First Report of Johnsongrass mosaic virus (JGMV) Infecting Pennisetum purpureum in Brazil

Tropical grass and legume species used as pasture grasses for cattle feeding cover over 25% of the agricultural area in Brazil. In recent years, plants showing virus-like symptoms have been observed in the main pasture grass growing areas. Plants of Pennisetum purpureum line CNPGL 00211 showing typical virus mosaic symptoms on leaves and growth reduction were collected in Bahia State, Brazil. Flexuous elongated potyvirus-like particles were observed in the leaf-dip preparation of diseased plants by electron microscopy. In addition, the virus was mechanically transmitted using a standard procedure for potyviruses (4) and produced similar symptoms in inoculated P. purpureum plants. For further molecular identification, total RNA was extracted from frozen symptomatic leaves following the guanidine thiocyanate method (3). cDNA synthesis was performed using oligonucleotide, OligodT50M10 and PCR was carried out using Potyvirus degenerate primers PY11 (5′-GGNAAYAAYAGYGGNCARCC-3′) (2) and M10 (5′-AAGCAGTGTTATCAACGCAGA-3′). The amplified fragments of the expected size (approximately 2 kb comprising part of the NIb protein gene, the entire coat protein [CP] gene, and the 3′ nontranslated region) were separated using agarose gel electrophoresis, excised, and cloned into plasmid vector pGEMT-Easy (Promega) according to the manufacturer's instructions. Four selected clones were sequenced (Macrogen, South Korea). The sequenced 2.0-kb fragment (GenBank Accession No. KC333416) was compared with sequences available in GenBank and the highest nucleotide identity of 79% was observed with Johnsongrass mosaic virus (JGMV) isolated in Australia (4). According to the Potyvirus species demarcation convention based on CP identity (1), the virus isolate from P. purpureum belongs to the JGMV species. However, the amino acid sequence of the N-terminus of the CP of the Bahia isolate is distinct from JGMV sequences reported in GenBank. The phylogenetic analysis of the CP confirmed the difference since this Bahia isolate was located in a clearly distinct branch separate from all JGMV isolates. To our knowledge, this is the first report of a JGMV in Brazil infecting tropical grass in the main pasture areas. References: (1) M. J. Adams et al. Arch. Virol. 150: 459, 2005. (2) J. Chen et al. Arch. Virol. 146:757. 2001. (3) P. Chomczynski and N. Sacchi. Nature Protocols 1:581, 2006. (4) H. K. Laidlaw et al. Arch. Virol. 149:1633, 2004.

Download Full-text

First Report of Yam mild mosaic virus in Yam in Guangxi Province, China

Plant Disease ◽

10.1094/pdis-04-11-0350 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1320-1320 ◽

Cited By ~ 4

Author(s):

C. Zou ◽

J. Meng ◽

Z. Li ◽

M. Wei ◽

J. Song ◽

...

Keyword(s):

