中药材储藏期主要害虫种群生态及气调毒理研究
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摘要
药材甲Stegobium paniceum、烟草甲Lasioderma serricorne和咖啡豆象Araecerusfasciculatus均是贵阳地区中药材储藏昆虫群落的优势种,三种害虫的发生和危害每年导致我国大量的中药材报废、药效降低或价值丧失,是中药材储藏期害虫中经济意义最为重要的代表类群。本论文以中药材为寄主食料研究三种中药材储藏期害虫实验种群生态学,探讨其发生危害规律以及环境因子对其生长发育及繁殖参数的影响,以便进行有效管理和调控;在种群生态学研究的基础上,进行了二氧化碳气调技术杀灭中药材储藏期害虫的实验研究,分析了三种害虫对二氧化碳气调胁迫的耐性差异,获得了一些指导气调杀虫实践的关键数据;论文重点研究了以上三种昆虫的羧酸酯酶、磷酸酯酶、乙酰胆碱酯酶、超氧化物岐化酶和谷胱甘肽转移酶的生物化学性质,探讨了以上酶系在二氧化碳气调胁迫下的应激反应,以便丰富气调杀虫机理及昆虫抗气性分析的理论体系。主要研究结果如下:
一、中药材储藏期主要害虫实验种群生态学研究
1、药材甲实验种群卵、幼虫和蛹的发育起点温度分别为5.05、6.00和8.36℃,有效积温分别为177.9、939.8和153.7日度;烟草甲实验种群卵、幼虫和蛹的发育起点温度分别为16.51、11.07和11.78℃,有效积温分别为89.59、669.53和137.12日度;咖啡豆象实验种群卵、幼虫和蛹的发育起点温度分别为9.60、11.48和7.60℃,有效积温分别为135.51、523.57和157.10日度。温度对三种昆虫的生长发育和繁殖参数影响显著。
2、在贵阳地区,药材甲一年发生3～4代,以第3代老熟幼虫或第4代低龄期幼虫越冬;烟草甲一年发生4～5代,越冬虫期为第4代老熟幼虫或第5代低龄幼虫;咖啡豆象一年发生3～4代,少数发生5代,多以幼虫越冬,极少见蛹或成虫越冬。
二、二氧化碳气调对中药材储藏期主要害虫的毒力作用
1、10%二氧化碳气调处理药材甲、烟草甲和咖啡豆象成虫,24 h内,三种昆虫死亡率均较低,分别为55.56±6.94、58.89±8.39和58.89±5.09%。30%、50%和80%二氧化碳气调则能很快的杀灭三种实验昆虫。
2、10%二氧化碳气调对药材甲、烟草甲和咖啡豆象毒力测定的半数致死时间分别为22.39,23.21和21.15 h,而致死99%的昆虫个体,理论上需要4～5 d。含CO_230%以上浓度气体作用以上三种昆虫,在25±1℃,rh70±5%条件下,45 h内能实现药材甲、烟草甲和咖啡豆象三种实验昆虫100%死亡。
三、中药材储藏期主要害虫部分酶学性质及二氧化碳气调对其性质的影响
1、药材甲经二氧化碳气调处理3、6和9 h后,羧酸酯酶比活力分别比对照升高了7.81%、14.96%和22.58%;烟草甲在3 h处理时羧酸酯酶比活力下降了3.18%,6 h处理时,升高了22.2%,9 h处理时基本恢复至对照水平;咖啡豆象在二氧化碳气调胁迫下处理3和6 h时活力分别比对照升高了31.75%和50.82%。三种昆虫的羧酸酯酶酶促动力学参数也受二氧化碳气调胁迫的影响。
2、在高浓度二氧化碳气调处理6和9 h时,药材甲和烟草甲AChE K_m值均分别由14.290μmol/L和30.392μmol/L持续降低至最低点1.830μmol/L和7.632μmol/L;而随着处理时间的延长,其K_m值不再降低,而逐步恢复至对照水平。高浓度CO_2对两种昆虫AChE的V_(max)有相似影响。
3、二氧化碳气调处理药材甲幼虫6 h时,其ACP酶活力显著升高78.59%;二氧化碳气调胁迫处理6 h时,咖啡豆象幼虫ACP活力升高了72.45%。
4、药材甲幼虫在二氧化碳气调胁迫下,碱性磷酸酯酶ALP的活力先是受到一定程度的抑制,经3 h气调降低了30.42%,而随着气调时间的延长,ALP活力逐渐恢复,至6 h处理时,又表现为诱导轻微激活。
5、二氧化碳气调胁迫处理3和6 h时,药材甲、烟草甲和咖啡豆象成虫SOD酶活力均有一定程度的持续升高,其中处理6h,其活力升高显著,分别有对照2.011±0.954,2.664±0.218和1.458±0.718上升到3.135±0.105,3.050±0.673和2.975±0.229酶活力单位/头/30min。
6、二氧化碳气调对三种昆虫GSTs的影响研究,发现仅在二氧化碳气调胁迫3 h时烟草甲GSTs酶比活力有稍微降低,其余在二氧化碳气调处理3、6和9 h时,药材甲、烟草甲和咖啡豆象成虫的GSTs酶比活力均有一定程度的升高,可见,二氧化碳气调亚致死处理对三种实验昆虫GSTs有一定的诱导激活作用。
Drugstore beetle Stegobium paniceum,cigarette beetle Lasioderma serricorne and coffee bean beetle Araecerus fasciculatus,the most important three members of pests to destroy stored Chinese medicinal materials(CMM),are dominant populations in stored CMM insect community. Breeding of these pests are responsible to the quantity discarding as useless of CMM,decreasing of medicine power even to become garbage.In the present paper,the population ecology of the three pests were investigated in laboratory with given CMM host.The habit and the influence of ecological factor(temperature) on