Paratlanticus ussuriensis enter prolonged diapause at an egg stage. Environmental conditions, such as temperature, can modify the diapause duration at initial diapause. Eggs enter initial diapause at 20℃, but continued early embryonic development at 30℃. Final diapause at a fully developed embryonic stage is obligatory regardless of temperature conditions. To determine temperature effects on diapause mechanism of P. ussuriensis eggs, we compared weights, DNA and RNA amounts of eggs incubated at either 20℃ or 30℃ for 50 days after oviposition. Both egg weight and total amount of DNA were constant at 20℃ but gradually increased at 30℃. However, total RNA level was rapidly increased at 15 days-old eggs at 30℃ and maintained high levels during further period whereas its level was constant at 20℃. In addition, we identified three heat shock protein 70 (hsp70a, hap70b, hsp70c) genes of P. ussuriensis and determined those expression levels at different temperature conditions. The levels of hsp70a and hsp70b was not detectable until 20 days after oviposition at both temperature conditions, but highly increased at 50 and 60 days when incubated at 30℃. In contrast, hsp70c level was rapidly peaked at 20 days after oviposition, which is the time of initial diapause entrance. Our results suggest that high temperature breakdown initial diapause and a certain hsp gene, such as hsp70c, may involve in the initial diapause mechanism of P. ussuriensis eggs.

The diapause duration P. ussuriensis eggs is known to either one year or prolonged to additional years. This complex life cycle is mainly caused by the interruption at two embryonic stages: early stage (initial diapause) and fully developed late stage (final diapause). The environmental cues, such as temperature, is critical to determine the diapause duration but the precise diapause mechanism of P. ussuriensis eggs is unclear. We demonstrated temperature effects on the entrance of initial diapause. Newly oviposited eggs were incubated at two temperature conditions (20℃and 30℃) for 60 days. When eggs were incubated at 20℃, egg weights were not significantly changed, but at 30℃ those were gradually increased to 1.5 times for 50 days. Genomic DNA contents of eggs were similarly increased at 30℃. Total RNA contents at 30℃ were highly increased from the 15-days-old but not at 20℃. In addition, we further analysed expression levels of two heat shock protein 70 (hsp70a, hap70b) genes during embryonic development. Our results suggest that initial diapause occurred between 10 and 15 days after oviposition and its entrance was blocked by high temperature incubation.

During the last several years, the population of the ussur brown katydid, Paratlanticus ussuriensis has been highly increased at mountain-side orchards in Youngdong province in Korea. Nymphs and adults emerged one time per year from early May to August and cause a serious demage on the crop production. Adults oviposit eggs from July to September under the ground and eggs overwinter at diapause stage. However, the diapause duration P. ussuriensis is known to either one year or prolonged to additional years. The environmental cues, such as temperature, could be a critical to determine the diapause duration but the precise diapause mechanism of P. ussuriensis eggs is unclear. We demonstrated to determine the stage of diapause initiation during embryogenesis. Newly oviposited eggs were incubated for 60 days at two temperature conditions (20℃and 30℃). When the eggs were incubated at 20℃ those weights were not changed, but when they incubated at 30℃ those were increased from 15-days-old. Total RNA contents of eggs incubated at 30℃ were highly increased from the 15-days-old when they compared with those of eggs incubated at 20℃. In addition, total DNA contents of eggs were greatly higher when they incubated at 30℃. Our results suggest that early embryonic development was sensitive to high temperature and it was stimulated at 15-days-old. To determine the temperature response of embryonic development at molecular level we identified heat shock protein 70 (hsp70) gene of P. ussuriensis. Further analysis would conduct the gene expression rate of diapausing eggs on different temperature conditions by using real time RT-PCR technique and determine the molecular characteristics of P. ussuriensis egg diapause.