First Report of Beet virus Q on Sugar Beet in Poland

The objective of this work was to determine whether Beet virus Q (BVQ), a member of the genus Pomovirus, is present in Poland. BVQ, like Beet necrotic yellow vein virus (BNYVV), is transmitted by Polymyxa betae Keskin. Earlier, BVQ was described as the Wierthe serotype of Beet soilborne virus (BSBV). Now, on the basis of its genomic properties (2), BVQ is recognized as a distinct virus species. BVQ is often found in fields where BSBV and BNYVV are present (4). During the fall of 2005, five plants of a cultivar susceptible to rhizomania (cv. Alyssa) and five resistant to rhizomania (cv. Henrietta) were collected from a field in the Wielkopolska Region of Poland, where BSBV and BNYVV had been previously identified, and tested for BVQ (1). All samples were analyzed by a double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) with antiserum against BNYVV (Bio-Rad, Hercules, CA). Rhizomania was identified only in sugar beet samples of the susceptible variety. The same samples were then tested using a triple antibody sandwich (TAS)-ELISA with commercial antisera against BSBV/BVQ (As-0576.2) and BSBV (As-0576.1) (DSMZ, Braunschweig, Germany). Nine sugar beet plants gave positive reactions with antiserum against BSBV/BVQ and negative reactions with antiserum specific to BSBV. Total RNA extracted from roots of 10 beet samples was then tested using a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) and specific primers designed to amplify a fragment of the RNA2 for BNYVV and BVQ (3). The primers specifically amplified fragments of 545 bp and 291 bp of the BNYVV and BVQ, respectively. BNYVV was detected in all five samples from susceptible sugar beet plants. The presence of BVQ was confirmed in nine of the sugar beer plants, and the RT-PCR products were sequenced. Sequence analysis of the 206-nt amplicon sequence of the Polish isolate of BVQ (GenBank Accession No. DQ309444) indicated 97% nucleotide and 94% amino acid sequence identity with the previously published sequence of BVQ (GenBank Accession No. AJ223596) (2). To my knowledge, this is the first report of the natural occurrence of BVQ on sugar beet in Poland. In Europe, it has been previously reported in Belgium, Bulgaria, France, Germany, Hungary, Italy, the Netherlands, Spain, and Sweden (3,4). References: (1) N. Borodynko et al. Plant Dis. 90:112, 2006. (2) R. Koenig et al. J. Gen. Virol. 79:2027, 1998. (3) A. Meunier et al. Appl. Environ. Microbiol. 69:2356, 2003. (4) C. Rubies Autonell et al. Plant Dis. 90:110, 2006.

Download Full-text

First Report of Beet virus Q in Spain

Plant Disease ◽

10.1094/pd-90-0110b ◽

2006 ◽

Vol 90 (1) ◽

pp. 110-110 ◽

Cited By ~ 3

Author(s):

C. Rubies Autonell ◽

C. Ratti ◽

R. Resca ◽

M. De Biaggi ◽

J. Ayala García

Keyword(s):

