Cucumber Mosaic Virus Transmission Research Articles

Investigating the Interactions of the Cucumber Mosaic Virus 2b Protein with the Viral 1a Replicase Component and the Cellular RNA Silencing Factor Argonaute 1.

The cucumber mosaic virus (CMV) 2b protein is a suppressor of plant defenses and a pathogenicity determinant. Amongst the 2b protein's host targets is the RNA silencing factor Argonaute 1 (AGO1), which it binds to and inhibits. In Arabidopsis thaliana, if 2b-induced inhibition of AGO1 is too efficient, it induces reinforcement of antiviral silencing by AGO2 and triggers increased resistance against aphids, CMV's insect vectors. These effects would be deleterious to CMV replication and transmission, respectively, but are moderated by the CMV 1a protein, which sequesters sufficient 2b protein molecules into P-bodies to prevent excessive inhibition of AGO1. Mutant 2b protein variants were generated, and red and green fluorescent protein fusions were used to investigate subcellular colocalization with AGO1 and the 1a protein. The effects of mutations on complex formation with the 1a protein and AGO1 were investigated using bimolecular fluorescence complementation and co-immunoprecipitation assays. Although we found that residues 56-60 influenced the 2b protein's interactions with the 1a protein and AGO1, it appears unlikely that any single residue or sequence domain is solely responsible. In silico predictions of intrinsic disorder within the 2b protein secondary structure were supported by circular dichroism (CD) but not by nuclear magnetic resonance (NMR) spectroscopy. Intrinsic disorder provides a plausible model to explain the 2b protein's ability to interact with AGO1, the 1a protein, and other factors. However, the reasons for the conflicting conclusions provided by CD and NMR must first be resolved.

Absence of Seed-Mediated Transmission of Cucumber Mosaic Virus in Espelette Pepper Crops despite Widespread and Recurrent Epidemics.

In the past decade, severe epidemics of cucumber mosaic virus (CMV) have caused significant damage to Espelette pepper crops. This virus threatens the production of Espelette pepper, which plays a significant role in the local economy and touristic attractiveness of the French Basque Country, located in southwestern France. In 2021 and 2022, CMV was detected via double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) in Gorria pepper seed lots harvested from naturally infected fields scattered throughout the entire Espelette pepper production area. These seed lots were used in greenhouse grow-out tests to determine whether CMV could be transmitted to seedlings from contaminated seeds, using visual symptom assessment, DAS-ELISAs, and reverse transcription-polymerase chain reaction (RT-PCR). Despite the widespread occurrence of CMV in seeds of field samples, the grow-out experiments on a total of over 5000 seedlings yielded no evidence of seed transmission of local CMV isolates in Gorria pepper. Therefore, rather than seeds from infected pepper plants, sources of CMV inoculum in Espelette are more likely to be alternative hosts present in and around pepper fields that can allow for the survival of CMV during the off-season. These results have important epidemiological implications and will guide the choice of effective measures to control current epidemics.

Combined Transcriptomic and Proteomic Analysis of Myzus persicae, the Green Peach Aphid, Infected with Cucumber Mosaic Virus.

Simple SummaryIn this study, an integrated analysis of the mRNA and protein was performed to identify important putative regulators involved in the transmission of CMV (cucumber mosaic virus) by aphids. At the level of transcription, a total of 20,550 genes (≥2-fold expression difference) were identified as being differentially expressed genes (DEGs) 24 h after healthy aphid transfer to infected tobacco plants using the RNA-seq approach. At the protein level, 744 proteins were classified as being differentially abundant between virus-treated and control Myzus persicae using iTRAQ (isobaric tags for relative and absolute quantitation) analysis. The combined mRNA and protein analysis enabled the identification of some viral putative regulators, such as cuticle proteins, ribosomal proteins, and cytochrome P450 enzymes. The results show that most of the key putative regulators were highly accumulated at the protein level. Based on those findings, we can speculate that the process by which aphids spread CMV is mainly related to post-translational regulation rather than transcription.Aphids transmit CMV (cucumber mosaic virus) in a non-persistent manner. However, little is known about the mechanism of CMV transmission. In this study, an integrated analysis of the mRNA and protein was performed to identify important putative regulators involved in the transmission of CMV by aphids. At the level of transcription, a total of 20,550 genes (≥2-fold expression difference) were identified as being differentially expressed genes (DEGs) 24 h after healthy aphid transfer to infected tobacco plants using the RNA-seq approach. At the protein level, 744 proteins were classified as being differentially abundant between virus-treated and control M. persicae using iTRAQ (isobaric tags for relative and absolute quantitation) analysis. The combined mRNA and protein analysis enabled the identification of some viral putative regulators, such as cuticle proteins, ribosomal proteins, and cytochrome P450 enzymes. The results show that most of the key putative regulators were highly accumulated at the protein level. Based on those findings, we can speculate that the process by which aphids spread CMV is mainly related to post-translational regulation rather than transcription.

