scholarly journals The spreading of Alfalfa mosaic virus in lavandin in Croatia

2014 ◽  
Vol 29 (2) ◽  
pp. 115-122 ◽  
Author(s):  
Ivana Stankovic ◽  
Karolina Vrandecic ◽  
Jasenka Cosic ◽  
Katarina Milojevic ◽  
Aleksandra Bulajic ◽  
...  
Keyword(s):  
Amino Acid ◽  
Mosaic Virus ◽  
Chenopodium Quinoa ◽  
Alfalfa Mosaic Virus ◽  
Amino Acid Identity ◽  
Acid Identity ◽  
Lavandula Stoechas ◽  
Pcr Products ◽  
Test Plants ◽  
Das Elisa

A survey was conducted in 2012 and 2013 to detect the presence and distribution of Alfalfa mosaic virus (AMV) in lavandin crops growing in continental parts of Croatia. A total of 73 lavandin samples from six crops in different localities were collected and analyzed for the presence of AMV and Cucumber mosaic virus (CMV) using commercial double-antibody sandwich (DAS)-ELISA kits. AMV was detected serologically in 62 samples collected at three different localities, and none of the samples tested positive for CMV. For further analyses, six selected samples of naturally infected lavandin plants originating from different localities were mechanically transmitted to test plants: Chenopodium quinoa, C. amaranticolor, Nicotiana benthamiana and Ocimum basilicum, confirming the infectious nature of the disease. Molecular detection was performed by amplification of a 751 bp fragment in all tested samples, using the specific primers CP AMV1/CP AMV2 that amplify the part of the coat protein (CP) gene and 3?-UTR. The RT-PCR products derived from the isolates 371-13 and 373-13 were sequenced (KJ504107 and KJ504108, respectively) and compared with the AMV sequences available in GenBank. CP sequence analysis, conducted using the MEGA5 software, revealed that the isolate 371-13 had the highest nucleotide identity of 99.5% (100% amino acid identity) with an isolate from Argentina originating from Medicago sativa (KC881010), while the sequence of isolate 373-13 had the highest identity with an Italian AMV isolate from Lavandula stoechas (FN667967) of 98.6% (99% amino acid identity). Phylogenetic analysis revealed the clustering of selected isolates into four molecular groups and the lavandin AMV isolates from Croatia grouped into two distinct groups, implying a significant variability within the AMV lavandin population.

scholarly journals Occurrence and molecular characterization of alfalfa mosaic virus in eggplant in Serbia

2021 ◽  
Vol 26 (51) ◽  
pp. 33-39
Author(s):  
Dragana Milošević ◽  
Maja Ignjatov ◽  
Zorica Nikolić ◽  
Gordana Tamindžić ◽  
Gordana Petrović ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Potato Virus Y ◽  
Chenopodium Quinoa ◽  
Alfalfa Mosaic Virus ◽  
Rt Pcr ◽  
Bright Yellow ◽  
Polymerase Chain ◽  
Test Plants ◽  
Das Elisa

In the 2018-19 growing season, a total of 51 leaves of eggplant plants grown under field conditions were collected randomly from nine private gardens at four different localities in the Province of Vojvodina. Eggplants with nearly 40% of plants showing bright yellow to white mosaic or mottling of leaves were found throughout the inspected fields (gardens). The collected samples were analyzed for the presence of alfalfa mosaic virus (AMV), cucumber mosaic virus (CMV) and potato virus Y (PVY) using commercial double-antibody sandwich (DAS)-ELISA kits. Serological analysis of eggplant samples revealed the presence of AMV in 80.39% collected samples. None of the analyzed samples was positive for CMV and PVY. The virus was successfully mechanically transmitted to test plants including Nicotiana benthamiana, Chenopodium quinoa, C. amaranticolor, as well as eggplant seedlings, confirming the infectious nature of the disease. The presence of AMV in eggplants was further verified by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing, using the primers CP AMV1 and CP AMV2 that amplify part of the coat protein (CP) gene. The phylogenetic analysis showed that Serbian AMV isolates grouped into a separate well-supported group together with AMV isolates from Italy, Croatia and previously characterized isolates from Serbia. To our knowledge, this is the first report of AMV infection of eggplant in Serbia.