Mosaic Virus ◽

Viral Genome ◽

Terminal Region ◽

Mild Mosaic ◽

Rt Pcr ◽

Degenerate Primers ◽

Germplasm Exchange ◽

First Report ◽

Pcr Products ◽

Mild Mosaic Virus

Yams (Dioscorea spp.) are widely grown in China as vegetables and herbal medicine. However, studies on viral diseases on yams are still limited. As a pilot project of a government initiative for improving yam productivity, a small study was conducted in Guangxi, a southern province of China, on viral disease in yams. Incidence of virus-like disease for the three extensively grown D. alata cultivars, GH2, GH5, and GH6, were 12 to 40%, 12 to 29%, and 11 to 25%, respectively, as found in a field survey with a five-plot sampling method in 2010. A total of 112 leaf samples showing mosaic or mottling or leaves without symptoms were collected from the cvs. GH2, GH5, GH6, and seven additional cultivars (D. alata cvs. GY2, GY23, GY47, GY69, GY62, GY72, and D. batatas cv. Tiegun). To determine if the symptoms were caused by Yam mild mosaic virus (YMMV; genus Potyvirus, family Potyviridae), total RNA was extracted from leaves with a commercial RNA purification kit (TIANGEN, Beijing, China), and reverse-transcription (RT)-PCR was conducted with a YMMV-specific primer pair (4) that amplifies the 3′-terminal portion of the viral genome. A PCR product with the predicted size of 262 bp was obtained from samples of GH5 (number testing positive of total number of leaves = 5 of 12), GH6 (24 of 42), and GY72 (1 of 1), but not from asymptomatic leaves. PCR products from a GH5 sample (YMMV-Nanning) and a GH6 sample (YMMV-Luzhai) were cloned and sequenced using an ABI PRISM 3770 DNA Sequencer. The two PCR products were 97% identical at nucleotide (nt) level and with the highest homology (89% identity) to a YMMV isolate (GenBank Accession No. AJ305466). To further characterize the isolates, degenerate primers (2) were used to amplify viral genome sequence corresponding to the C-terminal region of the nuclear inclusion protein b (NIb) and the N-terminal region of the coat protein (CP). These 781-nt fragments were sequenced and a new primer, YMMV For1 (5′-TTCATGTCGCACAAAGCAGTTAAG-3′) corresponding to the NIb region, was designed and used together with primer YMMV UTR 1R to amplify a fragment that covers the complete CP region of YMMV by RT-PCR. These 1,278-nt fragments were sequenced (GenBank Accession Nos. JF357962 and JF357963). CP nucleotide sequences of the YMMV-Nanning and YMMV-Luzhai isolates were 94% similar, while amino acid sequences were 99% similar. BLAST searches revealed a nucleotide identity of 82 to 89% and a similarity of 88 to 97% for amino acids to sequences of YMMV isolates (AF548499 and AF548519 and AAQ12304 and BAA82070, respectively) in GenBank. YMMV is known to be prevalent on D. alata in Africa and the South Pacific, and has recently been identified in the Caribbean (1) and Colombia (3). To our knowledge, this is the first report of the natural occurrence of YMMV in China and it may have implications for yam production and germplasm exchange within China. References: (1) M. Bousalem and S. Dallot. Plant Dis. 84:200, 2000. (2) D. Colinet et al. Phytopathology 84:65, 1994. (3) S. Dallot et al. Plant Dis. 85:803, 2001. (4) R. A. Mumford and S. E. Seal. J. Virol. Methods 69:73, 1997.

Download Full-text

First Report of Bean yellow mosaic virus in Alaska from Clover (Trifolium spp.)

Plant Disease ◽

10.1094/pdis-94-3-0372a ◽

2010 ◽

Vol 94 (3) ◽

pp. 372-372 ◽

Cited By ~ 1

Author(s):

N. L. Robertson ◽

K. L. Brown

Keyword(s):

Coat Protein ◽

Host Range ◽

Mosaic Virus ◽

Bean Yellow Mosaic Virus ◽

Yellow Mosaic Virus ◽

Degenerate Primers ◽

First Report ◽

Chlorotic Spot ◽

Putative Coat Protein ◽

Das Elisa

In mid-June 2008, distinct mosaic leaves were observed on a cluster of clover (Trifolium spp.) with light pink and white flowers growing at the edge of a lawn in Palmer, AK. Virus minipurification from leaves of affected clover and protein extractions on a polyacrylamide electrophoresis implicated a ~35-kDa putative coat protein (CP). Subsequent western blots and ELISA with a universal potyvirus antiserum (Agdia Inc., Elkhart, IN) confirmed potyvirus identity. Total RNA extracts (RNeasy Plant Mini Kit, Qiagen Inc., Valencia, CA) from the same plant were used for reverse transcription (RT)-PCR. Three sets of degenerate primers that targeted potyvirus-specific genes, HC-Pro (helper component protease) and CI (cylindrical inclusion protein) and the genomic 3′-terminus that included a partial NIb (nuclear inclusion), CP (coat protein), and UTR (untranslated region), produced the expected PCR segments (~0.7, ~0.7, and ~1.6 kbp, respectively) on 1% agarose gels (1). Direct sequencing of the HC-Pro (GenBank No. GQ181115), CI (GQ181116), and CP (GU126690) segments revealed 98, 97, and 99% nucleotide identities (no gaps), respectively, to Bean yellow mosaic virus (BYMV)-chlorotic spot (CS) strain, GenBank No. AB373203. The next closest BYMV percent identity comparisons decreased to 79% for HC-Pro (GenBank No. DQ641248; BYMV-W), 79% for CI (U47033; BYMV-S) partial genes, and 96% for CP (AB041971; BYMV-P242). Mechanical inoculations of purified virus preparations produced local lesions on Chenopodium amaranticolor Coste & A. Reyn. (2 of 5) and C. quinoa Willd. (6 of 7), and mosaic on Nicotiana benthamiana Domin (5 of 5). BYMV was specifically confirmed on tester plants using a double-antibody sandwich (DAS)-ELISA BYMV (strain 204 and B25) kit (AC Diagnostics, Inc., Fayetteville, AR) as directed. The absence of another potyvirus commonly found in clover, Clover yellow vein virus (ClYVV), was verified in parallel DAS-ELISA ClYVV assays (AC Diagnostics, Inc). The BYMV isolate was maintained in N. benthamiana, and virion or sap extracts inoculated to the following host range (number of infected/total inoculated plants [verified by BYMV ELISA]): Cucumis sativus L. ‘Straight Eight’ (0/5), Gomphrena globosa L. (1/4), Nicotiana clevelandii A. Gray (4/7), Phaseolus vulgaris L. ‘Bountiful’ (1/3), Pisum sativum L. (Germplasm Resources Information Network Accession Nos. -PI 508092 (8/12), -W6 17525 (13/13), -W6 17529 (0/13), -W6 17530 (13/14), -W6 17537 (0/12), -W6 17538 (0/12), and -W6 17539 (0/21), Tetragonia tetragoniodes (2/2), Trifolium pretense L. ‘Altaswede’ (6/10), T. repens L. ‘Pilgrim’ (0/8), and Vicia faba L. (1/3). All infected plants had symptoms ranging from systemic mosaic (T. pretense, P. sativum) to leaf distortions (N. clevelandii, Tetragonia tetragoniodes). Interestingly, the host range and genomic sequences of the BYMV Alaskan strain resemble the BYMV-CS (chlorotic spot) strain that was originally isolated from a diseased red clover (T. pretense) plant in Japan more than 40 years ago (2). Although BYMV occurs worldwide and has a wide host range in dictoyledonous and monocotyledonous plants (3), to our knowledge, this is the first report of a natural occurrence of BYMV in Alaska. The incidence and distribution of BYMV in clover and other plant species are not known in Alaska. References: (1) C. Ha et al. Arch. Virol. 153:36, 2008. (2) H. Kume et al. Mem. Fac. Agric. Hokkaido Univ. 7:449, 1970. (3) S. J. Wylie et al. Plant Dis. 92:1596, 2008.

