The complete genome sequences of two isolates of spiraea yellow leafspot virus (SYLSV) were determined. Spiraea (Spiraea x bumalda) 'Anthony Waterer' plants showing virus-like symptoms including yellow spotting and leaf deformation were used for sequencing. The viral genome of SYLSV-MN (Minnesota) and SYLSV-MD (Maryland) is 8,017bp in length. The sequences share 95% identity at the nucleotide level. Both isolates have the same genome organization containing three open reading frames (ORFs), with ORF3 being the largest, encoding a putative polyprotein of 232 kDa with conserved domains including a zinc finger, pepsin-like aspartate protease, reverse transcriptase (RT), and RNase H. Pairwise comparisons between members of the genus Badnavirus showed that gooseberry vein banding associated virus GB1 (HQ852248) and rubus yellow net virus isolate Baumforth's Seedling A (KM078034) were the closest related virus sequences to SYLSV, sharing 73% identity at the nucleotide level. Bacilliform virions with dimensions of 150 nm × 30 nm were observed in virus preparations from symptomatic, but not asymptomatic, plants.

Cutaneous T cell lymphomas (CTCL) represent rare extranodal non-Hodgkin’s lymphomas, which are characterised by pleomorphic skin lesions and distinct T-cell markers. CTCL is a relatively benign disease in its early stages, but survival rates decrease significantly with progression. Histone deacetylase inhibitors (HDACi) have recently emerged as a new class of targeted anticancer therapies for CTCL, which have been shown to induce growth inhibition, terminal differentiation and apoptosis in various cancers in vitro and in vivo. In addition to the intrinsic anticancer properties of HDACi, recent studies have demonstrated its ability to synergise with phototherapy. In particular, we examine the therapeutic potential of HDACi in combination with ultraviolet A (UV-A) phototherapy, employing a halogenated DNA minor groove binding ligand called UVASens as a photosensitiser. In vitro studies have demonstrated that UVASens is approximately 1000-fold more potent than current psoralens. The extreme photopotency of UVASens allows the use of lower radiation doses minimising the carcinogenic risks associated with the long-term use of phototherapy. Considering, previous findings using the photosensitiser UVASens and potential synergy of HDACi with phototherapy, it was hypothesised that HDACi will augment photochemotherapy-induced cytotoxicity in CTCL MyLa cells. The findings indicated that combinations of UVASens/UV-A photochemotherapy and HDACi significantly decreased cell viability and increased apoptosis and DNA double-strand breaks in MyLa cells.

Citrus tristeza virus (CTV) is a plant pathogenic virus first reported infecting citrus plants in the 1930’s in Brazil and Argentina and is now widely spread through all citrus growing areas. A CTV isolate from an infected Pakistani sweet orange tree was selected for sequencing to subsequently produce recombinant CTV coat protein. The purified recombinant CTV coat protein was used as an antigen for the production of polyclonal antisera in rabbits. The antisera were tested in direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA) against various CTV isolates from Pakistan and T-30 from the USA. The antisera reacted strongly in DAC-ELISA with both homologous and heterologous isolates of CTV from infected citrus leaf tissue. These antisera were compared with a commercially available ELISA kit and were more sensitive to Pakistani isolates of CTV than the commercial ELISA kit. An inexpensive virus-free testing and certification program using the antisera produced here would provide farmers with the knowledge to eliminate CTV infected and unproductive trees, which could then be replaced with healthy young trees. The implementation of such a scheme would reduce the incidence of CTV in citrus orchards, reduce yield losses caused by CTV, and could lead to the elimination of CTV from Pakistan

A previously undescribed badnavirus was isolated from Zamia fischeri showing symptoms of chlorosis, necrosis, and ringspot. The virus has bacilliform virions 30 nm in diameter and averaging 120 nm in length. The viral genome is 9227 bp in length and contains three open reading frames characteristic of members of the genus Badnavirus. The largest open reading frame (ORF3) encodes a putative polyprotein, with predicted domains including zinc finger, aspartic protease, reverse transcriptase (RT) and RNase H. The virus is tentatively named "cycad leaf necrosis virus" (CLNV). Within the genus Badnavirus, CLNV was most closely related to sugarcane bacilliform Guadeloupe D virus (FJ439817), sharing 69% identity at the nucleotide level in the RT + RNase H region. This virus is the first badnavirus reported to infect cycads, and it has the largest genome among the currently characterized badnaviruses.

