02561nam a2200205 a 450000100080000000500110000800800410001910000190006024501340007926001130021350002310032652016620055765000320221965000120225165000220226365300140228565300180229970000180231770000200233518645222023-10-19       2010    bl uuuu     u00u1 u    #d1 aANDRADE, E. C.  aDevelopment of degenerated primers to detect different viruses associated with mealybug wilt of pineapple.h[electronic resource]  aPineapple News, n. 17, p. 8, Aug. 2010. Addendum to Newsletter Pineapple Working Group of the I.S.H.S.c2010  aDisponível em: &lt http://www.ishs-horticulture.org/workinggroups/pineapple/PineNews17_addendum.pdf&gt Acesso em: 16 fev. 2011.  Edição  de Abstracts of the 7th International Pineapple Symposium, Johor Baru, Malaysia, 2010.  aPineapple (Ananas comosus var. comosus) is propagated vegetatively which facilitates the production of planting material but leads to dissemination of viruses. Mealybug wilt of pineapple (MWP) is a devastating disease of pineapple worldwide. The disease is characterized by severe leaf tip dieback, wilting of the leaves and can lead the plant to die. The MWP is associated with three Ampelovirus species, Pineapple mealybug wilt associated virus-1 (PMWaV-1), PMWaV-2, and PMWaV-3. Besides to the direct damage on plant production, there is great difficulty to select plantlets for planting because some infected plants exhibits no symptoms. The virus can be detected by RT PCR using specific primers; however for phytosanitary purposed, it's interesting to screen plant material for all viruses simultaneously in order to save time and cost. With this objective, degenerate primers were designed and tested to detect PMWaV-1, -2, -3 in pineapple hybrids of Embrapa breeding program. Using the degenerate primers in a RT PCR, it was possible to amplify a fragment of the expected size only in infected samples. Also, the reverse degenerated primer was combined with each of the forward specific primers in order to identify the PMWaV species present in infected plant. These two approaches led to detection of PMWaVs in 20/27 asymptomatic hybrids, with higher prevalence of PMWaV-1 (12 plants), PMWaV-2 (10 plants) and PMWaV-3 (7 plants). A total of 12 plants present mixed infections. The results show that degenerated primers provided a reliable alternative to plant material certification with about 30-40% on cost reduction and short period of indexing.  aAnanas comosus var. comosus  aAbacaxi  aDoença de Planta  aPineapple  aPlant disease1 aSANTOS, K. C.1 aMEISSNER, P. E.