scholarly journals A Polish Isolate of Zucchini yellow mosaic virus from Zucchini is Distinct from Other European Isolates

Plant Disease ◽  
2007 ◽  
Vol 91 (5) ◽  
pp. 639-639 ◽  
Author(s):  
H. Pospieszny ◽  
B. Hasiów ◽  
N. Borodynko
Keyword(s):  
Amino Acid ◽  
Mosaic Virus ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
Amino Acid Identity ◽  
Plant Viruses ◽  
Yellow Mosaic Virus ◽  
Parameter Method ◽  
Acid Identity ◽  
Conserved Sequence

Zucchini yellow mosaic virus (ZYMV) is a member of the Potyvirus genus in the Potyviridae family, the largest group of plant viruses. Different isolates of this virus have been found in infected cucurbits throughout the world, including localities in Europe, America, Australia, and Asia. In August 2005, mosaic and yellowing of leaves, as well as yellow spots on green fruits, were observed on zucchini (Cucurbita pepo cv. giromontiina) growing in commercial fields in the Kujawsko-Pomorskie Region of Poland. Flexuous virus particles (~750 nm long), typical of potyviruses, were observed in leaf-dip preparations from symptomatic zucchini plants. The virus in the sap from symptomatic plants was mechanically transmitted and systemic infections were produced on Citrullus lanatus, Cucumis melo, Cucumis sativus, C. pepo cvs. giromontiina and patissoniana, C. maxima, and Nicotiana benthamiana. Severe symptoms such as severe malformation of leaves and stunting of plants were observed on zucchini plants (cv. giromontiina) infected mechanically with the virus and grown in the greenhouse. Double-antibody sandwich (DAS)-ELISA using an anti-ZYMV polyclonal antiserum (AS-0234; DSMZ, Braunschweig, Germany) identified the presence of ZYMV in mechanically infected C. pepo cv. giromontiina and N. benthamiana plants. Subsequently, a reverse transcription (RT)-PCR using a universal primer, Sprimer, designed from the consensus sequences that code for the conserved sequence GNNSGQP in the NIb region of Potyviridae family members and the M4 primer was performed (1). The 1740-bp PCR fragments were cloned into the pGEM-T vector (Promega, Madison, WI) and three randomly selected clones were sequenced on an ABI automatic sequencer. An 837-bp sequence representing the full length coat protein gene (GenBank Accession No. EF178505) was compared with homologous sequences from other ZYMV isolates using BioEdit and Mega 3.1 softwares. Genetic distances were calculated by Kimura's two-parameter method (2). Surprisingly, the Polish ZYMV isolate (ZYMV-Zug) was more closely related to ZYMV isolates from Asia than those from Europe. Pairwise comparisons of ZYMV-Zug with several other European ZYMV isolates (GenBank Accession Nos. DQ645729, AJ420020, AJ459956, AJ420014, AJ420019, DQ124239, and AJ420018) indicated an 81 to 82% nucleotide and 91 to 92% amino acid identity, while there was a 94% nucleotide and 99% amino acid identity with the Shanxi (GenBank Accession No. AY074808) and Shandong isolates (GenBank Accession No. AF513552) from China. References: (1) J. Chen et al. Arch. Virol. 146:757, 2001. (2) S. Kumar et al. Brie. Bioinform. 5:150, 2004.

scholarly journals Detection of Poinsettia mosaic virus by RT-PCR in Euphorbia spp. in New Zealand

Plant Disease ◽  
2007 ◽  
Vol 91 (1) ◽  
pp. 110-110 ◽  
Author(s):  
B. S. M. Lebas ◽  
F. M. Ochoa-Corona ◽  
D. R. Elliott ◽  
J. Z. Tang ◽  
B. J. R. Alexander
Keyword(s):  
New Zealand ◽  
Mosaic Virus ◽  
Rna Extraction ◽  
Virus Detection ◽  
Amino Acid Identity ◽  
Rt Pcr ◽  
Taq Polymerase ◽  
Pcr Products ◽  
Area North ◽  
Das Elisa