Aglaonema bacilliform virus (ABV), a member of the genus Badnavirus in the family Caulimoviridae, is associated with leaf deformation and chlorosis in Aglaonema modestum. The complete genome sequence of a Minnesota isolate of ABV was determined. The ABV genome is 7,178 bp in length and similar in size and organization to those of the members of the genus Badnavirus, containing three open reading frames (ORFs) with the potential to encode three proteins of 14.92, 13.33 and 207.95 kDa, respectively. The last ORF (ORF3) encodes a putative polyprotein with conserved domains, including zinc finger, aspartic protease, reverse transcriptase (RT) and RNase H domains, in that order. Phylogenetic analysis using the amino acid sequence of the ORF3 polyprotein showed that ABV clusters with several isolates of taro bacilliform CH virus (TaBCHV). Pairwise alignment using the highly conserved RT/RNase H region reveals that ABV has the highest level of identity (71%) at the nucleotide level to a Hawaiian isolate of TaBCHV.

HomePlant DiseaseVol. 103, No. 7Detection and First Report of Beet Ringspot Virus in Ornamental Oxalis in the United States PreviousNext DISEASE NOTESDetection and First Report of Beet Ringspot Virus in Ornamental Oxalis in the United StatesR. Jordan, D. Mollov, M. A. Guaragna, and B. LockhartR. Jordan†Corresponding author: R. Jordan; E-mail Address: [email protected]http://orcid.org/0000-0003-1105-4066Floral and Nursery Plants Research Unit, U.S. National Arboretum, ARS, USDA, Beltsville, MDSearch for more papers by this author, D. MollovNational Germplasm Resources Laboratory, ARS, USDA, Beltsville, MDSearch for more papers by this author, M. A. GuaragnaFloral and Nursery Plants Research Unit, U.S. National Arboretum, ARS, USDA, Beltsville, MDSearch for more papers by this author, and B. LockhartDepartment of Plant Pathology, University of Minnesota, St. Paul, MNSearch for more papers by this authorAffiliationsAuthors and Affiliations R. Jordan1 † D. Mollov2 M. A. Guaragna1 B. Lockhart3 1Floral and Nursery Plants Research Unit, U.S. National Arboretum, ARS, USDA, Beltsville, MD 2National Germplasm Resources Laboratory, ARS, USDA, Beltsville, MD 3Department of Plant Pathology, University of Minnesota, St. Paul, MN Published Online:14 May 2019https://doi.org/10.1094/PDIS-09-18-1680-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat The plant genus Oxalis (Oxalidaceae) is morphologically diverse, cosmopolitan, and comprises at least 500 species distributed in North and South America and South Africa. Ornamental Oxalis triangularis (formerly O. regnellii), commonly known as false shamrock because of its triangular leaves, is grown as a potted plant in the United States, especially for marketing in the spring around St. Patrick’s Day. A potyvirus associated with chlorotic ringspot in ornamental O. regnellii was first described in Washington in 1981 (Coyier 1981). Plants showing similar symptoms and having potyvirus particles were reported in New York in 2009 and in Florida in 2012 (Baker 2013). The name Shamrock chlorotic ringspot virus (SCRV) was coined. Oxalis plants from Wisconsin (2012 to 2013) showing similar symptoms were submitted for analysis to the University of Minnesota Plant Disease Clinic. Potyvirus-like filamentous virus particles (720 to 780 nm) were observed in symptomatic leaf samples by transmission electron microscopy (TEM). Total RNA extracted from a symptomatic plant tested positive for the presence of a potyvirus in reverse transcription polymerase chain reaction (RT-PCR) using universal primers. The 3′-terminal 1,690-bp region was cloned and sequenced following established protocols (Jordan et al. 2011), found to be unique, and deposited in GenBank as SCRV-WI (GenBank no. KJ619376). The full genome sequence and phylogenetic analysis of this novel potyvirus will be reported elsewhere. Later, TEM analysis of partially purified virion preparations obtained as previously described (Mollov et al. 2013) from symptomatic tissue revealed filamentous and 25- to 30-nm spherical virus particles. Total RNA extracted from these preparations was used as a template for a random PCR using random primers to produce a cDNA library (Mollov et al. 2013). Cloned sequences revealed identity to SCRV-WI or, after BLASTN analysis, to beet ringspot virus (BRSV-S; GenBank nos. D00322 and X04602). Using primers