Alina Gospodaryk, Inga Moročko-Bičevska, Neda Pūpola and Anna Kāle
To evaluate the occurrence of nine viruses infecting Prunus a large-scale survey and sampling in Latvian plum orchards was carried out. Occurrence of Apple mosaic virus (ApMV), Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV), Apple chlorotic leaf spot virus (ACLSV), and Plum pox virus (PPV) was investigated by RT-PCR and DAS ELISA detection methods. The detection rates of both methods were compared. Screening of occurrence of Strawberry latent ringspot virus (SLRSV), Arabis mosaic virus (ArMV), Tomato ringspot virus (ToRSV) and Petunia asteroid mosaic virus (PeAMV) was performed by DAS-ELISA. In total, 38% of the tested trees by RT-PCR were infected at least with one of the analysed viruses. Among those 30.7% were infected with PNRSV and 16.4% with PDV, while ApMV, ACLSV and PPV were detected in few samples. The most widespread mixed infection was the combination of PDV+PNRSV. Observed symptoms characteristic for PPV were confirmed with RT-PCR and D strain was detected. Comparative analyses showed that detection rates by RT-PCR and DAS ELISA in plums depended on the particular virus tested. The results obtained in this study revealed that commonly grown plum cultivars in Latvia are infected with economically important stone fruit viruses and highlight the need to implement a programme to produce and propagate virus-free planting material.
Dora Krezhova, Antoniy Stoev, Nikolay Petrov and Svetla Maneva
Two hyperspectral remote sensing techniques, spectral reflectance and chlorophyll fluorescence, were used for the identification of biotic stress (sharka disease) in plum trees at an early stage without visible symptoms on the leaves. The research was focused on cultivars that are widely spread in Bulgaria: ‘Angelina’, ‘Black Diamond’ and ‘Mirabelle’. Hyperspectral reflectance and fluorescence data were collected by means of a portable multichannel fibre-optics spectrometer in the visible and near infrared spectral ranges (400-1000 nm). Statistical and deterministic analyses were applied for assessing the significance of the differences between the spectral data of healthy (control) and infected plum leaves. Comparative analyses were performed with complementary serological test DAS-ELISA, broadly implemented in plant virology. The strong relationship that was found between the results from the two remote sensing techniques and serological analysis indicates the applicability of hyperspectral reflectance and fluorescence techniques for conducting health condition assessments of vegetation easily and without damage before the appearance of visible symptoms.
Lindsey Y. K. Suh, Tayabaa Kartoon, Naiyana Gujral, Youngmee Yoon, Joo Won Suh and Hoon Sunwoo
Two enzyme-linked immunosorbent assay (ELISA) - based detection systems: indirect competitive ELISA and biotinylated double antibody sandwich ELISA (DAS-ELISA) were developed to determine the melittin concentration in honeybee (Apis mellifera) venom and the melittin concentration in cosmetics which contain bee venom. The indirect competitive ELISA employed chicken anti-melittin IgY. The biotinylated DAS-ELISA employed anti-melittin monoclonal antibody (MAb) and biotinylated anti-melittin IgY. To produce anti-melittin IgY; Sigma melittin was emulsified with Freund‘s incomplete adjuvant and immunised to Leghorn laying chickens intramuscularly at four different sites (50 μg/mL, 0.25 mL per site) of the breast muscles. After 5 to 8 weeks of the immunisation, anti-melittin IgY was extracted and analysed by ELISA. The anti-melittin IgY antibody produced was highly specific to melittin and did not cross-react with other bee venom proteins, as examined by ELISA and a western-blot assay. Indirect competitive ELISA demonstrated a higher range of melittin detection (2.5 to 80 μg/mL). Double antibody sandwich ELISA using MAb as the capture antibody and biotinylated polyclonal IgY as the detection antibody, provided a lower range of detection (2.5 - 40 ng/mL), which has a 1000 times higher sensitivity than that of indirect competitive ELISA. Therefore, indirect competitive ELISA is a useful tool to measure the concentration of melittin in bee venom as a raw material. Biotinylated DAS-ELISA, on the other hand, is more suitable for nanoscale quantification of melittin in commercial products.
The viruses infecting tulips have a big influence on the yield and the quality of bulbs and forced flowers. Commercial bulb production is based on clonal propagation, which leads to the accumulation of viruses. Among 22 viruses occurring in tulips, the most common and the most dangerous are Tulip breaking virus, TBV; Tobacco necrosis virus, TNV; Lily symptomless virus, LSV; Cucumber mosaic virus, CMV and Tobacco rattle virus, TRV. The aim of the research was to check which viruses occur most often on Polish tulip plantations. The research was done on two tulip (Tulipa L.) cultivars ‘Strong Gold’ and ‘Leen van der Mark’ grown at 3 farms situated in different parts of Poland (Warsaw Region, Pomerania and Podlasie) during 2006-2007, and then at 2 farms located in Warsaw Region during 2008-2011. Five of the most important viruses infecting tulips (TBV, TNV, LSV, CMV, and TRV) were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) in leaves and in bulbs during the period 2006-2010. In the last year of the research two different strains of TRV were detected (TRV-J and TRV-F) and Tulip virus X (TVX) as well. Search for viruses showed that most often TBV virus was detected both in the leaves and the bulbs regardless of the year and plantations. Yellow flowering cultivar ‘Strong Gold’ was infected by viruses more often than in bi-coloured (with red) ‘Leen van der Mark’, because of the difficulties with effective roguing of infected plants due to inconspicuous symptoms of virus infections on yellow coloured flowers. Other viruses were detected sporadically, however increasing occurrence of LSV and TRV was noticed from year to year. In 2011, TVX virus was detected in a few plants of ‘Strong Gold’ and it was the first case of detection of this virus in tulip in Poland.