Download Full-text

First Report of Alternanthera mosaic virus Infection in Angelonia in the United States

Plant Disease ◽

10.1094/pdis-92-10-1473b ◽

2008 ◽

Vol 92 (10) ◽

pp. 1473-1473 ◽

Cited By ~ 7

Author(s):

B. E. Lockhart ◽

M. L. Daughtrey

Keyword(s):

United States ◽

Nucleotide Sequence ◽

Mosaic Virus ◽

Genomic Sequence ◽

Nutrient Deficiency ◽

Leaf Tissue ◽

The United States ◽

Nucleotide Sequence Analysis ◽

Degenerate Primers ◽

First Report

Stunting, chlorosis, and light yellow mottling resembling symptoms of nutrient deficiency were observed in angelonia (Angelonia angustifolia) in commercial production in New York. Numerous, filamentous particles 520 to 540 nm long and spherical virus particles 30 nm in diameter were observed by transmission electron microscopy (TEM) in negatively stained partially purified extracts of symptomatic Angelonia leaf tissue. Two viruses, the filamentous potexvirus Alternanthera mosaic virus (AltMV) and the spherical carmovirus Angelonia flower break virus (AnFBV) were subsequently identified on the basis of nucleotide sequence analysis of amplicons generated by reverse transcription (RT)-PCR using total RNA isolated from infected leaf tissue. A 584-bp portion of the replicase-encoding region of the AltMV genome was obtained with the degenerate primers Potex 2RC (5′-AGC ATR GNN SCR TCY TG-3′) and Potex 5 (5′-CAY CAR CAR GCM AAR GAT GA-3′) (3). Forward (AnFBV CP 1F-5′-AGC CTG GCA ATC TGC GTA CTG ATA-3′) and reverse (AnFBV CP 1R-5′-AAT ACC GCC CTC CTG TTT GGA AGT-3′) primers based on the published AnFBV genomic sequence (GenBank Accession No. NC_007733) were used to amplify a portion of the viral coat protein (CP) gene. The nucleotide sequence of the amplicon generated using the potexvirus-specific primers (GenBank Accession No. EU679362) was 99% identical to the published AltMV (GenBank Accession No. NC_007731) sequence and the nucleotide sequence of the amplicon obtained using the AnFBV CP primers was 99% identical to the published AnFBV genomic sequence (GenBank Accession No. EU679363). AnFBV occurs widely in angelonia (1) and AltMV has been identified in phlox (2). These data confirm the presence of AltMV and AnFBV in diseased angelonia plants showing stunting and nutrient deficiency-like symptoms and substantiates, to our knowledge, this first report of AltMV in angelonia in the United States. References: (1) S. Adkins et al. Phytopathology 96:460, 2006. (2) J. Hammond et al. Arch. Virol. 151:477, 2006. (3) R. A. A. van der Vlugt and M. Berendeson. Eur. J. Plant Pathol. 108:367, 2002.