Arabidopsis thaliana Genes Associated with Cucumber mosaic virus Virulence and Their Link to Virus Seed Transmission.

Virulence, the effect of pathogen infection on progeny production, is a major determinant of host and pathogen fitness as it affects host fecundity and pathogen transmission. In plant–virus interactions, ample evidence indicates that virulence is genetically controlled by both partners. However, the host genetic determinants are poorly understood. Through a genome-wide association study (GWAS) of 154 Arabidopsis thaliana genotypes infected by Cucumber mosaic virus (CMV), we identified eight host genes associated with virulence, most of them involved in response to biotic stresses and in cell wall biogenesis in plant reproductive structures. Given that virulence is a main determinant of the efficiency of plant virus seed transmission, we explored the link between this trait and the genetic regulation of virulence. Our results suggest that the same functions that control virulence are also important for CMV transmission through seeds. In sum, this work provides evidence of a novel role for some previously known plant defense genes and for the cell wall metabolism in plant virus interactions.

Aphid endosymbiont facilitates virus transmission by modulating the volatile profile of host plants

BackgroundMost plant viruses rely on vectors for their transmission and spread. One of the outstanding biological questions concerning the vector-pathogen-symbiont multi-trophic interactions is the potential involvement of vector symbionts in the virus transmission process. Here, we used a multi-factorial system containing a non-persistent plant virus, cucumber mosaic virus (CMV), its primary vector, green peach aphid, Myzus persicae, and the obligate endosymbiont, Buchnera aphidicola to explore this uncharted territory.ResultsBased on our preliminary research, we hypothesized that aphid endosymbiont B. aphidicola can facilitate CMV transmission by modulating plant volatile profiles. Gene expression analyses demonstrated that CMV infection reduced B. aphidicola abundance in M. persicae, in which lower abundance of B. aphidicola was associated with a preference shift in aphids from infected to healthy plants. Volatile profile analyses confirmed that feeding by aphids with lower B. aphidicola titers reduced the production of attractants, while increased the emission of deterrents. As a result, M. persicae changed their feeding preference from infected to healthy plants.ConclusionsWe conclude that CMV infection reduces the B. aphidicola abundance in M. persicae. When viruliferous aphids feed on host plants, dynamic changes in obligate symbionts lead to a shift in plant volatiles from attraction to avoidance, thereby switching insect vector’s feeding preference from infected to healthy plants.

Genetic Diversity of Cucumber Mosaic Virus from Catharanthus roseus, Jasminum sambac, Patchouly, Cubeb and Java-tea

<p><em>Cucumber mosaic virus (CMV) symptoms are found in </em>Catharanthus roseus, patchouly (Pogostemon cablin), cubeb (Piper chaba), Jasminum sambac and Java-tea (Orthosiphon aristatus); however, their genetic characterization has not been studied.<em> The study aimed to molecularly characterize the CMV isolates from </em>Catharanthus roseus, patchouly, cubeb, Jasminum sambac and Java-tea.<em> </em><em>Disease plant samples showing mosaic and yellow mosaic symptom</em>s<em> were collected from Petak Pamer Garden, ISMCRI, Bogor. Molecular characterization was carried out by reverse transcription</em>-<em>polymerase chain reaction (RT-PCR) assay using a specific primer of CMV coat protein gene and DNA sequenced. Sequence analysis was performed using the BLAST, Bioedit, Genedoc, Mega 5 programs. The RT-PCR technique succeeded in amplifying a DNA band measuring 650 bp, according to the prediction of the primary design. BLAST analyses revealed that all of these CMV isolates belonged to subgroup IB. Nucleotide sequence homology of CMV from </em>C. roseus,<em> </em>patchouly, P. chaba, <em>and</em> J. sambac, <em>were more than 95.00%. Based on </em>the <em>phylogenetic tree, these four isolates were closely related to CMV isolate from Japan (AB070622). Homology of </em>the <em>nucleotide sequence of CMV from </em>Java-tea<em> </em><em>with </em>the <em>other four isolates w</em>as<em> below 95.00%. This isolate clustered with CMV isolate from Indonesia </em>(<em>AB042294</em>) <em>and</em> <em>was</em> <em>separated with another four isolates according to </em>the <em>phylogeny tree. In the amino acid sequence alignment, </em>Java-tea <em>isolates had five different amino acids compared to the other four isolates. This result indicates the poss</em>i<em>bility of CMV transmission between patchouly</em>, Java-tea, C. roseus <em>and</em><em> </em>J. sambac, <em>so it must be anticipated to prevent its spread. </em></p>

