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利用末龄幼虫蜕和蛹壳对单头昆虫基因的无损伤检测 被引量:1
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作者 郑美艳 朱冠恒 +3 位作者 陈严 韩琪 杨亦桦 董双林 《应用昆虫学报》 CAS CSCD 北大核心 2018年第2期304-309,共6页
【目的】在昆虫基因功能等相关研究中,通常需要利用单对交配策略来筛选纯合突变品系,如何在配对前确定个体基因型同时又不对昆虫造成损伤,显得尤为重要。本文旨在探讨利用末龄幼虫蜕和蛹壳进行单头昆虫的无损伤基因检测方法。【方法】... 【目的】在昆虫基因功能等相关研究中,通常需要利用单对交配策略来筛选纯合突变品系,如何在配对前确定个体基因型同时又不对昆虫造成损伤,显得尤为重要。本文旨在探讨利用末龄幼虫蜕和蛹壳进行单头昆虫的无损伤基因检测方法。【方法】针对大小不同的3种鳞翅目昆虫斜纹夜蛾Spodoptera litura Fabricius、二点委夜蛾Athetis lepigone M?schler和小菜蛾Plutella xyllostella Linnaeus,收集末龄幼虫蜕及蛹壳,利用常规分子生物学技术进行基因组DNA提取、靶标基因PCR扩增、琼脂糖凝胶电泳检测、连接转化和单克隆测序验证。【结果】从斜纹夜蛾、二点委夜蛾末龄幼虫蜕和蛹壳提取的基因组DNA,以其为模板对GOBP1基因进行PCR扩增,产物经琼脂糖凝胶电泳检测得到单一、明显的条带,进一步经连接转化和单克隆测序得到目的序列;但由于小菜蛾末龄幼虫蜕和蛹壳太小,以同样方法提取的基因组DNA浓度太低,PCR产物经电泳检测,未能得到目的条带。【结论】对于与斜纹夜蛾和二点委夜蛾相近或更大的昆虫,可以利用单头末龄幼虫蜕或蛹壳提取基因组DNA,通过常规PCR技术克隆特定基因序列,为突变品系筛选过程中昆虫个体的无损伤基因型检测提供了方法。 展开更多
关键词 幼虫蜕 蛹壳 基因组DNA PCR 突变品系
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Pesticidal character of phytoecdysteroids from Ajuga multiflora Bunge (Labiatae) on larvae of Cryptorrhynchus lapathi L. (Coleoptera: Curculionidae)
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作者 迟德富 孙明学 夏文富 《Journal of Forestry Research》 SCIE CAS CSCD 2002年第3期177-182,250,共6页
Eight kinds of phytoecdysteroids extracted from different parts of Ajuga multiflora Bunge (Labiatae) that were collected from different places at different time were tested for killing effects on the 2-instar larvae o... Eight kinds of phytoecdysteroids extracted from different parts of Ajuga multiflora Bunge (Labiatae) that were collected from different places at different time were tested for killing effects on the 2-instar larvae of Cryptorrhynchus lapathi L by adding them to the artificial diet of larvae. The experimental results indicated that adding 1-3-mL phytoecdysteroids to the artificial diet could lead 58%-100% of 2-instar larvae of C. lapathi to death within 24 days. The phytoecdysteroids extracted from the whole plant of A. multiflora which was collected before flowering time were much more effective than those extracted from the plants collected at flowering and after flowering periods, and the modified mortality rate of larvae reached 65.22%, 85.07%, and 98.11% at the dosage level of 1-mL, 2-mL, and 3-mL extracts, respectively. The extract made from root of A. multiflora plant was more effective in killing efficiency than those from stem and leaves, and the average death rates of larvae were up to 100%, 98.20% and 98.32% at dosage levels of 1-mL, 2-mL, and 3-mL extracts, respectively. The killing speed of the extracted phytoecdysteroids was slower than that of triflumuron, hexaflumuron or deltamethrin emulsifiable concentrate. The mortality rate of larvae is closely related to the feeding duration on the diets containing phytoecdysteroids. Feeding on the diets with addition of phytoecdysteroids for 16 days, more than 80% of treated 2-instar larvae of C. lapathi were led to death. The killing effect of the extracts was little affected by the growth areas of A. multiflora plant and the adding way to artificial diet. 展开更多
关键词 Ajuga multifora Bunge Cryptorrhynchus lapathi L. Larva Phytoecdysteroid
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HORMONAL CONTROL OF THE VITELLOGENESIS IN THE JAPANESE OAK SILKWORM, ANTHERAEA YAMAMAI (LEPIDOPTERA : SATURNIIDAE)
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作者 叶恭银 胡萃 龚和 《Entomologia Sinica》 CSCD 2002年第1期17-25,共9页