Download Full-text

First Report of Turnip ringspot virus in Field Mustard (Brassica chinensis) in Taiwan

Plant Disease ◽

10.1094/pdis-03-11-0224 ◽

2011 ◽

Vol 95 (8) ◽

pp. 1036-1036

Author(s):

Y.-K. Chen ◽

Y.-S. Chang ◽

H.-J. Bau

Keyword(s):

Amino Acids ◽

Mosaic Virus ◽

Amino Acid Sequences ◽

Cauliflower Mosaic Virus ◽

Ringspot Virus ◽

Coat Proteins ◽

Degenerate Primers ◽

Brassica Chinensis ◽

First Report ◽

Beet Western Yellows Virus

Crucifer crops (Brassica spp.) are important winter vegetables in Taiwan. Five viruses, including Turnip mosaic virus (TuMV), Cucumber mosaic virus (CMV), Radish mosaic virus (RaMV), Beet western yellows virus (BWYV), and Cauliflower mosaic virus (CaMV), have been detected in a range of domestic-grown crucifers during past decades (1). Field mustard plants (Brassica chinensis) showing mosaic in the leaves were collected in the ChiaYi area in December 2007. Spherical virus-like particles, approximately 30 nm in diameter, were readily observed in crude sap of symptomatic plants. Tests by ELISA failed to detect any of the aforementioned viruses. A spherical agent was isolated through mechanical inoculation onto Chenopodium quinoa, and a virus culture was established and inoculated mechanically back to the original host as well as other crucifers. Systemic mosaic appeared on inoculated B. campestris, B. chinensis, and B. juncea, whereas ringspots appeared on inoculated leaves of B. oleracea. Total RNA was extracted from symptomatic leaves and used for reverse transcription (RT)-PCR amplification using degenerate primers for comoviruses (2). Other successive fragments of RNAs 1 and 2 were amplified by specific or degenerate primers designed on the basis of sequences of published Turnip ringspot virus (TuRSV). The RNA 1 (GenBank Accession No. GU968732) and RNA 2 (No. GU968731) of the isolated virus consisted of 6,076 and 3,960 nucleotides, respectively. The number of nucleotides and the arrangement of open reading frames on both RNA 1 and RNA 2 were similar to those of comoviruses. Sequence analysis revealed that the nucleotide sequences of RNA 1 and RNA 2 shared 54.2 to 82.5% and 50.2 to 79.3% similarities, respectively, to those of comoviruses and were most similar to Turnip ringspot virus. The deduced peptides of large and small coat proteins (LCP and SCP) contain 375 amino acids (41.2 kDa) and 251 amino acids (28.5 kDa), respectively. The deduced amino acid sequences of RNA-dependent RNA polymerase (RdRp), LCP, and SCP share 92.0 to 94.5%, 93.1 to 93.3% and 87.3 to 89.6% similarity, respectively, to those of published TuRSV isolates, i.e., -B (GenBank Accession No. GQ222382), -M12 (No. FJ516746), and -Toledo (No. FJ712027) indicating that the newly isolated virus from field mustard in Taiwan is an isolate of TuRSV, hence TuRSV-TW. Comparison of LCP and SCP between current TuRSV-TW and Radish mosaic virus (RaMV; GenBank Accession No. AB295644) showed 74% similarity, which is below the species demarcation level of 75% (3), indicating its discrimination from RaMV. To our knowledge, this is the first report of the occurrence of TuRSV in Taiwan and in the subtropics. References: (1) T. H. Chen et al. Plant Pathol. Bull. 9:39, 2000. (2) V. Maliogka et al. J. Phytopathol. 152:404, 2004. (3) K. Petrzik and I. Koloniuk. Virus Genes 40:290, 2010.

Download Full-text

First report of wild tomato mosaic virus infecting chilli pepper in China

Plant Disease ◽

10.1094/pdis-01-21-0010-pdn ◽

2021 ◽

Author(s):

Yongliang Hu ◽

Liping Fa ◽

Xiaoxia Su ◽

Yuqin Chen ◽

Jiawei Huang ◽

...

Keyword(s):