both the development and propagating of the pests were investigated.Whilst,the practice of carbon dioxide enriched atmosphere was carried out in management of pests in CMM.Some key information which are important to kill stored CMM pests was supplied.What's the most important is that toxicologically and biological characteristic of Carboxylesterases(CarE),Acetylcholinesterase(ACHE),Acid Phosphatases(ACP) & Alkaline Phosphatases(ALP),Superoxide Dismutase(SOD) and Glutathione Transferase(GSTs) from the three pest above were investigated.The current research presented some important and basic information to the research in toxicology of control atmosphere and related fields.The main results are as followed.
The population ecology of pest in stored CMM
1.The threshold temperature of development of drugstore beetle egg,larvae and pupae was 5.05,6.00 and 8.36℃respectively,and the accumulation temperature was 177.9,939.8 and 153.7 day degree.The threshold temperature of development for cigarette beetle egg,larvae and pupae was 16.5172,11.07 and 11.78℃respectively,and the accumulation temperature was 89.59, 669.53 and 137.12 day degree.The threshold temperature of development for coffee bean beetle egg,larvae and pupae was 9.60,11.48 and 7.60℃respectively,and the accumulation temperature was 135.51,523.57 and 157.10 day degree.
2.In Guiyang,there are generally 3～4 generations per year in stored CMM for drugstore beetle.Overwintering may be passed in the older larval stage.There are generally 4～5 generation per year in stored CMM for cigarette beetle.Overwintering may be passed in the 4~(th) generation older larval stage or the 5~(th) generation younger larval stage.There are generally 3～4 generations per year in stored CMM for coffee bean beetle.Overwintering may be passed in the older larval stage with a few adults or pupae stages.
3.Temperature was a key ecological factor to the development of both drugstore beetle, cigarette beetle and coffee bean beetle.Three insects can finish the reproduced and developed at the temperature of 17 to 35℃.However,The more great powerful reproduced and the better development speed at higher temperature were noted in our research.
The toxicity of carbon dioxide enriched atmosphere to pest in stored CMM
The toxicity of control atmosphere with different CO_2 levels to pests in stored CMM were investigated.The LT_(50)(the half lethal exposure periods),LT_(99)(the complete lethal exposure periods) and toxicological regress equation were calculated which could be use in practice of pest control base on the Goodness-of-Fit chi square test.
