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低温对绿盲蝽成虫耐寒性及主要耐寒物质的影响
Effect of Low Temperature on Cold Hardiness and the Main Cold Hardiness Substances of Apolygus lucorum Adults
旨在明确低温下绿盲蝽的存活能力、过冷却点和结冰点以及体内主要耐寒物质含量的变化,探究低温下绿盲蝽的耐寒能力以及生理响应。设置不同低温处理,测定绿盲蝽成虫的存活率;采用热电偶法,测定不同低温和时间组合下绿盲蝽成虫的过冷却点和结冰点;利用生理生化测定法比较山梨醇、葡萄糖、海藻糖、果糖等生化物质含量的变化。结果表明,低温-5、-10℃处理24 h及以上时间后,绿盲蝽成虫存活率均为0;0、5、15℃处理24 h及以上时间后,绿盲蝽成虫存活率均达96%以上。低温处理能够降低绿盲蝽成虫的过冷却点和结冰点,以5℃处理24 h时下降幅度最大;但处理时间进一步延长至48 h和96 h后,不同低温均不能有效降低过冷却点和结冰点。低温下绿盲蝽成虫体内的耐寒物质含量发生变化:处理24 h后,果糖含量在15℃下增加;处理48 h后,在10~15℃的低温下,果糖、葡萄糖、海藻糖、山梨醇含量增加。研究结果为揭示绿盲蝽的耐寒机理及其预测预报工作提供有益参考。
This study aims to investigate the physiological response mechanism of A. lucorum adults to low temperature by clarifying the survival, the supercooling point (SCP) and freezing point (FP), as well as the main cold hardiness substances under low temperature. The survival rate of A. lucorum adults was measured by different low temperature treatments. Thermocouple method was used to determine the SCP and FP under different temperature and time. The content of sorbitol, glucose, trehalose and fructose were compared by physiological and biochemical assay. The results showed that after being treated at -5℃ and -10℃ for 24 h or more, the survival rate of A. lucorum adults was 0. After treatment at 0, 5 and 15℃ for 24 h or more, the survival rate was above 96%. After different temperature treatments at 0, 5 and 15℃ for 24 h, the SCP and FP of the adult were decreased. When treated at 5℃ for 24 hours, there was the largest decline. However, when the treatment time extended to 48 h and 96 h, the SCP and FP of the adult were no longer decline. The content of biochemical substances in A. lucorum changed at low temperature. After treatment at 15℃ for 24 h, the content of fructose was increased. After 48 h treatment at a low temperature of 10℃ to 15℃, the contents of fructose, glucose, trehalose and sorbitol were increased. These results provide a reference for revealing the cold tolerance mechanism of A. lucorum and its forecasting.
绿盲蝽 / 成虫 / 低温 / 耐寒性 / 耐寒物质 {{custom_keyword}} /
Apolygus lucorum / adult / low temperature / cold hardiness / cold hardiness substances {{custom_keyword}} /
表1 试验仪器及来源 |
试验仪器 | 来源 |
---|---|
生物显微镜BM2000 | 上海光学仪器厂,中国 |
PGX-150B智能光照培养箱 | 无锡沃信仪器有限公司,中国 |
