Rose C. Gergerich

Cowpea mosaic virus-based chimaeras: Effects of inserted peptides on the phenotype, host range, and transmissibility of the modified viruses

Expression of foreign peptides on the surface of cowpea mosaic virus particles leads to the creation of chimaeras with a variety of phenotypes and yields. Two factors were shown to be particularly significant in determining the properties of a given chimaera: the length of the inserted sequence and its isoelectric point. The deleterious effect of high isoelectric point on the ability of chimeras to produce a systemic infection occurs irrespective of the site of insertion of the peptide. Ultrastructural analysis of tissue infected with chimaeras with different phenotypes showed that all produced particles with a tendency to aggregate, irrespective of the size or isoelectric point of the insert. Host range and transmission studies revealed that the expression of a foreign peptide did not (1) alter the virus host range, (2) increase the rate of transmission by beetles or through seed, or (3) change the insect vector specificity. These findings have implications for both the utility and the biosafety of Cowpea mosaic virus-based chimaeras.

Localization of Transmissible and Nontransmissible Viruses in the Vector Nematode Xiphinema americanum.

ABSTRACT The inner lining of the food canal of nematodes that transmit plantinfecting viruses is regarded as the retention region of viruses. To characterize the location of transmissible and nontransmissible viruses in the vector nematode Xiphinema americanum, three nepoviruses, Tobacco ringspot virus (TRSV), Tomato ringspot virus(TomRSV), and Cherry leaf roll virus(CLRV), and one non-nematode-transmissible virus, Squash mosaic virus (SqMV), were evaluated for transmission efficiency and localization sites in the nematode. Transmission trials showed highest transmission efficiency for TomRSV (38% with 1 and 100% with 10 nematodes, respectively), intermediate efficiency for TRSV (27% with 1 and 65% with 10 nematodes, respectively), and no transmission for CLRV and SqMV. Electron microscopy and immunofluorescent labeling revealed that TRSV was primarily localized to the lining of the lumen of the stylet extension and the anterior esophagus, but only rarely in the triradiate lumen. Within a nematode population, particles of TRSV were no longer observed in these three regions 10 weeks after acquisition, and it is assumed that there was gradual and random loss of the virus from these areas. The percentage of nematodes that were labeled by virus-specific immunofluorescent labeling in a population of viruliferous nematodes decreased gradually after TRSV acquisition when the nematodes were placed on a nonhost of the virus, and the loss of immunofluorescent labeling paralleled the decrease in the ability of the nematode population to transmit the virus. TomRSV was localized only in the triradiate lumen based on thin-section electron microscopy. No virus-like particles were observed in any part of the food canal of nematodes that had fed on CLRV-infected plants. Virus-like particles that appeared to be partially degraded were observed only in the triradiate lumen of nematodes that had fed on SqMV-infected plants. These results clarified the status of localization of two nontransmissible viruses in X. americanum and presented evidence that two nematode-transmissible viruses, TRSV and TomRSV, are localized in different regions of the food canal of X. americanum.

Safeguarding Fruit Crops in the Age of Agricultural Globalization

The expansion of fruit production and markets into new geographic areas provides novel opportunities and challenges for the agricultural and marketing industries. Evidence that fruit consumption helps prevent nutrient deficiencies and reduces the risk of cardiovascular disease and cancer has assisted in the expansion of all aspects of the fruit industry. In todays competitive global market environment, producers need access to the best plant material available in terms of genetics and health if they are to maintain a competitive advantage in the market. An ever-increasing amount of plant material in the form of produce, nursery plants, and breeding stock moves vast distances, and this has resulted in an increased risk of pest and disease introductions into new areas. One of the primary concerns of the global fruit industry is a group of systemic pathogens for which there are no effective remedies once plants are infected. These pathogens and diseases require expensive management and control procedures at nurseries and by producers locally and nationally. Here, we review (i) the characteristics of some of these pathogens, (ii) the history and economic consequences of some notable disease epidemics caused by these pathogens, (iii) the changes in agricultural trade that have exacerbated the risk of pathogen introduction, (iv) the path to production of healthy plants through the U.S. National Clean Plant Network and state certification programs, (v) the economic value of clean stock to nurseries and fruit growers in the United States, and (vi) current efforts to develop and harmonize effective nursery certification programs within the United States as well as with global trading partners.