1.The mortality rate was 55.56±6.94、58.89±8.39 and 58.89±5.09%respectively when the three experimental pest drugstore beetle Stegobium paniceum,cigarette beetle Lasioderma serricorne and coffee bean beetle Araecerus fasciculatus was exposed to the control atmosphere with 10%CO_2 for 24 h.And The LT_(50) was 22.39,23.21 and 21.15 h respectively.However,the higher mortality rate was noted when the pest was exposed to the control atmosphere with 30% CO_2 or higher level within a shorter exposure periods.
2.Results showed that experimental pests exposed to control atmosphere with 30%CO_2 or more could be killed within 45 h.Tempreture and CO_2 levels or other composing and exposure period are some key factors to control pest with control atmosphere.
The toxicologically and biological characteristic of some key enzymes in pests exposed to the CO_2 enriched atmosphere
1.Compared to control,the specific activity of CarE from drugstore beetle which was exposed CO_2 enriched atmosphere for 3 h,6 h and 9 h increased 7.81%、14.96%and 22.58% respectively.That of cigarette beetle exposed for 3 h decreased 3.18%and for 6 h increased 22.2%. That of coffee bean beetle exposed for 3 h,6 h increased 31.75%and 50.82%respectively.Whilst, both the K_m and V_(max) of CarE from the pest mentioned above were affected by the press of CO_2 enriched atmosphere.
2.The specific activity of AChE from drugstore beetle and cigarette beetle which was exposed CO_2 enriched atmosphere for 6 h and 9 h decreased from 14.29μmol/L and 30.39μmol/L to 1.83μmol/L and 7.63μmol/L respectively.However,this treat did not continue with the prolonging of exposure periods.The K_m and V_(max) of AChE from dugstore beetle and cigarette beetle were affected by the press of CO_2 enriched atmosphere.
3、Compared to control,the specific activity of ACP from larvae of drugstore beetle and coffee bean beetle which were exposed to CO_2 enriched atmosphere for 6 h increased 78.59% and 72.45%respectively.The specific activity of ALP changed when the larvae of drugstore beetle were exposed to control atmosphere for 0～6 h.
4、The activity of SOD from drugstore beetle,cigarette beetle and coffee bean beetle which were exposed to CO_2 enriched atmosphere for 6 h increased from 2.011±0.954,2.664±0.218 and 1.458±0.718 to 3.135±0.105,3.050±0.673 and 2.975±0.229 enzyme unite/pest/30 min.
一、中药材储藏期主要害虫实验种群生态学研究
1、药材甲实验种群卵、幼虫和蛹的发育起点温度分别为5.05、6.00和8.36℃,有效积温分别为177.9、939.8和153.7日度;烟草甲实验种群卵、幼虫和蛹的发育起点温度分别为16.51、11.07和11.78℃,有效积温分别为89.59、669.53和137.12日度;咖啡豆象实验种群卵、幼虫和蛹的发育起点温度分别为9.60、11.48和7.60℃,有效积温分别为135.51、523.57和157.10日度。温度对三种昆虫的生长发育和繁殖参数影响显著。
