Rapid detection of cowpea aphid-borne mosaic virus in cowpea seeds

Four cultivars of cowpea (Vigna unguiculata [L]. Walp.) were infected with cowpea aphid-borne mosaic virus (CABMV) by natural infection in field plots. Seeds taken from these plants were tested for the presence of the virus by ELISA and symptom observation on the plantlets grown from the seeds. A biotin/ streptavidin ELISA technique was used and found to be more sensitive than a standard ELISA protocol for detecting CABMV infection in seed. There was a good correlation between the ELISA detection of CABMV in tissue taken from single cowpea seeds and subsequent development of infected plants grown from the same seeds. The ELISA technique is reliable for selecting CABMV-free stocks of cowpea seeds.     

Aphis craccivora settling behaviour and acquisition of cowpea aphid-borne mosaic virus in aphid-resistant cowpea lines

The settling behaviour of Aphis craccivora Koch, the vector of cowpea aphid-borne mosaic virus (CAMV) in cowpea, Vigna unguiculata (L.) Walp., on aphid-resistant, aphid-tolerant and aphid-susceptible cowpea lines was investigated. It was found that although apterae counts on the aphid-susceptible and aphid-tolerant lines were higher than on the aphid-resistant ones, apterae were more widely dispersed among the latter. Whereas there was a positive significant correlation between alatae numbers and incidence of CAMV in the aphid-susceptible and aphid-tolerant lines, this correlation was negative in the aphid-resistant ones. However, the incidence of CAMV was not significantly different from each other in all the lines. This indicated that aphid activity (e.g. wide dispersal) was more important in the spread of CAMV than the absolute number of viruliferous alatae, an effect which was manifested on the aphid-resistant lines.Aphids acquire CAMV more readily from aphid-susceptible and aphid-tolerant source plants than from aphid-resistant ones. The significance of this in relation to secondary spread of CAMV and the effect of noncolonising aphids are discussed.

---

Le comportement d'atterissage d'Aphis craccivora et l'acquisition du virus de la mosaïque transmis par puceron chez les variétés de niébe résistantes au puceron

Résumé Le comportement d'atterissage d'Aphis craccivora, vecteur du virus de la mosaique transmis par puceron (CAMV) sur Vigna unguiculata, a été examiné sur des variétés résistantes, sensibles ou tolérantes aux pucerons. On a observé que bien que les dénombrements d'aptères sur variétés sensibles ou tolérantes aient été plus élevés que sur variétés résistantes, les aptères étaient plus largement dispersés sur ces dernières. Tandis qu'il y avait une corrélation positive significative entre les nombres d'ailés et la présence de CAMV sur les variétés sensibles et tolérantes aux pucerons, cette corrélation était négative sur les variétés résistantes. Cependant, la fréquence de CAMV n'était pas significativement différente parmi toutes ces variétés. Ceci a montré l'activité des pucerons (c'est à dire leur dispersion) étail plus importante dans la diffusion du CAMV que le nombre absolu d'ailés virosés, effet qui était manifeste sur les variétés résistantes aux pucerons.Les pucerons contaminés au CAMV le sont plus par des plantes sensibles ou tolérantes aux pucerons que par des plantes résistantes. La discussion porte sur la signification de ceci en relation avec la diffusion secondaire de CAMV, et de l'effet des pucerons non colonisateurs.

     

The effect of aphid-resistance in cowpea on infestation and development of Aphis craccivora and the transmission of cowpea aphid-borne mosaic virus1

The virus-vector-host relationships of cowpea aphid-borne mosaic virus (CAMV) and its vector, Aphis craccivora, were studied in cowpea lines differing in resistance to aphid infestation. CAMV was acquired and inoculated by its vector during brief probes, confirming that it is non-persistently transmitted. On aphid-resistant cowpea lines, the abundance and the relative sizes of aphids was less than in aphid-tolerant and aphid-susceptible lines. However, aphids were observed to make more numerous probes and probes of shorter duration on aphid-resistant lines than on aphid-susceptible lines. Resistance to aphid infestation in cowpea did not provide resistance to infection with CAMV.     