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The effect of Chrysoperla carnea (Neuroptera: Chrysopidae) and Adalia bipunctata (Coleoptera: Coccinellidae) on the spread of cucumber mosaic virus (CMV) by Aphis gossypii (Hemiptera: Aphididae).

The effects of two aphidophagous predators, the larvae of Chrysoperla carnea and adults of Adalia bipunctata, on the spread of cucumber mosaic virus (CMV) transmitted in a non-persistent manner by the cotton aphid Aphis gossypii were studied under semi-field conditions. Natural enemies and aphids were released inside insect-proof cages (1 m × 1 m × 1 m) with a central CMV-infected cucumber plant surrounded by 48 healthy cucumber seedlings, and the spatiotemporal dynamics of the virus and vector were evaluated in the short and long term (1 and 5 days) in the presence and absence of the natural enemy. The spatial analysis by distance indices methodology together with other indices measuring the dispersal around a single focus was used to assess the spatial pattern and the degree of association between the virus and its vector. Both natural enemies significantly reduced the number of aphids in the CMV-source plant after 5 days but not after 1 day. The CMV transmission rate was generally low, especially after 1 day, due to the limited movement of aphids from the central CMV-source plant, which increased slightly after 5 days. Infected plants were mainly located around the central virus-infected source plant, and the percentage of aphid occupation and CMV-infected plants did not differ significantly in absence and presence of natural enemies. The distribution patterns of A. gossypii and CMV were only coincident close to the central plant. The complexity of multitrophic interactions and the role of aphid predators in the spread of CMV are discussed.

Comparative transmission of two cucumber mosaic virus isolates by two color morphs of Acyrthosiphon pisum (Harris)

Cucumber mosaic virus (CMV) is one of the most important legume-infecting viruses, which is transmitted effectively by pea aphid Acyrthosiphon pisum (Harris) (Hem: Aphididae). Transmission efficiency of two CMV isolates (As and Kh from cowpea and bean hosts, resp.) by red and green color morphs of pea aphid were evaluated on bean plants. Triple-antibody sandwich ELISA (TAS-ELISA) using CMV-specific monoclonal antibodies revealed that both CMV isolates belonged to the serotype II. Bean plants inoculated by viruliferous aphids were assayed by double-antibody sandwich ELISA (DAS-ELISA) at 16 days post inoculation (dpi). The results showed that the transmission rate of CMV-As by the red morph was significantly higher than by the green morph, resulting in significantly higher transmission rate of the virus (As + Kh) by the red morph than by the green morph, with p≤ 0.1. Similarly, the efficiency of CMV transmission by the red morph of A. pisum was higher than the efficiency of transmission by the green morph. The higher transmission rate and efficiency of CMV by red pea aphid would be important in the epidemiology. Based on these results, we hypothesize that the transmission efficiency of CMV is affected more by the difference in transmission determinants of A. pisum color morphs than by the sequence of virus coat protein determinants. Aphididae; Bromoviridae; color polymorphism; transmission efficiency.

Transmission of Cucumber Mosaic Virus (CMV) infecting banana by aphid and mechanical methods

Banana is the premier fruit of the Asia and pacific region. It is the fourth most important food crop after rice, wheat and maize. In Tamil Nadu, banana is grown in 92,200 hectares with an annual production of 48.56 million tones. Among all biotic constraints, viruses cause huge economic loss to the crop. Some of the major diseases are caused by viruses which include Banana Bunchy Top Virus (BBTV), Cucumber Mosaic Virus (CMV), Banana Streak Virus (BSV), Banana Bract Mosaic Virus (BBMV) have been reported in our country. CMV virus concentration in host (Musa) tissue is very less. So it is necessary to transfer this virus to good propagation host such as Nicotiana sp for purification. The purpose of the study was the transmission of CMV infecting banana into propagation host either by aphid or mechanically. The transmission was confirmed through Direct Antigen Coating Enzyme Linked Immuno Sorbent Assay (DAC-ELISA).

