The Existence of Papaya ringspot virus-Papaya Strain on Cucumber in Gianyar, Bali

Yellow mosaic symptoms were identified from cucumber production systems in Gianyar and were similar to symptoms of PRSV infection. Further research was conducted to determine diseases incidence and molecular characteristic of PRSV. Ninety leaf samples were collected from Gianyar by purposive sampling and disease incidence calculations were based on symptoms in the field. Detection and identification were done using a RT-PCR with specific primers of CP PRSV-P, CP PRSV-W and DNA sequencing. Disease incidences in the fields ranged between 5.81–66.87%. Specific DNA band 470 bp was successfully amplified from several cucumber leaf samples collected from Ubud, Payangan, Tegallalang, Sukawati, Gianyar, and Blahbatuh; but no DNA were amplified from all samples when using CP PRSV-W specific primer. Nucleotide and amino acid analysis showed nucleotides homology to isolates from Ubud, Payangan, Tegallalang, Sukawati, Gianyar, and Blahbatuh, i.e. 98.9–99.5% and 99.1–100%, respectively. Results indicated that genetic variation of PRSV-P from Gianyar was low. Furthermore, the nucleotides homology of PRSV-P isolates from Ubud, Payangan, Tegallalang, Sukawati, Gianyar, and Blahbatuh were with PRSV-P isolates which infected cucumbers from Nganjuk (LC311783) and Brebes (LC311784), while from native papaya collected in Bali Bali (LC223115) were 97.2–98.4% and 98.6–100%, respectively. Phylogenetic analysis confirmed that PRSV-P isolates from Indonesia were in the same cluster with Thailand isolates. The results showed that sources of PRSV-P inoculums spreading into new areas.

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Comparação da seqüência de nucleotídeos do gene da proteína capsidial de estirpes severas e premunizantes do Papaya ringspot virus

Fitopatologia Brasileira ◽

10.1590/s0100-41582003000600015 ◽

2003 ◽

Vol 28 (6) ◽

pp. 678-681 ◽

Cited By ~ 3

Author(s):

Marilia G. S. Della Vecchia ◽

Luis E. A. Camargo ◽

Jorge A. M. Rezende

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Core Region ◽

Amino Acid Sequences ◽

Ringspot Virus ◽

Papaya Ringspot Virus ◽

Specific Primers ◽

Sequence Comparisons ◽

The Core ◽

Cp Gene

This study compared three mild and three severe strains of Papaya ringspot virus - type W (PRSV-W), based on nucleotide and amino acid sequences of the capsid protein (CP) gene. The CP nucleotide sequences of the mild strains shared 98% to 100% identity. When compared to the severe strains the identity ranged from 93% to 95%, except in the case of PRSV-W-2R, which resulted from reversion of the mild strains PRSV-W-2. The CP sequence of the reverting strain showed 100% identity with the sequence of its parental strain. An insertion of six nucleotides in the core region of the CP gene, which reflected the addition of two amino acids (Asn and Asp) in the deduced sequence of the protein, was found in all mild strains. These sequence comparisons were used to design strain-specific primers that were used to specifically amplify regions for either the mild or severe strains.

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The application of sequence specific primer and RT-PCR to LRRK2 gene polymorphism typing

Discussion of Clinical Cases ◽

10.5430/dcc.v6n2p17 ◽

2019 ◽

Vol 6 (2) ◽

pp. 17

Author(s):

Biao G ◽

Gaisheng T ◽

Qinxue L ◽

Fengrui L

Keyword(s):

Dna Markers ◽

Disease Susceptibility ◽

Susceptibility Loci ◽

Rt Pcr ◽

Specific Primer ◽

Specific Primers ◽

Lrrk2 Gene ◽

Polymerase Chain ◽

Quantitative Fluorescence ◽

Detecting Method

Objective: To establish a new detecting method for disease susceptibility loci R1628P and G2385R of Parkinson’s disease (PD) related gene LRRK2.Methods: Sequence specific primers were designed to make a genotyping of DNA markers with known genotypes by use of quantitative fluorescence real-time PCR (RT-PCR). 100 cases of PD samples with unknown genotypes were tested, and verified by use of polymerase chain reaction linked restriction fragment length polymorphism (PCR-RLFP).Results: The genotyping results of DNA markers proved to be correct, and 100 cases of samples to be tested had a completely consistent genotyping result with PCR-RLFP genotyping result.Conclusions: Sequence specific primer and quantitative fluorescence RT-PCR can successfully make a genotyping for disease susceptibility loci R1628P and G2385R of LRRK2.