Euphorbia pulcherrima (poinsettias) are commonly infected with Poinsettia mosaic virus (PnMV), which resembles the Tymovirus genus in its morphology and viral properties (2) but is closer to the Marafivirus genus at the sequence level (1). Symptoms induced by PnMV range from leaf mottling and bract distortion to symptomless (2). The presence of PnMV in plants imported into New Zealand had never been proven. Leaves of 10 E. pulcherrima samples and six samples from other Euphorbia spp. (E. atropurpurea, E. lambii, E. leuconeura, E. mellifera, E. milii, and E. piscatorial) were collected in the Auckland area, North Island in 2002. Isometric particles of 26 to 30 nm in diameter were observed with electron microscopy in 3 of 10 E. pulcherrima samples. These three samples produced systemic chlorosis and crinkling symptoms on mechanically inoculated Nicotiana benthamiana, which tested PnMV positive by double-antibody sandwich (DAS)-ELISA (Agdia, Elkart, IN). No particles or symptoms on N. benthamiana were observed with the other Euphorbia spp., which were also PnMV-negative by DAS-ELISA. A reverse transcription-polymerase chain reaction (RT-PCR) was developed to further characterize PnMV. Specific primers were designed from the PnMV complete genome sequence (Genbank Accession No. AJ271595) using the Primer3 web-based software (4). Primer PnMV-F1 (5′-CCTGTATTGTCTCTTGCCGTCC-3′) and primer PnMV-R1 (5′-AGAGGAAAGGAAAAGGTGGAGG-3′) amplified a 764-bp product from nt 5291 of the 5′-end RNA polymerase gene to nt 6082 of the 3′-untranslated region (UTR). Total RNA was extracted from leaf samples using the Qiagen Plant RNeasy Kit (Qiagen Inc., Chastworth, CA). RT was carried out by using PnMV-R1 primer and MMLV reverse transcriptase (Promega, Madison, WI). The PCR was performed in a 20-μl volume reaction containing 2 μl cDNA, 1× Taq reaction buffer, 1.5 mM MgCl2, 0.2 mM dNTPs, 0.2 μM PnMV-F1 primer, and 1 U of Taq polymerase (Promega) with a denaturation step (94°C for 5 min), 30 amplification cycles (94°C for 30 s; 55°C for 30 s; 72°C for 1 min), and a final elongation (72°C for 5 min). The sequence of the RT-PCR product (Genbank Accession No. DQ462438) had 98.7% amino acid identity to PnMV. PCR products were obtained from two of three PnMV ELISA-positive E. pulcherrima and three of three PnMV ELISA-positive symptomatic N. benthamiana. The failure to amplify the fragment from all ELISA-positive PnMV is likely because of the presence of inhibitors and latex in E. pulcherrima (3) that make the RNA extraction difficult. Thus, while RT-PCR may be useful for further characterizing PnMV isolate sequences, ELISA may be more reliable for virus detection. In conclusion, to our knowledge, this is the first report of PnMV in E. pulcherrima but not in other Euphorbia spp. in New Zealand. E. pulcherrima plants have been imported into New Zealand for nearly 40 years, and the virus is probably widespread throughout the country via retail nursery trading. References: (1) B. G. Bradel et al. Virology 271:289, 2000. (2) R. W. Fulton and J. L. Fulton. Phytopathology 70:321, 1980. (3) D.-E. Lesemann et al. Phytopathol. Z. 107:250, 1983. (4) S. Rozen and S. Skaletsky. Page 365 in: Bioinformatics Methods and Protocols: Methods in Molecular Biology. S. Krawetz and S. Misener, eds. Humana Press, Totowa, NJ, 2000.

scholarly journals Triticum mosaic virus: A Distinct Member of the Family Potyviridae with an Unusually Long Leader Sequence

