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锈色粒肩天牛Apriona swainsoni为鞘翅目Coleoptera天牛科Cerambycidae昆虫,主要寄生于国槐Sophora japonica,云实Caesalpinia decapetala等豆科Leguminosae植物[1-3],是危害较为严重的蛀干害虫之一。灭幼脲(chlorbenzuron)是保幼激素的类似物[4-6],由咽侧体分泌,对昆虫的生长、发育和生殖起着重要作用[7]。成虫饲喂灭幼脲后体质量减轻,飞行能力减弱,产卵量和孵化率下降,寿命缩短等[8-9],近年来被广泛用于双翅目Diptera和鳞翅目Lepidoptera等害虫的防治,在鞘翅目昆虫防治上也取得了一定成效[10],但在天牛幼虫的防治上研究较少。虫酰肼(flubenzuron)是昆虫蜕皮激素类似物,常见蜕皮激素类似物类杀虫剂有虫酰肼(RH5992)、米满(methoxyfenozide,RH2485)和氯虫酰肼(halofenozide,RH-0345)等3种,有报道称RH5992和RH2485只对鳞翅目昆虫有活性,RH-0345只对鞘翅目有活性[5],但也有文献指出虫酰肼对双翅目摇蚊属Chironomus昆虫同样有效[11]。机体正常生理功能多是通过酶的调控实现的,抗氧化酶和解毒酶是机体内广泛存在的2类酶。McCORD等[12]发现超氧化物歧化酶(SOD)可以使自由基发生歧化反应,谷胱甘肽S转移酶(GSTs)能催化还原型谷胱甘肽形成硫醚氨酸[13],羧酸酯酶(CarE)可以催化羧酸酯生成酸和醇,从而降低细胞毒性,保护细胞膜。本研究以虫酰肼和灭幼脲处理后木屑喂饲锈色粒肩天牛幼虫,通过测定虫体内4种酶的变化,反映药物处理后天牛的生理状态,为天牛的防治提供理论依据。
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由表 1可以看出:灭幼脲处理组(T1组)过氧化氢酶活性与对照组无显著差异;虫酰肼处理组(T2组)过氧化氢酶活性下降,相较于对照组以及T1组差异显著。就各时段而言,对照组在72 h内各相邻时段之间无明显差异。T1组过氧化氢酶活性先上升,24 h后开始下降。T2组过氧化氢酶活性在12 h内已低于对照组,并且有持续下降的趋势。无论哪种药物处理,处理组超氧化物歧化酶活性均高于对照组,且差异显著;灭幼脲处理12 h虫体超氧化物歧化酶活性已高于对照组,48 h后继续上升;虫酰肼处理24 h后虫体超氧化物歧化酶活性迅速上升,之后下降,与对照组差异显著。观测羧酸酯酶活性,处理组和对照组并无显著差异。与对照相比,处理组虫体谷胱甘肽S转移酶活性均显著增高,但处理组间差异不显著;虫酰肼处理后谷胱甘肽S转移酶活性在24 h时达到顶峰,之后下降;而灭幼脲处理后虫体谷胱甘肽S转移酶酶活性在12 h时已显著升高,之后下降。
表 1 不同处理对天牛体内4种酶的影响
Table 1. Effects of 4 enzymatics activities under different treatments
处理 t/h 酶活性/(×16.67 μkat.L-1) CAT SOD CarE GSTs 对照 12 26.06±1.22aA 47.31±2.49aA 35.71±7.13aA 21.71±1.24aA 24 25.59±0.28aA 45.53±2.95aA 34.83±5.02aA 21.98±2.14aA 36 24.24±2.58aA 48.92±3.01aB 30.78±8.89 aA 21.70±2.10aA 48 23.41±1.38aA 52.46±6.21aC 34.95±7.32aA 21.97±0.43aA 60 23.76±2.53aA 50.34±2.99aC 31.51±6.01aA 21.47±1.53aA 72 21.96±3.14aA 49.88±4.40aC 28.76±3.28aA 22.32±2.25aA T1 12 28.05±4.76aB 53.62±4.85bD 35.93±14.56aA 24.81±0.97bB 24 36.49±3.16aC 52.34±5.40bD 37.37±16.25 aA 23.08±1.45bC 36 20.26±1.75aD 53.67±3.90bD 36.52±17.06aA 24.51±0.61bD 48 23.59±3.69aE 53.25±1.23bD 34.69±5.87aA 23.78±1.37bD 60 25.54±5.63aE 49.92±1.22bE 32.41±8.95aA 24.78±0.96bD 72 17.58±0.80aF 55.22±1.29bF 35.36±9.25aA 24.55±0.68bD T2 12 19.86±1.12bG 54.67±2.51bG 35.31±13.91aA 22.79±2.70bE 24 19.64±5.13bG 60.09±8.00bH 37.52±1.61aA 25.62±4.87bF 36 16.35±2.45bH 52.46±4.77bI 37.75±5.93 aA 24.27±1.60bF 48 18.16±0.66bH 52.55±0.92bI 35.40±5.63aA 23.10±1.10bF 60 14.37±1.35bI 51.82±2.27bI 33.90±4.55aA 23.54±1.34bF 72 12.93±4.57bI 45.47±6.61bJ 36.19±6.78aA 24.03±4.02bF 说明:表中数据为平均值±标准差;不同小写字母表示2种处理之间有显著差异(P<0.05),不同大写字母表示同一处理不同时间段间有显著差异(P<0.05)。
Oxidase and detoxifying enzyme activity of Apriona swainsoni (Hope) with diflubenzuron and flubenzuron
