荔枝蝽臭腺提取物的杀虫活性及其熏蒸作用机理研究
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摘要
荔枝蝽Tessaratoma papillosa Drury属半翅目Hemiptera蝽科Pentatomidae荔蝽亚科Tessaratolninae,是荔枝、龙眼最重要的害虫之一。该虫以具有臭腺、受干扰时能释放臭液而著称。为了解荔枝蝽臭腺提取物的生物活性,本文进行其杀虫活性及熏蒸毒杀作用机理的初步研究,该研究方向尚未见报道。本试验得到主要结果如下：
采用三角瓶密闭熏蒸法测定了荔枝蝽臭腺提取物对害虫的熏蒸作用,结果表明：荔枝蝽雌、雄成虫臭腺提取物对供试害虫具有明显的熏蒸作用。熏蒸12h时,荔枝蝽雌成虫臭腺提取物对豇豆蚜Aphis craccivora Koch、甘蔗蚜Ceratovacuna lanigera Zehntner、瓜蚜Aphis gossypii Gloyer、玉米蚜Rhopalosiphum maidis Fitch、螺旋粉虱Subtropical Agriculture Rehash、烟粉虱Bemisia tabaci Gennadius及椰心叶甲Brontispa longissima Gestro的致死中浓度LC50值分别为1.0743μL/L、1.5324μL/L、3.6802μL/L、1.4398μL/L1.9495μL/L、1.8862μL/L和0.9910μL/L；熏蒸12h时,荔枝蝽雄成虫臭腺提取物对豇豆蚜、甘蔗蚜、黄瓜蚜、玉米蚜、螺旋粉虱、烟粉虱及椰心叶甲的致死中浓度LC50值分别为1.4038μL/L、1.5327μL/L、3.6351μL/L、1.5858μL/L、0.9673μL/L、0.9904μL/L和0.9536μL/L；荔枝蝽雌、雄臭腺提取物熏蒸24 h,对仓储害虫米象Sitophilus oryzae Linnaeus、谷蠹Rhyzopertha dominica Fabricius、四纹豆象Callosobruchus maculatus Fabricius、黄斑露尾甲Carpophilus hemipterus Linnaeus致死中浓度LC50值分别为0.4803μL/L、6.3803μL/L、2.5936μL/L、3.7925μL/L和0.5247μL/L、6.3125μL/L、2.4068μL/L、3.7753μL/L。
采用浸液法、培养皿药膜法和点滴法分别测定了荔枝蝽雌、雄成虫臭腺提取物对供试试虫的触杀作用,结果表明：荔枝蝽雌、雄成虫臭腺提取物对供试试虫有良好的触杀作用。触杀处理12h时,荔枝蝽雌成虫臭腺提取物对豇豆蚜、甘蔗蚜、瓜蚜、玉米蚜、螺旋粉虱、烟粉虱及椰心叶甲的致死中浓度LC50值分别为11.2703 mL/L、6.2152 mL/L、4.7612 mL/L、23.9420 mL/L、1.1769mL/L、1.0323 mL/L和0.0172μL/头；触杀处理12h时,荔枝蝽雄成虫臭腺提取物对豇豆蚜、甘蔗蚜、瓜蚜、玉米蚜、螺旋粉虱、烟粉虱及椰心叶甲的致死中浓度LC50值分别为8.5769 mL/L、7.6450 mL/L、4.1751 mL/L、9.8741mL/L、1.4276 mL/L、1.3245 mL/L和0.0103μL/头。
分别采用叶片涂药法和滤纸药膜法进行荔枝蝽成虫臭腺提取物对常见蚜虫和仓储害虫的驱避作用研究,结果表明：荔枝蝽成虫臭腺提取物对供试试虫有较好的驱避作用。处理12h时,10mL/L荔枝蝽雌、雄成虫臭腺提取物对黄瓜蚜虫、豇豆蚜、玉米蚜的驱避率分别为83.98%、91.01%、69.19%和92.89%、77.96%、64.87%：处理12h时,10mL/L荔枝蝽雌、雄成虫臭腺提取物对米象、黄斑露尾甲、四纹豆象、谷蠹的驱避率分别为68.71%、96.27%、33.33%、97.62%和72.27%、94.09%、50.00%、96.39%。
采用叶片涂药法进行荔枝蝽成虫臭腺提取物对椰心叶甲有较好的选择性驱避作用和非选择性驱避作用,但随时间和浓度的增加,选择性驱避效果增加。当处理18h、36h、48h,浓度为0.001mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的相对驱避率分别为-24.91%、12.00%、31.07%和-15.07%、13.65%、18.77%；浓度为10mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的相对驱避率分别为82.22%、75.62%、66.36%和62.81%、82.22%、50.75%；而非选择性驱避效果下降,当处理18h、36h、48h,浓度为0.001 mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的非选择性驱避率最小,分别为55.56%、38.10%、35.71%和70.18%、47.62%、35.71%；浓度为10 mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的非选择性驱避率最大,分别为83.33%、56.79%、47.62%和90.23%、81.63%、71.43%。
选取米象成虫为对象,对荔枝蝽臭腺提取物的作用机理进行研究,测定米象被臭腺提取物处理后,体内酶原蛋白含量和酶活性指标。研究发现,荔枝蝽臭腺提取物对米象成虫的酶活性具有明显的抑制作用。各处理试虫体内可溶性蛋白质含量均低于空白对照,且可溶性蛋白质含量随处理浓度的增加而降低；处理后,米象成虫体内乙酰胆碱酯酶(AChE)、酸性磷酸酯酶(ACP)、碱性磷酸酯酶(ALP)、谷胱甘肽-S-转移酶(GSTs)、过氧化物酶(POD)比活力均低于空白组,且随处理的时间和浓度增加,各酶的比活力均呈下降的趋势,表现为抑制作用。
综上所述,荔枝蝽臭腺提取物对常见害虫具有较好的触杀、熏蒸和驱避活性。试虫体内各种酶活性受到抑制是荔枝蝽臭腺提取物的作用机理之一。可见,荔枝蝽臭腺提取物可以作为一类生物农药,有必要进一步对其进行活性物质的筛选等方面的研究。