specific to the oxalis-infecting BRSV (BRSV-Ox) sequences to generate multiple overlapping RT-PCR cDNA clones, coupled with 5′ and 3′ rapid amplification of cDNA ends cloning, the complete nepovirus genome sequence of BRSV-Ox was determined to be 7,354 and 4,632 nt (excluding the 3′ poly-A ends) for RNA 1 and RNA 2, respectively (GenBank nos. MH939189 and MH939190, respectively). Pairwise identities of the BRSV-Ox RNAs ranged from 93 to 96% with BRSV-S, 65 to 83% with tomato black ring virus (TBRV), 50 to 62% with grapevine chrome mosaic virus (GCMV), 30 to 39% with Cycas necrotic stunt virus (CNSV), and 19 to 25% identity with other nepoviruses. Pairwise identities of the BRSV-Ox RNA 1 and RNA 2 polyproteins also ranged from 94 to 96% with BRSV-S, 68 to 84% with TBRV, 57 to 63% with GCMV, 44 to 58% with CNSV, and 22 to 31% identity with other nepoviruses. In preliminary studies to fulfill Koch’s postulates, BRSV-Ox was singly transferred from coinfected oxalis to Nicotiana benthamiana and N. occidentalis by mechanical inoculation. Both exhibited chlorotic ringspots or mild mottling. BRSV-Ox was then successfully transferred from N. benthamiana back to virus-free healthy oxalis, albeit with asymptomatic infection. BRSV-Ox was detected in all these plants by RT-PCR using BRSV-Ox specific primers (BRSV-Ox-F, 5′-TAGATCACATGTCTGGCTTAGA-3′; and BRSV-Ox-R, 5′-GGCTTGTGTTGCCACCATACAGGGC-3′), which amplify a 644-bp amplicon in RNA 1. Noninoculated asymptomatic plants were virus-free. No single infection has yet been established for SCRV-WI, because attempts to mechanically transfer SCRV-WI from BRSV-Ox and SCRV-WI coinfected oxalis plants have not been successful. To our knowledge, this is the first report of BRSV in ornamental Oxalis. BRSV is a soil-borne virus shown to infect a wide range of plant species including potato, sugar beet, strawberry, turnip, wheat, oat, many weeds, and peach (Brunt et al. 1997). It has recently been reported in Begonia in Hungary (Kis et al. 2017) and in several Euonymus alatus woody ornamental shrub plants exhibiting virus-like symptoms in Minnesota (Bratsch and Lockhart 2018).The author(s) declare no conflict of interest.References:Baker, C. A. 2013. Plant Pathology Circular No. 412. FDACS-P-01868. Division of Plant Industry, Florida Department of Agriculture and Consumer Services, Gainesville, FL. Google ScholarBratsch, S. A., and Lockhart, B. E. 2018. Acta Hortic. 1191:69. https://doi.org/10.17660/ActaHortic.2018.1191.10 Crossref, Google ScholarBrunt, A. A. et al. 1997. Plant viruses online: Descriptions and lists from the VIDE database. http://sdb.im.ac.cn/vide/ Google ScholarCoyier, D. L. 1981. Plant Dis. 65:275. https://doi.org/10.1094/PD-65-275 Crossref, ISI, Google ScholarJordan, R., et al. 2011. Acta Hortic. 901:159. https://doi.org/10.17660/ActaHortic.2011.901.20 Crossref, Google ScholarKis, S., et al. 2017. Arch. Virol. 162:3559. https://doi.org/10.1007/s00705-017-3521-z Crossref, ISI, Google ScholarMollov, D., et al. 2013. Arch. Virol. 158:1917. https://doi.org/10.1007/s00705-013-1686-7 Crossref, ISI, Google ScholarThe author(s) declare no conflict of interest.Funding: Funding was provided by USDA, Agricultural Research Service (Research Project 8020-22000-042-00D).DetailsFiguresLiterature CitedRelated Vol. 103, No. 7 July 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionApple cultivar Joya Cripps Red lesions caused by Colletotrichum fructicola (Nodet et al.). Photo credit: P. Nodet. Symptoms of Lotus powdery mildew caused by Erysiphe takamatsui (Zhou et al.). Photo credit: C. Liang. Symptoms of tar spot (Phyllachora maydis) on maize leaves (Dalla Lana et al.). Photo credit: F. Dalla Lana. Metrics Article History Issue Date: 20 Jun 2019Published: 14 May 2019First Look: 10 Mar 2019Accepted: 2 Mar 2019 Page: 1800 InformationThis article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source. The American Phytopathological Society, 2019.FundingUSDA, Agricultural Research ServiceGrant/Award Number: Research Project 8020-22000-042-00DKeywordsvirusesornamentalsherbaceous/flowering plantspathogen detectionThe author(s) declare no conflict of interest.Cited byHistorical and recent tomato black ring virus and beet ringspot virus isolate genomes reveal interspecies recombination and plant health regulation inconsistencies18 November 2021 | Plant Pathology, Vol. 71, No. 3