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摘要: 为探究虫酰肼和灭幼脲对锈色粒肩天牛Apriona swainsoni(Hope)生理机制的影响,用经灭幼脲(T1)和虫酰肼(T2)处理的木屑饲喂锈色粒肩天牛幼虫,测定天牛幼虫的过氧化氢酶(CAT),超氧化物歧化酶(SOD),谷胱甘肽S转移酶(GSTs)和羧酸酯酶(CarE)活性,每12 h取样,持续72 h。结果显示:T1处理CAT活性先升高,24 h后降低;T2处理CAT活性降低;与对照组相比,T1和T2处理SOD活性、GSTs活性显著增高(P < 0.05),CarE活性则无明显差异。推测锈色粒肩天牛幼虫GSTs活性和虫体抗性的产生有关,GSTs活性可能是天牛幼虫的抗性标识物。Abstract: To research the influence of tebufenozide and chlorbenzuron on the physiological mechanism of Apriona swainsoni (Hope), A. swainsoni larvae were fed with sawdust treated by chlorbenzuron (T1) and tebufenozide (T2). These were then sampled to determine enzymatic activity of catalase (CAT), superoxide dismutase (SOD), glutathione S transferases (GSTs), and carboxylesterase (CarE) in larvae at intervals of 12 h to 72 h in succession. Results showed that after treatment with chlorbenzuron, the enzymatic activity of CAT increased first and then decreased 24 h later. Enzymatic activity of SOD maintained a high level. Also, after treatment by tebufenozide, the enzymatic activity of CAT decreased first with enzymatic activity of SOD being higher than the control. After treatment by two types of the pesticides, enzymatic activity of GSTs was higher than the control with no differences of CarE activity with the control. Therefore, enzymatic activity of GSTs in A. swainsoni was assumed to be associated with resistance generated in the larva and was likely to be a resistance marker of the larvae.
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Key words:
- forest protection /
- Apriona swainsoni (Hope) /
- chlorbenzuron /
- tebufenozide /
- enzymatic activity
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表 1 不同处理对天牛体内4种酶的影响
Table 1. Effects of 4 enzymatics activities under different treatments
处理 t/h 酶活性/(×16.67 μkat.L-1) CAT SOD CarE GSTs 对照 12 26.06±1.22aA 47.31±2.49aA 35.71±7.13aA 21.71±1.24aA 24 25.59±0.28aA 45.53±2.95aA 34.83±5.02aA 21.98±2.14aA 36 24.24±2.58aA 48.92±3.01aB 30.78±8.89 aA 21.70±2.10aA 48 23.41±1.38aA 52.46±6.21aC 34.95±7.32aA 21.97±0.43aA 60 23.76±2.53aA 50.34±2.99aC 31.51±6.01aA 21.47±1.53aA 72 21.96±3.14aA 49.88±4.40aC 28.76±3.28aA 22.32±2.25aA T1 12 28.05±4.76aB 53.62±4.85bD 35.93±14.56aA 24.81±0.97bB 24 36.49±3.16aC 52.34±5.40bD 37.37±16.25 aA 23.08±1.45bC 36 20.26±1.75aD 53.67±3.90bD 36.52±17.06aA 24.51±0.61bD 48 23.59±3.69aE 53.25±1.23bD 34.69±5.87aA 23.78±1.37bD 60 25.54±5.63aE 49.92±1.22bE 32.41±8.95aA 24.78±0.96bD 72 17.58±0.80aF 55.22±1.29bF 35.36±9.25aA 24.55±0.68bD T2 12 19.86±1.12bG 54.67±2.51bG 35.31±13.91aA 22.79±2.70bE 24 19.64±5.13bG 60.09±8.00bH 37.52±1.61aA 25.62±4.87bF 36 16.35±2.45bH 52.46±4.77bI 37.75±5.93 aA 24.27±1.60bF 48 18.16±0.66bH 52.55±0.92bI 35.40±5.63aA 23.10±1.10bF 60 14.37±1.35bI 51.82±2.27bI 33.90±4.55aA 23.54±1.34bF 72 12.93±4.57bI 45.47±6.61bJ 36.19±6.78aA 24.03±4.02bF 说明:表中数据为平均值±标准差;不同小写字母表示2种处理之间有显著差异(P<0.05),不同大写字母表示同一处理不同时间段间有显著差异(P<0.05)。 -
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https://zlxb.zafu.edu.cn/article/doi/10.11833/j.issn.2095-0756.2018.01.023