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During the summer of 2003, weed samples of Rapistrum rugosum and Sisymbrium loeselii showing severe mosaic, malformation, and stunting were collected from cauliflower fields in Tehran Province of Iran. Using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with specific polyclonal antibodies, the samples were tested for the presence of Beet western yellows virus, Cauliflower mosaic virus, Radish mosaic virus, Turnip crinkle virus, Turnip mosaic virus (TuMV) (DSMZ, Braunschweig, Germany), Cucumber mosaic virus, and Tobacco mosaic virus (Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France). Leaf extracts were used for mechanical inoculation and they produced chlorotic local lesions on Chenopodium amaranticolor, necrotic lesions on leaves and shoot apex necrosis on Nicotiana glutinosa, leaf deformation, mosaic, and stunting on Petunia hybrida, and severe mosaic, distortion, and stunting on Brassica rapa. These symptoms were similar to those that were described previously for TuMV (4). ELISA results showed that the original leaf samples and inoculated indicator plants reacted positively with TuMV antibodies, but not with antibodies for any of the other viruses listed above. Also, reverse transcription-polymerase chain reaction of total RNA extracted from the collected leaf samples using the universal primers for potyviruses (3) resulted in the amplification of two fragments of the expected sizes, approximately 700 and 1,700 bp. TuMV, a member of the genus Potyvirus in the family Potyviridae, is transmitted by aphids in a nonpersistent manner (4). This virus is geographically widespread with a wide host range that can infect 318 species in 156 genera of 43 plant families including, Brassicaceae, Chenopodiaceae, Asteraceae, Cucurbitaceae, and Solanaceae (2,4). R. rugosum and S. loeselii, two annual or biennial plants in the Brassicaceae family, were common and widely distributed in the fields surveyed. The presence of TuMV-infected weed hosts in cauliflower fields may impact disease management strategies. TuMV was first observed on stock plants (Matthiola sp.) in Iran (1). To our knowledge, this is the first report of natural occurrence of TuMV on weed hosts in Iran. References: (1) M. Bahar et al. Iran. J. Plant Pathol. 21:11, 1985. (2) J. R. Edwardson and R. G. Christie. The potyvirus group. Fla. Agric. Exp. Stn. Monogr. Ser. No. 16, 1991. (3) A. Gibbs and A. Mackenzie. J. Virol. Methods 63:9, 1997. (4) J. A. Tomlinson. Turnip mosaic virus. No. 8 in: Descriptions of Plant Viruses. CMI/AAB, Surrey, England, 1970.

