Differential accumulation of Phytophthora cambivora cox II gene transcripts in infected chestnut tissue.

Abstract:

:This study provides a novel qRT-PCR protocol for specific detection and proof of viability of Phytophthora in environmental samples based on differential accumulation of cox II transcripts. Chemical and physical treatments were tested for their ability to induce in vitro the accumulation of cytochrome oxidase genes encoding subunits II (cox II) transcripts in Phytophthora cambivora. Glucose 170 mM, KNO3 0.25 mM and K3 PO3 0.5 and 0.8 mM induced the transcription of cox II in P. cambivora living mycelium while no transcription was observed in mycelium previously killed with 0.5% (p/v) RidomilGold(®) R WG. Living chestnut tissue was artificially infected with P. cambivora and treated with inducers. In vivo experiments confirmed the ability of glucose to induce the accumulation of P. cambivora cox II transcripts. Based on these results, pretreatment of environmental samples with glucose prior to nucleic acid extraction increased the accumulation of specific cox II transcripts, and therefore the sensitivity of qRT-PCR assay for detection of P. cambivora in living tissues. Furthermore, differential accumulation of transcripts between treated and untreated samples represents an unequivocal proof of inoculum viability.

journal_name

FEMS Microbiol Lett

authors

Vannini A,Tomassini A,Bruni N,Vettraino AM

doi

10.1111/1574-6968.12398

subject

Has Abstract

pub_date

2014-04-01 00:00:00

pages

19-25

issue

1

eissn

0378-1097

issn

1574-6968

journal_volume

353

pub_type

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