Mosaic Virus ◽

Disease Incidence ◽

Tomato Mosaic Virus ◽

Nucleotide Identity ◽

Rt Pcr ◽

Degenerate Primers ◽

Chilli Pepper ◽

First Report ◽

Wild Tomato ◽

Pepper Plants

Chilli pepper is an important economic crop and virus diseases are constraints on its production. In 2018, disease surveys were conducted at a 15-ha chilli pepper plantation in Dehong, southwest of Yunnan Province, China. Throughout the chilli pepper growing season from March to September, pepper plants developed three different virus-like symptoms on leaves, including mosaic, yellow mottle and shrinkage (Fig. S1). Based on observation of virus-like symptomatic phenotypes, the field surveys indicated that the disease incidence ranged from 30% in March to a peak 100% in July, resulting in a significant loss of pepper fruit from 30 to 100% depending on plot of the field. Potyvirus-like filamentous particles, around 11*760 nm, were observed under electron microscopy in the sap of symptomatic leaves (Fig. S1). To further determine the viral species in these samples, total RNA was extracted from three symptomatic samples using a Trans ZolUp Plus RNA Kit (Trans Gene, Beijing, China). Complementary DNA (cDNA) was synthesized using oligo (dT) and M-MLV reverse transcriptase (Promega, Madison, Wisconsin, USA) according to the manufacturer’s instructions, and the polymerase chain reaction (PCR) was performed using degenerate primers specific to genus Potyvirus targeting HC-Pro region (HPFor: 5-TGYGAYAAYCARYTIGAYIIIAAYG-3; HPRev: 5-GAICCRWAIGARTCIAIIACRTG-3) (Ha et al. 2008) under the following conditions: an initial denaturation at 94°C for 4min, 30 cycles of denaturation at 94°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 30s, and a 10min final extension at 72°C. An expected 683-bp DNA fragment was amplified and cloned into the pMD 18-T Vector (Takara, Japan) for sequencing. Sequence analysis using BLAST revealed that the amplicons of phenotype I (Fig. S1a) shared highest nucleotide identity (85.6%) with wild tomato mosaic virus (WTMV) isolate from Vietnam (GenBank no. DQ851495) while the amplicons of phenotype III (Fig. S1c) showed the highest nucleotide identity (93%) with chilli veinal mottle virus (ChiVMV) isolate from Sichuan, China. (GenBank no. MK405594). Amplicons of phenotype II included both sequence of above WTMV and ChiVMV, indicating co-infection of phenotype II (Fig. S1b). Phenotype I sample was used for mechanical inoculation on chilli pepper as described previously (Yang et al.2013). After ten days, virus-like symptoms similar to phenotype I were observed on leaves, and WTMV infection, but not ChiVMV infection, was confirmed by RT-PCR tests on inoculated pepper plants (Fig. S1 e, f). To further ascertain the incidence of these two viruses in the field, primers WT-F: 5'-GTTGTTGAATGTGGTTTAGTT-3' and WT-R: 5'-AGATGTGCTTTGGAAGCGACC-3' were designed based on the WTMV sequence (GenBank no. DQ851495) to amplify a 476 bp product, and primers Ch-F/Ch-R (Ch-F: 5'-AAAGAAGAACAAGCGACAGAA-3', Ch-R: 5'-CATCACGCAAATATTCAAAGC-3') were designed based on ChiVMV sequence (GenBank no. MK405594.1) to amplify a 332 bp product. RT-PCR was conducted on 31 field-collected samples, and amplicons of expected sizes, 476bp and 332bp, corresponding to WTMV and ChiVMV, respectively, were obtained and sequenced to verify their identity. The results (Fig. S2) showed that 71% (22/31) of the samples tested positive for WTMV, 90% (28/31) tested positive for ChiVMV, and 65% (20/31) were co-infected with the two viruses. The WTMV was first reported infecting wild tomatoes in Vietnam in 2008 (Ha et al. 2008), and later reported in China in Nicotiana tabacum (Sun et al. 2015), Solanum nigrum (Zhang et al. 2019), and wild eggplant (Zhang et al. 2014). To our knowledge, this is the first report of WTMV infection on chilli pepper under natural conditions. Our study revealed that the chilli pepper disease in Dehong was caused by single or co-infection of WTMV and ChiVMV. It is necessary to find effective methods to control these two viruses.

Download Full-text

First Report of Turnip mosaic virus in Tomatillo (Physalis philadelphica) in California

Plant Disease ◽

10.1094/pdis-09-11-0751 ◽

2012 ◽

Vol 96 (2) ◽

pp. 296-296 ◽

Cited By ~ 2

Author(s):

H.-Y. Liu ◽

S. T. Koike ◽

D. Xu ◽

R. Li

Keyword(s):