Sugar Beet ◽

Soil Samples ◽

Root Tissue ◽

Research Centre ◽

Enzyme Linked Immunosorbent Assay ◽

Polymyxa Betae ◽

Virus Species ◽

Rt Pcr ◽

First Report ◽

Beet Virus Q

Beet virus Q (BVQ) is a member of the genus Pomovirus that is transmitted by Polymyxa betae Keskin. Initially described as the Wierthe serotype of Beet soilborne virus (BSBV), BVQ is now considered a distinct virus species based on its genomic properties (1). BVQ is commonly found in fields where BSBV and the causal agent of rhizomania disease, Beet necrotic yellow vein virus (BNYVV), are also present. Simultaneous infection of sugar beet plants with multiple virus species could affect disease symptom expression (4). For this reason, the pathogenicity of BVQ and its role in the epidemiology of rhizomania disease remain a subject of study. During 2004, six soil samples were collected from different sites in the Castilla-La Mancha Region in Spain (Albacete and Ciudad Real provinces) where rhizomania symptoms were observed in BNYVV-tolerant sugar beet cultivars. Soil from the Hainaut Region of Belgium, infected with BNYVV, BSBV, and BVQ and supplied by Prof. C. Bragard (Unité de Phytopathologie, Université Catholique de Louvain, Belgium) was used as a positive control. Sugar beet plants (cv. Asso) were grown in the soil samples for 45 days at 24°C and then root tissue was harvested. All samples were analyzed using enzyme-linked immunosorbent assay (ELISA) with commercial BNYVV antiserum (BIOREBA AG, Reinach, Switzerland) and BSBV/BVQ antisera (IC10 and 6G2) supplied by R. Koenig (Federal Biological Research Centre for Agriculture and Forestry, Braunschweig, Germany). Total RNA extracted from sugar beet roots as previously described (3) was tested using reverse transcription-polymerase chain reaction (RT-PCR). Primers BVQ3F (5′-GTT TTC AAA CTT GCC ATC CT-3′) and BVQ3R2 (5′-CCA CAA TGG GCC AAT AGA-3′), which amplify a 690-bp fragment of the triple gene block region of BVQ RNA 3, were designed based on the published sequence (GenBank Accession No. AJ223598). The presence of BSBV and BNYVV was assayed using RT-PCR with previously described primers (2,3). BVQ was detected from plants grown in soil collected from La Roda (Albacete) in Spain and from Hainaut in Belgium. The fragments amplified from Spanish sample with BVQ3F and BVQ3R2 (GenBank Accession No. AY849375) showed 95.9% nucleotide sequence identity with the previously published sequence of BVQ (1). The La Roda BVQ isolate was mechanically transmitted to Chenopodium quinoa from infected sugar beet root tissue. BVQ was detected using RT-PCR in local lesions that appeared approximately 5 days after inoculation and subsequently spread along veins. To our knowledge, this is the first report of BVQ in soil from Spain, although it has been previously reported in Belgium, Bulgaria, France, Germany, Hungary, and the Netherlands (2). BSBV and BNYVV (type A) were detected in all six Spanish samples, as well as in the Belgian soil. References: (1) R. Koenig et al. J. Gen. Virol. 79:2027, 1998. (2) A. Meunier et al. Appl. Environ Microbiol. 69:2356, 2003. (3) C. Ratti et al. J. Virol. Methods 124:41, 2005. (4) C. Rush Annu. Rev Phytopathol 41:567, 2003.

Download Full-text

First Report of Beet virus Q on Sugar Beet in Iran

Plant Disease ◽

10.1094/pd-89-1359b ◽

2005 ◽

Vol 89 (12) ◽

pp. 1359-1359 ◽

Cited By ~ 2

Author(s):

Sh. Farzadfar ◽

R. Pourrahim ◽

A. R. Golnaraghi ◽

A. Ahoonmanesh

Keyword(s):

Sugar Beet ◽

Type A ◽

Single Strand Conformation Polymorphism ◽

Natural Occurrence ◽

Polymyxa Betae ◽

Rt Pcr ◽

First Report ◽

Beet Root ◽

Almost All ◽

Beet Virus Q

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.

Download Full-text

First Report of Beet soilborne virus in Poland

Plant Disease ◽

10.1094/pd-90-0112b ◽

2006 ◽

Vol 90 (1) ◽

pp. 112-112 ◽

Cited By ~ 2

Author(s):

