Alfalfa mosaic and cucumber mosaic virus infection in chickpea and lentil: incidence and seed transmission

Samples collected in 1994 and 1995 from commercial crops of chickpeas and lentils growing in the agricultural region of south-west Western Australia were tested for infection with alfalfa mosaic (AMV) and cucumber mosaic (CMV) viruses, and for members of the family Potyviridae using enzyme-linked immunosorbent assay (ELISA). In 1994 no virus was detected in the 21 chickpea crops tested but in 1995, out of 42 crops, AMV was found in two and CMV in seven. With lentils, AMV and/or CMV was found in three out of 14 crops in 1994 and 4 out of 13 in 1995, both viruses being detected in two crops in each year. Similar tests on samples from chickpea and lentil crops and plots growing at experimental sites, revealed more frequent infection with both viruses. No potyvirus infection was found in chickpeas or lentils in agricultural areas either in commercial crops or at experimental sites. However, bean yellow mosaic virus (BYMV) was detected along with AMV and CMV in irrigated plots of chickpeas and lentils at a site in Perth. When samples of seed from infected crops or plots of chickpeas and lentils were germinated and leaves or roots of seedlings tested for virus infection by ELISA, AMV and CMV were found to be seed-borne in both while BYMV was seed-borne in lentils. The rates of transmission found through seed of chickpea to seedlings were 0.1–1% with AMV and 0.1–2% with CMV. Seed transmission rates with lentil were 0.1–5% for AMV, 0.1–1% for CMV and 0.8% for BYMV. Individual seed samples of lentil and chickpea sometimes contained both AMV and CMV. With both species, infection with AMV and CMV was sometimes found in commercial seed stocks or seed stocks from multiplication crops of advanced selections nearing release as new cultivars. Seed-borne virus infection has important practical implications, as virus sources can be re-introduced every year to chickpea and lentil crops or plots through sowing infected seed stocks leading to spread of infection by aphid vectors, losses in grain yield and further contamination of seed stocks.     

Cucumber mosaic virus genome is encapsidated in alfalfa mosaic virus coat protein expressed in transgenic tobacco plants

The expression of viral coat protein (CP) in transgenic plants has been shown to be very effective in virus plant protection. However, the introduction of CP genes into plants presents the potential risk of the encapsidation of a superinfecting viral genome in the transgenic protein, an event which could change the epidemiology of the disease. To detect the potential heterologous encapsidation of the cucumber mosaic virus (CMV) genome by alfalfa mosaic virus (AIMV) CP expressed in transgenic tobacco plants, a system of immunocapture (IC) and amplification by polymerase chain reaction (PCR) was optimized. This provided high sensitivity and reliable selection of the heterologously encapsidated CMV genome in the presence of natural CMV particles. As little as 2 pg of virus could be detected by immunocapture/polymerase chain reaction (IC/PCR) technique. Evidence for heterologous encapsidation of the CMV genome was found in 11 of the 33 transgenic plants tested two weeks after CMV inoculation. This demonstrates a significant rate of heterologous encapsidation events between two unrelated viruses in transgenic plants. Since CP is involved in the interactions of the virus particle with its vector, the release in the field of such transgenic plants could alter the transmission properties of some important viruses.     

侵染香蕉的黄瓜花叶病毒株系的形态和生物学特征

对香蕉花叶病广东三个代表分离物的形态和生物学特征进行了较为全面的研究。在六种鉴别寄主上,斑驳类（ＭＭ）和断续条纹类（ＢＳ）分离物侵染较多的寄主种类诱导较为严重的症状,连续条纹类（ＣＳ）分离物侵染能力较弱。电镜观察表明：ＢＳ、ＣＳ和ＭＭ三个代表分离物的粒子呈球状、其粒子直径分别为２２．５、２４．３和２７ｎｍ,纯化的病毒粒子电泳相对迁移率分别为０．７２、０．５６和０．５１,在西葫芦上的增殖速度：不管在子叶或真叶,都是ＭＭ最快、ＢＳ次之、ＣＳ最慢。从西葫芦接种叶运转出接种叶的速度：ＭＭ和ＢＳ相似、ＣＳ明显较慢。在烟草上,三个分离物的运转速度和增殖速度以ＭＭ最快、ＢＳ次之、ＣＳ最慢;而且在烟草上随着接种后时间的延长,三个分离物除ＭＭ维持高浓度的时间较长外,ＢＳ和ＣＳ的都很短。在西葫芦和烟草上提纯产量,ＭＭ类最多、ＢＳ类次之、ＣＳ类最低。     

Thymidine kinase activity in mouse embryo fibroblast cells infected with murine cytomegalovirus

Thymidine kinase activity was found in whole cell extracts of growing and stationary mouse embryo fibroblast cells after infection with murine cytomegalovirus. Determination of the kinetic constants and heat stability characteristics indicated that the enzyme activity from infected cells was different to that found in uninfected cells in the growth phase. The expression of thymidine kinase activity during virus replication was reflected by the incorporation of (6-³H) thymidine into acid precipitable fractions of infected cell cultures. Preliminary data from kinetic studies showed a reduction in the phosphorylation of thymidine by this enzyme activity in the presence of Acyclovir, a potent inhibitor of herpes virus replication.     

Indices of water deficit in susceptible and resistant cucumbers in response to infection by cucumber mosaic virus

Indices of water deficit were determined under conditions of non-limited water supply in cucumber mosaic virus (CMV)-infected seedlings of the susceptible cv. Bet Alpha. An increase in the concentration of soluble solids, decrease of water and osmotic potentials, and increase of proline concentration were found in the CMV-infected cotyledons. In the cv. Shimshon, which is resistant to CMV, virus infection caused only a slight change in the concentration of the soluble solids and in the osmotic potential; water potential and proline content were not affected. Concomitantly, infectivity of cotyledons by CMV was much lower in the tolerant cv. Shimshon than in the susceptible cv. Bet Alpha. The possible association of water deficit with virus-induced growth retardation is discussed.     