Hanna Berniak, Beata Komorowska and Dariusz Sochacki
A one-step RT-PCR procedure was developed for specific detection of Narcissus latent virus (NLV) isolates. Following alignment of RNA sequences of three NLV isolates, the conserved sequence fragments were identified in viral polyprotein gene and 3’UTR region. Based on those fragments, a forward NLs1 and two reverse: NLa1 and NLSCPR2 primers were designed. Primer pairs NLs1-NLa1 and NLs1- NLSCPR2 proved to be effective in amplification of a 669 bp and 1295 bp products, respectively. Sequence analysis of amplified products confirmed their specificity.
F. Hutton, J.H. Spink, D. Griffin, S. Kildea, D. Bonner, G. Doherty and A. Hunter
Virus diseases are of key importance in potato production and in particular for the production of disease-free potato seed. However, there is little known about the frequency and distribution of potato virus diseases in Ireland. Despite a large number of samples being tested each year, the data has never been collated either within or across years. Information from all known potato virus testing carried out in the years 2006–2012 by the Department of Agriculture Food and Marine was collated to give an indication of the distribution and incidence of potato virus in Ireland. It was found that there was significant variation between regions, varieties, years and seed classes. A definition of daily weather data suitable for aphid flight was developed, which accounted for a significant proportion of the variation in virus incidence between years. This use of weather data to predict virus risk could be developed to form the basis of an integrated pest management approach for aphid control in Irish potato crops.
foods. Trends Food Sci. Tech. 22, 561-569.
M owat W.P., 1995. Tulip. In: Virus and Virus-like Diseases of Bulb and Flower Crops. G. Loebenstein, R.H. Lawson and A.A. Brunt (Eds), John Wiley & Sons/Balaban, Chichester, UK, 352-383.
O zaslan M., B as B., A ytekin T., S igirci Z., 2006. Identification of pepper viruses by DAS-ELISA assays in Gaziantep-Turkey. Plant Path. J. 5, 11-14.
P older G., van der H eijden G.W.A.M., van D oorn J., C levers J.G.P.W., van der S choor R., B altissen A.H.M.C., 2010. Detection of the tulip breaking
Experiments were carried out to evaluate the reaction of cabbage cultivars to mechanical inoculation with selected isolates of the turnip mosaic virus (TuMV). Simultaneously we aimed for the assessment of TuMV pathogenicity towards cultivars chosen to be transformed in order to obtain the resistance trait. The TuMV-CAR37A and TuMV-CAR39 isolates from horseradish proved to be infective towards ‘Amager’ and ‘Langedijker’ B. oleracea subsp. capitata f. alba. The course of symptom expression was assessed and the results of virus detection in symptomless leaves, using DAS-ELISA, were documented. Both tested cultivars showed a similar level of susceptibility. TuMV-CAR37A and TuMV-CAR39 can be useful in the selection of cabbage lines with resistance to the turnip mosaic virus.
Tadeusz Kobyłko, Piotr Dańda, Beata Hasiów, Henryk Pospieszny and Natasza Borodynko
A virus was isolated from Lavandula angustifolia Mill. plants exhibiting yellow mottling and distortion of leaves. After mechanical inoculation it induced in the major part of used test plants symptoms characteristic for Cucumber mosaic virus (CMV). Its standard properties regarding the stability in crude plant sap were as follows: longevity in vitro 1-2 days, thermal inactivation point 55-60°C, dilution end point log10minus 3 - 4. The virus reacted positive with diagnostic antiserum against CMV in DAS-ELISA test. RT-PCR reaction revealed similarity between the investigated isolate and the isolate of CMV from the Netherlands belonging to subgroup II. In the light of the foregoing facts the isolated pathogen can be identified as the Cucumber mosaic virus and Lavandula angustifolia may be regarded as its natural host.
Natural occurrence of Cucumber mosaic virus infecting water mint (Mentha aquatica) in Antalya and Konya, Turkey
Avirus causing a disease in mint (the aromatic and culinary plant) has recently become a problem in the Taurus Mountains, a mountain range in the Mediterranean region of Turkey. To detect the virus and investigate its distribution in the region, mint leaf samples were collected from the vicinity of spring areas in the plateaus of Antalya and Konya in 2009. It was found that Cucumber mosaic virus (CMV) was detected in 27.08% of symptomatic samples tested by DAS-ELISA. To the best of our knowledge, this is the first report of CMV on mint plants in this region of Turkey.