Tessaratoma papillosa Drury (Hemiptera, Pentatomidae) is a serious pest of litchi and longan fruit and known for the scent glands, which released foul liquid when being disturbed. To understand the biological activity of the scent gland extracts from litchi stink bug, the common pests and pathogens were used to test the insecticidal activity of scent gland extracts, and there has no report about this research. The research results were as follows:
The fumigation action of the scent gland extracts was tested by flask sealed fumigation. The scent gland extracts of female and male adults have significant fumigation effect on the tested pests. After fumigating for 12h, the LC50 values of scent gland extracts of the female and male adults to Aphis craccivora Koch, Ceratovacuna lanigera Zehntner, Aphis gossypii Gloyer, Rhopalosiphum maidis Fitch, Subtropical Agriculture Rehash, Bemisia tabaci Gennadius and Brontispa longissima Gestro were 1.0743μL/L,1.5324μl/L,3.6802μL/L, 1.4398μL/L,1.9495μl/L,1.8862μL/L,0.9910μL/L and 1.4038μL/L,1.5327μL/L,3.6351μL/L,1.5858μL/L,0.9673μL/L,0.9904μL/L,0.9536μL/L, respectively. And after fumigating for 24h, the LC50 values of scent gland extracts of the female and male adults to Sitophilus oryzae (Linnaeus), Rhyzopertha dominica Fabricius, Callosobruchus maculatus Fabricius, Curpophilus hemipterus Linnaeus were 0.4803μL/L,6.3803μL/L,2.5936μL/L, 3.7925μL/L and 0.5247μL/L,6.3125μL/L,2.4068μL/L,3.7753μL/L, respectively.
The contact action of the scent gland extracts of stink bugs to pests were analyzed by methods of immersion, culture dish membrane and drop respectively. The scent gland extracts of female and male adults have remarkable action of contact poison on the tested pests. After treating for 12h, the LC50 values of the scent gland extracts of female and male adults to A. craccivora, C. lanigera, A. gossypii, R. maidis, S. Agriculture, B. tabaci and B. longissima were 11.2703 mL/L,6.2152 mL/L,4.7612 mL/L,23.9420 mL/L,1.1769mL/L,1.0323 mL/L 0.0172μL per insect dose and 8.5769 mL/L,7.6450 mL/L,4.1751 mL/L,9.8741 mL/L 1.4276 mL/L,1.3245 mL/L,0.0103μL per insect dose, respectively.
The repellent action of the scent gland extracts of stink bugs to the aphids and common storage pests were tested by blade applicator method and membrane filter method, the results showed that the scent gland extracts of female and male adults have remarkable repellent effect to the tested pests. After treating for 12h, the repellent rate of the scent gland extracts of female and male adults to A. gossypii, A. craccivora, R. maidis, were respectively 83.98%, 91.01%,69.19% and 92.89%,77.96%,64.87%; the repellent rate of the scent gland extracts of female and male adults to S. oryzae, C. hemipterus, C. maculates, R. dominica were 68.71%,96.27%,33.33%,97.62% and 72.27%,94.09%,50.00%,96.39%, respectively.
