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.     

Characterisation of a Quantitative Resistance to Vector Transmission of Tomato yellow leaf curl virus in Lycopersicon pimpinellifolium

Two wild genotypes from the same species Lycopersicon pimpinellifolium, WVA106 (susceptible) and INRA-Hirsute (so-called ‘resistant’), were compared with respect to their reaction to Tomato yellow leaf curl virus isolate Réunion (TYLCV-Mld[RE]), using both whitefly-mediated inoculation and graft inoculation. Disease incidence and symptom severity were scored. Presence and quantification of viral DNA were assessed by dot blot hybridisation. Upon insect inoculation, accession INRA-Hirsute showed a moderate resistance against TYLCV that was overcome by a high inoculation pressure obtained by increasing the cumulative number of inoculative whiteflies. Temporal analyses of the disease progress in relation to this criterion exhibited that the protection was quantitative, mainly reducing the TYLCV-Mld[RE] incidence by at maximum 50% at low inoculation pressure. When graft inoculated, the final TYLCV-Mld[RE] disease incidence was 100% in both susceptible and resistant genotypes with severe symptoms, suggesting a reduction of virus transmission by a vector resistance as a possible mechanism. Implications of using such type of resistance in breeding programmes are discussed.     

A Severe Hellenic CMV Tomato Isolate: Symptom Variability in Tobacco, Characterization and Discrimination of Variants

A severe strain of Cucumber mosaic virus (CMV) originating from an infected tomato plant (Gastouni-Olympia, Greece) was isolated in tobacco (Nicotiana tabacum cv. Xanthi nc), after three serial local lesion passages in Chenopodium quinoa and designated CMV-G. CMV-G induces yellow mosaic (YM) symptoms in tobacco. When CMV-G was passed mechanically through C. quinoa, phenotypic variants inducing YM or green mild mosaic (MM) in tobacco were isolated. Aphid transmission, from different hosts, appears to be an effective approach for separating MM variants of CMV-G from YM variants. In particular, aphid transmission from zucchini proved to be very efficient in selecting for MM variants. In contrast, aphids transmitted only YM variants from tomato plants. Molecular characterization of CMV-G and its progeny resulted in their classification in the CMV subgroup IB, free of satellite RNA, being the first discovery of the subgroup IB in Greece. In the Solanaceae family (tobacco, tomato, pepper) YM variants induced more severe symptoms than the MM variants. YM and MM phenotype was stable in tobacco for all seven passages tried using the obtained YM and MM variants. Cross-protection experiments showed that an isolated MM variant was able to protect tobacco plants against a challenge infection by a YM variant.     

Comparison ofPotato Virus Y andPlum Pox Virus transmission by two aphid species in relation to their probing behavior

Two different aphid species,Myzus persicae (Sulzer) andHyalopterus pruni (Geoffroy) (Homoptera: Aphididae), were used to analyze their ability to transmit two different potyviruses,Potato virus Y (PVY) andPlum pox virus (PPV), to pepper (Capsicum annuum) andNicotiana benthamiana plants, respectively. In parallel experiments,M. persicae consistently transmitted both viruses with high efficiency, whereasH. pruni always failed to transmit either virus. This is in contrast to previous reports describingH. pruni as a vector of these potyviruses. Different aphid probing behavior among individual aphids of each species was obtained in electrical penetration graph (EPG) experiments performed on pepper plants. This suggested thatH. pruni did not transmit these potyviruses due to behavioral differences during probing that impeded virus acquisition and/or inoculation. It was found thatM. persicae usually makes its first probe within the first 2 min, whereasH. pruni individuals remained for more than 10 min on the plant before starting to probe. Furthermore,M. persicae individuals displayed their first intracellular puncture during the first minute of probing whereasH. pruni needed ∼ 15 min to penetrate the cell plasmalemma with their stylets. In addition, intracellular stylet punctures occurred very frequently forM. persicae but was a rare event, never exceeding a single one, forH. pruni. The relevance of these findings for the epidemiological spread of potyviruses by different aphid species is discussed. http://www.phytoparasitica.org posting May 14, 2006.     