Mosaic Virus ◽

Green Peach Aphid ◽

Chenopodium Quinoa ◽

Turnip Mosaic Virus ◽

Causal Agent ◽

Western Hemisphere ◽

Degenerate Primers ◽

First Report ◽

Severe Stunting ◽

Access Period

Tomatillo is an important vegetable in Mexican cuisine. It is of Mesoamerica origin and now is grown widely in the Western Hemisphere. In 2011, 2% of commercially grown tomatillo plants in San Benito County, California exhibited severe stunting with foliage showing mosaic symptoms and leaf distortion. The fruits on infected plants were mottled and unmarketable. Flexuous filamentous-shaped virus particles of 800 to 850 nm long and 11 to 12 nm wide were observed from sap of the symptomatic plants with a transmission electron microscope. Sap from the diseased tomatillo plants reacted positively in an immunostrip assay for potyvirus (Agdia Inc., Elkhart, IN), indicating a potyvirus was associated with the disease. The causal agent was mechanically transmitted from the diseased field plants to six virus-free greenhouse tomatillo plants and all inoculated plants induced identical symptoms. The causal agent was also transmitted to Chenopodium quinoa and C. murale (chlorotic local lesions) and Nicotiana clevelandii, N. tabacum, and Physalis wrightii (systemic symptoms). The disease was also transmitted to tomatillo plants by the green peach aphid (Myzus persicae) in a nonpersistent manner (1-min acquisition access period and 1-min transmission access period with no latent period). To further identify the causal agent, total nucleic acids were extracted by a cetyltrimethylammoniumbromide (CTAB) method (2) and tested by reverse transcription-PCR using potyvirus degenerate primers CIFor and CIRev (1). An amplicon of approximately 700 bp from the diseased tomatillo was cloned and sequenced. Analysis of the 631-bp partial CI sequence (GenBank Accession No. JN601884) showed that the virus had 93.6% nucleotide identity and 100% amino acid identity with cognate regions of Turnip mosaic virus (TuMV) (GenBank Accession No. D10927). Our results indicated that the disease was caused by TuMV. To our knowledge, this is the first report of TuMV in tomatillo. Since TuMV has a wide host range and is readily transmitted by green peach aphids, TuMV could be a new threat to tomatillo production in California. References: (1) C. Ha et al. Arch. Virol. 153:25, 2008. (2) R. Li et al. J. Virol. Methods 154:48, 2008.

Download Full-text

First Report of Cucumber mosaic virus in Anemone sp. in the United States

Plant Disease ◽

10.1094/pdis-93-4-0431b ◽

2009 ◽

Vol 93 (4) ◽

pp. 431-431 ◽

Cited By ~ 1

Author(s):

I. E. Tzanetakis

Keyword(s):

United States ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Protein A ◽

The United States ◽

University Of Arkansas ◽

Degenerate Primers ◽

First Report ◽

Complete Coat Protein ◽

The University

In the spring of 2008, more than a dozen, aphid-infested, anemone plants (Anemone sp.) grown at the campus of the University of Arkansas in Fayetteville showed stunting and mosaic, whereas only two were asymptomatic. Leaf homogenates from four symptomatic plants were inoculated onto Nicotiana benthamiana that became stunted and developed severe mosaic approximately 7 days postinoculation, whereas buffer-inoculated plants remained asymptomatic. Double-stranded RNA (dsRNA) extraction (4) from symptomatic anemone revealed the presence of four predominant bands of approximately 3.2, 2.9, 2.2, and 0.9 kbp, a pattern indicative of cucumovirus infection. Cucumber mosaic virus (CMV) is the only cucumovirus reported in anemone in Europe (2) and Israel (3), and for this reason, anemone and N. benthamiana plants were tested by Protein A ELISA with antisera against CMV developed by H. A. Scott. ELISA verified the presence of CMV in symptomatic anemone and inoculated N. benthamiana, while asymptomatic plants were free of the virus. Using cucumovirus degenerate primers, essentially as described by Choi et al. (1), a region of approximately 940 bases that includes the complete coat protein gene of the virus was amplified from symptomatic anemone and N. benthamiana but not asymptomatic plants of either species. This anemone isolate (GenBank Accession No. FJ375723) belongs to the IA subgroup of CMV because it shares 99% nucleotide and 100% amino acid sequence identities with the Fny isolates of the virus. To my knowledge, this is the first report of CMV infecting anemone in the United States and an important discovery for the ornamental industry since anemone is commonly grown together with several ornamental hosts of CMV in nursery and garden settings. References: (1) S. K. Choi et al. J. Virol. Methods 83:67, 1999. (2) M. Hollings. Ann. Appl. Biol. 45:44, 1957 (3) G. Loebenstein. Acta Hortic. 722:31, 2006 (4) I. E. Tzanetakis and R. R. Martin, J. Virol. Methods 149:167, 2008.

Download Full-text

First Report of Basella rugose mosaic virus Infecting Four O'Clock (Mirabilis jalapa) in China

Plant Disease ◽

10.1094/pdis-07-11-0586 ◽

2012 ◽

Vol 96 (2) ◽

pp. 294-294 ◽

Cited By ~ 6

Author(s):

J. G. Wang ◽

J. J. Peng ◽

H. R. Chen ◽

S. Y. Chen

Keyword(s):

