Use of Mycoplasmalike Organism (MLO) Group-Specific Oligonucleotide Primers for Nested-PCR Assays to Detect Mixed-MLO Infections in a Single Host Plant

Lee, I. M.; Gundersen, D. E.; Hammond, R. W.; Davis, R. E.

doi:10.1094/phyto-84-559

Cited by 340 publications

(245 citation statements)

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“…Total nucleic acids were extracted from dissected single specimens, dissolved in 40 µl TE buffer, and maintained at -20 °C. Following the modified protocol described by Angelini et al (2001), phytoplasma detection in insects was carried out by nested PCR assays with universal phytoplasma primer pair R16F2/R2 (Lee et al, 1993), followed by R16(I)F1/R1 primers (Lee et al, 1994) in nested PCR on amplicons obtained with R16F2/R2 primers diluted 1:30 with sterile distilled water. Each 25 μl PCR reaction mix contained 1 μl of template DNA diluted 1: 30 with sterile distilled water, 12.5 μl 2X PCR master mix (Fermentas, Lithuania), and 0.4 μM each primer.…”

Section: Nucleic Acid Extraction and Pcr/rflp Analysesmentioning

confidence: 99%

Leafhoppers and cixiids in phytoplasma-infected carrot fields: Species composition and potential phytoplasma vectors

et al. 2010

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SUMMARYThe first molecular analysis of samples collected in southern Bačka (Serbia) confirmed the presence of aster yellows (16SrI) and stolbur phytoplasmas (16SrXII) in insects belonging to the family Cicadellidae, as well as in carrot plants where the insects were collected. A correct identification of the phytoplasmas and their vectors is essential to arrange effective control strategies to prevent diseases associated with phytoplasmas from spreading to carrots and other vegetable crops. In order to enhance knowledge about insect vectors of aster yellows and stolbur phytoplasmas in Serbia, Cicadellidae and Cixiidae (Homoptera Auchenorrhyncha), the most common vectors of these phytoplasmas, were monitored in southern Bačka during 2008. Adults leaf-and planthoppers were collected and identified at species level using standard entomological methods, and tested for phytoplasma presence by means of PCR/RFLP. A total of 13 insect species of Cicadellidae were identified, as follows: a) three species of the subfamily Agallinae: Anaceratagallia ribauti (Ossiannilsson), Anaceratagallia venosa (Fourcroy), and Anaceratagallia laevis (Ribaut); b) seven species of the subfamily Deltocephalinae: Psammotettix confinis (Dahlbom), Psammotettix striatus (Linnaues) Psammottettix alienus (Dahlbom), Macrosteles sexnotatus (Fallén), Ophiola decumana (Kontkanen), Errastunus ocellaris Fallén, and Scaphoideus titanus Ball; c) three species of the subfamily Typhlocibinae: Eupteryx atropunctata (Goeze), Eupteryx mellissae Curtis, Zyginidia pullula (Boheman). Female specimens of the genus Euscelis (Deltocephalinae) were also collected,

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Section: Nucleic Acid Extraction and Pcr/rflp Analysesmentioning

confidence: 99%

Leafhoppers and cixiids in phytoplasma-infected carrot fields: Species composition and potential phytoplasma vectors

et al. 2010

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“…Various PCR-based assays have been developed to detect FDP. A widely applied procedure is based on direct PCR amplification with phytoplasma-universal primer pair P1/P7 (Schneider et al, 1995) followed by nested, 16S rDNA V group-specific primers (Lee et al, 1994). To achieve the identification of 16S rDNA V-C and -D subgroups, as FDPs belong to these subgroups (see section 3.1.1), a restriction fragment length polymorphism analysis has been proposed (Davis and Dally, 2001) and is routinely applied.…”

Section: Detection and Identification Of Flavescence Dorée Phytoplasmamentioning

confidence: 99%

Scientific Opinion on pest categorisation of Grapevine Flavescence dorée

Rossi¹

2014

EFSA Journal

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The Panel on Plant Health performed a pest categorisation of Grapevine Flavescence dorée, also known as Flavescence dorée phytoplasma (FDP), for the European Union (EU) territory. FDP has not yet been defined as a phytoplasma species of the genus Candidatus Phytoplasma. Routine molecular detection assays are available. FDP is transmitted by grafting and vegetative propagation material as well as by insect vectors. FDP is only reported in Europe. Within Europe it is present in Serbia, in Switzerland and in 10 out of 21 countries of the grapevine producing EU countries. Besides grapevine, FDP is also commonly found in other hosts such as Ailanthus, Alnus and Clematis. FDP is included in the annexes II/A/II and II/B of the Council Directive 2000/29/EC. Grapevine, the main host plant, is included into annexes III, IV and V of the Directive 2000/29/EC. FDP is not expected to be affected by EU ecoclimatic conditions wherever its hosts are present and has the potential to establish largely within the EU territory. The specific leafhopper vector of FDP, Scaphoideus titanus, is an invasive insect that was introduced in Europe. It is only in areas where FDP and Scaphoideus titanus are associated that the direct and indirect impacts are considered to be high: yield reduction, death of grapevine plants, costs for disease and vector management procedures. Additional insect vectors are known, but are not directly associated with FDP epidemics. A major outcome of this pest categorisation has been to emphasise the role of S. titanus. Uncertainty lies mostly in the knowledge on specific FDP strain transmissibility, susceptibility of specific grapevine varieties and distribution in alternate hosts. It is not fully known how far the invasive vector insect S. titanus is still enlarging its distribution within the risk assessment area.

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“…Current detection methods have been facilitated by the systematic use of polymerase chain reaction (PCR): both real-time PCR (Christensen et al, 2004) for broad-range phytoplasma detection and nested PCR detection using phytoplasma-universal primers targeting 16S rDNA (Lee et al, 1994, Daire et al, 1997Boudon-Padieu et al, 2004) combined with a targeted sequencing to verify the presence of the CPu signature sequence. Identification can also be achieved by the use of A B C D E restriction fragment length polymorphism (RFLP) or through multilocus sequence analysis (Arnaud et al, 2007;Jović et al, 2011), targeting other genes such as secY, rpV or FD9.…”

Section: Detection and Identification Of Candidatus Phytoplasma Ulmimentioning

confidence: 99%

Scientific Opinion on the pest categorisation of Elm phloem necrosis mycoplasm

Rossi¹

2014

EFSA Journal

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Use of Mycoplasmalike Organism (MLO) Group-Specific Oligonucleotide Primers for Nested-PCR Assays to Detect Mixed-MLO Infections in a Single Host Plant

Cited by 340 publications

References 0 publications

Leafhoppers and cixiids in phytoplasma-infected carrot fields: Species composition and potential phytoplasma vectors

Leafhoppers and cixiids in phytoplasma-infected carrot fields: Species composition and potential phytoplasma vectors

Scientific Opinion on pest categorisation of Grapevine Flavescence dorée

Scientific Opinion on the pest categorisation of Elm phloem necrosis mycoplasm

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