In addition, the scent gland extracts have remarkable selective repellent effect and nonselective repellent effect to B. longissima by blade applicator method. The selective repellent effect increased with the raise of time and concentration. When dealing with 18h, 36h,48h, the concentration of 0.001mL/L, the relative repellency rates of the scent gland extracts of female and male adults to B. longissima were-24.91%,12.00%,31.07% and-15.07%,13.65%,18.77%. When the concentration of 10 mL/L, the relative repellent rates were 82.22%,75.62%,66.36% and 62.81%,82.22%,50.75%, respectively. But the nonselective repellent effect decreased with the raise of time and concentration. When dealing with 18h,36h,48h, the nonrelative repellency rates of the female and male adult stink bugs scent gland extracts to B. longissima were minimumat the concentration of 0.001mL/L, 55.56%,38.10%,35.71% and 70.18%,47.62%,35.71%, respectively. The nonrelative repellent rates were maximum at the concentration of 10 mL/L,83.33%,56.79%,47.62% and 90.23%,81.63%,71.43%, respectively.
The changes of zymogen protein's content and the main enzymes'activity of S. oryzae were tested after fumigation treated with the scent gland extracts of the litchi stink bugs. The results showed that the scent gland extracts of the litchi stink bugs have strong inhibitory action to the main enzymes'activity of S. oryzae. The contents of soluble protein in the insects treated with the scent gland extracts of the litchi stink bugs were lower than control, and the contents of soluble protein in the treated insects decreased with the increase of the concentration. The scent gland extracts of the litchi stink bugs had the effect of inhibiting all the specific activity of AChE, ACP, ALP, GSTs and POD of S. oryzae. After treated with the scent gland extracts of the litchi stink bugs, all the specific activity of AChE, ACP, ALP, GSTs and POD of S. oryzae were lower than control, and all the specific activity of AChE, ACP, ALP, GSTs and POD of S. oryzae decreased with the increase of the concentration and time.
The results above showed that the scent gland extracts of the litchi stink bugs had significantly insecticidal and bactericidal biological activity. The extracts had obvious contact, fumigation and repellent activities to pests be tested. Inhibiting the enzyme activity in the tested insects is one of the mechanisms of the scent gland extracts of litchi stink bugs. Therefore, the extracts of litchi stink bug scent gland could be used as a class of bio-pesticides, and it was necessary to do further research on screening its active substance.
采用三角瓶密闭熏蒸法测定了荔枝蝽臭腺提取物对害虫的熏蒸作用,结果表明：荔枝蝽雌、雄成虫臭腺提取物对供试害虫具有明显的熏蒸作用。熏蒸12h时,荔枝蝽雌成虫臭腺提取物对豇豆蚜Aphis craccivora Koch、甘蔗蚜Ceratovacuna lanigera Zehntner、瓜蚜Aphis gossypii Gloyer、玉米蚜Rhopalosiphum maidis Fitch、螺旋粉虱Subtropical Agriculture Rehash、烟粉虱Bemisia tabaci Gennadius及椰心叶甲Brontispa longissima Gestro的致死中浓度LC50值分别为1.0743μL/L、1.5324μL/L、3.6802μL/L、1.4398μL/L1.9495μL/L、1.8862μL/L和0.9910μL/L；熏蒸12h时,荔枝蝽雄成虫臭腺提取物对豇豆蚜、甘蔗蚜、黄瓜蚜、玉米蚜、螺旋粉虱、烟粉虱及椰心叶甲的致死中浓度LC50值分别为1.4038μL/L、1.5327μL/L、3.6351μL/L、1.5858μL/L、0.9673μL/L、0.9904μL/L和0.9536μL/L；荔枝蝽雌、雄臭腺提取物熏蒸24 h,对仓储害虫米象Sitophilus oryzae Linnaeus、谷蠹Rhyzopertha dominica Fabricius、四纹豆象Callosobruchus maculatus Fabricius、黄斑露尾甲Carpophilus hemipterus Linnaeus致死中浓度LC50值分别为0.4803μL/L、6.3803μL/L、2.5936μL/L、3.7925μL/L和0.5247μL/L、6.3125μL/L、2.4068μL/L、3.7753μL/L。