双光束紫外可见分光光度计 | 上海仪电分析仪器有限公司,中国 |
Master-QUT试验室超纯水机 | 上海和泰仪器有限公司,中国 |
PWC 124型分析天平 | 艾德姆衡器(武汉)有限公司,中国 |
浮游生物计数框 | 金坛市康华电子仪器制造厂,中国 |
移液器 | Eppendorf,德国 |
表2 稀释水培养基成分及含量 |
成分 | 浓度/(mg/L) |
---|---|
氯化钙(CaCl2) | 2.94 |
硫酸镁(MgSO4) | 1.23 |
碳酸氢钠(NaHCO3) | 0.65 |
氯化钾(KCl) | 0.0625 |
表3 BG-11培养基成分及含量 |
成分 | 浓度/(g/L) |
---|---|
硝酸钠(NaNO3) | 1.5 |
磷酸氢二钾(K2HPO4) | 0.04 |
七水硫酸镁(MgSO4·7H2O) | 0.075 |
柠檬酸(C6H8O7) | 0.036 |
枸椽酸铁铵(C6H10FeNO8) | 0.006 |
乙二胺四乙酸二钠(EDTANA2) | 0.001 |
碳酸钠(Na2CO3) | 0.02 |
硼酸(H2BO3) | 2.86 |
四水氯化锰(MnCl2·4H2O) | 1.86 |
七水硫酸锌(ZnSO4·7H2O) | 0.22 |
钼酸钠(NaMoO4) | 0.39 |
五水硫酸铜(CuSO4·5H2O) | 0.08 |
硝酸钴六水合物(Co(NO3)2·6H2O) | 0.05 |
表4 灭多威对小球藻的96 h抑制效应 |
浓度/(mg/L) | 0.05 | 0.19 | 0.7 | 2.65 | 10 | 37.2 |
---|---|---|---|---|---|---|
抑制效应/% | 9.60 | 14.70 | 19.80 | 32.40 | 51.30 | 78.67 |
回归方程 | y=0.7084x+4.4169 | |||||
相关系数 | 0.975* | |||||
EC50/(mg/L) | 6.76 |
注:*表示相关显著(P<0.05),下同。 |
表5 灭多威对大型溞的急性毒性 |
浓度/(μg/L) | 死亡率/% | ||
---|---|---|---|
24 h | 48 h | 96 h | |
空白组 | 0 | 0 | 0 |
25 | 10.0 | 53.3 | 80.0 |
32.9 | 26.7 | 73.3 | 90.0 |
43.43 | 36.7 | 90.0 | 93.3 |
57.24 | 63.3 | 93.3 | 96.7 |
75.44 | 66.6 | 96.7 | 100 |
99.44 | 70.0 | 100 | 100 |
回归方程 | y=3.0312x-0.3309 | y=3.5589x+0.2059 | y=2.441x+2.4863 |
相关系数 | 0.970* | 0.974* | 0.986* |
LC50 | 57.37 | 22.23 | 12.58 |
95%置信限 | 50.58~65.46 | 18.32~26.99 | 10.47~14.45 |
表6 灭多威对斑马鱼的急性毒性 |
浓度/(mg/L) | 死亡率/% | ||
---|---|---|---|
24 h | 48 h | 96 h | |
0 | 0 | 0 | 0 |
1 | 0 | 0 | 17 |
1.57 | 0 | 10 | 30 |
2.47 | 7 | 20 | 67 |
3.87 | 20 | 30 | 77 |
6.08 | 27 | 53 | 87 |
9.55 | 60 | 73 | 100 |
15 | 73 | 100 | 100 |
回归方程 | y=2.69x+2.4768 | y=2.165x+3.3674 | y=2.7664x+4.0835 |
相关系数 | 0.976* | 0.978* | 0.964* |
LC50 | 8.64 | 5.00 | 2.12 |
95%置信限 | 7.23~10.77 | 4.20~6.03 | 1.76~2.50 |
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【目的】研究重大入侵害虫三叶斑潜蝇(Liriomyza trifolii)对快速冷驯化和长时冷驯化的响应以及3个地理种群(湖北省武汉市、海南省海口市和安徽省安庆市)的耐寒性,为明确该虫在我国高纬度地区定殖的潜在可能性提供参考。【方法】将海南三叶斑潜蝇种群蛹和成虫在-12、-14、-16、-18、-20、-22和-24℃低温暴露30 min,选择存活率为15%—30%的温度为识别温度。设置5℃为快速冷驯化的驯化温度,12℃为长时冷驯化的驯化温度,将三叶斑潜蝇蛹和成虫置于5℃下1—6 h或置于12℃下1—6 d,再置于识别温度下30 min检测存活率,未驯化的蛹和成虫也置于识别温度下30 min作为对照,比较其冷驯化后低温存活率的差异。另外,设置5个靶标温度,分别为0、-5、-10、-15和-20℃,将3个地理种群的三叶斑潜蝇蛹在靶标温度下冷暴露2 h,比较其存活率差异。最后,利用过冷却点测定仪对3个地理种群蛹的过冷却点(supercooling point,SCP)进行测定。【结果】三叶斑潜蝇蛹和成虫暴露于-20℃下30 min后,其存活率分别为15.0%、19.6%、21.0% (介于15%—30%),因此将-20℃确定为三叶斑潜蝇的冷驯化识别温度。