Standardization of the ReverseTranscription Polymerase Chain Reaction for the diagnosis of Capsicum annuum L in Cuba

Abstract

Potyvirus cause diseases that cannot be controlled once the plant is infected, such as in other viral plant diseases. On the other hand, non persistent transmition by aphids is a challenge for the prevention and control strategies for viral diseases in crops with economic relevance. However, some integrated pest management measures could prevent the introduction of potyvirus into healthy gardens and plantations thus preventing their spread. In spite of all measures that have been taken, in the last years it has been observed a strong rise of the potyvirus related symptoms in solanaceus crops, which suggests either the presence of new potyvirus strains or the apparition of recombinant strains from the already circulating ones, behaviour rather frequent for this viral family. This work is aimed to standardize a technique for the molecular diagnosis of potyvirus through RTPCR using the generic primers CP1 and CP2. In silico analyses were carried out to detect the presence of potyvirus in pepper crop and to demonstrate the amplification of a conserved region containing a gene coding for the viral capsid protein. Then one step RTPCR standardization was done. The results showed that this technique is highly sensitive and specific for the generic diagnosis of potyvirus in pepper crop. This is the first report in Cuba of the use of a molecular diagnosis method to assist the genetic improvement program of pepper.