采用浸液法、培养皿药膜法和点滴法分别测定了荔枝蝽雌、雄成虫臭腺提取物对供试试虫的触杀作用,结果表明：荔枝蝽雌、雄成虫臭腺提取物对供试试虫有良好的触杀作用。触杀处理12h时,荔枝蝽雌成虫臭腺提取物对豇豆蚜、甘蔗蚜、瓜蚜、玉米蚜、螺旋粉虱、烟粉虱及椰心叶甲的致死中浓度LC50值分别为11.2703 mL/L、6.2152 mL/L、4.7612 mL/L、23.9420 mL/L、1.1769mL/L、1.0323 mL/L和0.0172μL/头；触杀处理12h时,荔枝蝽雄成虫臭腺提取物对豇豆蚜、甘蔗蚜、瓜蚜、玉米蚜、螺旋粉虱、烟粉虱及椰心叶甲的致死中浓度LC50值分别为8.5769 mL/L、7.6450 mL/L、4.1751 mL/L、9.8741mL/L、1.4276 mL/L、1.3245 mL/L和0.0103μL/头。
分别采用叶片涂药法和滤纸药膜法进行荔枝蝽成虫臭腺提取物对常见蚜虫和仓储害虫的驱避作用研究,结果表明：荔枝蝽成虫臭腺提取物对供试试虫有较好的驱避作用。处理12h时,10mL/L荔枝蝽雌、雄成虫臭腺提取物对黄瓜蚜虫、豇豆蚜、玉米蚜的驱避率分别为83.98%、91.01%、69.19%和92.89%、77.96%、64.87%：处理12h时,10mL/L荔枝蝽雌、雄成虫臭腺提取物对米象、黄斑露尾甲、四纹豆象、谷蠹的驱避率分别为68.71%、96.27%、33.33%、97.62%和72.27%、94.09%、50.00%、96.39%。
采用叶片涂药法进行荔枝蝽成虫臭腺提取物对椰心叶甲有较好的选择性驱避作用和非选择性驱避作用,但随时间和浓度的增加,选择性驱避效果增加。当处理18h、36h、48h,浓度为0.001mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的相对驱避率分别为-24.91%、12.00%、31.07%和-15.07%、13.65%、18.77%；浓度为10mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的相对驱避率分别为82.22%、75.62%、66.36%和62.81%、82.22%、50.75%；而非选择性驱避效果下降,当处理18h、36h、48h,浓度为0.001 mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的非选择性驱避率最小,分别为55.56%、38.10%、35.71%和70.18%、47.62%、35.71%；浓度为10 mL/L时,荔枝蝽雌、雄成虫臭腺提取物对椰心叶甲的非选择性驱避率最大,分别为83.33%、56.79%、47.62%和90.23%、81.63%、71.43%。
选取米象成虫为对象,对荔枝蝽臭腺提取物的作用机理进行研究,测定米象被臭腺提取物处理后,体内酶原蛋白含量和酶活性指标。研究发现,荔枝蝽臭腺提取物对米象成虫的酶活性具有明显的抑制作用。各处理试虫体内可溶性蛋白质含量均低于空白对照,且可溶性蛋白质含量随处理浓度的增加而降低；处理后,米象成虫体内乙酰胆碱酯酶(AChE)、酸性磷酸酯酶(ACP)、碱性磷酸酯酶(ALP)、谷胱甘肽-S-转移酶(GSTs)、过氧化物酶(POD)比活力均低于空白组,且随处理的时间和浓度增加,各酶的比活力均呈下降的趋势,表现为抑制作用。
综上所述,荔枝蝽臭腺提取物对常见害虫具有较好的触杀、熏蒸和驱避活性。试虫体内各种酶活性受到抑制是荔枝蝽臭腺提取物的作用机理之一。可见,荔枝蝽臭腺提取物可以作为一类生物农药,有必要进一步对其进行活性物质的筛选等方面的研究。
Tessaratoma papillosa Drury (Hemiptera, Pentatomidae) is a serious pest of litchi and longan fruit and known for the scent glands, which released foul liquid when being disturbed. To understand the biological activity of the scent gland extracts from litchi stink bug, the common pests and pathogens were used to test the insecticidal activity of scent gland extracts, and there has no report about this research. The research results were as follows:
The fumigation action of the scent gland extracts was tested by flask sealed fumigation. The scent gland extracts of female and male adults have significant fumigation effect on the tested pests. After fumigating for 12h, the LC50 values of scent gland extracts of the female and male adults to Aphis craccivora Koch, Ceratovacuna lanigera Zehntner, Aphis gossypii Gloyer, Rhopalosiphum maidis Fitch, Subtropical Agriculture Rehash, Bemisia tabaci Gennadius and Brontispa longissima Gestro were 1.0743μL/L,1.5324μl/L,3.6802μL/L, 1.4398μL/L,1.9495μl/L,1.8862μL/L,0.9910μL/L and 1.4038μL/L,1.5327μL/L,3.6351μL/L,1.5858μL/L,0.9673μL/L,0.9904μL/L,0.9536μL/L, respectively. And after fumigating for 24h, the LC50 values of scent gland extracts of the female and male adults to Sitophilus oryzae (Linnaeus), Rhyzopertha dominica Fabricius, Callosobruchus maculatus Fabricius, Curpophilus hemipterus Linnaeus were 0.4803μL/L,6.3803μL/L,2.5936μL/L, 3.7925μL/L and 0.5247μL/L,6.3125μL/L,2.4068μL/L,3.7753μL/L, respectively.