乳腺分泌物中体细胞的研究进展

不同物种的乳腺分泌物中含有的细胞成分被称为体细胞,其中包括淋巴细胞、白细胞、巨噬细胞和上皮细胞。物种、乳腺感染情况、不同生理阶段和饲养管理条件等因素均会影响乳中的体细胞数量和细胞类型。近年来,乳中体细胞得到了人们的关注和深入研究,显示出广阔的应用前景。人们利用从初乳和常乳中得到的乳腺上皮细胞已经成功进行了乳腺细胞的原代培养和建立了乳腺细胞系,为乳生成、被动免疫转移和乳腺癌的研究提供了良好平台。体细胞中提取的RNA代表了乳腺组织的基因表达,因此为研究乳腺组织的基因表达提供了方便、良好的来源。     

H9N2亚型禽流感病毒传播途径特性的比较及其表面基因序列分析

选择中国大陆最早分离的H9N2亚型禽流感病毒(avian influenza virus,AIV)A/Chicken/Guangdong/SS/94(H9N2)(缩写为SS株)和1998年大流行时期分离的H9N2亚型AIVA/Chicken/Shanghai/F/98(H9N2)(缩写为F株)为研究对象,对其在SPF鸡体内的复制能力和传播途径特性比较后发现,F株在4周龄SPF鸡气管中的复制能力高于SS株,F株可以经气溶胶传播途径传播,SS株不能经气溶胶传播途径传播;利用反转录-聚合酶链反应(RT-PCR)方法获取F株和SS株的HA和NA基因的cDNA,序列分析得知,F株和SS株的HA和NA基因的同源性分别是96.6%和98.1%;HA基因的裂解位点氨基酸序列都是PARSSR↓GL,但有5个氨基酸的差异,即166位N(F)→D(SS)、198位A(F)→V(SS)、217位V(F)→I(SS)、335位G(F)→R(SS)、504位L(F)→S(SS);2株病毒的NA基因在63~65位都存在氨基酸缺失,但在NA基因红细胞吸附位点的氨基酸序列不同,分别是IKKDSRSG(F)和IKEDLRSG(SS)。F株和SS株的传播特性差异是否与其表面基因序列有关,有待进一步研究。     

Partial characterization of phytoplasmas associated with lettuce and wild lettuce phyllodies in Iran

Phytoplasmas associated with lettuce phyllody (LP) and wild lettuce phyllody (WLP) in southern Iran were partially characterized by molecular analyses and host-range studies. Agents of both diseases were transmitted by Neoaliturus fenestratus , a leafhopper colonizing lettuce and wild lettuce, to lettuce, wild lettuce, sowthistle and periwinkle, but not to safflower, sunflower, calendula and sesame. Both phytoplasmas induced bud proliferation, virescence, phyllody and witches' broom in infected plants. Total DNA extracted from infected lettuce and wild lettuce or from vector tissues was subjected to PCR using phytoplasma-specific primer pair P1/P7 or nested PCR using P1/P7 followed by R16F2n/R16R2. PCR product of nested PCR (1·2 kbp) was subjected to restriction fragment length polymorphism (RFLP). RFLP analysis of nested PCR product identified the LP, WLP and N. fenestratus -associated phytoplasmas as members of the pigeon pea witches' broom group, 16SrIX. Phylogenetic analysis of the 16S rRNA gene sequence also clustered LP and WLP phytoplasmas with other known members of the 16SrIX group. While no significant differences could be detected between LP and WLP phytoplasmas, both isolates differed from Lebanese wild lettuce phyllody in molecular properties.     

Detection of lethal yellowing phytoplasma in embryos from coconut palms infected with Cape St Paul wilt disease in Ghana

This study investigated the potential of seed transmission of Cape St. Paul wilt disease (CSPWD) in coconuts. PCR amplification was used to assess the distribution of phytoplasmas in parts of West African Tall (WAT) palms infected with CSPWD. Employing phytoplasma universal primer pair P1/P7 in standard PCR, or followed with a nested PCR using CSPWD–specific primer pair G813f/AwkaSR, phytoplasma infection was detected in the trunks, peduncles, spikelets, male and female flowers of four infected WAT coconut palms. Through nested PCR, phytoplasma was also detected in four of 19 embryo DNA samples extracted individually from fruits harvested from three of the four infected palms and was confirmed as CSPWD by cloning and sequencing. Subsequently, CSPWD phytoplasma was again detected in five of 33 embryos from nine infected palms, and in one of eight fruits from two symptomless palms. Fruits from infected palms recorded higher percentage germinations in two field nurseries (average of 71·0%) compared to fruits from healthy palms (average of 57·6%), and matured fruits that had dropped from infected palms showed the same levels of germination as those harvested directly from the palms. This indicates that infected fruits retain the ability to germinate whether harvested or dropped. No phytoplasmas were detected in any of the resulting seedlings and plantlets obtained through embryo in-vitro culture. Therefore, although phytoplasma DNA can be detected in embryos, there is as yet no evidence that the pathogen is seed transmitted through to the seedling to cause disease in progeny palms.