Disruption of Two Defensive Signaling Pathways by a Viral RNA Silencing Suppressor

The Cucumber mosaic virus (CMV) 2b counter-defense protein disrupts plant antiviral mechanisms mediated by RNA silencing and salicylic acid (SA). We used microarrays to investigate defensive gene expression in 2b-transgenic Arabidopsis thaliana plants. Surprisingly, 2b inhibited expression of few SA-regulated genes and, in some instances, enhanced the effect of SA on certain genes. Strikingly, the 2b protein inhibited changes in the expression of 90% of genes regulated by jasmonic acid (JA). Consistent with this, infection of plants with CMV, but not the 2b gene-deletion mutant CMVDelta2b, strongly inhibited JA-inducible gene expression. JA levels were unaffected by infection with either CMV or CMVDelta2b. Although the CMV-Arabidopsis interaction is a compatible one, SA accumulation, usually considered to be an indicator of plant resistance, was increased in CMV-infected plants but not in CMVDelta2b-infected plants. Thus, the 2b protein inhibits JA signaling at a step downstream of JA biosynthesis but it primes induction of SA biosynthesis by another CMV gene product or by the process of infection itself. Like many plant viruses, CMV is aphid transmitted. JA is important in plant defense against insects. This raises the possibility that disruption of JA-mediated gene expression by the 2b protein may influence CMV transmission by aphids.

Capsid protein of cowpea chlorotic mottle virus is a determinant for vector transmission by a beetle

Cowpea chlorotic mottle virus (CCMV) is a bromovirus transmitted by species of chrysomelid beetles, including the spotted cucumber beetle, Diabrotica undecimpunctata howardii Barber. An experimental system was set up to identify the viral determinant(s) of the beetle transmission of CCMV. Nicotiana clevelandii was selected as an experimental plant host because it supports the replication and accumulation of both CCMV and a second member of the family Bromoviridae, cucumber mosaic virus (CMV). Using a reverse genetic system for CMV, a cDNA copy of the CCMV capsid protein (CP) gene was substituted for that of the CMV CP gene. The resulting 'CMV-hybrid' consisted of wild-type CMV RNA1, RNA2, and a chimeric CMV RNA3 expressing the CCMV structural protein. The CMV-hybrid replicated and formed virions in N. clevelandii; in electron micrographs the hybrid virus was indistinguishable from CCMV. In beetle feeding assays, both CCMV and the CMV-hybrid were transmitted by D. undecimpunctata, while beetle transmission of CMV was not observed. Conversely, only CMV was observed to be transmitted by the aphid Myzus persicae. Surprisingly, the CMV-hybrid was transmitted more efficiently than the parental CCMV, and a virus-induced alteration in beetle feeding behaviour is proposed to account for the difference. These results indicate that the CCMV CP is a viral determinant for beetle vector transmission.

Seed transmission of Cucumber mosaic virus in pepper

Infection caused by Cucumber mosaic virus (CMV) is one of the most important viral diseases of pepper worldwide. Young pepper seedlings were inoculated mechanically with CMV-Fny (Fast New York) isolate and were kept in growth chambers at 20–25 °C for symptom and fruit development. All inoculated plants developed severe mosaic symptoms and produced fruit except one. Mature seeds were isolated from fruits harvested from CMV-infected plants. Total RNA was extracted from pepper seeds and analyzed by reverse transcription-polymerase chain reaction (RT-PCR) using CMV sub-group IA specific primers. Analysis of individual whole seeds showed that seed-borne infection of CMV in pepper ranged from 95 to 100%. Further seed-growth tests were performed in Petri dishes and CMV was detected in both seed coat and embryo. Seed coat infection of CMV ranged from 53 to 83% while that of the embryo ranged from 10 to 46%. Seed-growth tests in pots were also performed and the rate of seed transmission was approximately 10 to 14%. This is the first report of CMV seed transmission in pepper.