During the 2001 growing season, a survey was conducted to determine the incidence of Beet necrotic yellow vein virus (BNYVV), Beet soilborne virus (BSBV), and Beet virus Q (BVQ) in Iran. A total of 2,816 random and 76 samples with rhizomania were collected from 131 fields in the main sugar beet cultivation areas of 13 provinces in Iran. All samples were tested using a tissue-blot immunoassay (TBIA) with commercial BNYVV (As-0799.1/CG6-F4), BSBV (As-0576.1), and BSBV/BVQ (As-0576.2) antisera provided by S. Winter (DSMZ, Braunschweig, Germany). For randomly collected samples, the highest incidence of virus infection was found for BNYVV (52.3%), followed by BSBV (9.5%) and BVQ (1.5%). Co-infection of BNYVV with BSBV or BVQ was 6.6% and 0.9%, respectively. Infection with both BSBV and BVQ was found in 16 (0.6%) samples. In addition, 0.4% (12) of the samples was infected with all three viruses. Our results indicated the presence of BVQ in samples from 10 fields located in Azarbayejan-e-gharbi, Esfahan, Fars, Kermanshah, Khorasan, Lorestan, and Semnan provinces of Iran, with or without rhizomania-like symptoms. The presence of viruses was confirmed using reverse transcription-polymerase chain reaction (RT-PCR) of RNA from 81, 19, and 14 root samples with positive reaction in TBIA to BNYVV, BSBV, and BVQ, respectively, with previously described primers (3,4). The primers specifically amplified fragments of 501 bp, 602 bp, 399 bp, and 291 bp of the BNYVV RNAs 1 and 4, BSBV RNA-2, and BVQ RNA-1, respectively. Our results indicated that the samples tested were also positive using RT-PCR. The putative vector for BNYVV, BSBV, and BVQ, Polymyxa betae, was also detected in 161 samples (from 127 fields) by amplification of a 170-bp fragment of the P. betae repetitive EcoRI-like fragments using previously described primers (4). RT-PCR products from 72 BNYVV-positive sugar beet root samples from 58 fields that also gave positive reactions in TBIA were analyzed using single-strand conformation polymorphism (SSCP) as previously described with extracts from root beards of the susceptible sugar beet cvs. OPUS and IC1 grown in the soils infested with BNYVV types A and B (provided by A. Meunier, Unite de Phytopathologie-UCL-AGRO-BAPA, Louvain-la-Neuve, Belgium) as positive controls (3). The patterns obtained with SSCP were uniform and showed widespread occurrence of BNYVV type A in almost all provinces surveyed. The fragments obtained for BNYVV RNAs 1 and 4 of an isolate from Qazvin (BNQ1) were sequenced (GenBank Accession Nos. AY703452 and AY703455) and compared with other sequences available in GenBank using Clustal W, which revealed 99.3 and 99.6% identity with the Japanese S (D84410) and Italian type A (AF197552) isolates, respectively. The economic importance of BVQ and its interactions with other sugar beet soilborne viruses remains a matter of debate. BNYVV and BSBV have been previously reported from Iran (1,2). To our knowledge, this is the first report of the natural occurrence of BVQ in sugar beets in Iran. References: (1) Sh. Farzadfar et al. Plant Dis. 86:187, 2002. (2) K. Izadpanah et al. Iran. J. Plant Pathol. 32:155, 1996. (3) R. Koenig et al. J. Gen. Virol. 76:2051, 1995. (4) A. Meunier et al. Appl. Environ. Microbiol. 69:2356, 2003.

The main areas for field-grown sugar beet (Beta vulgaris) production in Iran were surveyed to study the occurrence and incidence of Alfalfa mosaic virus (AlMV), Beet curly top virus (BCTV), Beet mosaic virus (BtMV), Beet western yellows virus (BWYV), Beet yellows virus (BYV), Chickpea chlorotic dwarf virus (CpCDV), Cucumber mosaic virus (CMV), and Turnip mosaic virus (TuMV) during the growing season of 2001. A total of 5,292 random leaf samples in addition to 1,294 symptomatic leaves were collected from nine commercial sugar beet growing provinces of Iran and tested by tissue-blot immunoassay (TBIA). Serological diagnoses were confirmed by electron microscopy and host range studies. The highest virus incidence among the surveyed provinces was recorded in Qazvin, followed by Fars, Esfahan, Azarbayejan-e-gharbi, Khorasan, Kermanshah, Semnan, and Hamedan. According to the TBIA results, viruses in decreasing order of incidence in sugar beet were BCTV (27.9%), followed by BWYV (17.4%), CpCDV (12.5%), BYV (10.6%), BtMV (7.4%), TuMV (2.9%), AlMV (1.3%), and CMV (1.2%). Nearly 35% of sugar beets in Iran were infected by one or both of the two leafhopper-transmitted viruses (BCTV and CpCDV). Moreover, about 28% were infected by at least one of the six aphid-transmitted viruses (AlMV, BWYV, BtMV, BYV, CMV, and TuMV). Overall, one or more of the eight viruses assayed were detected in 45.5% of the plants surveyed. Several plants (35%) displaying virus-like symptoms did not react with the virus antisera used, suggesting that more viruses or virus-like agents are infecting sugar beets in Iran. In reference to the earlier reports, this is the first report of AlMV and TuMV in sugar beet fields of Iran. Also, this is the first detection of CpCDV as a pathogen of sugar beet.