The contact action of the scent gland extracts of stink bugs to pests were analyzed by methods of immersion, culture dish membrane and drop respectively. The scent gland extracts of female and male adults have remarkable action of contact poison on the tested pests. After treating for 12h, the LC50 values of the scent gland extracts of female and male adults to A. craccivora, C. lanigera, A. gossypii, R. maidis, S. Agriculture, B. tabaci and B. longissima were 11.2703 mL/L,6.2152 mL/L,4.7612 mL/L,23.9420 mL/L,1.1769mL/L,1.0323 mL/L 0.0172μL per insect dose and 8.5769 mL/L,7.6450 mL/L,4.1751 mL/L,9.8741 mL/L 1.4276 mL/L,1.3245 mL/L,0.0103μL per insect dose, respectively.
The repellent action of the scent gland extracts of stink bugs to the aphids and common storage pests were tested by blade applicator method and membrane filter method, the results showed that the scent gland extracts of female and male adults have remarkable repellent effect to the tested pests. After treating for 12h, the repellent rate of the scent gland extracts of female and male adults to A. gossypii, A. craccivora, R. maidis, were respectively 83.98%, 91.01%,69.19% and 92.89%,77.96%,64.87%; the repellent rate of the scent gland extracts of female and male adults to S. oryzae, C. hemipterus, C. maculates, R. dominica were 68.71%,96.27%,33.33%,97.62% and 72.27%,94.09%,50.00%,96.39%, respectively.
In addition, the scent gland extracts have remarkable selective repellent effect and nonselective repellent effect to B. longissima by blade applicator method. The selective repellent effect increased with the raise of time and concentration. When dealing with 18h, 36h,48h, the concentration of 0.001mL/L, the relative repellency rates of the scent gland extracts of female and male adults to B. longissima were-24.91%,12.00%,31.07% and-15.07%,13.65%,18.77%. When the concentration of 10 mL/L, the relative repellent rates were 82.22%,75.62%,66.36% and 62.81%,82.22%,50.75%, respectively. But the nonselective repellent effect decreased with the raise of time and concentration. When dealing with 18h,36h,48h, the nonrelative repellency rates of the female and male adult stink bugs scent gland extracts to B. longissima were minimumat the concentration of 0.001mL/L, 55.56%,38.10%,35.71% and 70.18%,47.62%,35.71%, respectively. The nonrelative repellent rates were maximum at the concentration of 10 mL/L,83.33%,56.79%,47.62% and 90.23%,81.63%,71.43%, respectively.
The changes of zymogen protein's content and the main enzymes'activity of S. oryzae were tested after fumigation treated with the scent gland extracts of the litchi stink bugs. The results showed that the scent gland extracts of the litchi stink bugs have strong inhibitory action to the main enzymes'activity of S. oryzae. The contents of soluble protein in the insects treated with the scent gland extracts of the litchi stink bugs were lower than control, and the contents of soluble protein in the treated insects decreased with the increase of the concentration. The scent gland extracts of the litchi stink bugs had the effect of inhibiting all the specific activity of AChE, ACP, ALP, GSTs and POD of S. oryzae. After treated with the scent gland extracts of the litchi stink bugs, all the specific activity of AChE, ACP, ALP, GSTs and POD of S. oryzae were lower than control, and all the specific activity of AChE, ACP, ALP, GSTs and POD of S. oryzae decreased with the increase of the concentration and time.
The results above showed that the scent gland extracts of the litchi stink bugs had significantly insecticidal and bactericidal biological activity. The extracts had obvious contact, fumigation and repellent activities to pests be tested. Inhibiting the enzyme activity in the tested insects is one of the mechanisms of the scent gland extracts of litchi stink bugs. Therefore, the extracts of litchi stink bug scent gland could be used as a class of bio-pesticides, and it was necessary to do further research on screening its active substance.
引文
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