2009 ◽  
Vol 99 (8) ◽  
pp. 943-950 ◽  
Author(s):  
Satyanarayana Tatineni ◽  
Amy D. Ziems ◽  
Stephen N. Wegulo ◽  
Roy French
Keyword(s):  
Amino Acid ◽  
Mosaic Virus ◽  
Phylogenetic Analyses ◽  
Amino Acid Identity ◽  
Recent Common Ancestor ◽  
Acid Identity ◽  
Most Recent Common Ancestor ◽  
Nontranslated Region ◽  
The Family ◽  
Distinct Member

The complete genome sequence of Triticum mosaic virus (TriMV), a member in the family Potyviridae, has been determined to be 10,266 nucleotides (nt) excluding the 3′ polyadenylated tail. The genome encodes a large polyprotein of 3,112 amino acids with the “hall-mark proteins” of potyviruses, including a small overlapping gene, PIPO, in the P3 cistron. The genome of TriMV has an unusually long 5′ nontranslated region of 739 nt with 12 translation initiation codons and three small open reading frames, which resemble those of the internal ribosome entry site containing 5′ leader sequences of the members of Picornaviridae. Pairwise comparison of 10 putative mature proteins of TriMV with those of representative members of genera in the family Potyviridae revealed 33 to 44% amino acid identity within the highly conserved NIb protein sequence and 15 to 29% amino acid identity within the least conserved P1 protein, suggesting that TriMV is a distinct member in the family Potyviridae. In contrast, TriMV displayed 47 to 65% amino acid sequence identity with available sequences of mature proteins of Sugarcane streak mosaic virus (SCSMV), an unassigned member of the Potyviridae. Phylogenetic analyses of the complete polyprotein, NIa-Pro, NIb, and coat protein sequences of representative species of six genera and unassigned members of the family Potyviridae suggested that TriMV and SCSMV are sister taxa and share a most recent common ancestor with tritimoviruses or ipomoviruses. These results suggest that TriMV and SCSMV should be classified in a new genus, and we propose the genus Poacevirus in the family Potyviridae, with TriMV as the type member.

scholarly journals First Report of Pepper veinal mottle virus in Tomato and Pepper in Taiwan

Plant Disease ◽  
2009 ◽  
Vol 93 (1) ◽  
pp. 107-107 ◽  
Author(s):  
Y. H. Cheng ◽  
R. Y. Wang ◽  
C. C. Chen ◽  
C. A. Chang ◽  
F.-J. Jan
Keyword(s):  
Amino Acid ◽  
Mosaic Virus ◽  
Polyclonal Antibodies ◽  
Amino Acid Identity ◽  
Mottle Virus ◽  
Rt Pcr ◽  
Acid Identity ◽  
First Report ◽  
Cp Gene ◽  
Pepper Veinal Mottle Virus

