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意大利蝗卵发育过程中血蓝蛋白基因表达分析
引用本文:王香香,闫蒙云,张敏,余成敏,于非,王晗,扈鸿霞,袁亮,何岚,王伟亮,季荣,叶小芳. 意大利蝗卵发育过程中血蓝蛋白基因表达分析[J]. 昆虫学报, 2019, 62(8): 912-920. DOI: 10.16380/j.kcxb.2019.08.003
作者姓名:王香香  闫蒙云  张敏  余成敏  于非  王晗  扈鸿霞  袁亮  何岚  王伟亮  季荣  叶小芳
作者单位:(1. 新疆师范大学生命科学学院, 中亚区域跨境有害生物联合控制国际研究中心, 新疆特殊环境物种保护与调控生物学实验室, 乌鲁木齐 830054; 2. 新疆玛纳斯县蝗虫鼠害预测预报防治站, 新疆玛纳斯832200)
摘    要:【目的】意大利蝗Calliptamus italicus是新疆草原的主要优势危害种,以卵在土壤中越冬。呼吸代谢可反映蝗卵的生理状态,呼吸蛋白对于呼吸系统不完善的蝗卵尤为重要。本研究旨在明确意大利蝗卵发育过程中血蓝蛋白基因的表达情况。【方法】采用实时荧光定量PCR方法检测不同发育阶段的蝗卵以及1龄蝗蝻的血蓝蛋白2个亚基基因Hc1和Hc2的表达量。【结果】根据解剖形态观察,将意大利蝗越冬卵的整个发育过程分为10个阶段,包括9个卵发育阶段(C-Ⅰ-C-Ⅹ)和1龄蝗蝻阶段(C-Ⅹ)。Hc1和Hc2在越冬蝗卵各发育阶段以及1龄蝗蝻中均有表达。其中,在蝗卵早期发育阶段(C-Ⅰ, C-Ⅱ和C-Ⅲ),Hc1表达量逐渐增加,C-Ⅲ阶段表达量显著高于C-Ⅰ和C-Ⅱ阶段;滞育阶段(C-Ⅳ, C-Ⅴ和C-Ⅵ),胚胎发育停滞,Hc1表达量较C-Ⅲ,C-Ⅶ和C-Ⅷ阶段低;滞育后发育阶段(C-Ⅶ和C-Ⅷ),蝗卵解除滞育,快速发育,Hc1表达量较早期发育阶段和滞育阶段高,其中,C-Ⅷ阶段Hc1表达量最高(212.3156±10.5470),显著高于其他所有阶段;1龄蝗蝻(C-Ⅹ)的Hc1表达量最低,为0.4017±0.1010。Hc2表达量在C-Ⅴ阶段最高(679.7511±54.5719),显著高于其他所有阶段;除C-Ⅴ阶段外,其他各阶段之间Hc2表达量差异均不显著。Hc1在蝗卵滞育后阶段高表达,而Hc2在蝗卵滞育阶段高表达。【结论】血蓝蛋白亚基基因Hc1和Hc2在整个意大利蝗卵发育过程均有表达,且具有阶段特异性。Hc1与Hc2协同作用为蝗卵发育供氧,其中,Hc1主要负责蝗卵滞育后发育期间的氧气运载,而Hc2主要维持滞育期间的氧气运载,且载氧效率较低。研究结果可为进一步探讨意大利蝗卵的抗逆机制提供科学依据。

关 键 词:意大利蝗  越冬卵  胚胎发育  血蓝蛋白  基因表达  表达谱  

Expression profiling of hemocyanin gene in the developmental process of eggs of Calliptamus italicus (Orthoptera: Catantopidae)
WANG Xiang-Xiang,YAN Meng-Yun,ZHANG Min,YU Cheng-Min,YU Fei,WANG Han,HU Hong-Xia,YUAN Liang,HE Lan,WANG Wei-Liang,JI Rong,YE Xiao-Fang. Expression profiling of hemocyanin gene in the developmental process of eggs of Calliptamus italicus (Orthoptera: Catantopidae)[J]. Acta Entomologica Sinica, 2019, 62(8): 912-920. DOI: 10.16380/j.kcxb.2019.08.003
Authors:WANG Xiang-Xiang  YAN Meng-Yun  ZHANG Min  YU Cheng-Min  YU Fei  WANG Han  HU Hong-Xia  YUAN Liang  HE Lan  WANG Wei-Liang  JI Rong  YE Xiao-Fang
Affiliation:(1. International Center for the Collaborative Management of Cross border Pest in Central Asia, Xinjiang Key Laboratory of Special Species Conservation and Regulatory Biology, College of Life Sciences, Xinjiang Normal University, Urumqi 830054, China; 2. Manasi Station of Forecast and Control on Locusts and Rodents, Manasi, Xinjiang 832200, China)
Abstract:【Aim】 Calliptamus italicus is a major dominant pest in Xinjiang and overwinters in the soil with eggs. Respiratory metabolism can reflect the physiological state of locust eggs. Respiratory protein is especially important for C. italicus eggs, whose respiratory system is not well-developed. This study aims to clarify the expression profiles of hemocyanin gene in the developmental process of C. italicus eggs. 【Methods】 The expression levels of hemocyanin subunit genes Hc1 and Hc2 in different developmental stages of eggs and the 1st instar nymph of C. italicus were analyzed by quantitative real-time PCR. 【Results】 Based on anatomical observation, the developmental process of overwintering eggs of C. italicus is divided into ten stages, including nine egg stages (C-Ⅰ-C-Ⅸ) and the 1st instar nymph stage (C-Ⅹ). Hc1 and Hc2 were expressed in all the developmental stages of eggs and the 1st instar nymph of C. italicus. During the early stage of embryogenesis (C-Ⅰ, C-Ⅱ and C-Ⅲ), the expression level of Hc1 increased gradually, and was significantly higher at the stage C-Ⅲ than at the stages C-Ⅰ and C-Ⅱ. During the diapause stage (C-Ⅳ, C-Ⅴ and C-Ⅵ), the embryonic development was stagnant, and the expression level of Hc1 was lower than those at the stages C-Ⅲ, C-Ⅶ and C-Ⅷ. During the post-diapause developmental stage (C-Ⅶ and C-Ⅷ), the diapause of eggs were terminated and eggs developed rapidly, and the expression level of Hc1 was higher than those at the early developmental stage and diapause stage. Especially, the expression level of Hc1 in eggs at the stage C-Ⅷ was the highest (212.3156±10.5470), significantly higher than that in eggs at all of the other stages, and the expression level of Hc1 in the 1st instar nymph was the lowest (0.4017±0.1010). The expression level of Hc2 in eggs at the stage C-Ⅴ was the highest (679.7511±54.5719). The expression level of Hc2 was not significantly different at various developmental stages of eggs except stage C-Ⅴ. Hc1 was highly expressed during the post-diapause stage of eggs, while Hc2 was highly expressed during the diapause stage of eggs. 【Conclusion】 The hemocyanin subunit genes Hc1 and Hc2 are expressed throughout the developmental process of C. italicus eggs and show phase-specific expression. Hc1 and Hc2 synergistically provide oxygen for egg development. Hc1 is mainly responsible for oxygen transport during the post-diapause developmental stage of eggs, while Hc2 is mainly responsible for oxygen transport during the diapause stage of eggs with low oxygen carrying efficiency. The results can provide a scientific basis for further exploration of the stress resistance mechanism of C. italicus eggs.
Keywords:Calliptamus italicus  overwintering egg  embryonic development  hemocyanin  gene expression  expression profile  
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