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Abstract
Background
The state of Espirito Santo has in the fruit growing, one of its main economical activities.
One of them, the pineapple growth, represents a good economic potential to increase
income and job opportunities in the state. However, diseases such as fusariosis and
mealybug wilt of pineapple are responsible for the yield losses of up to 80% [1].
The mealybug wilt of pineapple is caused by the complex Pineapple mealybug wilt-associated
virus (PMWaV-1, PMWaV-2, PMWaV-3 and PMWaV-5), a single strand RNA [2], while fusariosis
on pineapple is caused by the fungus Fusarium guttiforme [3]. This study aimed to
develop methodologies for detection of these etiological agents in the pineapple plant.
Methods
For the analyzes, healthy and diseased pineapple plant were collected at the Experimental
Farm of Incaper in the city of Sooretama, in the state of Espirito Santo. The samples
were submitted to a superficial disinfection process, followed by nucleic acid extraction
protocols with Trizol® and SDS buffer [4]. The nucleic acids were treated with DNAse
and Reverse Transcriptase enzymes for samples infected with PMWaV. A RNAse enzyme
was used for samples infected with F. guttiforme. The diagnosis was realized by molecular
biology techniques, conventional PCR and real time PCR with specific primers for each
strain of PMWaV [5] and H3 primer for F. guttiforme. The samples were visualized by
electrophoreses on 1% agarose.
Results and conclusions
Through these techniques were diagnosed the F. guttiforme in fruits, and detected
the PMWaV in different plant organs. Conventional PCR does not proved to be a good
diagnostic tool due to poor sensitivity, the need for visualization on gels and the
high risk of contamination. The technique of Real Time PCR can generated diagnosis
with greater sensitivity, reproducibility, accuracy and speed and allowed the detection
in diseased plants and plants considered healthy. Therefore, this study created molecular
tools for a precise and rapid diagnosis of the etiological agents of the fusariosis
disease and mealybug wilt of pineapple that allow indexing plants and propagative
material the spread of disease to new areas. It is hoped that this study contribute
to raising the standards of quality and competitiveness of Brazilian fruit growing,
especially pineapple, the level of excellence required by national and international
markets, as well as implement actions for incorporation of technological methods,
techniques and processes based mainly on concepts of integrated disease management
and food security, with a view to the expansion of this crop, increased productivity
and income generation for farmers.
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Conference name:
5th Congress of the Brazilian Biotechnology Society (SBBIOTEC)