Abstract
BackgroundStrawberry diseases are a major limiting factor that severely impact plant agronomic performance. Regarding limitations of traditional techniques for detection of pathogens, researchers have developed specific DNA-based tests as sensitive and specific techniques. The aim of this review is to provide an overview of polymerase chain reaction (PCR)-based methods used for detection or quantification of the most widespread strawberry pathogens, such as Fusarium oxysporum f.sp. fragariae, Phytophthora fragariae, Colletotrichum acutatum, Verticillium dahliae, Botrytis cinerea, Macrophomina phaseolina, and Xanthomonas fragariae. An updated and detailed list of published PCR protocols is presented and discussed, aimed at facilitating access to information that could be particularly useful for diagnostic laboratories in order to develop a rapid, cost-effective, and reliable monitoring technique.MethodsThe study design was a systematic review of PCR-based techniques used for detection and quantification of strawberry pathogens. Using appropriate subject headings, AGRICOLA, AGRIS, BASE, Biological s, CAB s, Google Scholar, Scopus, Web of Knowledge, and SpringerLink databases were searched from their inception up to April 2014. Two assessors independently reviewed the titles, abstracts, and full articles of all identified citations. Selected articles were included if one of the mentioned strawberry pathogens was investigated based on PCR methods, and a summary of pre-analytical requirements for PCR was provided.ResultsA total of 259 titles and abstracts were reviewed, of which 22 full texts met all the inclusion criteria. Our systematic review identified ten different protocols for X. fragariae, eight for P. fragariae, four for B. cinerea, six for C. acutatum, three for V. dahlia, and only one for F. oxysporum. The accuracy and sensitivity of PCR diagnostic methods is the focus of most studies included in this review. However, a large proportion of errors in laboratories occur in the pre-analytical phase of the testing process. Due to heterogeneity, results could not be meta-analyzed.ConclusionsFrom a systematic review of the currently available published literature, effective detection assays to detect the major strawberry pathogens have been developed. These assays can function as a basis for clinical labs, regulatory personnel, and other diagnosticians to adapt or implement for detection of these six important strawberry pathogens.Electronic supplementary materialThe online version of this article (doi:10.1186/2046-4053-4-9) contains supplementary material, which is available to authorized users.
Highlights
Strawberry diseases are a major limiting factor that severely impact plant agronomic performance
Molecular-based techniques can overcome many of the shortcomings of the conventional assays, especially if they rely on polymerase chain reaction (PCR) assays
The primary objective of this review is to provide an exhaustive overview of the existing scientific literature available on PCR-based diagnostic techniques that is restricted to the detection and quantification of the seven most abundant strawberry pathogens: F. oxysporum f.sp. fragariae, P. fragariae, C. acutatum, V. dahliae, B. cinerea, M. phaseolina, and X. fragariae
Summary
Strawberry diseases are a major limiting factor that severely impact plant agronomic performance. Strawberry diseases are a major limiting factor that severely impact the plant agronomic performance and lead to economic losses. Conventional methods to detect and identify pathogens have often relied on isolating the pathogen onto selective media or through biochemical, chemical, and immunological analyses [7] These methods are fundamental to diagnose the presence of plant pathogens, but they rely on time-consuming and labor-intensive lab techniques and on skilled taxonomical expertise [8]. An increasing amount of diagnostic methods recommended by the European and Mediterranean Plant Protection Organization (EPPO) are based on PCR assays [11,12]. This technique is nowadays considered a routine technique in molecular diagnosis
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