditions, is not reliable. Therefore, there is a need to develop nucleic acid based identification markers. A 482bp random amplified polymorphic DNA (RAPD) fragment from P. foveata was isolated and sequenced. Polymerase chain reaction (PCR) primers, developed from the sequence of the RAPD product, amplified a 474bp fragment for P. foveata and P. exigua varieties exigua, diversispora, inoxydibilis and sambuci-nigrae. The similarity of the PCR fragments was demonstrated by sequence analysis and by using the restriction enzymes DdeI and DpnII. P. foveata was distinguished from the four varieties of P. exigua on the basis of the RFLP patterns of the PCR fragment. Ten isolates of P. foveata and nine of P. exigua var. exigua from different geographic locations were tested and all isolates but one showed the restriction digest pattern of the PCR fragment (PCR-RFLP) specific to each species. One isolate of P. foveata demonstrated a PCR-RFLP pattern similar to P. exigua var. exigua leading to the conclusion that the isolate had been previously misidentified as a strain of P. foveata lacking the ability to produce pigment.     

Laboratory assessments of the susceptibility of potato tubers to gangrene (Phoma exigua var. foveata)

Disease incidence was recorded in tubers of various cultivars following inoculation with macerated mycelium of Phoma exigua var. foveata and wounding in various ways. A method which produced predominantly crush wounds gave most rots and the most consistent results. The length of time between inoculation and wounding did not significantly affect the incidence of disease but disease severity did increase with damage infliction time. Inoculated, undamaged tubers produced rots but the incidence was low and discrimination between cultivars was poor when compared with damaged tubers. In some tests the resistance to gangrene could be measured by the proportion of tubers affected, in others all tubers had lesions but the extent of infection was different, The latent period of the disease differed between cultivars. Rate of lesion development was constant over a period of 12 wk within cultivars but differed between them.     

The effect of temperature before or after inoculation with Phoma exigua var. foveata on the development of gangrene in potato tubers

Potato tubers of five cultivais were incubated at five temperatures between 2 and 20°C for 1 month and then inoculated with Phoma exigua var. foveata and stored at 5°C. Larger rots usually developed on tubers previously incubated at the higher temperatures but cultivar differences were greatest where tubers had been incubated at 2°C. In another experiment, rot size was related to the duration of incubation at 20°C before inoculation.
In a series of experiments, tubers were stored at different temperatures after inoculation; other factors (e.g. wound type or r.h.) were also investigated in some of these. Combining the results of all experiments a linear decrease of % wounds infected (logit transformation) with storage temperature was demonstrated. The size of rots was not simply related to temperature, but tended to increase over the range 2–10°C and then stabilized or declined at higher temperatures. Relative humidity treatments of 75% v. 95% did not consistently affect either the incidence of rotting or rot size. The other factors investigated did not usually interact significantly with temperature.
The relevance of the results for resistance testing is discussed and a simple equation relating disease incidence to inoculum levels, type of damage and temperature is proposed.     

Mass spectrometric techniques as aids in the diagnosis of gangrene in potatoes caused by Phoma exigua var. foveata

Current diagnosis of gangrene in potato tubers, caused byPhoma exigua var.foveata, is based on isolation and identification of the causal organism or on thin-layer chromatographic (TLC) detection of the anthraquinone pigment pachybasin (1-hydroxy-3-methylanthraquinone). In this paper alternative diagnostic approaches are described based on detection of specific ions by pyrolysis mass spectrometry (Py-MS), direct probe mass spectrometry (DIP-MS) and gas chromatography-mass spectrometry (GC-MS). The advantages and disadvantages of these methods are discussed in comparison with the TLC approach. GC-MS is a promising technique because of its high specificity, sensitivity and suitability for automation. Moreover, the MS approaches may lead to a reduction of total expertise time.Samenvatting De huidige diagnostiek van gangreen in aardappelknollen, veroorzaakt doorPhoma exigua var.foveata, is gebasserd op de isolatie en identifikatie van de schimmel of op de dunnelaag-chromatografische (TLC) bepaling van het pigment pachybasine (1-hydroxy-3-methylanthrachinon).In dit artikel worden nieuwe diagnostische benaderingen beschreven die gebaseerd zijn op de detectie van specifieke ionen door middel van pyrolyse-massaspectrometrie (Py-MS), direct-probe-massaspectrometrie (DIP-MS) en gaschromatografie-massaspectrometrie (GC-MS). De voor- en nadelen van deze methoden worden behandeld en vergeleken met de dunnelaag-chromatografische methode.GC-MS is een veelbelovende methode vanwege de grote specificiteit, gevoeligheid en geschiktheid voor automatisering. Op den duur kunnen de MS-methoden leiden tot een verkorting van de totale expertisetijd.     

