Identification and quantification of Aphelenchoides besseyi from rice using qPCR

Çelik, Elvan Sert; Devran, Zübeyir

doi:10.1007/s10658-019-01692-4

Cited by 7 publications

(7 citation statements)

References 26 publications

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“…The results showed that a small number of immobile juveniles that had been stored for 1 year may have survived. Christoforou et al (2014) In conclusion, although molecular methods have been used for identification of A. besseyi (Devran et al 2017;Rybarczyk-Mydłowska et al 2012;Buonicontro et al 2018;Çelik and Devran 2019), there has been no study to investigate the discrimination of live and dead individuals of A. besseyi. In this study, Reagent-D and PMA have been successfully used in conjunction with qPCR to preferentially amplify from viable samples, thus enabling estimation of the number of live and dead A. besseyi individuals within a sample.…”

Section: Resultsmentioning

confidence: 99%

“…Aphelenchoides besseyi populations were initially isolated from infected rice seeds (Çelik and Devran 2019). They were then cultured on excised carrot discs in the laboratory according to Tülek et al (2009).…”

Section: Nematode Culturementioning

confidence: 99%

“…The reaction mixture had a total volume of 20 µl comprising 5 µl DNA, 500 nM of each primer (AbF11 and AbR22), 200 nM probe AbPr, 10 µl LightCycler® 480 Probe Master (Roche Diagnostics GmbH, Mannheim, Germany) and 3.8 µl water. qPCR was conducted using a LightCycler® 480 II according to the protocol of Çelik and Devran (2019). Each experiment was conducted with three replications and repeated twice.…”

Section: Real-time Pcr (Qpcr)mentioning

confidence: 99%

“…Molecular techniques have been widely used for detection and identification of A. besseyi (Ibrahim et al 1994;Sánchez-Monge et al 2017;Devran et al 2017;Rybarczyk-Mydłowska et al 2012;Buonicontro et al 2018;Çelik and Devran 2019;Çelik et al 2020). However, there has been no study for the discrimination of viable A. besseyi.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of promidium monoazide and Reagent D for discrimination of viable Aphelenchoides besseyi using real-time PCR

2020

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Aphelenchoides besseyi Christie, 1942 is a seed-borne nematode and causes economic yield losses in rice. Information on the viability of A. besseyi in stored rice seeds is important to manage this nematode. In this study, propidium monoazide (PMA) and Reagent D, combined with qPCR, were used for discrimination of viable A. besseyi individuals. A. besseyi individuals that were live, stored for 1 year or heat-treated (40 or 60°C, 15 min) were exposed to PMA and Reagent-D dyes. In addition, A. besseyi individuals not treated with any chemicals were used as a control. DNA was then isolated from all individuals and real-time PCR was carried out. Ct values for samples treated with Reagent-D and PMA were different from untreated samples. Results showed that live individuals of the rice white tipe nematode could be distinguished from nonviable individuals using Reagent-D or PMA combined with qPCR.

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