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Karakterisasi Molekuler Papaya ringspot virus tipe P pada Tanaman Mentimun di Jawa

Jurnal Fitopatologi Indonesia ◽

10.14692/jfi.14.3.75 ◽

2018 ◽

Vol 14 (3) ◽

pp. 75

Author(s):

Listihani Listihani ◽

Tri Asmira Damayanti ◽

Sri Hendrastuti Hidayat ◽

Suryo Wiyono

Keyword(s):

Ringspot Virus ◽

Mosaic Symptom ◽

Papaya Ringspot Virus ◽

Rt Pcr ◽

Chain Reaction ◽

Specific Primers ◽

Central Java ◽

Dna Fragment ◽

Polymerase Chain

Moleculer Characterization of Papaya ringspot virus type P on Cucumber in JavaInfection of Papaya ringspot virus (PRSV) on cucumber plants showing mosaic symptom was detected using specific antibody. Further investigation was conducted to determine molecular characters and status of PRSV infecting cucumber in Java. Infection of PRSV was detected from leaf samples collected from the field using dot immunobinding assay (DIBA). Disease frequency caused by PRSV infection reached 81.11%, 95.86%, 91.66%, and 92.3% in East Java, Central Java, Yogyakarta, and West Java, respectively. Characterization of PRSV isolates was conducted by reverse transcription polymerase chain reaction (RT-PCR) using specific primers for PRSV-P and PRSV-W, followed by cloning, and DNA sequencing. DNA fragment of 470 bp was successfully amplified using specific primers for PRSV-P from several samples from Nganjuk, Brebes, Kulon Progo, and Subang; but no amplification was achieved using specific primers for PRSV-W. Nucleotide and amino acid analysis showed high homology among PRSV-P isolates from Nganjuk, Brebes, Kulon Progo, and Subang, i.e. 98.6%-99.7% and 99.3%-100%, respectively. This is an indication of a low genetic variation among PRSV-P from Java. Further phylogenetic analysis indicated that PRSV-P isolate cucumber is in the same cluster with PRSV-P isolate papaya from Bali, Indonesia. This is the first report of PRSV-P infecting cucumber in Indonesia.

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First report of cucurbit chlorotic yellows virus (CCYV) infecting pumpkin in India

Plant Disease ◽

10.1094/pdis-07-21-1473-pdn ◽

2021 ◽

Author(s):

