TaqMan real-time PCR versus four conventional PCR assays for detection of apple proliferation phytoplasma

Baric, Sanja; Kerschbamer, Christine; Via, Josef Dalla

doi:10.1007/bf02914056

Cited by 37 publications

(26 citation statements)

References 23 publications

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“…P. mali' in inoculated apple trees that did not show visible disease symptoms until October 2009, root samples were collected for DNA extraction. DNA was isolated from 100 mg dissected phloem tissue using the DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) (Baric et al 2006) and tested with a highly sensitive real-time PCR method (Baric & Dalla Via 2004). Infected trees treated with Dormex and the corresponding infected control trees were sampled in July and October 2009 as well as in July and October 2010 in order to be tested for an effect of cyanamide treatment on the phytoplasma titre.…”

Section: Biomass Of Symptomatic Vs Non-infected Apple Plantletsmentioning

confidence: 99%

Resistance Inducers and Plant Growth Regulators Show Only Limited and Transient Effects on Infection Rates, Growth Rates and Symptom Expression of Apple Trees Infected With ‘candidatus Phytoplasma mali’

Schmidt

Baric

et al. 2015

J Plant Dis Prot

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The effects of four commercially available bio-active compounds on the infection rates, symptom expression and growth rates of apple trees (Malus × domestica Borkh.) cv. Golden Delicious infected with 'Candidatus Phytoplasma mali' (the so-called Apple Proliferation phytoplasma or AP) were tested over a three-year period under controlled conditions. Post-infection treatments using Bion ® (active ingredient: Acibenzolar-S-Methyl), Messenger ® (Harpin protein), Regalis ® (Prohexadione-Ca) and Dormex ® (Cyanamide) had no significant effect on infection rates. Terminal growth of apple trees (grown as one-shoot pruned trees) was increased significantly by AP infection; Prohexadione-Ca was the only compound which had a significant (inhibiting) effect on the growth of both infected and non-infected apple trees. Acibenzolar-S-Methyl and Harpin had no significant effects on symptom expression. AP symptoms were masked during summer by Prohexadione-Ca, which caused severe growth abnormalities. Cyanamide changed the seasonal appearance of AP symptoms: while symptoms were delayed compared to the untreated control the first two years (2008 and 2009), symptoms appeared earlier the third year (2010). Differences in symptom expression levelled off later in the vegetative season, and no significant difference was found in October.

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Section: Biomass Of Symptomatic Vs Non-infected Apple Plantletsmentioning

confidence: 99%

Resistance Inducers and Plant Growth Regulators Show Only Limited and Transient Effects on Infection Rates, Growth Rates and Symptom Expression of Apple Trees Infected With ‘candidatus Phytoplasma mali’

Schmidt

Baric

et al. 2015

J Plant Dis Prot

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“…PCR with primer pair AP5/AP4 was found to be the least costly test assay. As real-time PCR assay has a high potential for automation, it may be more suitable for large-scale routine diagnosis of AP disease (Baric et al 2006).…”

Section: Real-time Polymerase Chain Reactionmentioning

confidence: 99%

Detection of Bacterial and Phytoplasmal Pathogens

Narayanasamy¹

2010

Microbial Plant Pathogens-Detection and Disease Diagnosis:

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Bacterial pathogens classified as prokaryotes are much simpler in structure and smaller in size and they have less complex structural features compared to fungal plant pathogens. As the morphological characteristics of bacterial cells are less variable, biological, biochemical, physiological, immunological and genomic characteristics have to be determined for reliable identification and meaningful classification of bacterial pathogens. Detection and identification of bacterial plant pathogens present in whole plants and in propagative plant materials have been possible by employing isolation on cultural media and metabolic fingerprinting methods. But they are labor-intensive and require long time. Results often are inconclusive. Isozyme analysis, direct colony thin layer chromatography and gel electrophoresis techniques have been successfully applied for the detection of some bacterial pathogens. Immunoassays and nucleic acid-based assays have become widely accepted techniques, providing more sensitive and specific detection and quantification of bacterial pathogens affecting a wide range of host plant species. However, the major limitation of the molecular techniques is their inability to discriminate living cells from the dead ones. Attempts have been made to overcome this limiting factor by using DNA binding dyes and estimating pathogen RNAs as an indicator of cell viability. The diagnostic methods have both merits and demerits and hence, appropriate method has to be selected based on cost-effectiveness and possibility of obtaining reliable results rapidly to suit the requirements of the investigation concerned.Phytoplasmas are cell wall-less, nonculturable bacteria belonging to Mollicutes. Lack of cultural characteristics has made obligatory to study the morphological characteristics of phytoplasmas present in the phloem cells of infected plant hosts by observing under electron microscopes. As most of the phytoplasmas look alike in the ultrathin sections, the morphological characters have no diagnostic value. Histochemical methods using DNA binding dyes have been employed to localize the phytoplasmas in the host cells. Application of immunoassays and nucleic acidbased techniques has been effective in providing information for the identification and differentiation of phytoplasams. Polyclonal and monoclonal antibodies have been generated for detection of phytoplasmas infecting several plant species. Universal and species-specific primers and probes have been designed based on

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“…We found that the qPCR-based method can detect the pathogen in artificial and natural latent brown rot infections. Many authors have reported that qPCR has a higher sensitivity and test specificity than conventional PCR for detecting and quantifying the DNA of soilborne fungi, oomycetes, bacteria, nematodes, viruses, and phytoplasmas (Baric et al, 2006;Ippolito et al, 2004;Lievens et al, 2006;Schena et al, 2004;. PCR-based methods are also considered the most effective method for detecting infectious microorganisms with a low titer and an uneven distribution in plants, such as apple proliferation phytoplasma (Baric et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

“…Many authors have reported that qPCR has a higher sensitivity and test specificity than conventional PCR for detecting and quantifying the DNA of soilborne fungi, oomycetes, bacteria, nematodes, viruses, and phytoplasmas (Baric et al, 2006;Ippolito et al, 2004;Lievens et al, 2006;Schena et al, 2004;. PCR-based methods are also considered the most effective method for detecting infectious microorganisms with a low titer and an uneven distribution in plants, such as apple proliferation phytoplasma (Baric et al, 2006). Moreover, cross detection of M. fructigena and M. laxa was found when using the P_fc probe on this study and an additional allelic discrimination step had to be added to distinguish between M. fructicola and M. fructigena/ M. laxa while van Brouwershaven et al (2010) did not detect any cross identification.…”

Section: Discussionmentioning

confidence: 99%

“…Due to the nature of latent infections where a small fungal infection has arrested its development, a highly sensible method would be required to detect (Baric et al, 2006;Ippolito et al, 2004;Lievens et al, 2006;Schena et al, 2004;. PCR-based methods are also considered the most effective methods for detecting infectious microorganisms with a low titter and an uneven distribution in plants, such as apple proliferation phytoplasma (Baric et al, 2006).…”

Section: Introduction 43mentioning

confidence: 99%

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Epidemiología y control de la podredumbre parda del melocotón en postcosecha

Benítez¹

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TaqMan real-time PCR versus four conventional PCR assays for detection of apple proliferation phytoplasma

Cited by 37 publications

References 23 publications

Resistance Inducers and Plant Growth Regulators Show Only Limited and Transient Effects on Infection Rates, Growth Rates and Symptom Expression of Apple Trees Infected With ‘candidatus Phytoplasma mali’

Resistance Inducers and Plant Growth Regulators Show Only Limited and Transient Effects on Infection Rates, Growth Rates and Symptom Expression of Apple Trees Infected With ‘candidatus Phytoplasma mali’

Detection of Bacterial and Phytoplasmal Pathogens

Epidemiología y control de la podredumbre parda del melocotón en postcosecha

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