Development of a Pineapple Clonal Cleaning Process for Pineapple Mealybug Wilt Associated Virus (pmwav) Using Hydrothermotherapy

Name: Kellyn Joselyn Andino Lopez
Type: MSc dissertation
Publication date: 28/03/2018

Namesort descending Role
Antonio Alberto Ribeiro Fernandes Advisor *

Examining board:

Namesort descending Role
Antonio Alberto Ribeiro Fernandes Advisor *
David dos Santos Martins External Examiner *
Diolina Moura Silva Internal Examiner *
Flavia de Paula (M/D) Internal Alternate *
Helcio Costa External Examiner *
Jose Aires Ventura (M/D) Co advisor *

Summary: The pineapple mealybug wilt is a disease of high economic importance that occurs in all pineapple production regions in the world. That is caused by the viral complex Pineapple mealybug wilt associated virus (PMWaV- 1, PMWaV-2 and PMWaV-3) and the mealybug Dysmicoccus brevipes. In Brazil, it has been associated with the presence of PMWaV-2 but the control methods have been not effective controlling infected plants. In this study we developed a PMWaV-2 clonal cleaning methodology with hydrothermotherapy (48 °C, 50 °C, 52 °C) for 10, 20 and 30 minutes and 54 °C, 56 °C, 58 °C and 60 °C for 10, 20, 30, 40 and 50 minutes in pineapple slips of cv. Pérola. The viability was determined in each sample using different concentrations of tetrazolium chloride (0.01%, 0.05%, 0.075%, 0.1%, 0.5%, 1%) obtaining the best results using 0.075% and 0.05% of tetrazolium chloride. It was possible to obtain viable pineapple slips with hydrothermal treatment using temperatures from 54 ° C to 58 ° C for 50 minutes and 60 ° C for 30 minutes. Survival pineapple was drastically affected when were exposed to temperatures of 60 °C for more than 30 minutes. The molecular analysis using RT- PCR showed the presence of PMWaV-2 in the control samples of pineapple slips without hydrothermal treatment. The presence of PMWaV-2 was not detected in one pineapple leaf sample and three roots pineapple samples that were using the treatment of 56 °C for 30 minutes. In the other hand, pineapple leaf samples exposed to treatments at 54°C for 40 and 50 minutes and 60 °C for 10, 20 and 30 minutes were still diagnosed with the virus presence. The proposed method showed the possibility of obtaining pineapple slips of cv. Pérola free of PMWaV-2 using a hydrothermal treatment at 56°C for 30 minutes. These results show that hydrothermotherapy could be a viable and economical method used in control strategies for cleaning pineapple slips affected by Pineapple mealybug wilt associated virus caused by PMWaV- 2.

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