Use of intramolecular chimeras to map molecular determinants of symptom severity of potato spindle tuber viroid (PSTVd)

Nucleotide sequence comparison shows that sequence variations are mostly clustered in the P (pathogenicity) and V (variable) domains of the potato spindle tuber viroid (PSTVd) molecule. Although these comparisons suggest the P domain as the primary determinant of PSTVd symptom severity, the potential contribution of the V domain has never been analysed in detail. To investigate the relationship between the structure of these domains and pathogenicity, six intraspecific chimeric PSTVd variants were constructed by exchanging P and V domains between a mild and two different severe PSTVd isolates. Infectivity studies showed that the P domain is directly responsible for the severity of symptoms induced in tomato. The four recombinants containing a P domain from a severe isolate caused severe symptoms including severe epinasty, stunting and veinal necrosis: while the two chimeras containing the mild isolate P domain induced only mild symptoms. Quantitation of viroid accumulation in plants infected with the various recombinants suggests that, with the constructions used, symptom severity did not correlate with viroid accumulation, indicating that the P domain did not influence symptom production through this simple mechanism.