In May of 2006, samples from tomato plants (Solanum lycopersicum cv. Known-you 301) exhibiting necrotic symptoms on stems, petioles, and leaves were collected from Chiayi County, Taiwan. Double-antibody sandwich-ELISAs were performed using Cucumber mosaic virus, Tomato mosaic virus, Potato virus Y, Watermelon silver mottle virus, and Chilli veinal mottle virus (ChiVMV) polyclonal antibodies. Three of eight samples reacted with antibodies against ChiVMV but not with the others. Using the potyvirus degenerate primers (Hrp 5/Pot 1) (2), an expected 1.5-kb DNA fragment including the 3′-end of the NIb gene, the complete coat protein (CP) gene, and the 3′-nontranslatable region of the virus was amplified from total RNA isolated from these three samples by reverse transcription (RT)-PCR. A homology search in GenBank indicated that the new tomato-infecting virus in Taiwan belongs to Pepper veinal mottle virus (PVMV) since they shared >90% amino acid identity in the CP gene. A virus culture (Tom1) isolated from one of the diseased tomatoes was then established in Chenopodium quinoa and Nicotiana benthamiana and the CP gene was amplified and sequenced (GenBank Accession No. EU719647). Comparisons of the 807-nt CP gene with those of five PVMV isolates available in GenBank showed 81.5 to 93.1% nucleotide and 90.0 to 97.8% amino acid identity. Tom1 induced irregular necrotic lesions on stems, petioles, and leaves of tomato while inducing only mild mottle symptoms on pepper. Serological cross reaction between ChiVMV and PVMV has been observed previously (1,3) and also found in this study. To differentiate these two potyviruses by RT-PCR, primer pair CPVMVup/dw (5′-TATTC(T/C)TCAGTGTGG(A/T/C)T(T/C)CCACCAT and 5′-(T/C)C(A/T)C(A/T)(A/T/G)(A/T)AA(A/G)CCATAA(A/C)(A/C)ATA(A/G)T(T/C)T) was designed on the basis of the comparison of the CP gene and the 3′-nontranslatable region of the PVMV and ChiVMV. DNA fragments of 171 and 259 bp are expected to be amplified from ChiVMV and PVMV, respectively, by RT-PCR with primers CPVMVup/dw. In a field survey done in 2006, samples from diseased peppers (Capsicum annuum) that reacted with the polyclonal antibodies against ChiVMV were further identified by RT-PCR with primers CPVMVup/dw, indicating that both ChiVMV and PVMV infected pepper crops (Capsicum spp.) in Taiwan. A pepper isolate (Pep1) of PVMV was obtained from Nantou County through three times of single lesion passages on C. quinoa and then propagated on N. benthamiana. The CP gene of Pep1 was amplified and sequenced (GenBank Accession No. EU719646) and found to share 99.1% nucleotide and 100% amino acid identity with that of Tom1. Pep1 caused mild mottle symptoms on leaves of both tomato and pepper. To our knowledge, this is the first report of the presence of PVMV in Taiwan as well as in East Asia. References: (1) B. Moury et al. Phytopathology 95:227, 2005. (2) S. S. Pappu et al. Plant Dis. 82:1121, 1998. (3) W. S. Tsai et al. Plant Pathol. 58:408, 2008.

scholarly journals Identification of Tomato mosaic virus Infection in Lisianthus in Taiwan

Plant Disease ◽  
2003 ◽  
Vol 87 (12) ◽  
pp. 1537-1537 ◽  
Author(s):  
F.-J. Jan ◽  
C.-C. Chen ◽  
H. T. Hsu
Keyword(s):  
Amino Acid ◽  
Mosaic Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Systemic Infection ◽  
Amino Acid Identity ◽  
Tomato Mosaic Virus ◽  
Nucleotide Identity ◽  
Acid Identity ◽  
Cp Gene

In recent years, Lisianthus (Eustoma russellianum (Don.) Griseb) has become popular as potted plants and cut flowers in Taiwan. They are grown in the central and southern regions of the island. Since 1998, diseased plants with mosaic symptoms, followed by necrosis of leaf tissues, were observed in commercial greenhouses and field-grown lisianthus. Newly emergent leaves were curled and smaller compared with those on healthy plants. These symptoms greatly decreased the commercial value of the crop. Rigid rods similar to tobamoviruses that measured 300 × 18 nm were found consistently associated with symptomatic plants. In July 2002, a virus culture was isolated from diseased lisianthus from Chiayi County, Taiwan and established and maintained in systemic hosts Nicotiana tabacum L. and N. benthamiana Domin. Chlorotic and necrotic spots developed on lisianthus leaves 1 to 2 weeks after inoculation with the virus; symptoms eventually became systemic. Virions were purified from inoculated N. tabacum. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the virus contained one 18-kDa (Mr) polypeptide. The virus reacted positively in agar gel double diffusion tests and enzyme-linked immunosorbent assays with antisera prepared to Tobacco mosaic virus (TMV) or Tomato mosaic virus (ToMV) (gifts of S. D. Yeh, National Chung Hsing University, Taichung, Taiwan). A viral coat protein (CP) gene approximately 0.5 kb was amplified by reverse transcription-polymerase chain reaction from total RNA prepared from infected N. benthamiana using 5′-GCGAGCCATGGATTCTTACTCAATTACT as a forward primer and 5′-ACTCTCGGATCCTTAAGATGCAGGTGCAGA as a reverse primer. Comparison of the 480-nt CP gene region with that of ToMV-OM (GenBank Accession No. X02144) (3) revealed 99.2% nucleotide identity and 99.4% amino acid identity. It shares, however, 74.4% nucleotide identity and 83.9% amino acid identity with CP genes of TMV-U1 (GenBank Accession No. AX040174) and TMV-vulgare (GenBank Accession No. J02415) (1). The virus induced local lesion responses similar to ToMV on inoculated N. tabacum cv. White Burley, N. sylvestris Speg. & Comes, and Datura stramonium L. Inoculation of TMV, however, resulted in a systemic infection in these plants. Results from sequence analysis and diagnosis based on host reaction to the virus inoculation indicated that the tobamovirus infecting lisianthus in Taiwan is an isolate of ToMV. The virus is economically important to lisianthus and tomato in Taiwan. To our knowledge, this is the first report of ToMV causing disease on lisianthus in Taiwan. The disease was previously observed on lisianthus in Italy (2). References: (1) P. Goelet et al. Proc. Natl. Acad. Sci. USA 79:5818, 1982. (2) V. Lisa and A. Gera. Lisianthus. Pages 443–448 in: Virus and Virus-like Diseases of Bulb and Flower Crops. G. Loebenstein et al. eds. John Wiley and Sons, West Sussex, U.K., 1995. (3) T. Ohno et al. J. Biochem. 96:1915, 1984.