短时间的5℃低温暴露提高了三叶斑潜蝇蛹和成虫的耐寒性,成虫对快速冷驯化的响应更加积极。1 h和2 h的冷驯化效果最好,随着暴露时间的延长,其驯化效果会逐渐减弱直至消失。在6 d内,不同时长的12℃低温暴露均会提高三叶斑潜蝇蛹和成虫的耐寒性且经历不同时长冷驯化后其耐寒性差异较小。在5、0、-5℃的低温暴露下,海南种群与安徽、湖北种群蛹的存活率差异不显著,在-10、-15、-20℃的低温暴露下,海南种群蛹的存活率显著低于安徽和湖北种群。安徽(-22.19℃)和湖北(-22.19℃) 种群蛹的过冷却点显著低于海南种群(-21.06℃)。【结论】三叶斑潜蝇的耐寒性可以通过快速冷驯化或长时冷驯化获得增强,这可能是三叶斑潜蝇逐步向我国高纬度地区扩散的原因之一。湖北和安徽种群表现出的耐寒性较海南种群强。研究结果有助于预测三叶斑潜蝇在我国的越冬分布区域,指导其监测预警及防控。
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【目的】快速冷驯化能在短时间内迅速提高昆虫的耐寒性,是昆虫应对外界温度急剧变化以及短时低温胁迫的重要途径。本研究旨在探究入侵杂草豚草Ambrosia artemisiifolia生防天敌广聚萤叶甲Ophraella communa对不同强度快速冷驯化的生理响应机制。【方法】分别对广聚萤叶甲成虫进行了不同温度(-4, 0, 4和8℃)下4 h及0℃下不同时间(1, 4, 8和16 h)的快速冷驯化处理,并对其体内的生理物质含量和保护酶活性进行了测定。【结果】除8℃/4 h,0℃/1 h和0℃/8 h外,其余冷驯化处理均使广聚萤叶甲成虫过冷却点显著降低(PP≥0.05)。【结论】广聚萤叶甲快速冷驯化的诱导具有其临界强度值和最适条件,过大强度的驯化处理反而不利于其耐寒性的提高。本研究结果对于深入阐明广聚萤叶甲越冬策略以及人工培育耐寒种群的实践具有一定参考价值。
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意大利蝗Calliptamus italicus是新疆草原主要优势危害种,以滞育卵越冬。为弄清意大利蝗越冬卵发育及滞育过程的抗寒机制,本研究检测了自然条件下越冬卵的糖类、多元醇、脂肪和氨基酸等主要抗寒物质的含量变化,分析蝗卵发育过程不同阶段抗寒物质含量的变化趋势。结果表明,意大利蝗卵在越冬过程中,除脂肪含量无显著差异外,海藻糖、甘油、山梨醇以及氨基酸在低温滞育期作为主要抗寒物质在体内积累,有助于增强蝗卵的抗寒性;而糖原、葡萄糖、果糖、肌醇等则作为能源物质为蝗卵发育提供能量,且这些物质在蝗卵体内相互转化。研究结果可为深入研究意大利蝗卵的适应和发生机制奠定基础。
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赵丽娜, 张卫星, 秦明, 等. 外源添加山梨醇和海藻糖对意大利蜜蜂抗寒性能的影响[J]. 动物营养学报, 2021, 33(3):1652-1662.
本试验旨在通过测定意大利蜜蜂(简称意蜂)体内山梨醇、海藻糖等相关生化物质含量、抗氧化酶活性及抗寒基因表达的变化,探究外源添加山梨醇和海藻糖在蜜蜂抗寒过程中的作用机制。从意蜂姊妹蜂群中随机选取900只刚出房的蜜蜂,随机分成6组,每组3个重复,每个重复50只蜜蜂,分别饲喂蔗糖溶液(对照)、海藻糖溶液和山梨醇溶液。6组均于常温(30℃)下饲养12 d,其中3组在饲喂12 d后进行冷处理(4℃冷驯化2 h)。饲喂12 d后立即测定蜜蜂过冷却点、冰点,比较其抗寒性能差异。然后分别测定蜂体游离水、糖原、脂肪含量、山梨醇脱氢酶(SDH)、海藻糖酶(THL)活性以及海藻糖-6-磷酸合成酶(TPS)、SDH mRNA相对表达量的差异;另取900只意蜂,按照以上分组饲养,用于测定血淋巴中小分子糖醇含量。结果表明:1)与对照组相比,山梨醇组和海藻糖组过冷却点分别降低0.65(P<0.05)、0.41℃(P<0.05)。2)与对照组相比,常温以及冷处理条件下,山梨醇组和海藻糖组蜂体游离水含量变化不显著(P>0.05)。3)与对照组相比,常温以及冷处理条件下,海藻糖组和山梨醇组蜂体内脂肪含量变化不显著(P>0.05),而糖原含量显著提高(P<0.05)。4)与对照组相比,常温以及冷处理条件下,山梨醇组和海藻糖组血淋巴中葡萄糖含量显著增加(P<0.05);常温条件下,山梨醇组血淋巴中果糖含量显著提高(P<0.05),而冷处理条件下,海藻糖组血淋巴中果糖含量显著降低(P<0.05);常温条件下,山梨醇组血淋巴中山梨醇含量显著增加(P<0.05),而冷处理条件下,结果差异不显著(P>0.05)。5)与对照组相比,常温以及冷处理条件下,山梨醇组和海藻糖组蜂体SDH和THL活性均显著提高(P<0.05)。6)与对照组相比,常温以及冷处理条件下,山梨醇组SDH mRNA相对表达量显著上调(P<0.05);常温条件下,海藻糖组TPS mRNA相对表达量显著下调(P<0.05)。综上所述,在室内饲养的离群意蜂外源添加山梨醇、海藻糖显著降低了意蜂的过冷却点、冰点,进而提高了抗寒性。山梨醇和海藻糖通过调节意蜂血淋巴中小分子糖醇含量,以及与山梨醇、海藻糖代谢通路相关的基因表达量来提高蜜蜂的抗寒性。
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