The primary objective of this multicentric dose allocation and dose expansion study was to determine the MTD and the DLTs of the lucitanib (a tyrosine kinase inhibitor of the FGFR/VEGFR/PDFGR pathways)/fulvestrant combination. Postmenopausal women with ER+/HER2− mBC, who have relapsed during or after treatment with fulvestrant, were eligible. The study had a dose allocation part to assess the tolerability of the combination followed by a dose expansion part. Eighteen patients with ER+, mBC were enrolled; median age was 66 years, 50% had a PS: 0 and all had received previous endocrine treatment. The study was prematurely terminated after 18 patients (15 in part 1 and 3 in part 2) based on preclinical experiments that failed to confirm the hypothesis that addition of lucitanib would reverse sensitivity to endocrine treatments. Based on data of global lucitanib development, it was decided to stop the dose allocation at 12.5 mg and to start the dose expansion part at 10 mg/day. The most common grade ≥ 3 toxicities (> 10% of patients) were hypertension (78%) and asthenia (22%). All patients required at ≥ 1 interruption, 13 patients (72%) required ≥ 1 dose reduction. Three patients (72%) withdrew from the study for AEs (at 10 mg). Three patients achieved a confirmed PR (10 mg n = 1; 12.5 mg n = 2). Although the combination is feasible it requires close monitoring of the patients for the management of adverse events. Further investigation is required to better understand the potential role of FGFR inhibition in reversing resistance to endocrine treatment.

A previously undescribed badnavirus was identified in plants of Polyscias fruticosa (Ming aralia) showing symptoms of mild mosaic and leaf senescence. Characteristic bacilliform virions of the Polyscias badnavirus averaging 30 × 120 nm in size were observed by transmission electron microscopy in partially purified leaf tissue extracts from symptomatic but not asymptomatic plants collected in the USA and Nigeria. The isolate from the USA was complete sequenced. The genome is 7592 bp in length and contains three open reading frames with an arrangement similar to that of other members of the genus Badnavirus. The largest open reading frame (ORF3) encodes a putative polyprotein, with predicted domains including zinc finger, aspartic protease, reverse transcriptase (RT) and RNase H, in that order. The USA and Nigeria isolates of the virus had a high level (98%) of nucleotide sequence identity in the RT+RNase H region. Within the genus Badnavirus, these viruses were most closely related to schefflera ringspot virus (SRV), sharing 63% identity at the nucleotide level. Based on the ICTV species demarcation criteria for the genus Badnavirus (more than 20% nucleotide sequence divergence in the RT+RNase H region), the Polyscias virus is proposed to be a new member of the genus, and the name polyscias mosaic virus (PoMV) is proposed. The complete genome sequence was deposited in the NCBI GenBank database under accession no. MH475918.

The term “anthropocene” (Crutzen, 2002) refers to our current geological epoch and illustrates the manifold influences of human existence and actions on geology and evolution. Ornamentals are a true anthropogenic product solely manufactured to please the eye of the beholder. They are produced and traded all over the globe thus opening gateways for viruses. We investigated consequences of anthropogenic impact on virus spread and diversity in ornamentals. We focused on Tobamoviruses that are mechanically transmitted and are lacking a true natural vector. Our study spanned the period from 2000 to 2016 and identified Tobamovirus infections in host plants belonging to 15 different plant families. The ten identified virus species belonged mostly to Tobamovirus subgroup 1. In Solanaceae as well as in Cactaceae members of both Tobamovirus subgroup 1 and 3 were present. Different patterns of virus-host plant associations seem to exist when comparing members of both virus subgroups. Phylogenetic analysis of a partially sequenced Tobamovirus isolated from Hoya, Asclepiadaceae, in 2014 indicates the emergence of a new Tobamovirus species positioned separately from known viruses of the subgroup 3 cluster. Similar evidence has been reported independently from Florida (Schubert and Davison, 2012; Adkins et al., 2016) indicating manmediated global spread of this virus.

