| Papilio memnon L., P. xuthus L. and Kallima inachus Doubleday are famous enjoyableinsects, having a huge value in aesthetics and market. Using the climate-box, the effect ofphotoperiod and temperature on the development and growth, the inducement and terminationand development of diapause of the three butterflies are researched. Using spectrophotometerand capillary electrophoresis apparatus, the main biochemical substance and the kinds ofprotein in different stages during diapause are researched.1. The effect of photoperiod and temperature on development and growth of three butterflies.(1) To P. memnon. At 20℃and in 12～14h daylength, developmental period of larvae are39.3～48.5 days, and at 12～15h daylength, the developmental periods of diapause andnondiapause pupae are 124～146.6 and 24.4～28.6 days respectively; At 25℃and in 11～15hdaylength, developmental period of larvae are 25.4～36 days, and developmental periods ofdiapause and nondiapause pupae are 64.4～112 and 12.3～19.7 days respectively; At 30℃and in11～15h daylength, developmental period of larvae are 19.6～29.2 days, and developmentalperiods of nondiapause pupae are 7.5～16 days. The results show developmental periods oflarvae and pupae shorten when temperature rise in the scope of test temperature.At 20℃C and 25℃and 30℃, survival rates of larvae and pupae are 25～67, 20～65 and3～23 percent respectively. Analysis show survival rates having little difference between 20℃and 25℃, but falling at 30℃.In 20～30℃and 12～14h daylength, temperature effective thermal sum and developmentalthreshold temperature of larvae are 332.7～475.4 degree-days and 7.5～13.6℃respectively; in13～15h daylength, temperature effective thermal sum and developmental thresholdtemperature of nondiapause are 100.1～168.3 degree-days and 13.9～17℃respectively, anddiapause are 583.2～1160.6 degree-days and 11.3～16℃respectively. Results show photoperiodhave some affection on temperature effective thermal sum and developmental thresholdtemperature.During the scope of 20～30℃and 11～15h daylength, the results show that temperature isthe most important factor which affects development and growth of P. memnon. (2) To P. xuthus. At 20℃and in12～15h daylength, developmental period of larvae are32～45 days, developmental periods of diapause and nondiapause pupae are 123～196.5 and16～22 days respectively; Survival rates of larvae and pupae are 5～35 percent and low.(3) To K. inachus. At 20℃and in 12～15h daylength, developmental period of larvae andpupae are 39.3～48.5 days and 19.5～24.4 days respectively; At 25℃and in 12.5～14h daylength,developmental period of larvae and pupae are 26.3～27.4days and 12.6～13.9days respectively;At 30℃and in 12.5～14h daylength, developmental period of larvae and pupae are 21～21.5days and 9～9.3 days respectively. The results show developmental periods of larvae and pupaeshorten when temperature rise in the scope of test temperature. At 20℃, the affection on larvaeand pupae are obvious, but inconspicuous at 25℃and 30℃.At 20℃and 25℃and 30℃, survival rates of larvae and pupae are 64～92, 70～85 and55～85 percent severally. Analysis show survival rates having little difference between 20℃and 25℃, but falling at 30℃.In 20～30℃and 12.5～14h daylength, temperature effective thermal sum of larvae andpupae are 332.7～475.4 degree-days and 174～162.1, and developmental threshold temperatureare 0.8～5.1℃and 11.1～12.7℃respectively. Results show developmental thresholdtemperature of larvae rise and temperature effective thermal sum of larvae fall when daylengthshorten.During the scope of 20 to 30℃and 12～15h daylength, the results show temperature is themost important factor affecting development and growth of K. inachus.2. Diapause induction of three butterflies.(1) Temperature and critical photoperiod. At 20℃, Critical photoperiod of P. memnon, P.xuthus and K. inachus are LD13h11min:10h49min, LD13h18min:10h42min andLD14h19min:9h41min; at 25℃, P. memnon and K. inachus are LD12h49min:11h11min andLD13h22min:10h38min; At 30℃, K. inachus is LD12h: 12h. Results show short daylength andlow temperature benefit to diapause induction, and long daylength and high temperaturebenefit to development and restrain diapause.