Development of a species‐specific and sensitive detection assay for <i>Phytophthora infestans</i> and its application for monitoring of inoculum in tubers and soil

Hussain, Syed Ismat; Lees, A. K.; Duncan, J. M.; Cooke, D. E. L.

doi:10.1111/j.1365-3059.2005.01175.x

Cited by 54 publications

(32 citation statements)

References 42 publications

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“…The detection of P. infestans inoculum in soil samples was demonstrated using conventional PCR (Hussain et al . ), and later improved upon using quantitative PCR (Lees et al . ; Hussain et al .…”

Section: Introductionmentioning

confidence: 99%

Loop‐mediated isothermal amplification for detection of the tomato and potato late blight pathogen, P hytophthora infestans

et al. 2016

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Section: Introductionmentioning

confidence: 99%

Loop‐mediated isothermal amplification for detection of the tomato and potato late blight pathogen, P hytophthora infestans

et al. 2016

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“…1997; Judelson and Tooley 2000; Jyan et al. 2002; Wangsomboondee and Ristaino 2002; Hussain et al. 2005) and quantitative real‐time PCR (Böhm et al.…”

Section: Introductionmentioning

confidence: 99%

A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth

Llorente

Bravo-Almonacid

Cvitanich

et al. 2010

Letters in Applied Microbiology

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Aims: To establish a reliable and rapid protocol to simultaneously obtain high quality DNA from an infected host plant and the infecting pathogen. To develop an accurate and sensitive low‐cost assay for the quantification and in planta monitoring of Phytophthora infestans growth. Methods and Results: In this study, we describe a SYBR Green‐based quantitative real‐time PCR (qPCR) method for the quantification of P. infestans. The method is based on a simultaneous plant‐pathogen DNA purification followed by a qPCR in which the relative quantification of pathogen and plant DNA is performed. Besides assuring an accurate quantification, the use of a plant gene provides a reliable indicator of sample quality, allowing the exclusion of inappropriate samples. By applying this methodology, we were able to detect P. infestans in potato leaf and tuber tissue before the first symptoms of the disease were observed and to monitor the in planta growth of the pathogen for 6 days. Conclusions: This is a reliable low‐cost assay that provides rapid, accurate and sensitive quantification of the late blight pathogen, allowing the in planta monitoring of P. infestans growth. Significance and Impact of the Study: The quantitative nature of the assay described in this study may be useful in plant breeding programmes and basic research. The method is appropriate for the comparison of cultivars with different, and even subtle, degrees of pathogen resistance and in the screening of new anti‐oomycete compounds. The method can be easily adapted to tomato and the model plant Nicotiana benthamiana.

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“…It is now possible to test seed stocks for infection by P. infestans using a PCR assay specific to P. infestans (Hussain et al 2005). This technology can detect as little as 0.5 pg of P. infestans DNA from tubers (Hussain 2003) and allows infected stocks to be identified.…”

Section: Ukmentioning

confidence: 99%

Epidemiology and Integrated Control of Potato Late Blight in Europe

et al. 2011

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Phytophthora infestans, the causal agent of late blight, is a major threat to potato production in northwestern Europe. Before 1980, the worldwide population of P. infestans outside Mexico appeared to be asexual and to consist of a single clonal lineage of A1 mating type characterized by a single genotype. It is widely believed that new strains migrated into Europe in 1976 and that this led to subsequent population changes including the introduction of the A2 mating type. The population characteristics of recently collected isolates in NW Europe show a diverse population including both mating types, sexual reproduction and oospores, although differences are observed between regions. Although it is difficult to find direct evidence that new strains are more aggressive, there are several indications from experiments and field epidemics that the aggressiveness of P. infestans has increased in the past 20 years. The relative importance of the different primary inoculum sources and specific measures for reducing their role, such as covering dumps with plastic and preventing seed tubers from becoming infected, is described for the different regions. In NW Europe, varieties with greater resistance tend not to be grown on a large scale. From the grower's perspective, the savings in fungicide input that can be achieved with these varieties are not compensated by the higher (perceived) risk of blight. Fungicides play a crucial role in the integrated control of late blight. The spray strategies in NW Europe and a table of the specific attributes of the most important fungicides in Europe are presented. The development and use of decision support systems (DSSs) in NW Europe are described. In The Netherlands, it is estimated that almost 40% of potato growers use recommendations based on commercially available DSS. In the Nordic countries, a new DSS concept with a fixed 7-day spray interval and a variable dose rate is being tested. In the UK, commercially available DSSs are used for c. 8% of the area. The validity of Smith Periods for the new population of P. infestans in the UK is currently being evaluated.

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Development of a species‐specific and sensitive detection assay for Phytophthora infestans and its application for monitoring of inoculum in tubers and soil

Cited by 54 publications

References 42 publications

Loop‐mediated isothermal amplification for detection of the tomato and potato late blight pathogen, P hytophthora infestans

Loop‐mediated isothermal amplification for detection of the tomato and potato late blight pathogen, P hytophthora infestans

A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth

Epidemiology and Integrated Control of Potato Late Blight in Europe

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