N. Borodynko ◽

B. Hasiów ◽

H. Pospieszny

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Sugar Beet ◽

Amino Acid Sequence Identity ◽

Chenopodium Quinoa ◽

Enzyme Linked Immunosorbent Assay ◽

First Report ◽

Sequence Identity ◽

Pcr Products ◽

Soilborne Viruses

Beet necrotic yellow vein virus (BNYVV), the casual agent of rhizomania disease, was identified in sugar beet plants from several fields in the Wielkopolska Region of Poland (1). In greenhouse studies, sugar beets were grown in the soil from one of these fields to bait soilborne viruses. Of 200 sugar beet plants, three developed symptoms of vein clearing, vein banding, and mosaic. Crude sap from symptomatic plants was used for mechanical inoculation of various plants species. In Chenopodium quinoa, C. amaranticolor, and Tetragonia expansa only local lesions were observed. Electron microscope examination of negatively stained leaf-dip preparations from symptomatic sugar beet plants showed a mixture of rod-shape particles from 70 to 400 nm long. Using double-antibody sandwich enzyme-linked immunosorbent assay tests, two symptomatic sugar beet plants gave positive reactions with antiserum against BNYVV (Bio-Rad, Hercules, CA) and a third plant gave a positive reaction with antisera against BNYVV and Beet soilborne virus (BSBV). Total RNA was extracted from roots and leaves of the symptomatic plants and used in a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay. Specific primers were designed to amplify a fragment of the RNA1 for BSBV and RNA2 for BNYVV and Beet virus Q (BVQ) (2). Two mRT-PCR products amplified with the primers specific to BNYVV and BSBV were obtained and sequenced. A 274-nt amplicon sequence (GenBank Accession No. DQ012156) had 98% nucleotide sequence identity with the German BNYVV isolate F75 (GenBank Accession No. AF19754) and a 376-nt amplicon sequence (GenBank Accession No. AY999690) had 98% nucleotide and 98% amino acid sequence identity with the German BSBV isolate (GenBank Accession No. Z97873). The Polish BSBV isolate had 88% nucleotide and 62% amino acid sequence identity with BVQ, another pomovirus (GenBank Accession No. AJ 223596 formerly known as serotype Wierthe of BSBV (2). In 2005, mRT-PCR was used on samples collected from two fields of the Wielkopolska Region. Of 15 tested sugar beet plants, 12 gave positive reactions with primers specific for BSBV and nine with primers specific to BNYVV. To our knowledge, this is first report of BSBV in Poland. In Europe, BSBV was previously reported in England, the Netherlands, Belgium, Sweden, Germany, France, and Finland (2,3). References: (1) M. Jezewska and J. Piszczek. Phytopathol. Polonica, 21:165, 2001. (2) A. Maunier et al. Appl. Environ. Microbiol. 69:2356, 2003. (3) C. M. Rush and G. B. Heidel. Plant Dis. 79:868, 1995.

Download Full-text

Multiplex Reverse Transcription-PCR for Simultaneous Detection of Beet Necrotic Yellow Vein Virus, Beet Soilborne Virus, and Beet Virus Q and Their Vector Polymyxa betae KESKIN on Sugar Beet

Applied and Environmental Microbiology ◽

10.1128/aem.69.4.2356-2360.2003 ◽

2003 ◽

Vol 69 (4) ◽

pp. 2356-2360 ◽

Cited By ~ 51

Author(s):

Alexandre Meunier ◽

Jean-François Schmit ◽

Arnaud Stas ◽

Nazli Kutluk ◽

Claude Bragard

Keyword(s):

Sugar Beet ◽

Reverse Transcription ◽

Detection Threshold ◽

Simultaneous Detection ◽

Yellow Vein ◽

Enzyme Linked Immunosorbent Assay ◽

Polymyxa Betae ◽

Reverse Transcription Pcr ◽

Soilborne Viruses ◽

Beet Virus Q

ABSTRACT Three soilborne viruses transmitted by Polymyxa betae KESKIN in sugar beet have been described: Beet necrotic yellow vein virus (BNYVV), the agent of rhizomania, Beet soilborne virus (BSBV), and Beet virus Q (BVQ). A multiplex reverse transcription-PCR technique was developed to simultaneously detect BNYVV, BSBV, and BVQ, together with their vector, P. betae. The detection threshold of the test was up to 128 times greater than that of an enzyme-linked immunosorbent assay. Systematic association of BNYVV with one or two different pomoviruses was observed. BVQ was detected in samples from Belgium, Bulgaria, France, Germany, Hungary, Italy, Sweden, and The Netherlands but not in samples from Turkey.