Effects of ecdysteroid and juvenile hormone (JH) on vitellogenesis of the Japanese oak silkworm, Antheraea yamamai are reported in this article. After topical treatment with 20 hydroxyecdysone alone or JH analog ... Effects of ecdysteroid and juvenile hormone (JH) on vitellogenesis of the Japanese oak silkworm, Antheraea yamamai are reported in this article. After topical treatment with 20 hydroxyecdysone alone or JH analog (i.e. methoprene) alone and combined treatment with these two chemicals, vitellogenin (Vg) titers in the fat body and haemolymph at the pupal stage were mostly higher than those of the control, indicating that both ecdysteroid and JH exerted a promoting effect on the synthesis of Vg. In contrast, the Vg uptake was markedly inhibited by JH while stimulating effect of the ecdysteroid could be shown that vitellin (Vt) titer in the ovary was lower after methoprene treatments, but higher after 20 hydroxyecdyson treatments. Meanwhile, effects of these two hormones on Vg synthesis in the fat body were also tested with the incubation in vitro with Grace medium containing 3H leucine and the hormones. The results demonstrated that Vg synthesis was stimulated after treating with methoprene alone or 20 hydroxyecdysone alone and combined treating with these two chemicals, and particularly ecdysteroid had more marked positive effect. To comprehensively concluded our results, it could be regarded that ecdysteroid play the main role in the regulation of vitellogenesis for the Japanese oak silkworm. 展开更多
关键词 Antheraea yamamai VITELLOGENESIS juvenile hormone ECDYSTEROID
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Developmental changes in hemolymph ecdysteroid level and prothoracicotropic hormone activity during the fifth larval instar of the Eri silkworm, Samia cynthia ricini
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作者 RONG-JING JIANG HONG TENG +2 位作者 JAN KOOLMAN SHENG LI MEI-XUN CAO 《Insect Science》 SCIE CAS CSCD 2005年第4期241-247,共7页
Developmental changes in hemolymph ecdysteroid level, ecdysteroid synthesis by prothoracic glands (PGs) in vitro, prothoracicotropic hormone (PTTH) activity in brain extracts, and PTTH activity in the hemolymph we... Developmental changes in hemolymph ecdysteroid level, ecdysteroid synthesis by prothoracic glands (PGs) in vitro, prothoracicotropic hormone (PTTH) activity in brain extracts, and PTTH activity in the hemolymph were measured during the fifth larval instar of the Eri silkworm, Samia cynthia ricini. The changing patterns of hemolymph ecdysteroid level and ecdysteroid synthesis by laGs in vitro are similar to each other, with maximums on day 9. However, on this day, hemolymph ecdysteroid level was substantially higher than ecdysteroid synthesis by PGs in vitro suggesting a high PTTH activity in the hemolymph on day 9. Moreover, the changing pattern of PTTH activity in brain extracts is also similar to that of PTTH activity in the hemolymph, both peaking on day 9. However, on this day, activity in brain extracts was much smaller than PTTH activity in the hemolymph implying that most PTTH synthesized by the brain is secreted to the hemolymph and the brain stores a very little amount of PTTH. This study provides unique insights onto the hormonal regulation of ecdysteroid synthesis in the Eri silkworm and is useful for our future studies on signal transduction of insect neurolaelatides. 展开更多
关键词 Samia cynthia ricini ECDYSTEROIDS prothoracic gland prothoracicotropic hormone RADIOIMMUNOASSAY
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