Is period gene causally involved in the photoperiodic regulation of reproductive diapause in the linden bug, Pyrrhocoris apterus?

Earlier experiments demonstrated a strong up-regulation of per mRNA in wild-type (Wt) females of Pyrrhocoris apterus reared under diapause-inducing short days, while per mRNA levels were low in females of two non-diapause mutant strains (Nd), irrespective of photoperiod. In the present study, different sequences of per DNA in two strains of geographically different origin enabled us to analyse genetic linkage between the per gene and the Nd phenotype. Crosses between Wt females originating from C. Budejovice (Czech Republic) and Nd males originating from Lyon (France) resulted in F2 progeny where 411 females entered diapause under short days and 120 females were reproducing. Thus, the segregation was very close to the 3:1 ratio in favour of diapause females, suggesting that the Nd trait behaves as a single autosomal recessive. Analysis of DNA in 20 females of the F2 progeny revealed that their phenotype was not linked to the per genotype. We conclude that the per gene is not primarily responsible for the block to diapause photoresponsiveness in Nd mutants and its role, if any, is downstream from other gene(s) controlling diapause. This is the first attempt at genetic linkage analysis between a bona fide circadian clock gene and photoperiodism in a 'non-drosophilid' species.