基于MAXENT的大豆南北方茎溃疡病菌在中国适生区的预测

利用MAXENT生态位模型和GIS系统,对大豆南北方茎溃疡病菌在我国的适生区进行预测.模型分析结果表明,这两种病菌在我国的潜在适生区域广泛.大豆北方茎溃疡病菌除了我国的宁夏、海南省外,其它省市均有该菌的适生分布区,其中适生等级高的地区主要在江苏、安徽、浙江、上海、江西和湖北6省.大豆南方茎溃疡病菌除了宁夏、青海、西藏外,我国的其它地区均为该菌的潜在适生区域,其中适生等级高的地区主要集中在江西、上海、江苏、安徽、浙江、河南以及陕西南部、四川东部和重庆的部分地区.     

Spatiotemporal distribution of Ascochyta pinodes and Ascochyta pinodella during the winter growing season in France

Ascochyta blight of pea is caused by four related fungi, Ascochyta pisi, Phoma koolunga, Ascochyta pinodes and Ascochyta pinodella. The latter two taxa appear to be much more common and economically significant worldwide but the relative impact of each fungus on ascochyta blight epidemics is not well understood. To study the spatiotemporal distribution of A. pinodes and A. pinodella infecting pea in France, 368 isolates were sampled monthly, from February to May, at three locations (Rennes, Boigneville and Dijon) and molecular markers were used to genotype isolates. The aggressiveness of isolates from the fourth sampling date was estimated using a detached leaf assay on the winter cultivar Enduro. Disease was low during the sampling period as climatic conditions were generally not conducive to disease development (cold temperature, low rainfall). Population genetic analysis showed that 99% of the observed variation could be attributed to variation within populations compared to only 1% among populations. Both species were observed in each location, although A. pinodella was observed at a lower frequency (6–32%). Moreover, results showed that both species could develop on different nodes of the plant. Significant differences in aggressiveness were observed between species and among isolates within species with A. pinodes isolates being significantly more aggressive on average than A.  pinodella isolates. These results emphasize the necessity to study the components of disease complexes in order to understand the impact of pathogen species interactions on disease and yield reduction as well as the dynamics of disease epidemics during the cropping season.     

The influence of cultivar and mechanical damage on the relationship between inoculum concentration of Phoma exigua var. foveata on potato tubers and gangrene development

In an experiment to examine the influence of cultivar and tuber damage on the relationship between inoculum concentration and potato gangrene development, the extent to which cultivars differed in their response to increased inoculum concentration depended on the type of damage which the tubers received. There was considerable variation in cultivar response in terms of lesion frequency when tubers were inoculated by damaging them on a commercial grader, whereas when they were uniformly wounded or left undamaged cultivar differences were small. The relationship between the number of detectable wounds on tubers and gangrene incidence was not consistent for the three cultivars. Lesion size was shown to increase linearly with increasing concentrations of pycnidiospores when tubers of cultivars of different susceptibilities were point-inoculated.     

Genetics of virulence in Ascochyta rabiei

In order to critically test the hypothesis that virulence variation in the Ascochyta rabiei/chickpea pathosystem is a discrete character under simple genetic control, a genetic cross was made between a highly virulent isolate of A. rabiei from Syria and a less virulent isolate from the USA. Two independent virulence assays conducted by inoculating susceptible and resistant chickpea cultivars under controlled conditions with 77 independent progeny isolates from this cross revealed a continuous distribution of disease phenotypes. Bimodality, as would be predicted for the segregation of virulence under simple genetic control, was not supported by statistical tests of the progeny phenotype distribution. anova revealed highly significant pathogen‐genotype × host‐genotype interactions demonstrating the segregation of genes controlling specialization on the two cultivars tested. These interactions could be localized to two isolates that changed virulence rank on the cultivars. It was concluded that variation in virulence to these two cultivars is under quantitative genetic control. If this conclusion applies to other cultivars, it can be speculated that the discrete categories of virulence variation identified in previous studies were probably the result of incomplete sampling of host resistance or pathogen virulence variation and/or of selection for increased virulence in contemporary A. rabiei populations.     

利用甜菜夜蛾日本品系增殖核多角体病毒

比较了甜菜夜蛾3种品系增殖甜菜夜蛾核多角体病毒(SeNPV)的效果。结果表明,利用甜菜夜蛾日本品系幼虫增殖SeNPV产量最高,其平均单头病毒含量分别比广州品系和荷兰品系高19 6%和53 3%;病毒总含量分别高20 4%和116 7%。在此基础上,进一步研究了虫龄、感染浓度和饲养密度对用甜菜夜蛾日本品系幼虫增殖SeNPV的影响,确定了利用甜菜夜蛾日本品系增殖SeNPV时,以上3因子的最佳条件为:在27±1℃下用1×108PIB/ml的SeNPV感染5龄初幼虫,并按30头/dm2的密度饲养。     

大豆茎溃疡病菌TaqMan MGB探针实时荧光PCR检测

 The technique of TaqMan MGB real-time fluorescent PCR was established to detect Diaporthe phaseolorum var.caulivora (DPC) and D. phaseolorum var.meridionalis (DPM). The primers and TaqMan MGB probes were designed based on the ITS of DPC, DPM, D. phaseolorum var. sojae and Phomopsis longicolla. A series of genomic DNA dilution were used to detect sensitivity of the technique, the results showed that the limits of detection for DPM and DPC were 7 fg/μL and 6 fg/μL DNA respectively.