2、在贵阳地区,药材甲一年发生3～4代,以第3代老熟幼虫或第4代低龄期幼虫越冬;烟草甲一年发生4～5代,越冬虫期为第4代老熟幼虫或第5代低龄幼虫;咖啡豆象一年发生3～4代,少数发生5代,多以幼虫越冬,极少见蛹或成虫越冬。
二、二氧化碳气调对中药材储藏期主要害虫的毒力作用
1、10%二氧化碳气调处理药材甲、烟草甲和咖啡豆象成虫,24 h内,三种昆虫死亡率均较低,分别为55.56±6.94、58.89±8.39和58.89±5.09%。30%、50%和80%二氧化碳气调则能很快的杀灭三种实验昆虫。
2、10%二氧化碳气调对药材甲、烟草甲和咖啡豆象毒力测定的半数致死时间分别为22.39,23.21和21.15 h,而致死99%的昆虫个体,理论上需要4～5 d。含CO_230%以上浓度气体作用以上三种昆虫,在25±1℃,rh70±5%条件下,45 h内能实现药材甲、烟草甲和咖啡豆象三种实验昆虫100%死亡。
三、中药材储藏期主要害虫部分酶学性质及二氧化碳气调对其性质的影响
1、药材甲经二氧化碳气调处理3、6和9 h后,羧酸酯酶比活力分别比对照升高了7.81%、14.96%和22.58%;烟草甲在3 h处理时羧酸酯酶比活力下降了3.18%,6 h处理时,升高了22.2%,9 h处理时基本恢复至对照水平;咖啡豆象在二氧化碳气调胁迫下处理3和6 h时活力分别比对照升高了31.75%和50.82%。三种昆虫的羧酸酯酶酶促动力学参数也受二氧化碳气调胁迫的影响。
2、在高浓度二氧化碳气调处理6和9 h时,药材甲和烟草甲AChE K_m值均分别由14.290μmol/L和30.392μmol/L持续降低至最低点1.830μmol/L和7.632μmol/L;而随着处理时间的延长,其K_m值不再降低,而逐步恢复至对照水平。高浓度CO_2对两种昆虫AChE的V_(max)有相似影响。
3、二氧化碳气调处理药材甲幼虫6 h时,其ACP酶活力显著升高78.59%;二氧化碳气调胁迫处理6 h时,咖啡豆象幼虫ACP活力升高了72.45%。
4、药材甲幼虫在二氧化碳气调胁迫下,碱性磷酸酯酶ALP的活力先是受到一定程度的抑制,经3 h气调降低了30.42%,而随着气调时间的延长,ALP活力逐渐恢复,至6 h处理时,又表现为诱导轻微激活。
5、二氧化碳气调胁迫处理3和6 h时,药材甲、烟草甲和咖啡豆象成虫SOD酶活力均有一定程度的持续升高,其中处理6h,其活力升高显著,分别有对照2.011±0.954,2.664±0.218和1.458±0.718上升到3.135±0.105,3.050±0.673和2.975±0.229酶活力单位/头/30min。
6、二氧化碳气调对三种昆虫GSTs的影响研究,发现仅在二氧化碳气调胁迫3 h时烟草甲GSTs酶比活力有稍微降低,其余在二氧化碳气调处理3、6和9 h时,药材甲、烟草甲和咖啡豆象成虫的GSTs酶比活力均有一定程度的升高,可见,二氧化碳气调亚致死处理对三种实验昆虫GSTs有一定的诱导激活作用。
Drugstore beetle Stegobium paniceum,cigarette beetle Lasioderma serricorne and coffee bean beetle Araecerus fasciculatus,the most important three members of pests to destroy stored Chinese medicinal materials(CMM),are dominant populations in stored CMM insect community. Breeding of these pests are responsible to the quantity discarding as useless of CMM,decreasing of medicine power even to become garbage.In the present paper,the population ecology of the three pests were investigated in laboratory with given CMM host.The habit and the influence of ecological factor(temperature) on both the development and propagating of the pests were investigated.Whilst,the practice of carbon dioxide enriched atmosphere was carried out in management of pests in CMM.Some key information which are important to kill stored CMM pests was supplied.What's the most important is that toxicologically and biological characteristic of Carboxylesterases(CarE),Acetylcholinesterase(ACHE),Acid Phosphatases(ACP) & Alkaline Phosphatases(ALP),Superoxide Dismutase(SOD) and Glutathione Transferase(GSTs) from the three pest above were investigated.The current research presented some important and basic information to the research in toxicology of control atmosphere and related fields.The main results are as followed.