scholarly journals First Report of Tomato spotted wilt virus in Sweet Pepper in Taiwan

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 920-920 ◽  
Author(s):  
Y.-X. Zheng ◽  
C.-H. Huang ◽  
Y.-H. Cheng ◽  
F.-Y. Kuo ◽  
F.-J. Jan
Keyword(s):  
Amino Acid ◽  
Mosaic Virus ◽  
Sweet Pepper ◽  
Amino Acid Identity ◽  
N Gene ◽  
Rt Pcr ◽  
Acid Identity ◽  
Tomato Spotted Wilt ◽  
Wilt Virus ◽  
Pepper Plants

A new disorder on pepper showing symptoms of chlorosis and chlorotic spots on leaves was observed in sweet pepper (Capsicum annuum cv. Andalus) fields in Ren-Ai Township, Nantou County in July, 2009. The disorder occurred in more than 30% of the pepper plants, with a height of approximately 40 cm (1.5 feet), which was approximately one-half the size of the asymptomatic ones. Symptomatic plants bore much smaller fruits with abnormal shapes. Three symptomatic sweet pepper plants were collected and tested for potential viruses. Reverse transcription (RT)-PCR was performed for the detection using three degenerate primer pairs, gL3637/gL4435c for tospoviruses (2), Hrp5/Pot1 for potyviruses (1,3), and Tob-Uni1/Tob-Uni2 for tobamoviruses (4), and specific primers, FJJ2001-7/FJJ2001-8 (5′-TATGTCCATGGACAAATCCGAATCA and 5′-TCTCTGGATCCACGAGTTCAAACTGGGAG) for the coat protein gene of Cucumber mosaic virus (CMV). An 819-nt DNA fragment containing the partial L RNA of tospovirus was amplified from the total RNA isolated from each of these three samples by RT-PCR with primer pair gL3637/gL4435c. One amplified fragment was cloned and sequenced. A homology search in GenBank indicated that the new pepper-infecting virus in Taiwan was Tomato spotted wilt virus (TSWV) since the partial L RNA shared more than 94.5% nucleotide and 98.2% amino acid identity with five TSWV isolates (Accession Nos. AB190813, AB198742, AY070218, D10066, and NC_002052). No DNA fragment was obtained by RT-PCR using primer pairs for CMV, potyviruses, or tobamoviruses. A virus culture (TwPep1) isolated from one of the symptomatic sweet pepper plants was then established in Nicotiana tabacum cv. White Burley and N. benthamiana through triple single-lesion isolation. TWPep1 reacted positively only to the antiserum against TSWV by indirect-ELISA but not to those of Watermelon silver mottle virus, Capsicum chlorosis virus, Tobacco mosaic virus, Tomato mosaic virus, and CMV. Partial L RNA and the full-length nucleocapsid (N) gene of TWPep1 were obtained by RT-PCR with primer pairs gL3637/gL4435c and FJJ2002-74/FJJ2002-75 (5′-GCGCGCGGATCCTAATTTAACTTACARCTGCT 5′-TGCTGCCTCGAGCATACGGTCAAAGCATATAA), respectively. The 819-nt L RNA conserved region of TwPep1 (Accession No. GU222652) shared 94.4 to 97.7% nucleotide and 98.2 to 100% amino acid identity with those available in GenBank. The 777-bp N gene of TwPep1 (Accession No. GU222651) shared 96.7 to 99.1% nucleotide and 97.3 to 99.6% amino acid identity with 37 TSWV isolates available in GenBank. Sequence comparisons indicated that TwPep1 is an isolate of TSWV. TSWV was later detected by RT-PCR in all 10 symptomatic samples of sweet pepper plants collected from five fields in August 2009. To our knowledge, this is the first report of TSWV in sweet pepper in Taiwan. This is also the first demonstration of isolation and characterization of TSWV in Taiwan although TSWV was once detected in lisianthus (Eustoma rusellianum) by RT-PCR (1) but the isolation was not successful then. The occurrence of TSWV in pepper will have a direct economic impact on the important vegetable and floral industry in Taiwan because TSWV reportedly comprises a wide host range. References: (1) C. C. Chen et al. Bot. Stud. 947:369, 2006. (2) F. H. Chu et al. Phytopathology 91:361, 2001. (3) D. Colinet and J. Kummert. J. Virol. Methods 45:149, 1993. (4) B. Letschert et al. J. Virol. Methods 106:1, 2002.