Ashwini Kumar ◽

Bichhinna Maitri Rout ◽

Shakshi Choudhary ◽

Amish K. Sureja ◽

V. K. Baranwal ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Amino Acid Sequence Identity ◽

Rt Pcr ◽

Specific Primers ◽

First Report ◽

Virus Particles ◽

Sequence Identity ◽

Cp Gene ◽

Cucurbit Chlorotic Yellows Virus

Pumpkin (Cucurbita moschata), a member of the family Cucurbitaceae, is widely cultivated throughout the world including India. During August 2020 to January 2021, stunted pumpkin plants (cv. Pusa Vishwas), showing chlorotic patches, mosaic, and vein banding on leaves (e-Xtra Fig.1), were observed in the experimental fields of the Indian Agricultural Research Institute (IARI), New Delhi, India. Leaf-dip electron microscopy (EM) of the symptomatic plants (12 out of 37 samples) revealed the association of long flexuous virus particles measuring 650-950nm×10-12nm, suggestive of the presence of either crinivirus or potyvirus or both. Subsequently, a reverse transcription-polymerase chain reaction (RT-PCR) was performed on RNA extracted from the samples that had long flexuous virus particles using generic primers for criniviruses i.e. CriniPol-F: GCY CCS AGR GTK AAT GA and CriniPol-R: ACC TTG RGA YTT RTC AAA targeting partial RNA-dependent RNA polymerase coding region (Martin et al. 2003) and specific primers for papaya ringspot virus (PRSV) targeting a part of 3’ NIb and full coat protein (CP) gene (Basavaraj et al., 2019) separately. All tested samples were positive for both crinivirus and PRSV as expected size amplicons were obtained, accounting for about 32% prevalence. As PRSV is a well-studied virus infecting cucurbits, further work was not carried on this virus and only the RT-PCR amplicon indicative of crinivirus (~515 bp) was cloned into the pGEM-T easy cloning vector (Promega, Madison, WI) and sequenced for further confirmation of the virus presence. The obtained sequence (GenBank accession No MZ318672) shared up to 90% nucleotide and 100% amino acid sequence identity with the corresponding genomic region of a cucurbit chlorotic yellows virus (CCYV) isolate from Greece (LT841297). To confirm the identity of the crinivirus species present in the same pumpkin sample, the CP gene (753bp) was amplified and sequenced using CCYV CP gene-specific primers CP-F (5’-ATG GAG AAG ACY GAC AAT AAA CAA AAT GAT GA-3’) and CP-R (5’-TTA TTT ACT ACA ACC TCC CGG TGC CAA C-3’) (modified from Kheireddine et al. 2020). Sequence analysis using the BioEdit tool (version 2.0) revealed that the crinivirus present in pumpkin (KC577202) shared 95 to 100% nucleotide (and 98 to 100% amino acid) sequence identity with the corresponding gene sequences of CCYV isolates originating from cucurbitaceous hosts from diverse locations. The presence of CCYV was further validated by a whitefly transmission-based bioassay followed by RT-PCR confirmation. The bioassay was performed by the whitefly species Bemisia tabaci (biotype Asia II7) using the acquisition access period and inoculation access period of 24 hours each. Six whitefly individuals per plant were used for inoculating ten pumpkin plants (cv. Pusa Vishwas) at the first true leaf stage grown in pots containing soilrite as the medium in insect-proof cages. All ten plants inoculated using whiteflies exhibited chlorosis and stunting symptoms 12-15 days post-inoculation (e-Xtra Fig.2) and were found positive for CCYV in RT-PCR assay performed using CCYV CP gene-specific primers. Though CCYV had been reported worldwide (Tzanetakis et al. 2013), its occurrence had not been reported from India. Results of the present study confirm the infection of pumpkin plants by CCYV and constitute the first report of its presence in India. Further, there is a need to investigate the extent of its spread and impact of this virus on the production of cucurbitaceous crops in the country.

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Detection of Citrus Bent Leaf Viroid in Citrus Orchards of Sargodha, Pakistan

Arab Journal Of Plant Protection ◽

10.22268/ajpp-039.2.159163 ◽

2021 ◽

Vol 39 (2) ◽

pp. 159-163

Author(s):

Faheema Bakhtawar ◽

◽

Yasir Iftikhar ◽

Muhammad Ahmed Zeshan ◽

Muhammad Imran Hamid ◽

...

Keyword(s):

Molecular Detection ◽

Disease Incidence ◽

Mean Value ◽

Rt Pcr ◽

Specific Primers ◽

Citrus Orchards ◽

The Mean ◽

Citrus Varieties ◽

Near Future ◽

First Time

A study was conducted to monitor the Citrus bent leaf viroid (CBLVd) in citrus growing areas of district Sargodha, Pakistan during 2017-2018. Collected samples were tested by RT-PCR using specific primers. PCR positive samples were used to confirm the CBLVd incidence and severity on different citrus varieties grown at different regions of Sargodha. Maximum disease incidence was recorded in Kot Momin with the mean value of 24%, with severe symptoms of bark cracking, backward leaf bent and stunting. Minimum disease incidence was recorded in in Sillanwali region with the mean value of 3.33%. The symptoms in Sillanwali were only yellowing and slight leaf bent. Maximum severity was observed in Kot momin (0.60%). Molecular detection of CBLVd by RT-PCR confirmed the diagnosis of the viroid. This survey was carried out for the first time in Sargodha district to monitor the occurrence of citrus bent leaf viroid following the first report of its detection in Pakistan in 2009. Since many declining citrus trees were found negative to CBLVd testing, other causal agents can be involved, and extensive surveys are still required in the near future. Keywords: Citrus, RT-PCR, CBLVd, Disease incidence, viroid, Sargodha, Pakistan

