Isolation, identification and molecular characterization of physiological races of apple powdery mildew (Podosphaera leucotricha)

The development of apple varieties displaying durable resistance against powdery mildew is one of the major aims in apple breeding programmes worldwide. For a reliable judgment of the resistance of different Malus genotypes, an extended knowledge about the virulence of the pathogen is necessary. To prove the existence of physiological races of Podosphaera leucotricha , 31 monoconidial isolates of the obligate biotrophic fungus representing five locations within Europe have been established and maintained over a period of 3–4 years. The isolates were maintained on in vitro shoots of the highly susceptible apple cv. Gibb's Golden Gage. An AFLP-based DNA fingerprinting protocol was developed and, using 54 stably reproducible AFLP markers, a dendrogram revealed genetic variability among different isolates of P. leucotricha . Although the molecular characterization of the isolates showed an overall low level of genetic variability, the high phenotypic diversity among European isolates suggest that sexual reproduction may also be involved in the disease cycle of the pathogen in Europe. Phytopathological tests using detached leaves of a collection of 36 Malus genotypes allowed the differentiation of five selected isolates by their virulence patterns. A high level of diversity in terms of virulence was obtained in P. leucotricha. From the present study, based on apple breeding germplasm, cultivars and Malus species, it can be concluded that physiological races of P. leucotricha do indeed exist in Europe.     

In Planta - Complementation of Clavibacter Michiganensis Subsp. sepedonicus Strains Deficient in Cellulase Production or HR Induction Restores Virulence

Clavibacter michiganensis subsp. sepedonicus, a Gram positive bacterium that causes bacterial ring rot of potato, was studied in eggplant, an alternate host, using strains that differed in phenotype. Two factors affecting virulence, the ability to induce a hypersensitive response (HR) and cellulase production, were studied. A plasmid-free isolate of C. michiganensis subsp. sepedonicus that causes HR on tobacco but is unable to produce cellulase multiplied efficiently in planta, but caused only weak symptoms. In contrast, a strain that is unable to induce HR on tobacco but produces cellulase was impaired in the ability to multiply in the host and caused no symptoms. When the two non-virulent strains were coinoculated into eggplants, typical disease symptoms developed. This enhancement was not due to formation of a new phenotype or significant increases in population density of either of the strains. Our results suggest that both cellulase production and the ability to induce HR are required for a successful infection process and disease induction by C. michiganensis subsp. sepedonicus. Our results additionally suggest that the ability to induce HR on non-host plants is required for multiplication in the host plant, whereas cellulase expression is necessary for induction of disease symptoms.     

北京地区西瓜枯萎病生理小种分化研究

对在北京大兴、顺义、通县等西瓜主产区采集，经单孢分离得到的８个西瓜枯萎病菌株进行了生理小种分化研究。应用国际通用的３个鉴别寄主ＳｕｇａｒＢａｂｙ、ｃｈａｒｌｅｓｔｏｎＧｒａｙ、ｃａｌｈｏｕｒＧｒａｙ和国内京欣１号、苏蜜１号２个品种，采用全国协作组统一的苗期抗病鉴定方法，进行人工接种鉴定。结果表明北京地区西瓜枯萎病菌株均系同一生理型，为西瓜枯萎病生理小种１号。     

影响朱砂叶螨自然种群消长的气象因子的模糊划分

根据１９８７－１９９２年年茄子田内朱砂叶螨自然种群数量动态及同期气象资料，利用条件频率法建立了气象因子对叶螨种群消长的隶属函数，详细讨论了温度、湿度、降雨对群消长的影响，用隶属函数对叶螨种群消长进行了拟合，拟合率为９３．５％。结果表明，能很好地对气象因子进行模糊划分。     

The 1997-1998 classical swine fever epidemic in The Netherlands--a survival analysis

The aim of this analysis was to characterise the temporal pattern of infection during the 1997/98 classical swine fever (CSF) epidemic in The Netherlands and hence identify and quantify risk factors for infection in different enterprise types and areas. Survival analysis and Cox proportional hazards regression were used to describe the epidemic. Substantial differences in temporal survival patterns (herd breakdown rate) were found between areas where different control policies operated. Factors with a significant influence on the infection hazard of individual herds included: sow numbers as a percentage of total sows and fatteners (HR = 3.38 for mixed herds (0.1–60% sows) vs. fattening herds (0% sows) and HR = 2.74 for breeding herds (60–100% sows) vs. fattening herds), the number of ‘transport contacts per month’ (>0.3 vs. <0.3; HR = 4.11), pig density (pigs/km²) in the area (HR_{1000 pigs} 1.48) and herd size (HR_{100 pigs} = 1.01).