Download Full-text

First report of Beet virus Q in Belgium

Plant Disease ◽

10.1094/pdis.2001.85.12.1288b ◽

2001 ◽

Vol 85 (12) ◽

pp. 1288-1288 ◽

Cited By ~ 5

Author(s):

A. Stas ◽

A. Meunier ◽

J.-F. Schmit ◽

C. Bragard

Keyword(s):

Sugar Beet ◽

Blue Staining ◽

Enzyme Linked Immunosorbent Assay ◽

Polymyxa Betae ◽

Rt Pcr ◽

Economic Significance ◽

Pcr Product ◽

Bait Plants ◽

Soilborne Viruses ◽

Beet Virus Q

Beet necrotic yellow vein virus (BNYVV), the causal agent of rhizomania disease on sugar beet, has been reported in Belgium for more than 16 years. Other soilborne viruses belonging to the genus Pomovirus, such as Beet soilborne virus (BSBV) (3) and Beet virus Q (BVQ) (1), are suspected pathogens of sugar beets grown in Belgium. During the 2000 growing season, more than 20 fields showing rhizomania-like and yellowing symptoms on sugar beet leaves were investigated for the presence of BVQ, BNYVV, and BSBV. All samples were checked by enzyme-linked immunosorbent assay (ELISA) using commercial BNYVV (Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France) and BSBV/BVQ (DSMZ, Braunschweig, Germany - AS-0576 polyclonal, AS-0576/2 MAb) antisera. RNA was extracted from sugar beet rootlets using an RNeasy extraction kit (Qiagen, Hilden, Germany), before performing a reverse transcription-polymerase chain reaction (RT-PCR) using primers (5′-GCTGGAGTATATCACCGATGAC-3′ and 5′-AAAATC TCGGATAGCATCCAAC-3′) designed to specifically amplify a 510-bp region of BVQ RNA-1. The presence of BSBV and BNYVV was also checked by RT-PCR using previously described primers (1,2). The BVQ-derived PCR product was sequenced and proved to be more than 99% identical to the Wierthe BVQ isolate nucleotide sequence. Soil transmission of BVQ was demonstrated through a bioassay using soil dilutions with quartz and sugar beet cv. Cadyx as bait. After 6 weeks, BVQ was detected by RT-PCR in bait plants. The putative vector, Polymyxa betae, was identified by lactophenol-cotton blue staining of the roots followed by microscopic examination. BVQ produces irregularly shaped local lesions that appear ≈5 days after mechanical inoculation and tend to spread along veins. BVQ was detected in six fields located in the Polders Region and Brabant Province of Belgium. BVQ was always found in sugar beet samples coinfected with BNYVV and BSBV. The economic significance of BVQ and its interaction with other viruses is not known. References: (1) R. Koenig et al. J. Gen. Virol. 79:2027, 1998. (2) M. Saito et al. Arch. Virol. 141:2163, 1996. (3) M. Verhoyen and M. Van den Bossche. Parasitica. 44:71, 1987.

Download Full-text

First Report on the Natural Occurrence of Beet chlorosis virus in Poland

Plant Disease ◽

10.1094/pdis-91-3-0326c ◽

2007 ◽

Vol 91 (3) ◽

pp. 326-326 ◽

Cited By ~ 2

Author(s):

A. Kozlowska-Makulska ◽

M. S. Szyndel ◽

J. Syller ◽

S. Bouzoubaa ◽

M. Beuve ◽

...

Keyword(s):

