Protocols for extraction of Brevipalpus phoenicis total RNA and prospecting for RNA interference (RNAi) genes

• Grant number: 14/12514-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2014
• End date: August 31, 2015
• Field of knowledge: Agronomical Sciences - Agronomy - Plant Health
• Principal Investigator: Valdenice Moreira Novelli
• Grantee: Marcelo Campos dos Santos
• Host Institution: Instituto Agronômico (IAC). Agência Paulista de Tecnologia dos Agronegócios (APTA). Secretaria de Agricultura e Abastecimento (São Paulo - Estado). Campinas , SP, Brazil

Abstract

The citrus industry, of high economic importance to Brazil, is affected by various diseases and pests, especially citrus leprosy, caused by Citrus leprosy virus C (CiLV-C), and transmitted by B. phoenicis mite (Acari: Tenuipalpidae ). This mite is polyphagous, cosmopolitan, and relevant in agriculture due to its high vectorial capacity is responsible for the transmission of many other phytovirus in diverse cultures. In citrus orchards, acaricides are expensive and are used extensively in an attempt to control this mite vector. Understanding related to the interaction between plants, virus, and B. phoenicis are extremely desirable and still need to be clarified. One possible strategy is to identify the functional genomics (transcriptome) mite, expanding the information about the vector potential generating capacity and information for new methods of management and control. While there have been described several protocols for RNA extraction from mites, specifically for this species has not yet been established. Previous experience of our group in order to extract genomic DNA to other studies has shown to be difficult and laborious work to obtain a standard protocol with quality and concentration of the appropriate sequencing material. Whereas RNA molecules are much more unstable, it is necessary to standardize and optimize the process of obtaining good quality RNA before starting transcriptome studies of this species. Therefore, this proposal aims to test different extraction protocols to B. phoenicis RNA total, in order to get the quality and quantity of desirable material for these gene expression studies. A standardized method of extraction, candidate genes will be explored for application of RNA interference technology, using as reference the genomic databases of Tetranychus urticae, considered the standard for mite genomic studies of B. phoenicis.(AU)