HomePlant DiseaseVol. 100, No. 7First Report of Sugarcane mosaic virus Infecting Columbus Grass (Sorghum almum) in the United States PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Sugarcane mosaic virus Infecting Columbus Grass (Sorghum almum) in the United StatesD. Mollov, M. N. Tahir, C. Wei, C. Kaye, B. Lockhart, J. C. Comstock, and P. RottD. Mollov, M. N. Tahir, C. Wei, C. Kaye, B. Lockhart, J. C. Comstock, and P. RottAffiliationsAuthors and Affiliations D. Mollov M. N. Tahir , USDA-ARS, National Germplasm Resources Laboratory, Beltsville, MD 20705 C. Wei , University of Florida, IFAS, Plant Pathology Department, Everglades Research & Education Center, Belle Glade 33430 C. Kaye , US Sugar Corporation, Clewiston, FL 33440 B. Lockhart , University of Minnesota, Department of Plant Pathology, St. Paul 55108 J. C. Comstock , USDA-ARS, Sugarcane Field Station, Canal Point, FL 33438 P. Rott , University of Florida, IFAS, Plant Pathology Department, Everglades Research & Education Center, Belle Glade 33430. Published Online:4 Apr 2016https://doi.org/10.1094/PDIS-01-16-0093-PDNAboutSections ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Mosaic symptoms in sorghum can be caused by several potyviruses (family Potyviridae), including Sorghum mosaic virus and Sugarcane mosaic virus (SCMV). Both are responsible for global economic losses in sorghum, maize, and sugarcane. During spring 2015, in a patch of Columbus grass (Sorghum almum) growing near a sugarcane field at Canal Point in Florida, patterns of contrasting shades of green were observed on leaves of several plants. Ten of these plants with mosaic-looking symptoms were collected for further investigation. Partial virus purification was performed with 20 g of leaves from one plant, and filamentous virus particles (∼750 nm long) were observed by transmission electron microscopy (TEM). This preparation was used for immunosorbent electron microscopy (ISEM) with Maize dwarf mosaic virus (MDMV) antibody A and MDMV antibody B, which react with SCMV (Pirone 1972). Maize dwarf mosaic virus B antibody trapped and decorated virus particles, but the serological reaction with the MDMV A antibody was not readily observed. Five of the 10 plants with mosaic symptoms all tested positive by ELISA using a broad-spectrum potyvirus antibody (Agdia, Elkhart, IN). These plants also reacted in ELISA with SCMV-specific antibodies (Agdia). Total RNA was extracted from leaves of all 10 S. almum plants using RNeasy Plant mini Kit (Qiagen, Hilden, Germany) and used as a template for RT-PCR. Poaceae potyvirus-specific primer pair oligo 1n (ATGGTHTGGTGYATHGARAAYGG) and oligo 2n (TGCTGCKGCYTTCATYTG) (Marie-Jeanne et al. 2000) produced the expected 327-bp amplicon in all 10 samples. Five amplicons were sequenced revealing 93 to 95% nucleotide identity with SCMV isolates in GenBank (BLASTn). Additional cDNA was generated from two plants using primer M4T (GTTTTCCCAGTCACGAC-(T)15), and PCR amplified using universal primers Poty S (GGNAAYAAYAGYGGNCARCC) and M4 (GTTTTCCCAGTCACGAC) (Chen et al. 2001) yielding an approximately 1.8-kb product. The products were cloned into pGEM-T Easy Vector System (Promega, Madison, Wisconsin), and complete sequences were determined for three cloned fragments. These 1.8-kb sequences were most similar to isolates of SCMV and had 92% identity to GenBank Accession No. U57356 (SCMV strain D from sugarcane) at the nucleotide level, and 81% similarity to Accession No. CAX36842 (SCMV from Saccharum officinarum) at the amino acid level. TEM, ISEM, ELISA, RT-PCR, and sequence analysis from multiple plants confirmed the presence of SCMV in S. almum exhibiting mosaic symptoms in Florida. SCMV has been previously reported to infect S. almum in Australia (Teakle and Grylls 1973), but to our knowledge, this is the first report in the United States. This federal and state noxious weed is widely distributed in sugarcane-growing areas in Florida where sugarcane also occasionally exhibits mosaic symptoms. It may be an alternative host for SCMV. The relationship between SCMV strains occurring in S. almum and strains infecting sugarcane requires investigation to determine the importance of S. almum in the epidemiology and management of SCMV in sugarcane.References:Chen, J., et al. 2001. Arch. Virol. 