Fitness costs associated with resistance to insecticides have been well documented, usually at normal temperature conditions, in many insect species. In this study, using chlorpyrifos-resistant homozygote (RR) and chlorpyrifos-susceptible homozygote (SS) of resistance ace1 allele of Plutella xylostella (DBM), we confirmed firstly that high temperature experience in pupal stage influenced phenotype of wing venation in insecticide-resistant and insecticide-susceptible Plutella xylostella, and SS DBM showed significantly higher thermal tolerance and lower damages of wing veins under heat stress than RR DBM. As compared to SS DBM, RR DBM displayed significantly lower AChE sensitivity to chlorpyrifos, higher basal GSTs activity and P450 production at 25°C, but higher inhibitions on the enzyme activities and P450 production as well as reduced resistance to chlorpyrifos under heat stress. Furthermore, RR DBM displayed significantly higher basal expressions of hsp69s, hsp72s, hsp20,hsp90,Apaf-1, and caspase-7 at 25°C, but lower induced expressions of hsps and higher induced expressions of Apaf-1,caspase-9, and caspase-7 under heat stress. These results suggest that fitness costs of chlorpyrifos resistance in DBM may partly attribute to excess consumption of energy caused by over production of detoxification enzymes and hsps when the proteins are less demanded at conducive environments but reduced expressions when they are highly demanded by the insects to combat environmental stresses, or to excess expressions of apoptotic genes under heat stress, which results in higher apoptosis. The evolutionary and ecological implications of these findings at global warming are discussed.

Abstract.To gain further insight into the molecular features of the ubiquitous Hsp70 family of conserved heat shock proteins, total nine full-length cDNA sequences of inducible hsp70s and one constitutive hsc70 (Px-hsc70(C)) were isolated and characterized in the diamondback moth (DBM), Plutella xylostella, collected from Fuzhou, China. The nine Px-hsp70s cDNAs encoded the protein of between 629-669 amino acids with molecular weight ranging from 69.00-72.58 kDa and were derived from four hsp70 genes in the genome of DBM. The Px-hsc70(C) cDNA contained 1,953 bp of open reading frame (ORF), which produced a putative protein comprising 650 amino acids with a calculated molecular weight of 71.18 kDa. Whether in adults or larvae of chlorpyrifos-resistant (R R ) and chlorpyrifos-susceptible (S S ) strains of DBM, the basal level (at 25°C) of Px-hsc70(C) mRNA expression was high, but no significant up-regulation expression was found under heat stress. However, heat stress facilitated up-regulation expressions of Px-hsp70s, and S S DBM displayed higher up-regulation expression of Px-hsp70s than R R DBM. We suggest that higher up-regulation expression of Px-hsp70s in S S DBM is probably involved in their higher thermal tolerance.

A caspase gene in Plutella xylostella (DBM) was identified firstly and named Px-caspase-1. It had a full-length of 1172 bp and contained 900 bp open reading frame that encoded 300 amino acids with 33.6 kDa. The deduced amino acid of Px-caspase-1 had two domain profile including caspase_p20 (position 61-184) and caspase_p10 (position 203-298) (i.e. the big and small catalytic domains), and the highly conserved pentapeptide QACQG in caspase_p20 domain (the recognized catalytic site of caspases). Being highly homologous to effector caspase genes in other insect and mammalian species, Px-caspase-1 was thought to be an effector caspase gene. Heat stress could result in significant mortality increase on adult DBM. Px-caspase-1 mRNA expression and caspase-3 enzyme activity (a effector caspase) were elevated with age and heat treatment. And, heat stress facilitated the procession of Px-caspase-1 expression. Significantly higher mRNA transcription levels were found in a chlorpyrifos-resistant DBM strain, as compared to those in insecticide-susceptible DBM. The results indicated that high temperature could significantly promote apoptosis process resulting in an the increased DBM mortality rate, and that insecticide-susceptible DBM had a significantly higher physiological fitness at high temperatures than insecticide-resistant DBM.