From a total of 8,135 potato leaves collected from 132 fields in 11 provinces of Iran, the incidence and distribution of Alfalfa mosaic virus (AlMV), Eggplant mottled dwarf virus (EMDV), Potato leafroll virus (PLRV), Potato virus A (PVA), Potato virus M (PVM), Potato virus S(PVS), Potato virus X (PVX), Potato virus Y (PVY), and Tomato yellow fruit ring virus (TYFRV) were assessed using serological and biological methods. Based on enzyme-linked immunosorbent assay (ELISA) results, viruses in decreasing order of incidence in potato were PVS (35.9%), PVY (34.4%), PVA (27.0%), PVX (20.8%), PLRV (13.9%), PVM (9.0%), AlMV (7.0%), TYFRV (5.9%), and EMDV (5.1%). All 132 fields surveyed had some degree of virus infection, ranging from 28.8 to 98.6%, with an overall incidence of 75.2%. The highest and lowest incidence of virus infections among the surveyed provinces occurred in Kerman (93.2%) and Ardabil (56.7%), respectively. Overall, 25.0 and 50.2% of the collected potato samples had single or mixed infections, respectively. High levels of mixed infections were found between PVX and PVS (8.6%), and PVX and PVY (7.6%). Moreover, co-infection of samples with PVS and PVY, PVA and PVS, and PVA and PVY, the aphid-vectored virus/virus combinations, occurred at the highest incidence in almost all provinces surveyed, 15.3, 13.8, and 12.8%, respectively. In this study, Beet curly top virus was detected in symptomatic potato samples collected from some fields in the Kermanshah province.

A distinct isolate (TY-PF36) of Tomato yellow fruit ring virus (TYFRV) was obtained from potato (Solanum tuberosum) in Iran. Chlorosis and necrotic spots on leaves associated with leaf and stem necrosis symptoms appear on the affected plants. Of 32 plant species and cultivars mechanically inoculated, 24 were susceptible to the virus isolate. The isolate strongly reacted with TYFRV antibodies in enzyme-linked immunosorbent assay (ELISA), but not with the specific antibodies of other tospoviruses tested. TYFRV-specific N gene primers described previously, however, failed to produce DNA fragments from the total RNA extracts of the infected plants in reverse transcription–polymerase chain reaction (RT-PCR). Nucleotide sequencing of the complete N gene and partial L gene of this isolate revealed considerable differences to those reported for TYFRV with identities ranging from 83.9 to 84.2% and 84.9 to 85.4%, respectively. Two specific primers were designed for detecting TY-PF36 using RT-PCR; TY-PF36 was detected in symptomatic field samples of potato, peanut, soybean, and two weed species, Datura metel and D. stramonium.

A survey was conducted from 2001 to 2004 in the major cucurbit-growing areas in Iran to reassess the relative incidence of cucurbit viruses. Severe yellowing symptoms were observed frequently on older leaves of cucurbit plants in various regions in outdoor crops, suggesting the presence of Cucurbit aphid-borne yellows virus (CABYV, genus Polerovirus, family Luteoviridae) (1,2). Leaf samples (n = 1019) were collected from plants of melon (Cucumis melo L.), cucumber (C. sativus L.), squash (Cucurbita sp.), and watermelon (Citrullus lanatus L.) showing various virus-like symptoms (mosaic, leaf deformation, yellowing). All samples, collected from 15 provinces, were screened for the presence of CABYV by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with IgGs and alkaline phosphatase-conjugated IgGs against a CABYV reference isolate (1). Of the 1,019 samples tested, 471 were positive for CABYV using DAS-ELISA. Some of the positive samples had typical severe yellowing symptoms while symptoms in other samples were masked by mosaic or leaf deformations caused by other viruses frequently found in mixed infections (data not shown). During the entire survey, CABYV was detected by DAS-ELISA in 201 of 503 melon samples, 72 of 129 cucumber samples, 158 of 249 squash samples, and 40 of 138 watermelon samples. These results indicate that CABYV is widely distributed on four cucurbit species in the major growing areas of Iran. In order to confirm CABYV identification, total RNA extracts (TRI-Reagent, Sigma Chemical, St Louis, MO) were obtained from 25 samples that were positive using DAS-ELISA originating from Khorasan (n = 4), Esfahan (n = 6), Teheran (n = 3), Hormozgan (n = 4), Azerbaiejan-E-Sharqi (n = 4), and Kerman (n = 4). Reverse transcription-polymerase chain reactions (RT-PCR) were carried out using forward (5′-CGCGTGGTTGTGG-TCAACCC-3′) and reverse (5′-CCYGCAACCGAGGAAGATCC-3′) primers designed in conserved regions of the coat protein gene according to the sequence of a CABYV reference isolate (3) and three other unpublished CABYV sequences. RT-PCR experiments yielded an expected 479-bp product similar to the fragment amplified with extracts from the reference isolate. No amplification of the product occurred from healthy plant extracts. To our knowledge, this is the first report of the occurrence of CABYV in Iran on various cucurbit species. The high frequency (46.2%) with which CABYV was detected in the samples assayed indicates that this virus is one of the most common virus infecting cucurbits in Iran. References: (1) H. Lecoq et al. Plant Pathol. 41:749, 1992 (2) M. A. Mayo and C. J. D'Arcy. Page 15 in: The Luteoviridae. H. G. Smith and H. Barker, eds. CAB International Mycological Institute, Wallingford, UK, 1999. (3) H. Guilley et al. Virology 202:1012, 1994.