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Karakterisasi Genetik Fragmen Gen Penyandi RNA Polimerase Infectious Myonecrosis Virus (IMNV) yang Menginfeksi Udang Vannamei (Litopenaeus vannamei Boone.) dari Lampung, Gresik dan Pontianak

Bioma Berkala Ilmiah Biologi ◽

10.14710/bioma.16.1.18-25 ◽

2014 ◽

Vol 16 (1) ◽

pp. 18

Author(s):

Yason Lukman Sudjito ◽

Christina Retna Handayani ◽

Hermin Pancasakti Kusumaningrum ◽

Anto Budiharjo

Keyword(s):

Genetic Variation ◽

Amino Acid ◽

Rna Polymerase ◽

Litopenaeus Vannamei ◽

Genetic Characterization ◽

Rdrp Gene ◽

Rt Pcr ◽

Infectious Myonecrosis Virus

A massive death of vannamei shrimp (Litopenaeus vannamei Boone.) due to Infectious Myonecrosis Virus (IMNV) infection has occurred in Indonesia recently and still cannot be eradicated efficiently. The fast reproduction of IMNV depends on the RdRp gene that encodes for RNA polymerase. Genetic characterization of IMNV RdRp gene from Indonesia is important in order to compare with other IMNV to find out genetic variation as a base for combating this virus. IMNV-infected vannamei were taken from major aquaculture region in Indonesia (Lampung, Gresik and Pontianak). RNA polymerase coding genes (12 and 13 region) from infected vannamei were amplified using RT-PCR with appropriate primer. Amplification products were sequenced and the results were analyzed using BioEdit 7.1.3.0, ClustalW2, CLC free workbench 6.6.2. and ClustalX programs. Results showed that homology value of IMNV RdRp gene from Lampung and Gresik were 98,04-9958% compared with IMNV from Brazil (Acc. No. AY570982). Amino acid analysis revealed homology value of IMNV RdRp gene from Lampung and Gresik were 100% and 99.04% compared with IMNV from Brasil. IMNV RdRp gene from Pontianak cannot be analysed due to low quality of RNA. Key words: vannamei, IMNV, RdRp, genetic characterization

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Biological and Molecular Characterization of the Cucurbit aphid-borne yellows virus Affecting Cucurbits in Tunisia

Plant Disease ◽

10.1094/pdis-93-10-1065 ◽

2009 ◽

Vol 93 (10) ◽

pp. 1065-1072 ◽

Cited By ~ 22

Author(s):

M. Mnari-Hattab ◽

N. Gauthier ◽

A. Zouba

Keyword(s):

Amino Acid ◽

Production Systems ◽

Citrullus Lanatus ◽

Aphis Gossypii ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Sequence Comparisons ◽

Virus Identification ◽

Large Populations

Surveys of yellowing viruses under nonheated and geothermal heated plastic tunnels and in open field crops of melon (Cucumis melo), cucumber (C. sativus), zucchini (Cucurbita pepo), squash (C. maxima), watermelon (Citrullus lanatus), and ware cucurbit (Ecballium elaterium) were carried out year-round during 2000–2001, 2003, and 2004 in the major cucurbit-growing areas in Tunisia. Severe yellowing symptoms on older leaves of cucurbits were observed in open fields and under plastic-tunnel production systems. These yellowing symptoms and large populations of aphids (Aphis gossypii) on a diversity of cucurbit crops in Tunisia support the hypothesis of a viral cause of the disease. Virus identification using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), followed by reverse transcription–polymerase chain reaction (RT-PCR) and immunocapture (IC)-RT-PCR showed that Cucurbit aphid-borne yellows virus (CABYV) was largely distributed in melon, cucumber, zucchini, squash, and watermelon crops. Ware cucurbit (E. elaterium) and lettuce (Lactuca sativa) crops were identified as potential CABYV reservoirs. The RT-PCR-amplified partial coat protein (CP) and P4 genes were cloned and sequenced from nine Tunisian CABYV isolates. CP and P4 gene nucleotide and amino acid sequence comparisons as well as phylogenetic reconstructions showed that the Tunisian isolates clustered into two major subgroups. Comparisons with CABYV sequences retrieved from GenBank showed high nucleotide and CP amino acid identities, and close relationships of the Tunisian isolates with Italian and French isolates.