Transmission Efficiency of Cucumber mosaic virus by Aphids Associated with Virus Epidemics in Snap Bean

Cucumber mosaic virus (CMV) is a major component of the virus complex that has become more pronounced in snap bean in the midwestern and northeastern United States since 2001. Multiple-vector-transfer tests were done to estimate the CMV transmission efficiencies (p) of the main aphid species identified in commercial snap bean fields in New York and Pennsylvania. The four most efficient vectors (p > 0.05) were Aphis gossypii, A. glycines, Acyrthosiphon pisum, and Therioaphis trifolii, which were all significant species in the migratory aphid populations in snap bean. Moderately efficient vectors (0.01 < p < 0.04) were A. spiraecola, A. craccivora, Macrosiphum euphorbiae, and Rhopalosiphum maidis. Poor vectors (p < 0.01) included A. fabae, Nearctaphis bakeri, and Myzus persicae. Only one species, Sitobion avenae, failed to transmit CMV in replicated tests. Estimates of p were consistent between different clones of the same aphid species and among three different field isolates of CMV tested. Single-vector-transfer test results for a subset of the species supported those obtained via the multiple-vector-transfer approach. Our results are consistent with the notion that A. glycines is a major vector of recent CMV epidemics in snap bean, but that species is only one of several that are involved.

Ability of Aphis gossypii and Myzus persicae to Transmit Cucumber mosaic virus in Single and Mixed Infection with Two Potyviruses to Zucchini Squash

The main objective of this work was to investigate the ability of Aphis gossypii and Myzus persicae to transmit Cucumber mosaic virus (CMV) singly and mixed with two potyviruses (Papaya ringspot virus - type W, PRSV-W and Zucchini yellow mosaic virus, ZYMV), to zucchini squash plants (Cucurbita pepo). The results showed that the potyviruses in general were more efficiently transmitted by both species of aphids as compared to CMV. The transmission of PRSV-W, ZYMV and CMV separately was more efficient than in mixture.

Dalmatian Toadflax (Linaria dalmatica): New Host for Cucumber Mosaic Virus

To date, there have been no reports of Dalmatian toadflax serving as a host for cucumber mosaic virus (CMV). Infestations of Dalmatian toadflax may serve as a reservoir of CMV, thereby facilitating aphid transmission of CMV to both agricultural crops and native plants. The goal of this study was to determine whether Dalmatian toadflax is a host for CMV. Dalmatian toadflax seedlings were randomly assigned to two treatments (18 replicates/treatment): no inoculation (control) and inoculation with CMV (Fast New York strain). The Dalmatian toadflax seedlings were inoculated by standard mechanical methods and tested for the presence of CMV using enzyme-linked immunosorbent assay (ELISA). Ten of the 18 CMV-inoculated toadflax plants tested positive for the virus; 6 of the 18 displayed systemic mosaic chlorosis and leaf curling. All control plants tested negative. Transmission electron microscopy obtained from CMV-positive plants confirmed the presence of CMV based on physical properties. To verify CMV infestation, tobacco plants were assigned to the following treatments (six replicates/treatment): no inoculation (control), CMV-negative (control) inoculation, and a CMV-positive inoculation. Plants were inoculated by standard methods. Five of the 6 tobacco plants treated with the CMV-positive inoculum tested positive for CMV using ELISA. All control plants tested negative for the virus.

Analysis of genetic bottlenecks during horizontal transmission of Cucumber mosaic virus.

Genetic bottlenecks may occur in virus populations when only a few individuals are transferred horizontally from one host to another, or when a viral population moves systemically from the infection site. Genetic bottlenecks during the systemic movement of an RNA plant virus population were reported previously (H. Li and M. J. Roossinck, J. Virol. 78:10582-10587, 2004). In this study we mechanically inoculated an artificial population consisting of 12 restriction enzyme marker mutants of Cucumber mosaic virus (CMV) onto young leaves of squash plants and used two aphid species, Aphis gossypii and Myzus persicae, to transmit the virus populations from infected source plants to healthy squash plants. Horizontal transmission by aphids constituted a significant bottleneck, as the population in the aphid-inoculated plants contained far fewer mutants than the original inoculum source. Additional experiments demonstrated that genetic variation in the artificial population of CMV is not reduced during the acquisition of the virus but is significantly reduced during the inoculation period.

