Adaptation of RT-PCR technique for the diagnosis of potato yellow vein virus in Venezuela

Bioagro

Instituto Nacional de Investigaciones Agrícolas (INIA), Maracay, Venezuela¹  Instituto de Estudios Avanzados (IDEA), Caracas¹  Osorio, Martha¹  Demey, Jhonny¹  Vegas, Ariadne¹  Marques, Alexis¹  Roa, Sara¹  Romay, Gustavo¹ 

关键词: Detection;    PYVV;    Solanumtuberosum;    standardization;   

DOI  :  

学科分类：农业科学（综合）

来源: Universidad Centroccidental Lisandro Alvarado * Decanato de Agronomia

【 摘 要 】

 Potato yellow vein virus (PYVV)belongs to the genusCrinivirus, familyClosteroviridae, and is transmittedby thegreenhousewhitefly (Trialeurodesvaporariorum). It is an important problem forAndean countries because it causesloss ofproduction by reducingthe number and sizeof tubers. The diagnosisbased on symptom expression isinaccuratebecause it can infect plants asymptomatically, andserological diagnosis is hampered bythe lack ofcommercial kitsavailable in the country. The objective of this study was the adaptation of theRT-PCRtechnique for the PYVV diagnosis and subsequent viraltrackingin producing areas. Tensymptomatic samples were collected in Táchira and Trujillo States, Venezuela. RNA extraction was performed using the RNeasy Plant Minikit (Qiagen). Deep freezingof the tissue (-60 °C) and addition of sodium metabisulfite (0,05 mg·g-1) were incorporated as additional steps.Theretrotranscripción was executed by using the kitImPron-II Reverse-Transcription System (Promega). Specific primers to amplify CP and CPm genes were utilized, as well as different anneling and extension temperatures, and various amplification mixtures.Such adaptations allowed theamplification ofspecific fragments, and confirmed virus infection in 100 %of the samples.It is the first time that PYVV is diagnosed by molecular techniques in Venezuela, thus it representsa diagnostic procedure that can be implemented to prevent the spreadof the virus from seed tuber. 

【 授权许可】

Unknown