Effects of cypermethrin and deltamethrin on the feeding behaviour of Aphis craccivora and transmission of cowpea aphid-borne mosaic virus

The effects of two synthetic pyrethroids, cypermethrin and deltamethrin, on the feeding behaviour of Aphis craccivora and its transmission of the nonpersistent cowpea aphid-borne mosaic virus (CAMV) were compared to those of an organophosphate (dimethoate) and carbamate (pirimicarb) in greenhouse and laboratory tests. Cypermethrin at 100, 150 and 200 mg a.i. litre^-1 and deltamethrin at 75, 100 and 150 mg a.i. litre^-1 restricted the acquisition and inoculation of CAMV, whereas dimethoate and pirimicarb had no significant effect. Cypermethrin protected against transmission of CAMV longer after application than deltamethrin and there was a strong interaction between The duration of the first probe was shorter on pyrethroid-treated plants than on dimethoate-or pirimicarb-treated plants, and on pyrethroid-treated plants aphids probably did not probe long enough to acquire or inoculate CAMV. It is concluded that the use of synthetic pyrethroids is potentially valuable in controlling CAMV in the field until resistance against the virus has been bred into cultivars.     

The replication of cowpea mosaic virus

Cowpea mosaic virus (CPMV) is the type member of the comovirus group, which contains 14 different plant viruses that have the same structural organization of genomic RNAs and virions and use the same mechanism for expression of the viral RNAs. The combined structure and organization of the two CPMV genomic RNAs is strikingly similar to that of the single genome of animal picornaviruses. This suggests a common ancestry and similar replication mechanisms. CPMV is by far the best-studied comovirus and we shall limit this review to some recent data on this virus. For additional general information the reader is referred to other recent reviews on CPMV and comoviruses^1,2.     

Relationships among strains of bean common mosaic virus and blackeye cowpea mosaic virus - members of the potyvirus group

In host-range studies, bean common mosaic virus strains (BCMV-NL1, -NL3 and -NY 15) usually induced distinct systemic symptoms in susceptible bean cultivars and latent infection in several Vigna genotypes (except NY15 which gave mosaic symptoms in the latter), while blackeye cowpea mosaic virus (B1CMV-W) caused distinct systemic symptoms in several Vigna genotypes and only weak systemic symptoms in a few bean genotypes only. Biologically, B1CMV-W was closest to BCMV-NY15 and less close to -NL1. When using antisera to the three BCMV strains and five strains of B1CMV (including a strain originally considered cowpea aphid-borne mosaic virus CAMV-Mor) in SDS-immunodiffusion and ELISA, BCMV-NL1 and -NY15 were found to be closely related to each other and to BICMV-Fla, -NR and -W, and less closely to BICMV-Ind and -Mor. Serological relationships of BCMV-NL1 and -NY15 to BCMV- NL3 were more distant, which is in line with the biological distinction of NL3 in causing temperature-independent necrosis in bean cultivars with the necrosis gene I. PAGE analysis of coat proteins revealed that the three strains of BCMV and B1 CMV-W have similar but non-identical molecular masses. Although molecular hybridisation may further elucidate quantitative relationships between potyvir-uses, variation within and among the potyviruses may continue to pose problems in their classification and identification.     

Expression of bottom component RNA of cowpea mosaic virus in cowpea protoplasts

Upon inoculation of cowpea protoplasts with the bottom component of cowpea mosaic virus, at least six virus-induced proteins (with sizes of 170, 110, 87, 84, 60, and 32 kilodaltons) are synthesized, but not the capsid proteins (37 and 23 kilodaltons). These bottom-component-induced proteins were studied with respect to their genetic origin and mode of synthesis. The analyses were based on their electrophoretic peptide patterns resulting from partial digestion with Staphylococcus aureus protease V8. Comparison of the peptide patterns of the virus-induced proteins with those of the cowpea mosaic virus RNA-coded polypeptides produced in rabbit reticulocyte lysate showed that the 170- and 32-kilodalton polypeptides, which are the first viral products in cowpea mosaic virus-infected cells, were actually coded by the bottom component RNA of the virus. The 110-, 87-, and 84-kilodalton polypeptides, and possibly the 60-kilodalton polypeptide, appeared to have amino acid sequences in common with the 170-kilodalton polypeptide, demonstrating that they were virus coded as well. The results indicated that cowpea mosaic virus bottom component RNA was translated in vivo into a single 200-kilodalton polyprotein from which probably all bottom-component-specific proteins arose by three successive cleavages.     