Effects of various oils on survival of Myzus persicae Sulzer and its transmission of cucumber mosaic virus on pepper

SummaryMineral oils are widely used to control non-persistently transmitted viruses by aphid vectors, but mineral oils cause a number of problems including significant phytotoxicity. This study comprises three series of assays to compare a mineral oil, a fish oil and four vegetable oils for: a) insecticidal effects on the aphid Myzus persicae; b) inoculation of cucumber mosaic virus (CMV) by M. persicae to oil-sprayed pepper plants; and c) acquisition of CMV by M. persicae from oil-sprayed pepper plants. The vegetable oils tested were raw and refined rapeseed oil, and raw and refined soya oil. The oil showing the strongest insecticidal activity was raw soya oil (about 50% mortality at 72 h). Refined soya oil, both rapeseed oils and fish oil also showed appreciable insecticidal activity (30–40% mortality at 72 h). The oils most effectively suppressing inoculation were the mineral oil and the refined rapeseed oil, both of which suppressed inoculation entirely. All the oils except the raw soya oil and raw rapeseed oil suppressed acquisition entirely. These results point to the possible utility of vegetable oils, particularly refined rapeseed oil, for control of aphid-transmitted viruses in horticultural crops.

Cucumoviruses

Research on the molecular biology of cucumoviruses and their plant-virus interactions has been very extensive in the last decade. Cucumovirus genome structures have been analyzed, giving new insights into their genetic variability, evolution, and taxonomy. A new viral gene has been discovered, and its role in promoting virus infection has been delineated. The localization and various functions of each viral-encoded gene product have been established. The particle structures of Cucumber mosaic virus (CMV) and Tomato aspermy virus have been determined. Pathogenicity domains have been mapped, and barriers to virus infection have been localized. The movement pathways of the viruses in some hosts have been discerned, and viral mutants affecting the movement processes have been identified. Host responses to viral infection have been characterized, both temporally and spatially. Progress has been made in determining the mechanisms of replication, gene expression, and transmission of CMV. The pathogenicity determinants of various satellite RNAs have been characterized, and the importance of secondary structure in satellite RNA-mediated interactions has been recognized. Novel plant genes specifying resistance to infection by CMV have been identified. In some cases, these genes have been mapped, and one resistance gene to CMV has been isolated and characterized. Pathogen-derived resistance has been demonstrated against CMV using various segments of the CMV genome, and the mechanisms of some of these forms of resistances have been analyzed. Finally, the nature of synergistic interactions between CMV and other viruses has been characterized. This review highlights these various achievements in the context of the previous work on the biology of cucumoviruses and their interactions with plants.

Investigation on seed transmission of cucumber mosaic virus in cowpea

Cowpea breeding lines were infected with cucumber mosaic virus (CMV) by mechanical inoculation to investigate seed transmission rates for this virus. Transmission rates ranging from 0% to 6% were scored by symptom assessment. However, when cowpeas grown from seeds of infected mother plants were tested by ELISA, up to 30% of the plants were infected with CMV. Evaluating ELISA for CMV detection in cowpea revealed that virus concentration was highest in flowers and lowest in primary leaves. To compare the capacity of ELISA for CMV detection, the serological test was compared with RT-PCR; ELISA proved sensitive and reliable for detection of CMV in infected tissue. For seed analysis, a non-destructive assay for CMV detection based on tissue blotting of hypocotyls tissue was developed. RESUME Des varietes selectionnees du niebe etaient infectees par inoculation mecanique avec le virus de la mosaique de concombre (CMV) afin d'etudier la transmission par semence de ce virus. Les taux de transmission de l'ordre de 0 % a 6 % etaient enregistres apres l'evaluation des symptomes. Cependant, lorsque les plantes de niebe issues de semences provenant des parents infectes ont ete testees par ELISA, jusqu'a 30 % des plantes etaient infectees par le CMV. L'evaluation de L'ELISA pour la detection du CMV chez le niebe a revele que la concentration du virus etait la plus elevee dans les fleurs et le plus faible dans les feuilles primaires. Afin d'evaluer la capacite d'ELISA de detecter CMV, le test serologique etait compare a la RT-PCR, ou l'ELISA s'est montre sensible et fiable pour la detection du CMV dans des tissus infectes. Pour l'analyse de semence, une methode non-destructive de detection de CMV basee sur le blotting de tissus d'hypocotyle etait developpe. (Af Crop Science and Production: 2001 9(4): 677-684)