Sugar Beet ◽

Beta Vulgaris ◽

Amino Acid Sequences ◽

Enzyme Linked Immunosorbent Assay ◽

Natural Occurrence ◽

Rt Pcr ◽

First Report ◽

Beet Western Yellows Virus ◽

Access Period ◽

Commercial Sugar

Yellowing symptoms on sugar beet (Beta vulgaris L.) are caused by several viruses, especially those belonging to the genus Polerovirus of the family Luteoviridae, including Beet mild yellowing virus (BMYV) and Beet western yellows virus (BWYV), and recently, a new species, Beet chlorosis virus (BChV), was reported (2). To identify Polerovirus species occurring in beet crops in Poland and determine their molecular variability, field surveys were performed in the summer and autumn of 2005. Leaves from symptomatic beet plants were collected at 26 localities in the main commercial sugar-beet-growing areas in Poland that included the Bydgoszcz, Kutno, Lublin, Poznań, Olsztyn, and Warszawa regions. Enzyme-linked immunosorbent assay (ELISA) tests (Loewe Biochemica GmbH, Sauerlach, Germany) detected poleroviruses in 23 of 160 samples (approximately 20 samples from each field). Multiplex reverse-transcription polymerase chain reaction (RT-PCR) (1) (GE Healthcare S.A.-Amersham Velizy, France) confirmed the presence of poleroviruses in 13 of 23 samples. Nine of twenty sugar beet plants gave positive reactions with BChV-specific primers and three with primers specific to the BMYV P0 protein. Two isolates reacted only with primer sets CP+/CP, sequences that are highly conserved for all beet poleroviruses. Leaf samples collected from three plants infected with BChV were used as inoculum sources for Myzus persicae in transmission tests to suitable indicator plants including sugar beet, red beet (Beta vulgaris L. var. conditiva Alef.), and Chenopodium capitatum. All C. capitatum and beet plants were successfully infected with BChV after a 48-h acquisition access period and an inoculation access period of 3 days. Transmission was confirmed by the presence of characteristic symptoms and by ELISA. Amino acid sequences obtained from each of four purified (QIAquick PCR Purification kit, Qiagen S.A., Courtaboeuf, France) RT-PCR products (550 and 750 bp for CP and P0, respectively) were 100% identical with the CP region (GenBank Accession No. AAF89621) and 98% identical with the P0 region (GenBank Accession No. NP114360) of the French isolate of BChV. To our knowledge, this is the first report of BChV in Poland. References: (1) S. Hauser et al. J. Virol. Methods 89:11, 2000. (2) M. Stevens et al. Mol. Plant Pathol. 6:1, 2005.

Download Full-text

First Report of Barley yellow mosaic virus in Barley in Spain

Plant Disease ◽

10.1094/pd-89-0105a ◽

2005 ◽

Vol 89 (1) ◽

pp. 105-105 ◽

Cited By ~ 3

Author(s):

M. A. Achon ◽

M. Marsiñach ◽

C. Ratti ◽

C. Rubies-Autonell

Keyword(s):

Mosaic Virus ◽

Yellow Mosaic Virus ◽

Complete Agreement ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

First Report ◽

Barley Yellow Mosaic Virus ◽

Pcr Products ◽

Mild Mosaic Virus ◽

Polymerase Chain

Recently, the presence of Barley mild mosaic virus (BaMMV) and the weakly serological detection of Barley yellow mosaic virus (BaYMV) were reported in Spain (1); both viruses are members of the genus Bymovirus (family Potyviridae). Random and symptomatic surveys were conducted during February and March of 2003 in barley fields in northeastern Spain to determine the occurrence of BaMMV and BaYMV. Leaves from 316 samples collected in 15 fields were analyzed using enzyme-linked immunosorbent assay (ELISA) with commercial antisera specific for BaYMV and BaMMV (Loewe Biochemica, Munich) as well as antisera against both viruses (provided by T. Klumen). Positive ELISA samples were further analyzed using reverse transcription-polymerase chain reaction (RT-PCR) with specific primers that amplify 445 bp of BaMMV and 433 bp of BaYMV (2). Complete agreement was observed between the ELISA and RT-PCR results. Mixed infections of BaYMV and BaMMV were detected in 10 samples, BaYMV in 5 samples and BaMMV in 3 samples. Samples positive for both viruses that exhibited clear mosaic symptoms were collected in two fields. RT-PCR products from five BaYMV-infected samples were cloned and sequenced and showed 96 to 98% identity to BaYMV isolates previously reported from Europe (Genbank Accession Nos. AJ1515479-85 and X95695-7) and 92 to 95% identity with isolates reported from Asia (GenBank Accession Nos. AB023585-96, AJ132268, AJ224619-22, AJ224624-28, AF536944-46, AF536948-58, D01091, D00544, and Z24677). Sequence identity of Spanish isolates were 96 to 99%. To our knowledge, this is the first report of BaYMV infecting barley in Spain and illustrates the association of both Bymoviruses infecting barley. References: (1) M. A. Achon et al. Plant Dis. 87:1004, 2003. (2) D. Hariri et al. Eur. J. Plant Pathol. 106:365, 2000.