The population ecology of pest in stored CMM
1.The threshold temperature of development of drugstore beetle egg,larvae and pupae was 5.05,6.00 and 8.36℃respectively,and the accumulation temperature was 177.9,939.8 and 153.7 day degree.The threshold temperature of development for cigarette beetle egg,larvae and pupae was 16.5172,11.07 and 11.78℃respectively,and the accumulation temperature was 89.59, 669.53 and 137.12 day degree.The threshold temperature of development for coffee bean beetle egg,larvae and pupae was 9.60,11.48 and 7.60℃respectively,and the accumulation temperature was 135.51,523.57 and 157.10 day degree.
2.In Guiyang,there are generally 3～4 generations per year in stored CMM for drugstore beetle.Overwintering may be passed in the older larval stage.There are generally 4～5 generation per year in stored CMM for cigarette beetle.Overwintering may be passed in the 4~(th) generation older larval stage or the 5~(th) generation younger larval stage.There are generally 3～4 generations per year in stored CMM for coffee bean beetle.Overwintering may be passed in the older larval stage with a few adults or pupae stages.
3.Temperature was a key ecological factor to the development of both drugstore beetle, cigarette beetle and coffee bean beetle.Three insects can finish the reproduced and developed at the temperature of 17 to 35℃.However,The more great powerful reproduced and the better development speed at higher temperature were noted in our research.
The toxicity of carbon dioxide enriched atmosphere to pest in stored CMM
The toxicity of control atmosphere with different CO_2 levels to pests in stored CMM were investigated.The LT_(50)(the half lethal exposure periods),LT_(99)(the complete lethal exposure periods) and toxicological regress equation were calculated which could be use in practice of pest control base on the Goodness-of-Fit chi square test.
1.The mortality rate was 55.56±6.94、58.89±8.39 and 58.89±5.09%respectively when the three experimental pest drugstore beetle Stegobium paniceum,cigarette beetle Lasioderma serricorne and coffee bean beetle Araecerus fasciculatus was exposed to the control atmosphere with 10%CO_2 for 24 h.And The LT_(50) was 22.39,23.21 and 21.15 h respectively.However,the higher mortality rate was noted when the pest was exposed to the control atmosphere with 30% CO_2 or higher level within a shorter exposure periods.
2.Results showed that experimental pests exposed to control atmosphere with 30%CO_2 or more could be killed within 45 h.Tempreture and CO_2 levels or other composing and exposure period are some key factors to control pest with control atmosphere.
The toxicologically and biological characteristic of some key enzymes in pests exposed to the CO_2 enriched atmosphere
1.Compared to control,the specific activity of CarE from drugstore beetle which was exposed CO_2 enriched atmosphere for 3 h,6 h and 9 h increased 7.81%、14.96%and 22.58% respectively.That of cigarette beetle exposed for 3 h decreased 3.18%and for 6 h increased 22.2%. That of coffee bean beetle exposed for 3 h,6 h increased 31.75%and 50.82%respectively.Whilst, both the K_m and V_(max) of CarE from the pest mentioned above were affected by the press of CO_2 enriched atmosphere.
2.The specific activity of AChE from drugstore beetle and cigarette beetle which was exposed CO_2 enriched atmosphere for 6 h and 9 h decreased from 14.29μmol/L and 30.39μmol/L to 1.83μmol/L and 7.63μmol/L respectively.However,this treat did not continue with the prolonging of exposure periods.The K_m and V_(max) of AChE from dugstore beetle and cigarette beetle were affected by the press of CO_2 enriched atmosphere.
3、Compared to control,the specific activity of ACP from larvae of drugstore beetle and coffee bean beetle which were exposed to CO_2 enriched atmosphere for 6 h increased 78.59% and 72.45%respectively.The specific activity of ALP changed when the larvae of drugstore beetle were exposed to control atmosphere for 0～6 h.
4、The activity of SOD from drugstore beetle,cigarette beetle and coffee bean beetle which were exposed to CO_2 enriched atmosphere for 6 h increased from 2.011±0.954,2.664±0.218 and 1.458±0.718 to 3.135±0.105,3.050±0.673 and 2.975±0.229 enzyme unite/pest/30 min.
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