20-hydroxyecdysone and juvenile hormone analog prevent precocious metamorphosis in recessive trimolter mutants of Bombyx mori.

The trimolter mutants of Bombyx mori have four instead of five larval instars of normal tetramolters. Here, we show that the tetramolter was induced in the recessive trimolter European No.7 mutant (rt-E7) by application of either the juvenile hormone analog (JHA) or 20-hydroxyecdysone (20E). However, treatments with JHA or 20E did not change the number of larval instars of the dominant trimolter Si Chuan mutant (DT-SC). Krüppel-homolog 1 (Kr-h1) is an early JH-response gene that mediates the anti-metamorphic action of JH. In the wing disc of tetramolter B. mori, Kr-h1 RNAs decreased shortly after ecdysis to the fifth instar, while pupal specifier gene, Broad Complex Z1 (BR-Z1) RNAs slightly increased and coincided with the onset of metamorphic competence of wing discs. Analysis of the developmental profile of Kr-h1 in the wing disc of rt-E7 showed that its transcript slightly increased from 12 to 24 h and gradually decreased between 24 and 72 h in the fourth (last) larval instar, while Kr-h1 mRNA decreased rapidly between 12 and 72 h in DT-SC. In addition, the expression of BR-Z1 in DT-SC during the early fourth (last) larval instar is relatively higher than that in rt-E7. These results indicated that the occurrence of pupal commitment of the wing disc in DT-SC was much earlier than that in rt-E7. In the early fourth larval instar of rt-E7, feeding on 20E or treatments with exogenous JHA caused up-regulation of Kr-h1, suppressed premature induction of BR-Z1, and then induced an additional larval instar. By contrast, in DT-SC mutant, since pupal commitment immediately occurred after third ecdysis, precocious metamorphosis was not successfully rescued. The results suggest that Kr-h1 and BR-Z1 involved in the prevention of precocious metamorphosis in recessive trimolter mutants by application of 20E and JHA. The result indicated that Kr-h1 and BR-Z1 expression reflected larval-pupal transition of the recessive trimolter of B. mori.