scholarly journals First Report of Zantedeschia mosaic virus Infecting a Zantedeschia sp. in New Zealand

Plant Disease ◽  
2008 ◽  
Vol 92 (8) ◽  
pp. 1253-1253 ◽  
Author(s):  
T. Wei ◽  
M. N. Pearson ◽  
D. Cohen ◽  
J. Z. Tang ◽  
G. R. G. Clover
Keyword(s):  
New Zealand ◽  
Amino Acid ◽  
Mosaic Virus ◽  
Amino Acid Identity ◽  
Cut Flowers ◽  
Rt Pcr ◽  
Acid Identity ◽  
Virus Particles ◽  
Blast Search ◽  
Cp Gene

In February 2004, leaf yellowing, mottling, and mosaics were observed on a few plants of a Zantedeschia sp. (calla lily) growing in Rangiora, Canterbury, New Zealand. Zantedeschia spp. are known to be susceptible to at least 13 virus species (1). No symptoms were observed on Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Gomphrena globosa, Nicotiana benthamiana, N. clevelandii, N. occidentalis, or N. tabacum when inoculated with sap from symptomatic plants. However, electron microscopy of crude sap preparations from a symptomatic Zantedeschia sp. and inoculated N. clevelandii plants revealed the presence of flexuous, filamentous virus particles approximately 700 nm long and 12 nm wide. No virus particles were seen in the other inoculated indicator species. Nucleic acid was extracted from leaves of the infected Zantedeschia sp. and N. clevelandii plants and tested in reverse transcription (RT)-PCR using published potyvirus-specific primers (4). PCR amplicons of the expected size (327 bp) were obtained from both plant species and sequenced directly. The products were identical, and a BLAST search in GenBank showed 99% nucleotide identity with a Taiwanese isolate of the species Zantedeschia mosaic virus (ZaMV) (GenBank Accession No. AY026463). A product of 1,531 bp (GenBank Accession No. EU544542) was amplified from symptomatic Zantedeschia by RT-PCR using novel forward (5′-GCACGGCAGATAAACACGAC-3′) and reverse (5′-GTGGGCAACCTTCAACTGTG-3′) primers designed to amplify the 3′ untranslated region (3′UTR), coat protein (CP), and partial nuclear inclusion b protein (NIb) genes. The product was sequenced and had 94% nucleotide identity with a South Korean ZaMV isolate (GenBank Accession No. AB081519), with 95% nucleotide (97% amino acid) identity in the CP gene. A second crop of Zantedeschia spp. in Tauranga, New Zealand (approximately 700 km north of Rangiora) was observed to have similar disease symptoms. Symptomatic plants tested positive in ELISA using a potyvirus-specific monoclonal antibody (Agdia Inc., Elkhart, IN). Nucleic acid was extracted from leaves of symptomatic plants and tested in RT-PCR using potyvirus-specific primer pairs, PV2I/T7 and D335 and U335 and PV1/SP6, which amplify overlapping regions within the 3′UTR, CP, and NIb genes (2,3). The products were sequenced and a consensus sequence of 1,793 bp was generated (GenBank Accession No. EU532065). A BLAST search showed that the sequence had 78% nucleotide (88% amino acid) identity with Zantedeschia mild mosaic virus (ZaMMV) (GenBank Accession No. AY626825). However, the sequences had only 73% nucleotide (79% amino acid) identity in the CP gene, and therefore, this second virus may be a distinct species. To our knowledge, this is the first report of ZaMV in New Zealand. Cut flowers are an increasingly important commodity in New Zealand and Zantedeschia is one of the most important crops; in 2005, exports of rhizomes and cut flowers of the genus were worth NZ$10.9 million. These viral diseases may require management to ensure that the quality of production is maintained. References: (1) C. H. Huang et al. Plant Pathol. 56:183, 2007. (2) S. A. Langeveld et al. J. Gen. Virol. 72:1531, 1991. (3) A. M. Mackenzie et al. Arch. Virol. 143:903, 1998. (4) V. Marie-Jeanne et al. J. Phytopathol. 148:141, 2000.

