重庆地区柑桔衰退病毒多态性研究

柑桔衰退病毒(CTV)存在着复杂的株系分化现象.在使用弱毒株交叉保护防治柑桔衰退病时需要对田间病毒株系组成进行分析,并对选用的株系进行单蚜分离纯化.作者运用限制性片段长度多态性和单链构象多态性对田间获得的168个CTV样品和2个蚜传毒株的外壳蛋白基因进行分析,了解重庆市田间CTV组群构成情况,发现田间CTV以多株系混合发生为主,其中主要是CP/HinfⅠRFLP第1、3和6组群,约占总数的90%,并以强毒株混合感染为主.甜橙中CP/HinfⅠRFLP的组群构成最为复杂.单头褐色桔蚜从柚类至甜橙传播CTV的效率低(不足1%),该蚜的取食可改变CTV的株系组成.     

Identification and properties of a deviant isolate of the broad bean yellow band serotype of pea early-browning virus from faba bean (Vicia faba) in Algeria

A virus, isolated from faba bean (Vicia faba) obtained from Algeria, was readily recognized as a tobravirus by its particle sizes and morphology. Pea (Pisum sativum) and French bean (Phaseolus vulgaris) characteristically reacted to the isolate like pea early-browning virus (PEBV), but faba bean,Antirrhinum majus, Nicotiana rustica, andN. tabacum reacted with line-pattern symptoms which were unusually brilliant on theNicotiana species. In electronmicroscope decoration tests, the isolate did not react with an antiserum to the Dutch type strain of PEBV, but with one to the broad bean yellow band (BBYB) serotype from Italy. It resembles this serotype in reaction on faba bean, but seems to differ appreciably onN. rustica, N. tabacum, andPetunia hybrida. It is described as a deviant isolate of the BBYB serotype of PEBV.All thirteen faba-bean genotypes tested were found to be susceptible to the Algerian isolate and two Dutch type strain isolates of the virus, and to react with erratic line-pattern symptoms to the Algerian isolate only. All ten genotypes of chickpea (Cicer arietinum) tested reacted hypersensitively, and four out of ten genotypes of lentil (Lens culinaris) were susceptible to the virus but reacted differentially to the three isolates. Seed transmission of PEBV, including the new isolate, in faba bean is confirmed (9% for the Algerian isolate, and over 45% for one of the Dutch type strain isolates), and seed transmission of the virus in a non-legume (N. rustica, 4%) is herewith first reported. This is the first report on the occurrence of the BBYB serotype of PEBV outside Italy, and of PEBV outside Morocco in North Africa.     

Detection of the German grapevine yellows (Vergilbungskrankheit) MLO in grapevine,alternative hosts and a vector by a specific PCR procedure

A polymerase chain reaction procedure was developed which enables specific amplification of a ribosomal sequence from the mycoplasmalike organism (MLO) associated with German grapevine yellows (Vergilbungskrankheit, VK) and stolbur-related diseases of solanaceous plants. Successful amplification from all samples prepared from various cultivars collected in different viticultural areas indicates that the causal agent is a relatively homogeneous organism. Amplification was also achieved with template DNA prepared from naturally infected weeds in vineyards such asConvolvolus arvensis andSolanum nigrum, and from the planthopperHyalesthes obsoletus that was collected in the vineyards. Feeding of insects of this species on grapevine seedlings resulted in the development of typical yellows symptoms by the grapes.H. obsoletus could therefore be identified as a vector of Vergilbungskrankheit.Abbreviations FD Flavescence dorée - GY Grapevine yellows - MLO Mycoplasmalike organism - PCR Polymerase chain reaction - RFLP restriction fragment length polymorphism - VK Vergilbungskrankheit (German grapevine yellows)     

涩宁兰输气管道工程进度控制

运用项目工程的控制理论，结合国内管道工程建设的经验，创建了涩宁兰管道工程建设管理模式，从对材料的采办、发放、现场调拔到对进度数据统计分析、预测及计划执行情况进行分析，及时、合理地调配资源，确保月计划的完成，以实现工期控制目标。     

实验室人工感染诱发猪瘟野毒垂直传播

利用2头人工感染中等毒力猪瘟野毒耐过猪进行配种,诱发了猪瘟野毒垂直传播.带毒母猪于带毒后171 d产下9头仔猪,其中3头为死胎,6头为木乃伊.直接免疫荧光抗体试验和RT-PCR检测,9头均为阳性.测序结果表明,3头测序仔猪中,2头仔猪所分离病毒E2基因主要抗原编码区序列与公猪所接种病毒的一致;另1头仔猪所分离病毒E2基因主要抗原编码区序列与公猪所接种病毒的同源性高于与母猪所接种病毒的同源性.母猪在与公猪配种前后,其所分离病毒E2基因主要抗原编码区序列发生了变化,配种后与公猪所分离病毒的一致,说明猪瘟病毒在猪体内的繁殖存在一定的优势选择现象.     