International audience; The genomes of two Watermelon chlorotic stunt virus (WmCSV) isolates, one from the Sudan and one from Iran, were cloned and sequenced. Sequence relationship with other geminiviruses characterizes WmCSV as a typical Eastern Hemisphere geminivirus with a bipartite genome. The two geographically distant WmCSV isolates from Africa and the Middle East share a very high overall sequence similarity: 98% between their DNA-A and 96% between their DNA-B components, and their respective capsid proteins are identical. A single amino acid change in the capsid protein (N131D) renders WmCSV whitefly nontransmissible. This region of the capsid is also implicated in transmission by Bemisia tabaci of Tomato yellow leaf curl virus.

Several viral diseases are responsible for significant economic losses in commercial cucurbit production worldwide. During a survey conducted in July 2002 in cucurbit growing areas in southern Iran, vein-clearing symptoms and leaf chlorosis on older leaves were observed on a cucumber plant near Jiroft (Kerman Province). These symptoms were similar to those caused by Cucumber vein yellowing virus (CVYV, genus Ipomovirus, family Potyviridae), a virus first described in Israel (1) and now widespread in cucurbit crops in the Middle East and Mediterranean Regions (2). The identification of CVYV was established through differential host range reaction and immunosorbent electron microscopy (IEM) experiments. Typical vein-clearing symptoms were observed following mechanical inoculation of cucumber and melon plantlets, but no symptoms were observed in Chenopodium quinoa, C. amaranticolor, Nicotiana tabacum, or Vigna sinensis. Numerous, slightly flexuous, elongated virus particles were observed in infected plant extracts. The particles were decorated by a polyclonal antiserum raised against a Sudanese isolate of CVYV. To confirm CVYV identification, total RNA extracts (TRI-Reagent, Sigma Chemical, St. Louis, MO) were obtained from the original cucumber sample. Reverse transcription-polymerase chain reactions (RT-PCR) were carried out using CVYV-specific primers CVYV-CP-5′: 5′-GCTTCTGGTTCTCAAGTGGA-3′ and CVYV-CP-3′: 5′-GATGCATCAGTTGTCAGATG-3′ designed according to the partial sequence of the coat protein gene of CVYV-Isr (GenBank Accession No. AF233429) (2). A 540-bp fragment corresponding to the central region of CVYV coat protein was obtained from extracts of infected plants but not from healthy plant extracts. Additional watermelon (n = 6) and melon (n = 4) leaf samples collected from plants growing in the same farm were tested for the presence of CVYV using RT-PCR. All samples reacted positively for CVYV. However, a sample of Citrullus colocynthis, a wild relative of watermelon growing nearby, was negative. CVYV was not detected using RT-PCR in 123 additional cucurbit samples collected from the eastern and central regions of Iran during a survey conducted in 2002. To our knowledge, this is the first report of the occurrence of CVYV in Iran. Additional surveys in southern regions where Bemisia tabaci, the vector of CVYV, is abundant are required to better estimate the prevalence of this virus in cucurbit crops in Iran. References: (1) S. Cohen and F. E. Nitzany. Phytopathol. Mediterr. 1:44, 1960 (2) H. Lecoq et al. J. Gen. Virol. 81:2289, 2000.