Download Full-text

A PCR-Based Method of Detection and Differentiation of K88+ Adhesive Escherichia Coli

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879600800410 ◽

1996 ◽

Vol 8 (4) ◽

pp. 460-463 ◽

Cited By ~ 14

Author(s):

Mark A. Franklin ◽

David H. Francis ◽

Diane Baker ◽

Alan G. Mathew

Keyword(s):

Escherichia Coli ◽

Enzyme Linked Immunosorbent Assay ◽

Antigenic Variant ◽

Variable Regions ◽

E Coli ◽

Clinical Assay ◽

Pcr Product ◽

Structural Subunit ◽

Pcr Products ◽

Polymerase Chain

The objective of this study was to develop a polymerase chain reaction (PCR)-based method to detect and differentiate among Escherichia coli strains containing genes for the expression of 3 antigenic variants of the fimbrial adhesin K88 (K88ab, K88ac, and K88ad). Five primers were designed that allowed detection of K88+ E. coli, regardless of antigenic variant, and the separate detection of the ab, ac, and ad variants. Primers AM005 and AM006 are 21 base pair (bp) oligomers that correspond to a region of the K88 operon that is common to all 3 antigenic variants. Primers MF007, MF008, and MF009 are 24-bp oligomers that matched variable regions specific to ab, ac, and ad, respectively. Using primers AM005 and AM006, a PCR product was obtained that corresponds to a 764-bp region within the large structural subunit of the K88 operon common to all 3 antigenic variants. Primer AM005 used with MF007, MF008, or MF009 produced PCR products approximately 500-bp in length from within the large structural subunit of the K88 operon of the 3 respective antigenic variants. Fragments were identified by rates of migration on a 1% agarose gel relative to each other as well as to BstEII-digested lambda fragments. This PCR-based method was comparable to the enzyme-linked immunosorbent assay and western blot test in the ability to differentiate between the antigenic variants. K88+ E. coli were differentiated from among laboratory strains and detected in ileal samples taken from cannulated pigs challenged with a known K88+ variant. K88+ E. coli were also detected from fecal swabs taken from newly weaned pigs, thus confirming that this PCR-based test could provide a convenient clinical assay for the detection of K88+ E. coli. Detection and differentiation of K88+ E. coli using general and specific primers was successful. PCR methods of detection should permit identification of K88+ antigenic variants regardless of the level of expression of the antigen.

Download Full-text

First Report of a Group 16SrII Phytoplasma Infecting Chickpea in Oman

Plant Disease ◽

10.1094/pd-90-0973c ◽

2006 ◽

Vol 90 (7) ◽

pp. 973-973 ◽

Cited By ~ 8

Author(s):

N. A. Al-Saady ◽

A. M. Al-Subhi ◽

A. Al-Nabhani ◽

A. J. Khan

Keyword(s):

Nested Pcr ◽

Animal Feed ◽

Restriction Enzymes ◽

Universal Primers ◽

Polymorphism Analysis ◽

Disease Symptoms ◽

First Report ◽

Pcr Products ◽

Polymerase Chain ◽

Group 2

Chickpea (Cicer arietinum), locally known as “Dungo”, is grown for legume and animal feed mainly in the interior region of Oman. During February 2006, survey samples of chickpea leaves from plants showing yellows disease symptoms that included phyllody and little leaf were collected from the Nizwa Region (175 km south of Muscat). Total nucleic acid was extracted from asymptomatic and symptomatic chickpea leaves using a cetyltrimethylammoniumbromide method with modifications (3). All leaf samples from eight symptomatic plants consistently tested positive using a polymerase chain reaction assay (PCR) with phytoplasma universal primers (P1/P7) that amplify a 1.8-kb phytoplasma rDNA product and followed by nested PCR with R16F2n/R16R2 primers yielding a product of 1.2 kb (2). No PCR products were evident when DNA extracted from healthy plants was used as template. Restriction fragment length polymorphism analysis of nested PCR products by separate digestion with Tru9I, HaeIII, HpaII, AluI, TaqI, HhaI, and RsaI restriction enzymes revealed that a phytoplasma belonging to group 16SrII peanut witches'-broom group (2) was associated with chickpea phyllody and little leaf disease in Oman. Restriction profiles of chickpea phytoplasma were identical with those of alfalfa witches'-broom phytoplasma, a known subgroup 16SrII-B strain (3). To our knowledge, this is the first report of phytoplasma infecting chickpea crops in Oman. References: (1) A. J. Khan et al. Phytopathology, 92:1038, 2002. (2). I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998 (3) M. A. Saghai-Maroof et al. Proc. Natl. Acad. Sci. USA. 81:8014, 1984.