146:757. https://doi.org/10.1007/s007050170144 Crossref, ISI, Google ScholarMarie-Jeanne, V., et al. 2000. J. Phytopathol. 148:141. https://doi.org/10.1046/j.1439-0434.2000.00473.x Crossref, ISI, Google ScholarPirone, T. P. 1972. C.M.I./A.A.B. Description of Plant Viruses No. 88. CMI, Kew, Surrey, UK. Google ScholarTeakle, D. S., and Grylls, N. E. 1973. Aust. J. Agric. Res. 24:465. https://doi.org/10.1071/AR9730465 Crossref, Google ScholarDetailsFiguresLiterature CitedRelated Vol. 100, No. 7 July 2016SubscribeISSN:0191-2917e-ISSN:1943-7692 Metrics Article History Issue Date: 7 Jun 2016Published: 4 Apr 2016First Look: 22 Feb 2016Accepted: 12 Feb 2016 Pages: 1510-1510 Information© 2016 The American Phytopathological SocietyCited bySugarcane mosaic virus (sugarcane mosaic)CABI Compendium, Vol. CABI CompendiumBiotechnological Approaches to Improve Sugarcane Quality and Quantum Under Environmental Stresses1 January 2023Eight Species of Poaceae Are Hosting Different Genetic and Pathogenic Strains of Sugarcane Mosaic Virus in the Everglades Agricultural AreaMartha Hincapie, Sushma Sood, Dimitre Mollov, D. Calvin Odero, Michael Grisham, and Philippe Rott3 November 2021 | Phytopathology®, Vol. 111, No. 10Sugarcane Mosaic Disease: Characteristics, Identification and Control17 September 2021 | Microorganisms, Vol. 9, No. 9Screening for sugarcane yellow leaf virus in sorghum in Florida revealed its occurrence in mixed infections with sugarcane mosaic virus and a new marafivirusCrop Protection, Vol. 139Sorghum almum (Columbus grass)6 June 2020Detection and Response of Sugarcane against the Infection of Sugarcane Mosaic Virus (SCMV) in Indonesia24 July 2017 | Agronomy, Vol. 7, No. 3

Cranberry plants bearing disfigured, scarred fruit were reported by growers in the major cranberry-growing region of central Wisconsin in July 2012. Plants bearing scarred fruit have since been observed in Massachusetts and New Jersey. Three complementary methods provided evidence of Tobacco streak virus (TSV) in symptomatic plants: (i) leaves and scarred berries tested positive for TSV by double-antibody sandwich enzyme-linked immunosorbent assay; (ii) quasi-isometric particles approximately 33 nm in diameter were extracted from leaves of symptomatic plants and visualized using transmission electron microscopy; and (iii) coat protein gene sequence analysis revealed 94 to 99% nucleotide similarity with reference TSV sequences. In newer cultivars, 99% of uprights with scarred berries tested positive for TSV. In older cultivars, 31% of uprights with scarred berries tested positive for TSV and the remaining 69% of uprights with scarred berries tested positive for Blueberry shock virus. TSV overwintered in cranberry plants, and leaves, pollen, and fruit tested positive for TSV the year following symptom occurrence. Attempts to inoculate cranberry using infected pollen or sap as inoculum failed, but several herbaceous hosts tested TSV positive following mechanical inoculation. Phylogenetic analysis of the coat protein gene of 26 TSV isolates from various cultivars of cranberry in Wisconsin, New Jersey, and Massachusetts revealed diversity. This work provides information that will be useful in understanding the epidemiology of TSV in cranberry and in the development of management strategies.

