Development of polyclonal antisera for the detection of begomovirus and crinivirus and attempts to identify the causative agent of a new yellowing in tomato

• Grant number: 17/26393-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: June 01, 2018
• End date: January 24, 2019
• Field of knowledge: Agronomical Sciences - Agronomy - Plant Health
• Principal Investigator: Jorge Alberto Marques Rezende
• Grantee: Monica Alves de Macedo
• Host Institution: Escola Superior de Agricultura Luiz de Queiroz (ESALQ). Universidade de São Paulo (USP). Piracicaba , SP, Brazil

Abstract

Begomovirus disease affects several crops with economic importance in Brazil, especially tomato crop. In 2008, a new disease, crinivirus disease (yellowing of tomato), was reported in tomato plants in Sumaré-SP and rapidly the causal agent, Tomato chlorosis virus, was disseminated to other tomato production regions in the country. Crinivirus disease also affects pepper and potato crops. Currently the diagnosis of begomovirus and crinivirus disease is carried out mainly by molecular methods, which are expensive and require specialized technicians and equipment. Serological methods are a good alternative for diagnosis, they are cheaper and do not require specialized equipment. However, there are no antisera available for serological detection of these viruses in Brazil. The aim of this proposal is to obtain polyclonal antisera for the detection of begomovirus and crinivirus. Obtaining these antisera will make the diagnosis of these diseases cheaper and more accessible, besides to open a range of applications, like studies of protein interaction and virus localization in the vector and in the plant host. Since 2015, another yellowing symptom, similar to the caused by crinivirus, has been observed in tomato plants in Sumaré-SP. The incidence of this disease reaches more than 20% in certain tomato production areas and it have been worrying the tomato growers. To date, the causal agent of this disease is unknown, which makes the development of management strategies unfeasible. Therefore, another objective of this proposal is to identify the causal agent of this new yellowing in tomato crops. To attempt to isolate the causal agent, symptomatic tomato plants will be collected and used to inoculated in heathy tomato plants by mechanical, grafting and vector methods. Simultaneously, total RNA and DNA of these samples will be extracted and send to next generation sequencing (NGS). It is expected to identify the causal agent of this new yellowing of tomato, allowing the development of disease control strategies.