Viral Interactions Lead to Decline of Blackberry Plants

Blackberry yellow vein disease (BYVD) poses a new threat to the blackberry industry in the United States. Blackberry yellow vein associated virus (BYVaV) was originally thought to be the sole cause of this disease. However, BYVaV has been found in several asymptomatic blackberry cultivars. An unusual member of the family Potyviridae was identified recently from symptomatic plants and named Blackberry virus Y (BVY). BVY has been shown to spread in the field and cause BYVD when co-infected with BYVaV. Both viruses are asymptomatic in single infections but are readily detectable in asymptomatic plants by reverse transcription-polymerase chain reaction (RT-PCR). However, in mixed infections, the titer of BYVaV is repressed, sometimes to levels undetectable by RT-PCR, while the concentration of BVY is increased several fold. Electron microscopy revealed a variety of viral inclusions in symptomatic leaf samples, but none could be found in single infections with either BVY or BYVaV. Although BYVaV has been consistently associated with BYVD in different geographical regions, the detection of BVY has thus far been restricted to northwest Arkansas. It has been hypothesized that BYVaV is the synergistic determinant of BYVD that causes symptoms in different cultivars at various locations during co-infection with other viruses.

Incidence and Ecology of Blackberry yellow vein associated virus

Symptoms of leaf vein yellowing and bush decline in blackberry were attributed to infection by a novel crinivirus named Blackberry yellow vein associated virus (BYVaV). The disease is an emerging threat to blackberry production because it can cause substantial yield loss. The objective of this study was to identify the source and means of spread of BYVaV. A survey of blackberry plants for BYVaV from wild, cultivated, and nursery stocks was conducted. Insect traps and healthy blackberry sentinel plants were placed among symptomatic plants in a production field throughout two growing seasons to monitor the occurrence of potential vectors and virus spread. Virus indicator plants were grafted with BYVaV-infected blackberry because this virus was latent in some blackberry cultivars, but indicator plants failed to express symptoms when infected with BYVaV. Reverse-transcription polymerase chain reaction detection revealed the occurrence of BYVaV in blackberry nurseries in the United States, in production fields in Arkansas, South Carolina, and North Carolina, and in wild blackberry populations in Arkansas. Whiteflies (Trialeurodes packardii and T. ruborum), potential vectors of BYVaV, were observed on sticky traps placed in blackberry fields and were found colonizing blackberry plants; however, transmission studies failed to produce whitefly-mediated transmission of BYVaV. Further understanding of the disease etiology is needed to devise viable management strategies for this disease.

Determinants in the Specificity of Virus Transmission by Leaf-feeding Beetles

Vector specificity is a well-recognized characteristic of plant viruses. The plant viruses that are transmitted by leaf-feeding beetles are in four virus groups and cannot be transmitted by other commonly recognized virus vectors such as aphids, leafhoppers, or nematodes. The basis for vector specificity has been elucidated for some virus-vector combinations, and in many cases the interaction between the vector and virus is determined by virus coat proteins or by nonstructural proteins, as with the aphid-transmitted potyviruses and caulimoviruses (16, 17). These viral proteins have been shown to mediate a specific, definitive interaction of the virus particle with some surface of the vector. Sites of virus binding to vector surfaces that play a role in vector specificity have been identified for nematode, fungal, aphid, and leafhopper vectors (2, 16, 17). In some cases, virus coat proteins are involved in interactions with and transport across vector membranes as in the case with luteovirus passage through the aphid gut wall and accessory salivary gland cells (15).

A novel emaravirus is associated with redbud yellow ringspot disease.