Amino Acid ◽

Mosaic Virus ◽

Leaf Extracts ◽

Chenopodium Amaranticolor ◽

First Report ◽

Chlorotic Spot ◽

Cp Gene ◽

Potyvirus Species ◽

Mirabilis Jalapa ◽

Rugose Mosaic

Four o'clock (Mirabilis jalapa) and M. himalaica var. chinensis are members of the family Nyctaginaceae and are widely distributed weeds in Yunnan Province, China. In 2009, mosaic and malformation symptoms were observed on leaves of the four o'clock on the campus of Yunnan Agricultural University and in the Black Dragon Pool Park in Kunming City, China. More than 30% of the four o'clock plants showed symptoms of the disease. Sap from leaves of symptomatic four o'clock plants caused local chlorotic and necrotic lesions in inoculated Chenopodium amaranticolor after 7 to 10 days and systemic mosaic symptoms in C. quinoa and Nicotiana benthamiana after 10 to 12 days. No symptoms were observed following inoculation of sap from asymptomatic plants. A pure virus isolate (MJ) was obtained after three successive single-lesion transfers from C. amaranticolor. Following mechanical inoculation of the MJ isolate, seedlings of indicator plants, N. benthamiana, displayed mosaic symptoms. Moreover, back transmission to healthy four o'clock seedlings by leaf extracts from systemically infected N. benthamiana plants caused similar mosaic and malformation symptoms. Flexuous, filamentous particles (650 to 700 nm long and 13 nm wide) and cytoplasmic laminar aggregates and pinwheel inclusions typical of members of the genus Potyvirus were observed in infected four o'clock leaves by electron microscopy. No other virus particles were observed. Serological testing of 10 symptomatic and healthy plants using a monoclonal antibody specific for Potyvirus group members in an indirect ELISA (Agdia Inc., Elkhart, IN) also resulted in positive reactions in infected leaves, however, all healthy seedlings tested were negative. Total RNAs were extracted from infected four o'clock leaves with the RNeasy Plant Mini Kit (QIAGEN, Hilden, Germany) and the 3′-terminal portion of the viral genome (including part of the NIb polymerase, the entire coat protein (CP), and 3′-UTR) was then amplified by reverse transcription-PCR with a universal Potyviridae primer Sprimer/M4 and an M4T as the initial primer (2). A fragment of 1,720 nucleotides long were separated, purified, and cloned and three independent clones were sequenced (GenBank Accession No. JN250997). Nucleotide and amino acid sequence analysis of the putative CP gene, respectively, revealed 75.1 to 76.3% and 80.3 to 82.1% identity with the Basella rugose mosaic virus (BaRMV) (GenBank Accession Nos. DQ821938, DQ394891, and DQ821939), 77.4 and 81.0% identity with Peace lily mosaic virus (GenBank Accession No. DQ851494), and 76.0 and 81.7% identity with the Phalaenopsis chlorotic spot virus (GenBank Accession No. HM021142). However, on the basis of the CP gene sequence analyses, these three viruses shared high (>88.5 and >94.3%) CP nucleotide and amino acid identity and should be classified as the same Potyvirus species. According to the species demarcation criteria for the Potyviridae (1), the pathogen causing mosaic and malformation symptoms on four o'clock was BaRMV (3). To our knowledge, this is the first report of BaRMV in four o'clock. References: (1) M. J. Adams et al. Arch. Virol. 150:459, 2005. (2) J. Chen et al. Arch. Virol. 146:757, 2001. (3) C. H. Hung and Y. C. Chang. Plant Pathol. 55:819, 2006.

Download Full-text

First Report of Cucumber mosaic virus Infecting Blephilia hirsuta in North America

Plant Disease ◽

10.1094/pdis-94-8-1070c ◽

2010 ◽

Vol 94 (8) ◽

pp. 1070-1070 ◽

Cited By ~ 1

Author(s):

B. Poudel ◽

A. G. Laney ◽

I. E. Tzanetakis

Keyword(s):

North America ◽

Endangered Species ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Chenopodium Quinoa ◽

