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The three current genera in the family Closteroviridaeare supported by vector and phylogeneticdata. Dendrogramwas generated using TreePuzzle5.2 with coat protein sequence data in a maximum likelihood model. Numbers represent branch support in percentage following10,000 puzzling steps.

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  1. Orchids
  2. Detection, Characterization, and Management of Pineapple Mealybug Wilt-Associated Viruses John Hu University of Hawaii
  3. Pineapple in Hawaii Hawaii’s number one agricultural commodity Hawaii Agricultural Statistics Service (2002)
  4. Symptoms of MWP Healthy • Reddening of the leaves • Downward curling of the leaf margins • Loss of turgidity, leaves MWP reflex downwards • Leaf tip dieback • Plants either recover or endure further leaf tip dieback resulting in death
  5. Association of Mealybugs with the Disease • In 1931 Illingworth directly associated mealybugs with wilting pineapple plants • Psuedococcus brevipes: Dysmicoccus brevipes (pink) Dysmicoccus brevipes (pink) Dysmicoccus neobrevipes (gray) Dysmicoccus neobrevipes (gray)
  6. Association of Ants with the Disease • Caretakers of mealybugs • Protection against predators
  7. Search for the Latent Virus • In 1989, U.B. Gunasinghe and T.L. German isolated a closterovirus from MWP- affected pineapple • Named the Pineapple mealybug wilt-associated virus (PMWaV) • Based on mealybug transmissibility, placed in Ampelovirus genus
  8. Control Strategies • Amdro ®, applied as a broadcast bait (ants) • Diazinon ● Pre-plant dip (mealybugs) ● Overhead application (mealybugs)
  9. Potential Problems • Amdro® – Inactivated by moisture – Not effective against some ant species such as Technomyrmex albipes • Diazinon – Use in pre-planting dips has been eliminated
  10. Research Areas • Detection • Epidemiology • Etiology • Management
  11. Detection Assays 1. dsRNA analyses 2. EM & ISEM 3. ELISA 4. Tissue blot Immunoassay* 5. RT-PCR*
  12. Multiple Closteroviruses in Pineapple? • ISEM revealed that not all virus particles were being decorated by monoclonal antibodies • At least two serotypes exist 200nm
  13. Tissue blot immunoassay: - distinct signal - robust - minimal sample preparation - can process 100’s of samples per day
  14. PMWaV-Specific RT-PCR Assays RT-PCR Products Southern Hybridization PMWaV-1 PMWaV-2
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