Identification of divergent variants of Grapevine virus A

Eight isolates of Grapevine virus A (GVA), which induced different symptoms in leaves of Nicotiana benthamiana, were recovered from various grapevines. The dsRNA patterns of two isolates, which consistently induced mild vein clearing (referred here as mild isolates of GVA) were similar, but different from those of other isolates of GVA. Analysis based on overall nucleotide (nt) sequence identity in the 3 terminal part of the GVA genome, comprising part of ORF3 (putative movement protein, MP), entire ORF4 (capsid protein, CP), entire ORF5 and part of 3 UTR, revealed that GVA isolates separate into three groups (I, II, III), sharing 91.0–99.8% nt sequence identity within groups and 78.0–89.3% nt sequence identity between groups. Mild isolates of the virus were group III and shared only 78.0–79.6% nt sequence identity with the other isolates. The comparison of predicted amino acid sequences for MP and CP revealed many amino acid alterations, revealing distinct local net charges of these proteins for mild isolates of the virus. Based on both conserved and divergent nt regions in the CP and ORF5, oligonucleotide primers were designed for the simultaneous RT-PCR detection of all GVA isolates and for the specific detection of the most divergent virus variants represented here by mild isolates of the virus.     

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.     

Transmission of Theileriosis in the Traditional Farming Sector in the Southern Province of Zambia during 1995–1996

The incidence of first contact with the protozoan Theileria parva was determined in three traditional cattle herds in the Southern Province of Zambia in 1995 and 1996. The majority of first contacts occurred during the dry season in June, July and August, at a time of nymphal activity and in the absence of Rhipicephalus appendiculatus adults, indicating that larva to nymph transmission plays a more prominent role than nymph to adult transmission under the prevailing conditions.     

Assessment of Transmission Ability of Barley Yellow Dwarf Virus-PAV Isolates by Different Populations of Rhopalosiphum padi and Sitobion avenae

Populations of two aphid species from different geographic regions of Morocco were tested for their ability to transmit five barley yellow dwarf virus-PAV (BYDV-PAV) type isolates using Clintland 64 oat as the test plant. Transmission efficiencies were determined for 10 sub-populations of Rhopalosiphum padi and 12 sub-populations of Sitobion avenae. After a short acquisition access period (AAP) of 4h, all populations transmitted the virus but with different efficiencies. R. padi (Rp-S) and S. avenae (Sa-S) collected in the Settat region were the most efficient vectors, with transmission rates of 38% and 27%, respectively. R. padi (Rp-C) collected at Chaouen and S. avenae (Sa-B) at Berkane, were poor transmitters with respective vectoring abilities of 20% and 16%. These four sub-populations were chosen to study the acquisition of BYDV-PAV and the retention of virus within aphids in more detail. The transmission after two AAPs of 4h and 48h were compared. Starved aphids given a 4h AAP had significantly higher transmission efficiencies than non-starved aphids. However, after a 48h AAP, no difference was observed in the transmission between starved and non-starved aphids. Intraspecific variability was also detected by means of serial transfers of individual aphids after the given AAP. Following the first day of serial transfers, no differences were observed in transmission efficiency and virus titers for sub-populations within each species acquiring the virus during 48h, but there was significant variation when the virus was acquired in 4h. The levels of PAV antigen retained by aphids fed on healthy plants declined rapidly during the first day after acquisition, but remained fairly constant during the next 5–7 days depending on the length of the AAP. Virus antigen could be detected by ELISA in Rp-S and Sa-S for up to 11 days of serial transfer, but it was shown that aphids could retain and transmit BYDV-PAV for at least 3 weeks.     

Precision of estimated QTL positions in granddaughter designs using combined haplotype sharing TDT and linkage analysis

The aim of this study was to develop the linear haplotype sharing transmission disequilibrium test (LHS-TDT) method and combine this method with the simple regression method to estimate the precision of QTL positions in granddaughter designs. This precision was determined by Monte Carlo simulation in granddaughter designs. A single bi-allelic QTL at the midpoint of a linkage group and 26 markers with 1 cM intervals and with two alleles each were simulated. Three linear models, (i.e. the simple regression model, the linear haplotype sharing TDT method and the combination of these two models) were compared. The mean of absolute differences (A) between the estimated and true QTL position of each method was considered for six different scenarios consisting of combinations of a number of markers and the most frequent haplotypes. The mean of A, using the simple regression method, was 4.38 centimorgan (cM). The means of A using the LHS-TDT method were less than the simple regression method in all scenarios and ranged from 1.86 to 3.82 cM depending on the scenario. The mean of A using the combined method was more than the LHS-TDT method and less than the simple regression method. The means of A using the combined method ranged from 2.32 to 4.36 cM. Therefore, for populations similar to those population simulated in this study, the LHS-TDT was better than the simple regression method and the combined method for precision of estimated QTL position in granddaughter designs.