Dot Blot ◽

Degenerate Primers ◽

First Report ◽

Wide Range ◽

Symptomatic Tissue

Blephilia hirsuta (Pursh) Benth. var. hirsuta, an ornamental plant known as hairy pagoda or hairy wood mint (Lamiaceae), is native to eastern North America and is listed as an endangered species or a species of special concern in several northeastern states ( http://www.ct.gov/dep/cwp/view.asp?a=2702&q=323482&depNav_GID=1628 and http://www.mass.gov/dfwele/dfw/nhesp/species_info/mesa_list/mesa_list.htm ). B. hirsuta, grown as an ornamental on the University of Arkansas campus in Fayetteville, exhibited mottling symptoms indicative of viral infection. Double-stranded RNA extractions (3) yielded four bands of approximately 3.2, 2.9, 2.2, and 0.9 kb, a pattern identical to that of Cucumber mosaic virus (CMV [2]). Nicotiana benthamiana and Chenopodium quinoa seedlings were mechanically inoculated with sap from symptomatic tissue. N. benthamiana inoculated plants were stunted and developed systemic mosaic and C. quinoa inoculated plants developed local lesions, whereas mock inoculated plants remained symptomless. Dot-blot and indirect ELISA using antisera against CMV (developed by H. A Scott) gave strong reactions when testing symptomatic tissue from B. hirusta, N. benthamiana, and C. quinoa compared with no reaction for symptomless plants. Total nucleic acid extractions (4) from symptomatic tissue was subjected to reverse transcription-PCR using Cucumovirus degenerate primers (1). An amplicon of approximately 940 bases was obtained and sequenced. The sequence, deposited in GenBank under Accession No. GU453918, confirmed the results of the immunological assays that B. hirsuta was infected with CMV. The nucleotide identities between the B. hirsuta isolate and those of the Fny CMV group exceeded 98%. To our knowledge, this is the first report of CMV infecting B. hirsuta, not only in North America, but globally. This finding has major implications for the ornamental industry and the viability of the endangered species. Given the wide range of CMV, B. hirsuta may act as a reservoir for the virus and facilitate transmission to ornamentals and other plants. In addition, the virus may reduce host fitness and undermine the efforts to preserve the species in areas that is threatened. References: (1) S. K. Choi et al. J. Virol. Methods 83:67, 1999. (2) I. E. Tzanetakis. Plant Dis. 93:431, 2009. (3) I. E. Tzanetakis and R. R. Martin. J. Virol. Methods 149:167, 2008. (4) I. E. Tzanetakis et al. Virus Res. 127:26, 2007.

Download Full-text

First Report of Konjac mosaic virus in Zamioculcas zamiifolia

Plant Disease ◽

10.1094/pdis-05-13-0537-pdn ◽

2013 ◽

Vol 97 (11) ◽

pp. 1517-1517 ◽

Cited By ~ 5

Author(s):

M. A. V. Alexandre ◽

L. M. L. Duarte ◽

E. B. Rivas ◽

E. W. Kitajima ◽

R. Harakava

Keyword(s):

Mosaic Virus ◽

Ornamental Plants ◽

Cytoplasmic Inclusion ◽

Viral Disease ◽

Rt Pcr ◽

Chenopodium Amaranticolor ◽

Serological Tests ◽

Thin Sections ◽

First Report ◽

Potyvirus Species

Zamioculcas zamiifolia (Lodd.) Engl. (“Zanzibar Gem,” “ZZ plant”) is the monotypic species of the genus belonging to the family Araceae. It is a stemless perennial plant native to Africa, from Kenya to South Africa, that produces succulent rhizomes at the base of its attractive dark green and glossy foliage. Symptoms of mosaic and foliar distortion were observed on a plant purchased at an ornamental plants shop in São Paulo state, Brazil. In order to identify the causal agent, transmission and serological tests, as well as electron microscopy (EM) observations, reverse transcription (RT)-PCR, and sequencing were carried out. EM observations revealed the presence of elongated, flexuous viral particles in foliar extracts and cytoplasmic lamellar aggregates of type II lamellar inclusions (Edwardson's classification), in thin sections. No symptoms were induced following mechanical inoculation on Chenopodium amaranticolor, C. murale, Gomphrena globosa, Nicotiana megalosiphon, N. debneyii, nor on the aroids Philodendron scandens, P. selloum, Dieffenbachia amoena, Colocasia esculenta, and Z. zamiifolia. Up to 2 months after inoculation, plants were still symptomless, and the virus was not detected by RT-PCR. The indirect ELISA tests were negative with antisera against Dasheen mosaic virus (gift from F. W. Zettler, University of Florida) and Turnip mosaic virus (gift from P. Roggero, IFA, Turin, Italy). RT-PCR performed on the original purchased ornamental plant with potyvirus-specific primers (CI-R = ACICCRTTYTCDATDATRTTIGTIGC and CI-F = GGIVVIGTIGGIWSIGGIAARTCIAC) targeting the cytoplasmic inclusion protein cistron of the potyvirus genome produced a fragment of approximately 650 bp (GenBank Accession No. KC990386). The sequence was similar to those of potyvirus species with nucleotide identity, determined by PAUP v.4.0b10 for Macintosh, ranging from 64% for Pokeweed mosaic virus (JQ609065) to 93% for Konjac mosaic virus KoMV-F (NC007913). KoMV has been detected in aroid species in Taiwan, India, Korea, Japan (1,2), Germany, and The Netherlands (3,4). This is the first report of a viral disease on Z. zamiifolia and of KoMV in the Americas. Such information along with the vegetative propagation of ZZ plants strongly suggests that KoMV is spread worldwide. References: (1) P. Manikonda et al. J. Phytopathol. 159:133, 2011. (2) M. Nishiguchi et al. Arch Virol. 151:1643, 2006. (3) D.-E. Lesemann and S. Winter. Acta Hort. 568:135, 2002. (4) K. Pham et al. Acta Hort. 568:143, 2002.

Download Full-text