Yellow ringspot is the only virus-like disease reported in redbud (Cercis spp.) with symptoms including vein clearing, chlorotic ringspots and oak-leaf pattern. A putative new emaravirus was present in all trees displaying typical yellow ringspot symptoms and the name redbud yellow ringspot associated virus is proposed. The virus genome is composed of at least five RNA segments. Two coding regions were studied to determine isolate diversity with results pointing to a homogeneous virus population. Host range was evaluated using graft transmission and by testing species found in close proximity to infected trees. Mite transmission with Aculops cercidis, the predominant species found in redbud trees in the epicenter of the disease, was evaluated but was not found to be a vector of the virus. Based on this study and the accumulated knowledge on emaravirus evolution we propose that speciation is allopatric, with vectors being a major component of the process.

Temperature Affects Expression of Symptoms Induced by Soybean Mosaic Virus in Homozygous and Heterozygous Plants

Seven strains (G1 to G7) of soybean mosaic virus (SMV) and 3 resistance loci (Rsv1, Rsv3, and Rsv4) have been identified in soybean. The interaction of SMV strains and host resistance genes results in resistant (symptomless), susceptible (mosaic), or necrotic (leaf and stem necrosis) reactions. The necrotic reaction may be gene dosage dependent and influenced by temperature. Using a set of soybean isolines and hybrids containing homozygous or heterozygous alleles of rsv, Rsv1, Rsv1-n, Rsv3, or Rsv4, this study has explored the relationship of SMV-induced symptoms and resistance gene dosage at different temperatures. Results showed that SMV-inoculated plants carrying Rsv3 or Rsv4 were symptomless at both homozygous and heterozygous states at all temperature regimes. Threshold temperatures for symptoms changing from stem tip necrosis (STN) to mosaic were 30, 33, and 33 degrees C in G7-inoculated homozygous genotypes V94-3971(Rsv1) and PI 96983 (Rsv1) and G1-inoculated V262 (Rsv1-n), respectively. However, at the heterozygous state, threshold temperature was 30 degrees C in G7-inoculated V94-3971 x Essex F(1) for the symptom change from STN to mosaic, 31 degrees C in G7-inoculated Essex x PI 96983 F(1) from STN to mixture of necrosis and mosaic (N-M), and 32 degrees C in G1-inoculated V262 x Essex F(1) from N-M to mosaic. Incomplete necrosis was observed in the heterozygous state in G1-inoculated V262 x Essex F(1) and G7-inoculated PI 96983 x Essex F(1) where necrotic and mosaic symptoms were mixed. High temperature (37 degrees C) tends to mask the expression of mosaic symptoms in both homozygous and heterozygous plants. STN expression in response to temperature was affected by resistance gene, gene dosage, host genetic background, and specific SMV strains. Thus, Rsv3 and Rsv4 are a better choice as source of genetic resistance for breeding SMV-resistant cultivars.

Reovirus-like particles and their vertical transmission in the Mexican bean beetle, Epilachna varivestis (Coleoptera: Coccinellidae)

Abstract Examination of thin sections of organs and tissues from the Mexican bean beetle, Epilachna varivestis , adults and larvae revealed the consistent presence of isometric, reovirus-like particles in the cytoplasm and rarely in the nuclei. Cytoplasmic inclusions having a dense and finely granular matrix with virus-like particles at their periphery were noticed occasionally. These inclusions were, however, very frequent in the nurse cells in the germarium of the ovariole. Virus-like particles were found in the cytoplasm of oocytes and eggs as well as embedded in the nuclei of sperm, thus suggesting vertical transmission of these particles, and explaining the 100% infection of the Mexican bean beetle colony maintained at the Virology and Biocontrol Laboratory of the University of Arkansas. Partially purified extracts from ovarioles or entire beetles contained particles which, in negatively stained preparations, resembled reoviruses without the external protein coat. It is suggested that these particles represent a latent virus of the Mexican bean beetle.

Taxonomy of the family Arenaviridae and the order Bunyavirales: update 2018

In 2018, the family Arenaviridae was expanded by inclusion of 1 new genus and 5 novel species. At the same time, the recently established order Bunyavirales was expanded by 3 species. This article presents the updated taxonomy of the family Arenaviridae and the order Bunyavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV) and summarizes additional taxonomic proposals that may affect the order in the near future.