scholarly journals A50 Whole-genome sequencing of African swine fever isolates from Sardinia

2019 ◽  
Vol 5 (Supplement_1) ◽  
Author(s):  
C Torresi ◽  
F Granberg ◽  
L Bertolotti ◽  
A Oggiano ◽  
B Colitti ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sequence Data ◽  
Temporal Distribution ◽  
African Swine Fever ◽  
Amino Acid Identity ◽  
Genotype I ◽  
Acid Identity ◽  
Wide Range ◽  
Intergenic Sequences ◽  
Genes Encoding

Abstract In order to assess the molecular epidemiology of African swine fever (ASF) in Sardinia, we analyzed a wide range of isolates from wild and domestic pigs over a 31-year period (1978–2009) by genotyping sequence data from the genes encoding the p54 and the p72 proteins and the CVR. On this basis, the analysis of the B602L gene revealed a minor difference, placing the Sardinian isolates into two clusters according to their temporal distribution. As an extension of this study, in order to achieve a higher level of discrimination, three further variable genome regions, namely p30, CD2v, and I73R/I329L, of a large number of isolates collected from outbreaks in the years 2002–14 have been investigated. Sequence analysis of the CD2v region revealed a temporal subdivision of the viruses into two subgroups. These data, together with those from the B602L gene analysis, demonstrated that the viruses circulating in Sardinia belong to p72/genotype I, but since 1990 have undergone minor genetic variations in respect to its ancestor, thus making it impossible to trace isolates, enabling a more accurate assessment of the origin of outbreaks, and extending knowledge of virus evolution. To solve this problem, we have sequenced and annotated the complete genome of nine ASF isolates collected in Sardinia between 1978 and 2012. This was achieved using sequence data determined by next-generation sequencing. The results showed a very high identity with range of nucleotide similarity among isolates of 99.5 per cent to 99.9 per cent. The ASF virus (ASFV) genomes were composed of terminal inverted repeats and conserved and non-conserved ORFs. Among the conserved ORFs, B385R, H339R, and O61R-p12 showed 100 per cent amino acid identity. The same was true for the hypervariable ORFs, with regard to X69R, DP96R, DP60R, EP153R, B407L, I10L, and L60L genes. The EP402R and B602L genes showed, as expected, an amino acid identity range of 98.5 per cent to 100 per cent and 91 per cent to 100 per cent, respectively. In addition, all of the isolates displayed variable intergenic sequences. As a whole, the results from our studies confirmed a remarkable genetic stability of the ASFV/p72 genotype I viruses circulating in Sardinia.

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