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Comparison of promidium monoazide and Reagent D for discrimination of viable Aphelenchoides besseyi using real-time PCR
European Journal of Plant Pathology ( IF 1.7 ) Pub Date : 2020-07-27 , DOI: 10.1007/s10658-020-02090-x
Elvan Sert Çelik , Tevfik Özalp , Zübeyir Devran

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 non-viable individuals using Reagent-D or PMA combined with qPCR.



中文翻译:

实时荧光定量PCR比较单叠氮化pro和试剂D鉴别活的苯丙氨酸

Aphelenchoides besseyi Christie,1942年是种子传播的线虫,造成水稻的经济产量损失。有关存储的稻米种子中贝氏拟南芥生存力的信息对于控制该线虫很重要。在这项研究中,单叠氮丙啶(PMA)和试剂d,用定量PCR结合,被用于可行的歧视A.干尖线虫的个体。将活着,存放1年或进行热处理(40或60°C,15分钟)的贝西曲霉个体暴露于PMA和Reagent-D染料中。此外,贝西未使用任何化学药品治疗的个体用作对照。然后从所有个体中分离出DNA并进行实时PCR。用Reagent-D和PMA处理的样品的Ct值与未处理的样品不同。结果表明,使用Reagent-D或PMA结合qPCR,可以将稻白线虫的活体和不活的个体区分开。

更新日期:2020-07-27
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