Electronic Resource
Improved PCR detection of potyviruses in Allium species
| العنوان: | Improved PCR detection of potyviruses in Allium species |
|---|---|
| المؤلفون: | Lunello, Pablo [0000-0002-9237-4901], Lunello, Pablo, Ducasse, D., Conci, V. |
| بيانات النشر: | Springer Nature 2005 |
| نوع الوثيقة: | Electronic Resource |
| مستخلص: | Protocols for producing virus-free Allium plants require an indexing system that is more sensitive than DAS-ELISA and can detect low virus concentrations in infected plants. In the present work, degenerate primers were designed and a one-step IC-RT-PCR protocol was developed to differentiate between Leek yellow stripe virus (LYSV) and Onion yellow dwarf virus (OYDV) in single and mixed infections in several Allium spp. A 566-bp band was observed for LYSV, a 489-bp band for OYDV in single infections, and two bands of the same sizes in mixed infections in different species of Alliaceae. A 508-bp band of Shallot yellow stripe virus and a 594-bp band of Turnip mosaic virus were also amplified with the same primers. RT-nested-PCR was also conducted directly in microtitre plate wells after negative or questionable reactions were produced in an ELISA experiment. The detection limit of the DAS-ELISA for LYSV was 16.5-27.3 ng ml-1. The RT-nested-PCR done after DAS-ELISA was 102 times more sensitive than the DAS-ELISA alone. In parallel, an IC-RT-nested-PCR in microcentrifuge tubes was 104 times more sensitive than the DAS-ELISA. The DAS-ELISA-RT-nested-PCR enables the initial screening of samples by DAS-ELISA to eliminate a high percentage of virus-positive plants, considerably reducing the number of plants to analyze further by RT-PCR. © Springer 2005. |
| مصطلحات الفهرس: | Alliaceae, Garlic, Indexing, Leek yellow stripe virus, Molecular tests, Onion yellow dwarf virus, RT-PCR, Shallot yellow stripe virus, Turnip mosaic virus, artículo |
| URL: | Departamento de Biotecnología Sí |
| الاتاحة: | Open access content. Open access content closedAccess |
| ملاحظة: | English |
| Other Numbers: | CTK oai:digital.csic.es:10261/293387 European Journal of Plant Pathology 112: 371-378 (2005) 0929-1873 10.1007/s10658-005-6232-3 1573-8469 1373156216 |
| المصدر المساهم: | CSIC From OAIster®, provided by the OCLC Cooperative. |
| رقم الانضمام: | edsoai.on1373156216 |
| قاعدة البيانات: | OAIster |
| الوصف غير متاح. |