Pre-emptive slaughter in an area appeared to be associated with lower subsequent disease levels. Higher frequency of transport contacts for welfare slaughter during the epidemic, however, well regulated and controlled, was associated with a substantially higher risk of becoming infected. The positive association of a higher pig density with CSF indicates the potential importance of local spread as a factor in disease transmission and emphasizes that dilution of the pig population can contribute to reduction in CSF occurrence. This analysis suggests however, that if pre-emptive slaughter can promptly be applied effectively in an area after initial diagnosis, pig density is then not a significant factor. Mixed and breeding herds had a higher probability of becoming infected than fattening herds, possibly due to different types and frequencies of inter-herd contacts. These contacts continue to some extent during the epidemic, despite the standstill of animal movements.     

葡萄叶片光合速率日间降低内外因调控的研究

土壤水分充足时,葡萄日间光合速率的下降主要归因于CO2叶肉阻力的升高;光照、叶温和大气湿度可以影响气孔对CO2的阻力,但主要通过影响叶肉阻力来影响光合速率。‘巨峰’和‘长相思’葡萄具有不同的光合作用光响应曲线及其特征参数。     

反刍动物采食量的概念与研究方法

本研究对反刍家畜采食量的研究发展与现状进行了回顾与总结,采食量及其相关术语的定义主要基于饲草料或家畜,其测定方法和精确含义有差异;反刍家畜采食是由生物和非生物因素相互作用共同影响的复杂动态过程,采食量影响因子主要分为家畜的(胃肠蠕动力、选择性采食等)、饲草料的(粗纤维含量、抗营养成分等)和饲养条件的(草层结构、草地饮水点分布等)因素3个方面;采食量的测定方法有基于牧草测定的直接法和基于家畜测定的间接法,后者较多地应用于放牧家畜;预测方法有基于数学函数和生物学原理的模型法,基于消化试验的经验法和结合家畜属性的改进经验法。舍饲条件下可准确地测定和预测家畜的采食量,但对放牧家畜还没有精确的方法。通过测定与饲草水平密切相关的家畜尿液、血液或粪便中的代谢物水平将成为预测放牧家畜采食量的准确方法。     

Effects of growth factors (EGF,PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing

Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 10⁵/well) and epithelial cells (2 × 10⁵/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using ³H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.     

Neighbourhood infections of classical swine fever during the 1997-1998 epidemic in The Netherlands

Data of the 1997–1998 epidemic of classical swine fever (CSF) in The Netherlands were analysed in survival analysis to identify risk factors that were associated with the rate of neighbourhood infections. The study population consisted of herds within 1000 m of exclusively one previously infected herd. Dates of virus introduction into herds were drawn randomly from estimated probability distributions per herd of possible weeks of virus introduction. (To confirm the insensitivity of the results for this random data-selection procedure, the procedure was repeated 9 times (resulting in 10 different datasets).) The dataset had 906 non-infected and 59 infected neighbour herds, which were distributed over 215 different neighbourhoods. Neighbour herds that never became infected were right-censored at the last date of the infectious period of the infected source herd. Neighbour herds that became empty within the infectious period or within the following 21 days due to preventive depopulation or due to the implemented buying-out programme were right-censored 21 days before the moment of becoming empty. This was done as a correction for the time a herd could be infected without being noticed as such.

The median time to identified infection of neighbour herds was 2 weeks, whereas the median time to right censoring of non-infected neighbour herds was 3 weeks. The risk factors, radial distance ≤500 m, cattle present on source herd and increasing herd size of the neighbour herd were associated multivariably with the hazard for neighbour herds to become infected. We did not find an association between time down wind and infection risk for neighbour herds. Radial dispersion of CSFV seemed more important in neighbourhood infections than dispersion along the road on which the infected source herd is situated. The results of this study support the strategy of preventive depopulation in the neighbourhood of an infected herd. Recommendations are presented to adapt the applied control strategy for neighbourhood infections.     

猪肉品质的评价指标及其影响因素

本文对猪肉品质的评价指标及其影响因素进行探讨。