Litterarische Berichte

Ohne Zusammenfassung     

Development of specific PCR primers for identification and detection of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> Leon

A PCR-based method was developed for the identification and detection of Phytophthora capsici in pepper plants. Three PCR primers (CAPFW, CAPRV1 and CAPRV2) specific for P. capsiciwere designed based on the sequence of its internal transcribed spacer regions. CAPFW/CAPRV1 amplify a 452 bp product from P. capsici DNA whereas CAPFW/CAPRV2 a 595 bp fragment; neither set amplifies DNA from pepper or several fungi pathogenic to pepper. In conventional (single-round) PCR, the limit of detection was 5 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was of 0.5 fg. However, when the dilution series of target DNA were spiked with plant DNA, amplification declined two-fold in both conventional and nested PCR. The CAPFW/CAPRV2 set in conventional PCR was used to detect P. capsici DNA in inoculated plants. Detection occurred as soon as 8h post-inoculation in stem samples from infected but still symptomless plants. The method was also tested to detect fungal DNA in infected soils.     

黑龙江省大豆疫霉根腐病调查与病原分离

１９９６年对黑龙江省东部和中部大豆产区２３个市、县的大豆苗期疫霉根腐病进行了调查、研究,应用ＰＢＮＩＣ疫霉选择性培养基分离大豆疫霉根腐病病原菌,从牡丹江、穆棱、林口和佳木斯豆田具疫霉根腐症状的大豆植株上分离到大豆疫霉根腐病菌,并从根腐病株上单独或与大豆疫霉菌同时分离到终极腐霉菌,研究进一步证实我国黑龙江省有大豆疫霉根腐病。调查发现,大豆疫霉根腐病和终极腐霉根腐病主要发生在土质粘重、土壤含水量高或易积水的田块。     

Use of real-time PCR to examine the relationship between disease severity in pea and <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> DNA content in roots

Aphanomyces euteiches causes severe root rot of peas. Resistance is limited in commercial pea cultivars. Real-time fluorescent PCR assay specific for A. euteiches was used to study the relationship between disease severity and pathogen DNA content in infected peas. Five pea genotypes ranging in levels of resistance were inoculated with five isolates of A. euteiches. Plants were visually rated for disease development and the amount of pathogen DNA in roots was determined using the PCR assay. The susceptible genotypes Genie, DSP and Bolero tended to have significantly more disease and more pathogen DNA than the resistant genotypes 90-2079 and PI 180693. PI 180693 consistently had less disease, while 90-2079 had the lowest amount of pathogen DNA. The Spearman correlation between pathogen DNA quantity and disease development was positive and significant (P < 0.05) for three isolates, but was not significant for two other isolates. This suggests that the real-time PCR assay may have limited application as a selection tool for resistance in pea to A. euteiches. Its utility as a selection tool would be dependent on the correlation between disease development and pathogen DNA content for a given pathogen isolate. The accuracy and specificity of the real-time PCR assay suggests considerable application for the assay in the study of mechanisms of disease resistance and the study of microbial population dynamics in plants.     

Intra-specific variation and inheritance of BYDV-PAV transmission in the aphid <Emphasis Type="Italic">Sitobion avenae</Emphasis>

Vector efficiency of 44 clonal lines (clones) of Sitobion avenae belonging to 31 different genotypes (distinct patterns for five microsatellite loci) originating from Western France was evaluated by transmitting the isolate PAV4 of BYDV-PAV to barley seedlings. Variation in transmission rates from 3.7% to 92.5% was observed, with significant effects of the aphid clone, of the plant species on which clones were collected, and of the reproductive mode of the clones. When genotypes are considered instead of clones, a continuum in transmission rates was observed. A subset of S. avenae clones was tested for transmission of one (10 clones) and 13 (4 clones) other BYDV-PAV isolates, and a clear clone effect modulated by an isolate effect was observed. Crosses were made between clones with different vectoring phenotypes and their F1 progeny were tested for PAV4 transmission. The narrow sense heritability of the PAV transmission character was rather high in the F1 families (h²=0.5) and the segregation analyses suggested an oligo/polygenic determinism of this character. The possibility of generating new transmission variants by sexual reproduction and its consequences on transmission mechanism studies and on BYD epidemics are discussed.     

鸡马立克氏病肿瘤组织细胞端粒长度的测定

用马立克氏病病毒 ( MDV)京 -1株血毒对 SPF鸡攻毒 ,7周后扑杀 ,采取各内脏组织 ,一部分组织做病理切片 ,发现有肿瘤细胞浸润者即判定为阳性 ;另一部分组织用于端粒长度测定。以 DIG标记的探针( TTAGGG) 4,通过 Southern Blot方法测定细胞端粒长度 ,结果表明 ,阳性肾脏、肝脏、脾脏、神经、卵巢等组织细胞端粒长度较正常组织细胞的明显缩短     

Outbreak of clinical listeriosis in sheep: evaluation from possible contamination routes from feed to raw produce and humans

We report the results of clinical and microbiological investigations on Listeria monocytogenes infections in a flock of 55 sheep and describe the implications for the safety of the raw milk and raw-milk cheeses produced in the on-farm dairy. The outbreak was caused by feeding grass silage, which was contaminated with 5 log10 CFU L. monocytogenes/g. Clinically, although having been fed from the same batch of silage, abortive (nine ewes), encephalitic (one ewe) and septicaemic (four ewes) forms of listeriosis were observed during the outbreak phase. As the starting point of feeding the contaminated silage was known we could calculate an incubation period of 18+/-2 and 26 days for the abortive and the encephalitic form of listeriosis, respectively. Pathologically, the septicaemic cases suffered from Listeria accumulation at comparable numbers in visceral organs but not in the brain. Only a single ewe developed central nervous symptoms and a rhomb-encephalitis was immunohistologically confirmed. In this case the infection proceeded from the nasal mucosa into the brain, with no infections of the liver, spleen and other visceral organs. Sampling of the cheese production chain, the farm environment and the persons living at the farm revealed the exposure of a farm-worker to an isolate genetically indistinguishable from the outbreak clone, obviously through the consumption of faecally contaminated bovine raw milk. The cheese under processing was free of Listeria because, as a result of intensive consultations, the farmer ensured a proper acidification of the cheese. The epidemiological findings suggest that food safety matters should be assessed in any case where infection of food-producing animals with potential human pathogens is observed.