Insect vectors of cowpea mosaic virus in Nigeria

A yellow strain of cowpea mosaic virus (CPMV) was transmitted in cowpea by two thrips, Sericothrips occipitalis and Taeniothrips sjostedti; two chrysomelid beetles, Ootheca mutabilis and Paraluperodes quaternus; a curculionid beetle, Nematocerus acerbus; and two acridid grasshoppers, Catantops spissus spissus and Zonocerus variegatus. Summarizing trials with single insects, the efficiency of transmission of CPMV averaged 18—21% for N. acerbus and the two grasshoppers, 55% for P. quaternus, and 71% for O. mutabilis. Twenty-two and 40% of the plants exposed to large populations of S. occipitalis and T. sjostedti, respectively, were infected. In three trials with an aphid, Aphis craccivora, 4 of 49 plants were infected with CPMV, but these infections were considered spurious because no infections occurred in any of 63 plants exposed to this insect in four other trials. A coreid bug, Riptortus dentipes, did not transmit CPMV. Mosaic symptoms in infected plants appeared 5—39 days after they were exposed to vectors. Infective virus was recovered from fresh faecal pellets of each grasshopper vector.     

Distinction of strains of bean common mosaic virus and blackeye cowpea mosaic virus using antibodies to N- and C- or N-terminal peptide domains of coat proteins

Earlier attempts to discriminate serologically strains NL1, NL3 and NY15 of bean common mosaic virus (BCMV) and strain W of blackeye cowpea mosaic virus (B1CMV) had been unsuccessful. Antibodies directed towards N- and C-, or N-terminal peptide regions of the coat proteins of the above strains enabled the distinction between B1CMV-W, BCMV-NY15 and BCMV-NL3 in electroblot immunoassay and in ELISA. The distinction was better with antibodies directed towards N-termini than with those to N- and C-termini. Strain NL1 of BCMV cross-reacted with both B1CMV-W and BCMV-NY15, but not with BCMV-NL3. Taxonomic implications of these findings are discussed.     

The structure of cucumber mosaic virus and comparison to cowpea chlorotic mottle virus

The structure of cucumber mosaic virus (CMV; strain Fny) has been determined to a 3.2-A resolution using X-ray crystallography. Despite the fact that CMV has only 19% capsid protein sequence identity (34% similarity) to cowpea chlorotic mottle virus (CCMV), the core structures of these two members of the Bromoviridae family are highly homologous. As suggested by a previous low-resolution structural study, the 305-A diameter (maximum) of CMV is approximately 12 A larger than that of CCMV. In CCMV, the structures of the A, B, and C subunits are nearly identical except in their N termini. In contrast, the structures of two loops in subunit A of CMV differ from those in B and C. These loops are 6 and 7 residues longer than the analogous regions in CCMV. Unlike that of CCMV, the capsid of CMV does not undergo swelling at pH 7.0 and is stable at pH 9.0. This may be partly due to the fact that the N termini of the B and C subunits form a unique bundle of six amphipathic helices oriented down into the virion core at the threefold axes. In addition, while CCMV has a cluster of aspartic acid residues at the quasi-threefold axis that are proposed to bind metal in a pH-dependent manner, this cluster is replaced by complementing acids and bases in CMV. Finally, this structure clearly demonstrates that the residues important for aphid transmission lie at the outermost portion of the betaH-betaI loop and yields details of the portions of the virus that are hypothesized to mediate binding to aphid mouthparts.     

Geminate structures of African cassava mosaic virus

Two types of geminate structures were purified from African cassava mosaic geminivirus (ACMV)-infected Nicotiana benthamiana plants and analyzed by electron cryomicroscopy and image reconstruction. After cesium sulfate density gradient centrifugation, they were separated into lighter top (T) and heavier bottom (B) components. T particles comigrated with host proteins, whereas B particles were concentrated in a cesium density typical for complete virions. Both particles were composed of two incomplete icosahedra of 11 capsomers each, but T particles were slightly larger (diameter, 22.5 nm) and less dense in the interior than B particles (diameter, 21.5 nm). T particles were frequently associated with small globules of approximately 14 nm diameter of unknown origin. The overall structure of ACMV, a begomovirus transmitted by whiteflies, was similar to that of Maize streak virus (MSV), a mastrevirus transmitted by leafhoppers, although the vertices of the icosahedra were less pronounced. Models of ACMV coat proteins based on Satellite tobacco necrosis virus support the exposure of parts of the molecule essential for transmission specificity by whiteflies and provide possible structural explanations for the smaller protrusion of the ACMV capsid relative to MSV. The differences of ACMV and MSV virion shapes are discussed with reference to their different animal vectors.     