Four previously undescribed viruses infecting cultivated roses were identified and fully characterized in Minnesota. These four viruses were transmitted by grafting from infected to healthy roses and found to be the likely causal agents of the diseases with which they are associated. Viruses were provisionally named after the characteristic symptoms in infected plants as follows: Rose yellow vein virus (RYVV), Rose yellow mosaic virus (RoYMV), Rosa rugosa leaf distortion virus (RrLDV), and Rose yellow leaf virus (RYLV). Unlike the currently known viruses that affect rose, the ilarviruses and the nepoviruses, that only show symptoms and are detected early in the growing season, these new viruses exhibit symptoms throughout the season and can be detected readily during the entire year. Based on virion and genome properties it was determined that RYVV is a member of the family Caulimoviridae, RoYMV is a member of the family Potyviridae, and RrLDV and RYLV are members of the family Tombusviridae. Reliable diagnostic protocols were developed for each virus: PCR for RYVV; RT-PCR for RoYMV, RrLDV, and RYLV; and immunosorbent electron microscopy (ISEM) and indirect enzyme-linked immunosorbent assay (ELISA) for RoYMV detection.

Pachysandra terminalis Siebold & Zucc. (Japanese pachysandra, spurge) is widely used as a groundcover. In early 2012, Japanese pachysandra plants from Missouri, which originated in Pennsylvania, showed symptoms of light and dark green mosaic, leaf deformation, concentric ringspots, and stunting. Initial screening of symptomatic leaf tissue by transmission electron microscopy (TEM) using partially purified extracts confirmed the presence of spherical (~28 nm) and bacilliform (18-nm diameter, 35- to 58-nm length) virus particles. Immunosorbent electron microscopy (ISEM) using antisera to a clover isolate of Alfalfa mosaic virus (AMV) (PVAS 92) and to Cucumber mosaic virus (CMV) (ATCC PVAS-30) obtained from the American Type Culture Collection, Manassas, VA, confirmed the presence of AMV and CMV. No other type of virus-like particles were observed by TEM. After 6 months, nearly 20% of the 4,000 pachysandra cuttings exhibited the described symptoms. However, it is possible that more than 20% of the cuttings were infected with both viruses and not yet exhibiting symptoms. Reverse-transcription PCR (RT-PCR) was done using total RNA extracted with a Qiagen RNeasy kit and Ready-To-Go RT-PCR beads (GE Healthcare, UK Limited, UK). The primer pair CMV-1 (5′-GCCGTAAGCTGGATGGACCA) and CMV-2 (5′-TATGATAAGAAGCTTGTTTTCGCG) were used (3) to obtain a 502-bp amplicon from the coat protein (CP) region of CMV RNA 3. The product was ligated and cloned (pGEM-T Easy Vector System; Promega, USA). Three clones were sequenced (UMGC, USA), and the consensus sequence (Sequencher 5.1, Gene Codes Corp., USA) was deposited in GenBank (Accession No. JX227938). The sequence obtained had 100% identity with a homologous CP CMV sequence (AFQ94058) and 99% identity with several other homologous CP CMV sequences (CAX62443, CCK24369, and 15 others). It also contained an EcoRI site at nucleotides 332 to 337, characteristic of CMV Type II isolates (3). The primer pair AMV1F (5′-ATCCACCGATGCCAGCCTTA) and AMV1R (5′-TTCCGCCTCACTGCTGCTG) generated a 1,047-bp product from AMV RNA1 that was deposited in GenBank (JX227937). This product had 100% identity with a homologous AMV sequence (AFQ94057), and 99% identity with several other homologous AMV sequences (AGV15824, ADO85715, CBX36144). From the data presented here, it was concluded that the pachysandra had a mixed infection of AMV and a Type II isolate of CMV. Occurrence of AMV in pachysandra was first reported in New Jersey in 1982 (2) and reported for the first time in France and Germany in 2000 (1). The presence of CMV infection in pachysandra has not been reported in the present literature. Some of the symptoms associated with AMV infection in pachysandra in New Jersey (2) and Europe (1) were similar to the symptoms produced by pachysandra plants infected with both viruses (ring spots, mosaic, and line patterns). However, some symptoms were unique to the mixed infection in pachysandra by AMV and CMV (leaf deformation, stunting). A potential source of this co-infection could occur when plants are grown near alfalfa fields (AMV infection by aphids) and undergo vegetative propagation (CMV infection by contaminated tools). This is the first report of pachysandra co-infected by AMV and CMV in the United States. References: (1) L. Cardin and B. Moury. Plant Dis. 84:594, 2000. (2) D. E. Hershman and E. H. Varney. Plant Dis. 66:1195, 1982. (3) S. Wylie et al. Aust. J. Agric. Res. 44:41, 1993.