(2) Sensitive stage. The fifth instar is the sensitive stage of P. memnon. The second andthird instars are the sensitive stage of P. xuthus. The sensitive stage of K. inachus is in the fewdays following emergence. During the scope of 20 to 30℃and 11 to 15h daylength, results show diapause is affectedby temperature and photoperiod; the affection of photoperiod is bigger than temperature on P.memnon, but smaller on K. inachus.3. Release from diapause of P. memnon and P. xuthus(1) Temperature and diapause. At 15℃, 20℃, 25℃and 30℃, developmental period ofdiapause pupae of P. memnon are 71.2, 40.5, 22 and 16.8 days, and P. xuthus are 60.3, 26, 15.2and 11.6 days respectively. Results show higher temperature, earlier the eclosion time, andshorter developmental period; High temperature has some harm to developmental heath, andmortality rate rises when temperature rises.(2) Daylength and diapause. The affection of photoperiod on diapause development issmall. Analysis shows the developmental periods of diapause pupae are close at 15h and 12hdaylength.(3) Low temperature and diapause. At 20℃and after dealing with low temperature 20and 40 days, the developmental period of diapause pupae of P. memnon are 43.3 and 34.6 days,and P. xuthus are 44.6 and 29.5 days respectively; Not dealing with low temperature, thedevelopmental period of diapause pupae of P. memnon is 65.3 days, and P. xuthus is 76.8 days.At 25℃the developmental period of diapause pupae of P. memnon are 24.5and 21.3 days, andP. xuthus are 28.5 and 19days respectively; Not dealing with low temperature, thedevelopmental period of diapause pupae of P. memnon is 35.8 days, and P. xuthus is 60.9 days.Results show dealing with low temperature benefiting to release from diapause.Result of orthogonal experimental show the R~2 of temperature, photoperiod and dealingwith low temperature of P. memnon are 1621.342, 143.183 and 11.635, and P. xuthus are936.211, 154.7 and 21.264. The order of affection on diapause is temperature, photoperiod anddealing with low temperature.4. Diapause base on biochemistry.Test results show content of water, trehalose, hepatin, fat, protein, DNA and RNA aresignificantly different from nondiapause, and the kinds of protein have a significant varietybetween diapause and nondiapause individuals.Compared to nondiapause pupae, water content of diapause pupae of P. memnon and P.xuthus declined 2.3243～3.2783% and 6.3926～13.0439%. Compared to nondiapause adult of K.inachus, the lowest water content decline 7.7968%, and the highest water content rise8.1191%. Compared to nondiapause pupae, trehalose, hepatin and fat content of P. memnon rise0.57～6.32, 0.66～1.26 and 0.067～43.15 times; P xuthus rise 5.35～134.26, 6～16.37 and 1.16～1.23times respectively. Compared to nondiapause adult of K. inachus, trehalose, hepatin and fatcontent rise 14.24～17.73, 14.24～17.73 and 1.8～5.6 times.Compared to nondiapause pupae, the lowest protein content of P. memnon decline27.33%, and the highest protein content rise 14.42%; protein content of P. xuthus rise6.67～11.44%. Compared to nondiapause adult, protein content decline 7.16～48.01%.Compared to nondiapause pupae, RNA content of P. memnon decline 6.24～25.94%; thelowest content of RNA rise 22.98%, and the highest decline 0.09%; RNA content of K. inachusdecline 52.08～55.18%Compared to nondiapause pupae, DNA content of P. memnon and P. xuthus rise0.74～54.82% and 8.59～103.08% respectively. Compared to nondiapause adult of K. inachus,DNA content rise 17.09～115.42%.Compared to nondiapause pupae, water and RNA content of P. memnon are lower, andprotein are lower in most time; water content of P xuthus are lower, and RNA content are lowerin most time; RNA content of K. inachus are lower, and water content are lower in most time.Trehalose, hepatin and fat content are higher during the diapause, and DNA content are higherin most time of diapause stage of the three butterflies. Results show the characteristic ofdiapause stage are high energy substance and DNA content, low RNA content and metabolismlevel.There are 27～28 and 32～40 apexes in nondiapause stage, 25 and 32 apexes of adultscoming from diapause, and 25～42 and 18～45 apexes in diapause stage of P. memnon and P.xuthus respectively. There are 21～31 apexes in nondiapause adult and 17～34 apexes in thewinter adult of K. inachus. The kinds of protein in diapause stages of three butterflies are morethan nondiapause stages. The kinds of protein of P. memnon and P. xuthus add more at the lastten days of December, and K. inachus add more at the last ten days of November. The addingof protein maybe is related to the temperature falling, and maybe there are manydiapause-associated proteins produced, including antifreeze proteins, during diapause stage.
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