Comparison of barley stripe mosaic virus strains

BSMV (barley stripe mosaic virus) particles were obtained in a pure state from infected host plant tissues of Hordeum vulgare. The three genomic parities (alpha, beta and gamma) were amplified by PCR using specific primers for each particle; each was cloned. Partial sequence of the alpha, beta and gamma segments was determined for the Egyptian isolate of barley stripe mosaic virus (BSMV AE1). Alignment of nucleotide sequences with that of other known strains of the virus, BSMV type strains (CV17, ND18 and China), and the generation of phylogenetic trees was performed. A low level of homology was detected comparing 467 bp of the a and 643 bp of the segments to that of the other strains, and thus BSMV alpha and beta segments were in separate clusters. However, 1154 bp of the gamma segments of BSMV AE1 showed a high level of homology especially to strain BSMV ND18, as they both formed a distinct cluster. Northern blotting of pure BSMV AE1 virus and H. vulgare-infected tissue were compared using an alpha ND18 specific probe. Western blotting using antibodies specific for the coat protein (CP) and the triple gene block 1 (TGB1) protein, which are both encoded by the beta ND18 segment, still indicated a high level of similarity between proteins produced by BSMV ND18 and AE1. We suggest that the BSMV AE1 isolate is a distinct strain of BSMV which reflects the genetic evolutionary divergence among BSMV strains and members of the Hordeivirus group.     

Homologous sequences in non-structural proteins from cowpea mosaic virus and picornaviruses

Computer analyses have revealed sequence homology between two non-structural proteins encoded by cowpea mosaic virus (CPMV), and corresponding proteins encoded by two picornaviruses, poliovirus and foot-and-mouth disease virus. A region of 535 amino acids in the 87-K polypeptide from CPMV was found to be homologous to the RNA-dependent RNA polymerases from both picornaviruses, the best matches being found where the picornaviral proteins most resemble each other. Additionally, the 58-K polypeptide from CPMV and polypeptide P2-X from poliovirus contain a conserved region of 143 amino acids. Based on the homology observed, a genetic map of the CPMV genome has been constructed in which the 87-K polypeptide represents the core polymerase domain of the CPMV replicase. These results have implications for the evolution of RNA viruses, and mechanisms are discussed which may explain the existence of homology between picornaviruses (animal viruses with single genomic RNAs) and comoviruses (plant viruses with two genomic RNAs).     

Distinct functional domains in the cowpea mosaic virus movement protein.

Cell-to-cell movement of cowpea mosaic virus particles in plants takes place with the help of tubules that penetrate presumably modified plasmodesmata. These tubules, which are built up by the virus-encoded 48-kDa movement protein (MP), are also formed on single protoplast cells. To determine whether the MP contains different functional domains, the effect of mutations in its coding region was studied. Mutations between amino acids 1 and 313 led to complete abolishment of the tubule-forming capacity, while a deletion in the C-terminal region resulted in tubules that could not take up virus particles. From these observations, it is concluded that the MP contains at least two distinct domains, one that is involved in tubule formation and that spans amino acids 1 and 313 and a second that is probably involved in the incorporation of virus particles in the tubule and that is located in the C terminus between amino acids 314 and 331.     

Position-dependent processing of peptides presented on the surface of cowpea mosaic virus

The plant virus cowpea mosaic virus (CPMV) has been developed as an epitope-presentation system. Numerous epitopes have been expressed in the betaB-betaC loop of the CPMV small coat protein, all of which undergo a cleavage reaction between their two carboxy-terminal residues. Although many peptides presented in this manner give an authentic immune response, this was not the case for the NIm-1A epitope from human rhinovirus-14. Crystallography revealed significant differences between the structure of NIm-1A on CPMV compared with its native configuration. The 3D structure of C PMV expressing NIm-1A was used to design alterations to the context of the NIm-1A graft.     

Protein composition of unusual tobacco mosaic virus strains

Amino acid analyses have been made of the proteins of single-lesion isolates of five strains of tobacco mosaic virus (TMV) differentiated by Lycopersicon hosts. These hosts differed in their genetical control of resistance to TMV, and the virus strains had therefore survived specific selection pressures. Two of the five strains differed in their amino acid composition from type TMV and from all other tomato strains of TMV previously examined. Symptoms induced by the five strains in four tomato lines and in Nicotiana tabacum cvs White Burley and Kawala are described.