Biodiversity within virosphere comprising virus-host, virus-vector, virusvirus and virus-viroid interactions is illustrated by the genus Carlavirus. They belong to the family of Betaflexiviridae with the type member Carnation latent virus (CLV). 43 members are listed by the latest ICTV taxonomy report (Adams et al., 2012). Samples obtained in Germany in 2006 to 2012 included monocot host plants comprising the families Alliaceae and Convallariaceae as well as dicot host plants of the families of Cactaceae, Ericaceae, Passifloraceae, Ranunculaceae, Scrophulariaceae and Solanaceae. Twelve different carlaviruses have been identified, for two of which no published record for Germany existed so far. In Alliaceae, Cactaceae, Ranunculaceae and Solanaceae mixed infections were frequent. Those consisted of two different carlaviruses or of Carlavirus accompanied by Poty- or Allexi-, or Potex- or Tobamo-, or Cucumovirus infection. Thus, symptom expression was variable and visual diagnosis was limited. Additional methods were employed to increase reproducibility and to generate reliable results. Synergy of electron microscopy and molecular biology tools were used to improve virus detection. Doing so we obtained evidence for the presence of three different isolates of Shallot latent virus in Allium sativum and for at least three Potato virus M isolates in Solanaceae. In addition, we identified the presence of Helleborus net necrosis virus and Blueberry scorch virus in Germany.

Mandevilla (Apocynaceae) is an ornamental tropical vine popular for its bright and attractive flowers. During 2012 to 2013, 12 Mandevilla sp. samples from Minnesota and Florida nurseries were submitted for analysis at the University of Minnesota Plant Disease Clinic. Plants showed mosaic symptoms, leaf deformation, premature leaf senescence, and vine dieback. Filamentous virus particles with modal lengths 700 to 900 nm were observed by transmission electron microscopy (TEM) in partially purified preparations from symptomatic leaves. Partially purified virions were obtained using 30% sucrose cushion centrifuged at 109,000 gmax for 2 h at 10°C (5). No other virus particles were observed in these samples, nor were any observed in non-symptomatic samples. One sample was submitted as potted plant (Mandevilla 'Sunmandeho' Sun Parasol Giant White) and was kept under greenhouse conditions for subsequent analyses. Total RNA (Qiagen) was extracted from this sample, and Potyvirus was detected using the universal primers Poty S (5'-GGN AAY AAY AGY GGN CAR CC-3') and PV1 (5'-20(T)V-3') (1) by reverse transcription (RT)-PCR (3). The amplified product was the expected ~1.7-kb, corresponding to the partial nuclear inclusion body gene, the coat protein (CP) gene, and the 3' end untranslated region. The RT-PCR amplicon was cloned (NEB) and sequenced, and the 1,720-bp consensus sequence was deposited in GenBank (Accession No. KM243928). NCBI BLAST analysis at the nucleotide level revealed highest identity (83%) with an isolate of Catharanthus mosaic virus (CatMV) from Brazil (Accession No. DQ365928). Pairwise analysis of the predicted 256 amino acid CP revealed 91% identity with the CatMV Brazilian isolate (ABI94824) and 68% or less identity with other potyviruses. Two potyviruses are usually considered the same species if their CP amino acid sequences are greater than 80% identical (2). Serological analysis of the infected sample Mandevilla 'Sunmandeho' Sun Parasol Giant White using a CatMV specific antiserum (4) resulted in positive indirect ELISA reactions. CatMV has been previously reported in periwinkle (Catharanthus roseus) in Brazil (4). Based on the analyses by TEM, RT-PCR, nucleotide and amino acid sequence identities, and serological reactivity, we identify this virus as a U.S. Mandevilla isolate of CatMV. To our knowledge, this is the first report of Catharanthus mosaic virus both in the United States and in Mandevilla. References: (1) J. Chen et al. Arch Virol. 146:757, 2001. (2) A. Gibbs and K. Ohshima. Ann. Rev. Phytopathol. 48:205, 2010. (3) R. L. Jordan et al. Acta Hortic. 901:159, 2011. (4) S. C. Maciell et al. Sci. Agric. Piracicaba, Brazil. 68:687, 2011. (5) D. Mollov et al. Arch Virol. 158:1917, 2013.