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First Report on the Occurrence of Grapevine leafroll-associated virus 7 and 9 in Chilean Grapevines

Plant Disease ◽

10.1094/pdis-92-8-1252c ◽

2008 ◽

Vol 92 (8) ◽

pp. 1252-1252 ◽

Cited By ~ 10

Author(s):

E. A. Engel ◽

P. Escobar ◽

C. Montt ◽

S. Gómez-Talquenca ◽

P. D. T. Valenzuela

Keyword(s):

Amino Acid ◽

Mixed Infection ◽

Amino Acid Identity ◽

Microarray Hybridization ◽

Rt Pcr ◽

Specific Primers ◽

Hybridization Pattern ◽

Acid Identity ◽

Pcr Product ◽

Plant Pathol

Grapevine is one of the oldest horticultural crops and represents a highly valuable agricultural commodity. So far, nine distinct Grapevine leafroll-associated viruses (GLRaVs) within the Closteroviridae family have been found to be associated with grapevine leafroll disease (3). Previous studies have demonstrated a high incidence of GLRaV-1, -2, and -3 in Chile (2). To determine if other GLRaVs were present, 21 dormant cane samples were screened with a comprehensive 70-mer oligonucleotide microarray designed to simultaneously detect all grapevine viruses with total or partial genomic sequence available. The array contained 570 unique probes designed against specific regions of more than 40 viral genomes (E. Engel et al., 15th ICVG [Abstr.], 2006). One sample (cv. Black Seedless) showing a microarray hybridization pattern compatible with a mixed infection of GLRaV-7 and GLRaV-1 was analyzed by ELISA using GLRaV-7 specific antibodies (Agritest, Valenzano, Italy) and reverse transcription (RT)-PCR using virus-specific primers LR7-F: 5′- TAT ATC CCA ACG GAG ATG GC -3′ and LR7-R: 5′- ATG TTC CTC CAC CAA AAT CG -3′ (based on GenBank Accession No. Y15987). The serological analysis confirmed the presence of GLRaV-7 with further confirmation by the RT-PCR product of 502 bp corresponding to a fragment of the HSP70h gene that was cloned and sequenced. The Chilean GLRaV-7 sequence (GenBank Accession No. EU334662) showed 94% nucleotide and 95% amino acid identity when compared with a corresponding region of another GLRaV-7 isolate from Albania (GenBank Accession No. Y15987). GLRaV-1 infection was confirmed by ELISA (Bioreba AG, Reinach, Switzerland) and RT-PCR. A second sample (cv. Tintorera) showing microarray hybridization pattern compatible with a mixed infection of GLRaV-9 and Grapevine virus A (GVA) was analyzed by RT-PCR using virus-specific primers LR9-F: 5′- CGG CAT AAG AAA AGA TGG CAC -3′ and LR9-R: 5′- TCA TTC ACC ACT GCT TGA AC -3′ (1). The RT-PCR product of 393 bp corresponding to a fragment of the HSP70h gene was cloned and sequenced (GenBank Accession No. EU334663), showing 94% nucleotide and 95% amino acid identity when compared with a corresponding region of another GLRaV-9 isolate from the United States (GenBank Accession No. AY297819). Since there are no commercial antibodies available for GLRaV-9 detection, a second pair of primers, LR9-F1: 5′- AAA GGT TTC TGC TGG TTA CC -3′ and LR9-R1: 5′- CTT TCA GAA CAG TCC TCC TC -3′ that amplified a fragment of ORF1a was also used. The 301-bp product was cloned and sequenced (GenBank Accession No. EU588989) showing 93.7% nucleotide and 98% amino acid identity when compared with a corresponding region of another GLRaV-9 isolate (GenBank Accession No. AY297819). GVA infection was confirmed by ELISA (Bioreba AG) and RT-PCR. To our knowledge, this is the first report of GLRaV-7 and GLRaV-9 in Chile. Further studies will help determine the effect and incidence of these viruses in Chilean grapevines. References: (1) R. Alkowni et al. J. Plant Pathol. 86:123, 2004. (2) N. Fiore et al. J. Plant Pathol. 90:125, 2008. (3) G. P. Martelli and E. Boudon-Padieu. Options Méditerr. B55, 2006.