Download Full-text

First Report of Rhizomania Disease of Sugar Beet Caused by Beet necrotic yellow vein virus in the Great Lakes Production Region

Plant Disease ◽

10.1094/pdis.2003.87.2.201c ◽

2003 ◽

Vol 87 (2) ◽

pp. 201-201 ◽

Cited By ~ 2

Author(s):

William M. Wintermantel ◽

Teresa Crook ◽

Ralph Fogg

Keyword(s):

Sugar Beet ◽

Great Lakes ◽

Beta Vulgaris ◽

Yellow Vein ◽

Enzyme Linked Immunosorbent Assay ◽

Great Lakes Region ◽

Polymyxa Betae ◽

Mechanical Inoculation ◽

Production Region ◽

Das Elisa

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV) and vectored by the soilborne fungus Polymyxa betae Keskin, is one of the most economically damaging diseases affecting sugar beet (Beta vulgaris L.). The virus likely originated in Europe and was first identified in California in 1983 (1). It has since spread among American sugar beet production regions in spite of vigorous sanitation efforts, quarantine, and disease monitoring (3). In the fall of 2002, mature sugar beet plants exhibiting typical rhizomania root symptoms, including proliferation of hairy roots, vascular discoloration, and some root constriction (2) were found in several fields scattered throughout central and eastern Michigan. Symptomatic beets were from numerous cultivars, all susceptible to rhizomania. Two to five sugar beet root samples were collected from each field and sent to the USDA-ARS in Salinas, CA for analysis. Hairy root tissue from symptomatic plants was used for mechanical inoculation of indicator plants. Mechanical inoculation produced necrotic lesions on Chenopodium quinoa and systemic infection of Beta vulgaris ssp. macrocarpa, both typical of BNYVV and identical to control inoculations with BNYVV. Symptomatic sugar beet roots were washed and tested using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) for the presence of BNYVV using standard procedures and antiserum specific for BNYVV (3). Sugar beet roots were tested individually, and samples were considered positive when absorbance values were at least three times those of greenhouse-grown healthy sugar beet controls. Samples were tested from 16 fields, with 10 confirmed positive for BNYVV. Positive samples had mean absorbance values ranging from 0.341 to 1.631 (A405nm) after 30 min. The mean healthy control value was 0.097. Fields were considered positive if one beet tested positive for BNYVV, but in most cases, all beets tested from a field were uniformly positive or uniformly negative. In addition, soil-baiting experiments were conducted on seven of the fields. Sugar beet seedlings were grown in soil mixed with equal parts of sand for 6 weeks and were subsequently tested using DAS-ELISA for BNYVV. Results matched those of the root sampling. Fields testing positive for BNYVV were widely dispersed within a 100 square mile (160 km2) area including portions of Gratiot, Saginaw, Tuscola, and Sanilac counties in the central and eastern portions of the Lower Peninsula of Michigan. The confirmation of rhizomania in sugar beet from the Great Lakes Region marks the last major American sugar beet production region to be diagnosed with rhizomania disease, nearly 20 years after its discovery in California (1). In 2002, there were approximately 185,000 acres (approximately 75,00 ha) of sugar beet grown in the Great Lakes Region, (Michigan, Ohio, and southern Ontario, Canada). The wide geographic distribution of infested fields within the Michigan growing area suggests the entire region should monitor for symptoms, increase rotation to nonhost crops, and consider planting rhizomania resistant sugar beet cultivars to infested fields. References:(1) J. E. Duffus et al. Plant Dis. 68:251, 1984. (2) J. E. Duffus. Rhizomania. Pages 29–30 in: Compendium of Beet Diseases and Insects, E. D. Whitney and J. E. Duffus eds. The American Phytopathological Society, St. Paul, MN, 1986. (3) G. C. Wisler et al. Plant Dis. 83:864, 1999.

Download Full-text