Improved Molecular Methods for Detection of European Stone Fruit Yellows (Esfy) Phytoplasmas From in Vitro Shoots of Fruit Trees

Bertaccini, Assunta; Paltrinieri, Samanta; Caprara, L.; Laimer, Margit; Hanzer, V.; Balla, I.

doi:10.17660/actahortic.2004.657.80

Cited by 4 publications

(3 citation statements)

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“…Although the PCR detection procedure proved more critical than the extraction method in the case of periwinkle samples, a quicker and less expensive DNA extraction method was selected for routine detection in grapevine samples. In another study, two extraction methods involving the use of chloroform/phenol and silica gel were compared for detection of European stone fruit phytoplasma in in vitro samples (Bertaccini et al, 2004): phytoplasmas were detected in some samples after silica gel extraction, while the other method yielded negative results.…”

Section: Discussionmentioning

confidence: 99%

“…As fruit tree phytoplasmas usually occur at low titres and with uneven distribution in infected plants, their detection is difficult. In this context, the development of sensitive but also quick, reliable, and cost-effective methods for detecting fruit-tree phytoplasmas is essential to production of certified propagation materials (Bertaccini et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of ‘Candidatus Phytoplasma mali’

Aldaghi

Massart

Dutrecq

et al. 2009

Journal of Virological Methods

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a b s t r a c tDifferent PCR protocols have been established for detection of European fruit trees phytoplasmas; however the majority of the procedures for extracting phytoplasma DNA are complex, time consuming, and expensive, with a risk of contamination or loss of target DNA. In present study, a crude extract preparation method previously used to detect other plant pathogens was adapted to samples from apple trees infected by 'Candidatus Phytoplasma mali'. End-point and real-time PCR detection of 'Ca. P. mali' were used to compare this extraction procedure with an established method for efficient extraction of purified DNA. The crude extract proved fully adequate for phytoplasma detection in samples from 86 in vitro and 35 in vivo apple shoots or plants and 10 periwinkle plants. High inter-and intra-run reproducibility was obtained for phytoplasma detection with different TaqMan MGB-or SYBR Green-based real-time PCR protocols applied to the crude extracts. Real-time PCR applied to serially diluted crude and purified extracts revealed the same phytoplasma detection limit (dilution up to 10 5 ). All results confirm the suitability of this simple, quick, efficient extraction technique for accurate detection of 'Ca. P. mali' in different types of apple and periwinkle samples.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of ‘Candidatus Phytoplasma mali’

Aldaghi

Massart

Dutrecq

et al. 2009

Journal of Virological Methods

Self Cite

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“…Since the beginning of the research of this economically dangerous phytoplasma (Morvan, 1977), many studies have been carried out focusing on optimising its detection (Seemüller, 1976;Lee et al, 1991;Ahrens et al, 1992;Gibb et al, 1994;Bertaccini et al, 2004), on understanding of possibile ways of its transmission (Carraro et al, 1998;Jarausch et al, 1998;Jarausch et al, 1999;Pastore et al, 2001;Thébaud et al, 2009) and on the phytoplasma genome characteristics (Ahrens et al, 1993;Schneider et al, 1995;Lee et al, 1998;Marcone et al, 2001;Seemüller et al, 2004). The risk factors of phytoplasma ESFY occurrence in different regions and orchards were assessed by Thébaud et al (2006), as well as by Ulubaş et al (2007) who reported the differences in ESFY phytoplasma occurrence in extensive and intensive orchards, when in the extensive orchard, the ESFY phytoplasma was detected in 54.8% of the analysed samples, while in the intensive orchard the ESFY phytoplasma was detected in only 3.2% of analysed samples.…”

Section: Introductionmentioning

confidence: 99%

The Effect of Phytoplasma Disease Caused by ‘Candidatus Phytoplasma prunorum’ on the Phenological and Pomological Traits in Apricot Trees

Nečas

Kiss

Eichmeier

et al. 2018

Not Bot Horti Agrobo

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Abstract'Candidatus Phytoplasma prunorum', the causal agent of European stone fruit yellows (ESFY), is one of the most important pathogens causing considerable economic losses in stone fruit orchards. This study evaluated trees infected and noninfected by phytoplasma ESFY of 16 apricot varieties grown in an orchard in Lednice (Czech Republic) between the years 2008-2014. Pomological traits, phenophases, pollen germination and seed viability were analysed as well as the presence of 'Ca. P. prunorum' in pollen, flower organs, fruit flesh, immature seeds and seedlings by nested PCR. One of the most detrimental impacts was the decreased fruit set of infected trees which occurred in 12 out of 16 studied varieties reaching an average fruit set decrease of 16.1%. Pollen germination rates also suffered, showing an average decrease by 11.78% in infected trees. In the analysis of some traits, overall significant differences were observed between the infected and noninfected trees. However, for all analysed traits in different varieties, both positive and negative influences of phytoplasma ESFY were observed. The presence of 'Ca. P. prunorum' in infected trees was confirmed in flower parts (only in peduncle in 70.2% of cases) and fruit parts (immature fruit flesh in 42.0% and milky kernels in 26.2% of cases), however, neither in seedlings nor in pollen.

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Micropropagation and Maintenance of Phytoplasmas in Tissue Culture

Bertaccini

Paltrinieri

Martini

et al. 2012

Methods in Molecular Biology

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Maintenance of phytoplasma strains in tissue culture is achievable for all strains transmitted to periwinkle (Catharanthus roseus), and also for other naturally infected plant host species. Shoots of 1-3 cm length are grown in a solid medium containing Murashige and Skoog (MS) micro- and macroelements and 0.12 mg/L benzylaminopurine. The continued presence of phytoplasmas in infected shoots of periwinkle that have been maintained in micropropagation for up to 20 years can be shown by diagnostic methods such as nested PCR tests using the 16S rDNA gene (see Chapters 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,and 26 for phytoplasma diagnostic methods).

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Improved Molecular Methods for Detection of European Stone Fruit Yellows (Esfy) Phytoplasmas From in Vitro Shoots of Fruit Trees

Cited by 4 publications

References 0 publications

A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of ‘Candidatus Phytoplasma mali’

A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of ‘Candidatus Phytoplasma mali’

The Effect of Phytoplasma Disease Caused by ‘Candidatus Phytoplasma prunorum’ on the Phenological and Pomological Traits in Apricot Trees

Micropropagation and Maintenance of Phytoplasmas in Tissue Culture

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