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Species-specific RT-PCR amplification of human enteroviruses: a tool for rapid species identification of uncharacterized enteroviruses

Journal of General Virology ◽

10.1099/vir.0.81179-0 ◽

2006 ◽

Vol 87 (1) ◽

pp. 119-128 ◽

Cited By ~ 142

Author(s):

M. Steven Oberste ◽

Kaija Maher ◽

Alford J. Williams ◽

Naomi Dybdahl-Sissoko ◽

Betty A. Brown ◽

...

Keyword(s):

Pcr Amplification ◽

Pcr Primers ◽

Human Enterovirus ◽

Rt Pcr ◽

Specific Primer ◽

Specific Primers ◽

Typical Application ◽

Partial Sequencing ◽

Specific Pcr ◽

Species Specific

The 65 serotypes of human enteroviruses are classified into four species, Human enterovirus (HEV) A to D, based largely on phylogenetic relationships in multiple genome regions. The 3′-non-translated region of enteroviruses is highly conserved within a species but highly divergent between species. From this information, species-specific RT-PCR primers were developed that can be used to rapidly screen collections of enterovirus isolates to identify species of interest. The four primer pairs were 100 % specific when tested against enterovirus prototype strains and panels of isolates of known serotype (a total of 193 isolates). For evaluation in a typical application, the species-specific primers were used to screen 186 previously uncharacterized non-polio enterovirus isolates. The HEV-B primers amplified 68·3 % of isolates, while the HEV-A and HEV-C primers accounted for 9·7 and 11·3 % of isolates, respectively; no isolates were amplified with the HEV-D primers. Twelve isolates (6·5 %) were amplified by more than one primer set and eight isolates (4·3 %) were not amplified by any of the four primer pairs. Serotypes were identified by partial sequencing of the VP1 capsid gene, and in every case sequencing confirmed that the species-specific PCR result was correct; the isolates that were amplified by more than one species-specific primer pair were mixtures of two (11 isolates) or three (one isolate) species of viruses. The eight isolates that were not amplified by the species-specific primers comprised four new serotypes (EV76, EV89, EV90 and EV91) that appear to be unique members of HEV-A based on VP1, 3D and 3′-non-translated region sequences.

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A Description of the Possible Etiology of the Cilantro Yellow Blotch Disease

Plant Disease ◽

10.1094/pdis-09-19-1958-sc ◽

2020 ◽

Vol 104 (3) ◽

pp. 630-633

Author(s):

Olufemi J. Alabi ◽

Brianna C. Gaytán ◽

Maher Al Rwahnih ◽

Cecilia Villegas

Keyword(s):

High Throughput Sequencing ◽

Disease Incidence ◽

Natural Infection ◽

Alfalfa Mosaic Virus ◽

Leaf Tissue ◽

Rt Pcr ◽

Specific Primers ◽

Tissue Samples ◽

Pcr Assays ◽

Hidalgo County

A virus-like disease characterized by foliar yellow blotch symptoms and resembling those described for cilantro yellow blotch disease in California was observed in a 4.05-ha cilantro (Coriandrum sativum) cv. Santo field in Hidalgo County, Texas during spring 2019. Disease incidence at harvest was estimated at ∼20%, and the affected plants were rendered unmarketable. Foliar systemic chlorosis symptoms were observed on sap-inoculated Nicotiana occidentalis plants (n = 3) using inocula from symptomatic cilantro. Total RNA aliquots from 11 randomly collected leaf tissue samples (symptomatic = 7, asymptomatic = 4) were pooled into a composite cilantro RNA sample which was analyzed by high throughput sequencing (HTS). Analyses of the obtained 15.7 million raw reads (76 nt each) yielded virus-specific contigs that mapped to the genomes of alfalfa mosaic virus (AMV), beet pseudoyellows virus (BPYV), and lettuce chlorosis virus (LCV). Virus-specific primers designed from the HTS-derived sequences were used to screen the samples in two-step RT-PCR assays, resulting in the detection of AMV+BPYV in 3 of 7 symptomatic cilantro samples, AMV+LCV in 4 of 7 symptomatic cilantro samples, and AMV alone in the 4 asymptomatic cilantro and sap-inoculated N. occidentalis samples. The results represent the first reports of the natural infection of cilantro by BPYV and LCV and implicate the mixed infection of a Crinivirus and AMV in cilantro yellow blotch disease.

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