During 2001, a survey was conducted in vineyards in northwestern Iran and the eastern and western provinces of Azarbaijan, Zanjan, and Qazvin to detect the presence of Arabis mosaic virus (ArMV) and Grapevine leaf roll associated virus-3 (GLRaV-3). From December 2001 through March 2002, 5,352 dormant stem cuttings were collected. A portion of all stem cuttings was callused, rooted, potted, and grown in a greenhouse. Each sample was tested for the presence of ArMV and GLRaV-3 with specific antisera (Bioreba, Basel, Switzerland). Extracts of bark scrapings were prepared from the remaining portion of the dormant cuttings. After bud break of rooted cuttings, leaf extracts were prepared by the method used by Rowhani et al. (2). Dormant bark and leaf extracts were used with double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Among the samples tested, ArMV and GLRaV-3 were found in 4.7 and 2.3% of the collection, respectively. Leaf extracts that had tested positive for ArMV using ELISA, were mechanically inoculated on the indicator host plants of Chenopodium amaranticolor, Cucumis sativus, and Petunia × hybrida. All plants developed local lesions that subsequently developed systemic chlorosis that is reported for ArMV. Biological assays confirmed the results of ArMV using ELISA. To confirm testing, a number of the samples that were found positive for GLRaV-3 in ELISA tests were tested by reverse transcription-polymerase chain reaction (RT-PCR) technique using previously described specific primers (1). The PCR reaction resulted in the specifically amplification of a 300-bp fragment of GLRaV-3 RNA. In cvs. White Keshmesh, Ghezel Ozum, Red Lal, Askari, and Red Kehsmesh, symptoms associated with GLRaV-3 were reduced growth with smaller leaves and shoots. By late summer, the leaves rolled downward and the interveinal areas of the leaves turned to red, while the principal veins remained green in cvs. Red Lal and Red Keshmesh. Symptoms associated with ArMV were reduced growth, shoots with short internodes, and leaf chlorosis and distortion. To our knowledge, this is the first report of ArMV and GLRaV-3 from grapevines in Iran. References: (1) A. Nassuth et al. J. Virol. Methods 90:37, 2000. (2) A. Rowhani et al. Plant Dis. 81:799, 1997.

Severe leaf and stem necrosis before flowering was observed in potato (Solanum tuberosum) fields of Firouzkoh Province, Iran, during the summer of 1998. Infected plants died before the end of the growing season. Necrosis was more severe in cv. Agria than in cvs. Ajaxs and Arinda. A high population of Thrips tabaci was observed in August and September. Tomato spotted wilt virus (TSWV) (1) was detected in affected potatoes by using specific TSWV-IgG (from Bioreba) in double-antibody sandwich enzyme linked immunosorbent assay and by indicator plant reactions. Mechanical inoculation of indicator plants with leaf extracts of symptomatic potatoes produce necrotic local lesions in Chenopodium quinoa, C. amaranticolor, Gomphrena globosa, Vicia faba, Vigna sinensis, Phaseolus aureus var. Gohar, P. vulgaris, and Petunia hybrida. The virus caused systemic necrosis in Capsicum frutescens, Datura stramonium, D. metel, Nicotiana glutinosa, N. rustica, and Trapaeolum majus, preceded by systemic chlorotic spots. TSWV was reported from ornamental crops in Tehran and Absard areas near to Firouzkoh province (2), but this is the first report of TSWV occurrence on potatoes in Iran. References: (1) T. S. Ie. Descriptions of Plant Viruses. No. 39, 1970. (2) A. A. Moeini, et al. Iran. J. Plant Pathol. (In press.)