Integration of high-throughput analytics and cell imaging enables direct early productivity and product quality assessment during Chinese Hamster ovary cell line development for a complex multi-subunit vaccine antigen

Mammalian cell line generation typically includes stable pool generation, single cell cloning and several rounds of clone selection based on cell growth, productivity and product quality criteria. Individual clone expansion and phenotype-based ranking is performed initially for hundreds or thousands of mini-scale cultures, representing the major operational challenge during cell line development. Automated cell culture and analytics systems have been developed to enable high complexity clone selection workflows; while ensuring traceability, safety, and quality of cell lines intended for biopharmaceutical applications. Here we show that comprehensive and quantitative assessment of cell growth, productivity, and product quality attributes are feasible at the 200–1,200 cell colony stage, within 14 days of the single cell cloning in static 96-well plate culture. The early cell line characterization performed prior to the clone expansion in suspension culture can be used for a single-step, direct selection of high quality clones. Such clones were comparable, both in terms of productivity and critical quality attributes (CQAs), to the top-ranked clones identified using an established iterative clone screening approach. Using a complex, multi-subunit antigen as a model protein, we observed stable CQA profiles independently of the cell culture format during the clonal expansion as well as in the batch and fed-batch processes. In conclusion, we propose an accelerated clone selection approach that can be readily incorporated into various cell line development workstreams, leading to significant reduction of the project timelines and resource requirements.     

Die Bedeutung elektrophoretischer Proteinanalysen bei phytopathogenen Pilzen als Hilfsmittel zu ihrer taxonomischen Charakterisierung

Sunflower is one of the most important oilseed crops, which is grown in many countries. Increasing demand to use sunflower oil has expanded the under cultivation area throughout the world. Sunflower viruses have been reported as one of the pathogens that reduce the quantity and the quality of this product in a number of countries. In our research to study the occurrence and distribution of viruses in sunflower fields of Iran, 562 samples were collected from different fields in Kerman and Isfahan provinces during growing seasons from 2009 to 2011. Potato virus Y (PVY), Cucumber mosaic virus (CMV) and Tomato spotted wilt virus (TSWV) were detected by double antibody sandwich enzyme-linked immunosorbent assay. Also, numbers of samples were positive for infection to potyviruses (except PVY) in antigen-coated-plate enzyme-linked immunosorbent assay test using potyviruses general antiserum. Infected plants had symptoms like mosaic, yellowing, deformation, necrotic and chlorotic lesions and mottling on leaves and stunting. The infection rates of potyviruses, PVY, CMV and TSWV in Isfahan province were 33, 22, 4.18 and 3.25% of collected samples, respectively. The corresponding rates for samples from Kerman province were 15, 5, 0.8 and 0.4% of collected samples. According to these results, viral infection in Isfahan province was more than surveyed in Kerman province during the mentioned period of three years. Furthermore, generally there was a decrease in the percentage of viral infection during this three growing seasons in both provinces. This is the first report of the detection of CMV and TSWV in sunflower fields of Isfahan and Kerman and first report of PVY in Isfahan.     

Effect of cucumber mosaic virus infection on morphology,yield and phenolic contents of tomato

Ten tomato genotypes were screened for their resistance against cucumber mosaic virus (CMV) and its vector Myzus persicae under natural infection in field, using aphids M. persicae under net-house and mechanical inoculation under greenhouse. Large differences were observed among genotypes for infection percentage (IP) and severity index (SI) among the testing methods used. All genotypes showing tolerance to CMV in the field or through aphid inoculation, however, become susceptible and highly susceptible after mechanical inoculation. All the test genotypes also showed susceptibility to the aphid M. persicae population. Plants inoculated with CMV showed substantial decrease in yield and yield-contributing parameters which varied with cultivars that probably depended upon its genetic make up. All the test genotypes exhibited 0.97–30.19% decrease in plant height, 11.47–52.65% decrease in root length, 46.56–95.56% decrease in fresh plant weight, 65.78–92.84% decrease in root fresh weight, 19.97–87.65% decrease in the dry weight of plants, 75.63–95.43% decrease in dry root weight, 69.51–95.65% reduction in the number of fruits and 89.04–99.89% decrease in yield per plants. After 15 days of inoculation, the quantitative analysis using double beam spectrophotometer showed an increase in total phenolics in CMV-inoculated plants as compared to un-inoculated plants among genotypes. Similarly the thin layer chromatography (TLC) on silica gel G indicated that the number of phenolic compounds was increased in most of the inoculated genotypes while in others they were either decreased or remained same.     

Serological detection of yam viruses in farmers' fields in Ogun State,Nigeria

A field survey was conducted in eight local government areas (LGA) of Ogun state, Nigeria to assess the incidence of viral diseases of yams in the areas. Leaf samples were collected from 90 yam plants which were either symptomatic or asymptomatic. These were bulked into 45 during serological tests and the viruses indexed include yam mosaic virus (YMV); Dioscorea alata bacilliform virus (DaBV) and cucumber mosaic virus (CMV). DaBV was the most prevalent virus on the field with incidence of 48.9% (22/45) followed by YMV which occurred in 42.2% (19/45). CMV had the lowest percentage of incidence; 2.2% (1/45). Of all the LGAs visited, Abeokuta north and Abeokuta south had the highest incidence of YMV and DaBV, respectively. Mixed virus infections were also detected.