北京地区二斑叶螨不同种群的药剂敏感性

doi:10.3864/j.issn.0578-1752.2014.15.009

摘要/Abstract

摘要： 【目的】明确北京地区作物上二斑叶螨（Tetranychus urticae Koch）不同种群对杀螨剂的敏感性水平，了解二斑叶螨对药剂敏感性与体内4种解毒酶活力的相关性。【方法】在实验室内采用改进的玻片浸渍法（slide-dip method）检测北京房山、怀柔、昌平、延庆和平谷5个地区二斑叶螨田间种群对联苯肼酯、阿维菌素、螺螨酯和哒螨灵4种杀螨剂的敏感性；采用PCR技术对单头叶螨的CYTB基因进行扩增和测序，检测二斑叶螨对联苯肼酯的抗性突变位点；并采用酶标仪微量板法，检测二斑叶螨体内与抗性相关的多功能氧化酶、乙酰胆碱酯酶、羧酸酯酶和谷胱甘肽S-转移酶的活力。【结果】5个田间种群对联苯肼酯最敏感，其LC50分别为2.4880、6.4693、6.2398、0.7882和14.7783 mg•L-1。对阿维菌素的敏感性次之，其LC50分别为22.4712、35.4431、14.5260、15.4904和14.0023 mg•L-1。对螺螨酯和哒螨灵的敏感性非常低，螺螨酯的LC50分别为49.6833、81.8826、72.9609、204.4609和1 433.5137 mg•L-1，哒螨灵的LC50分别为202.6902、806.8324、375.3518、188.3234和2 310.9040 mg•L-1。延庆地区二斑叶螨乙酰胆碱酯酶、羧酸酯酶和谷胱甘肽S-转移酶的活性显著高于其他地区，其活力分别为14.9508 U/(mg protein)、0.2271 μmol•mg-1 protein•30 min-1和58.2962 U/(mg protein)。怀柔地区多功能氧化酶的活性显著低于其他地区，其活力为1.4272 μmol•mg-1 protein•30 min-1。联苯肼酯、阿维菌素、螺螨酯和哒螨灵的敏感性水平与乙酰胆碱酯酶、羧酸酯酶、谷胱甘肽S-转移酶和多功能氧化酶的酶活力之间没有直接的线性关系；采用PCR技术对5个地区的288个二斑叶螨个体进行CYTB基因检测后，仅发现怀柔地区的一个个体CYTB基因第126位氨基酸密码子的第一位核苷酸由G突变为A导致氨基酸由G突变为S，其余地区均无突变，这与生物测定方法中各地区对联苯肼酯较敏感的结果基本一致。【结论】北京地区二斑叶螨不同种群对联苯肼酯敏感性最高，对阿维菌素的敏感性次之，对螺螨酯和哒螨灵敏感性非常低；二斑叶螨对4种杀螨剂的敏感性水平与乙酰胆碱酯酶、羧酸酯酶、谷胱甘肽S-转移酶和多功能氧化酶的酶活力之间没有直接的线性关系；通过检测到的基因突变个体，说明怀柔地区二斑叶螨种群中存在着对联苯肼酯产生高抗性风险的个体。PCR方法能够更加快速准确地检测种群抗性发展程度。基于以上结果，在农业防治二斑叶螨中应降低联苯肼酯使用频率，交替使用阿维菌素，避免使用螺螨酯，停止使用哒螨灵，以延缓和降低二斑叶螨产生抗性的风险。

关键词: 二斑叶螨 , 生物测定 , 酶活力 , 联苯肼酯

Abstract: 【Objective】The objectives of this study are to survey the sensitivity of different populations of Tetranychus urticae Koch to the acaricides in Beijing area, as well as the relationship between their sensitivity to the acaracides and the activity of resitance-related enzymes. 【Method】 Bioassays of four frequently used acaricides of bifenazate, abamectin, spirodiclofen and pyridaben to five field populations of the T. urticae, i.e. Fangshan, Huairou, Changping, Yanqing and Pinggu of Beijing area, were conducted by using modified slide-dip method. Bifenazate resistance-related mutations were detected by PCR amplification and sequencing of the CYTB gene from individual specimens. Additionally, the activities of four resistance-related enzymes of cytochrome P450 monooxygenases, acetylcholinesterase (AChE), carboxylesterase (CarE) and glutathione S-transferase (GST) were tested by microplate assay in microplate reader for the five populations.【Result】All of the five field populations of Fangshan, Huairou, Changping, Yanqing and Pinggu were sensitive to bifenazate, with LC50 of 2.4880, 6.4693, 6.2398, 0.7882 and 14.7783 mg•L-1, respectively, followed by the abamectin, with LC50 of 22.4712, 35.4431, 14.5260, 15.4904 and 14.0023 mg•L-1, respectively. T. urticae showed low sensitivity to the spirodiclofen and pyridaben in the five tested populations. In populations of Fangshan, Huairou, Changping, Yanqing and Pinggu, the LC50 of spirodiclofen was 49.6833, 81.8826, 72.9609, 204.4609 and 1 433.5137 mg•L-1, respectively, whereas that of pyridaben was 202.6902, 806.8324, 375.3518, 188.3234 and 2 310.9040 mg•L-1, respectively. The sensitivity of the AChE, CarE and GST of Yanqing population was higher than that of other populations, with a value of 14.9508 U/(mg protein), 0.2271 μmol?mg-1 protein?30 min-1 and 58.2962 U/(mg protein), respectively. The cytochrome P450 monooxygenases of Huairou population was significantly lower than that of other populations with a value of 1.4272 μmol?mg-1 protein?30 min-1. There was no linear relationship between the toxicity of bifenazate, abamectin, spirodiclofen and pyridaben and the activities of cytochrome P450 monooxygenases, AChE, CarE and GST. One individual from Huairou population with one site mutation on the CYTB gene (the first codon position of the 126th amino acid was changed from G to A) was detected in 288 individuals of five populations, which lead the amino acid Gly mutated into Ser. The low level of resistance-related mutation in the detected populations of the T. urticae was consistent with their high sensitivity to the bifenazate. 【Conclusion】 T. urticae in Beijing area showed highest sensitivity to the bifenazate, followed by the abamectin, and lastly to spirodiclofen and pyridaben. The sensitivity of T. urticae to the four tested acaricides is not linearly related to the activity of cytochrome P450 monooxygenases, AChE, CarE and GST. The PCR detection of resistance-related mutation indicated that the Huairou population has high risk to generate resistant individuals to the bifenazate. PCR method could fastly and accurately detect the development of acaracide resistance than the bioassay method. In the controlling practice, the pyridaben should not be used, the spirodiclofen should be avoided, and abamectin and bifenazate could be used alternatively.

Key words: Tetranychus urticae Koch , bioassay , enzyme activity , bifenazate

宫亚军1, 王泽华1, 石宝才1, 崔文夏1, 金桂华1, 孙艳艳2, 魏书军1. 北京地区二斑叶螨不同种群的药剂敏感性[J]. 中国农业科学, 2014, 47(15): 2990-2997.

GONG Ya-Jun-1, WANG Ze-Hua-1, SHI Bao-Cai-1, CUI Wen-Xia-1, JIN Gui-Hua-1, SUN Yan-Yan-2, WEI Shu-Jun-1. Sensitivity of Different Field Populations of Tetranychus urticae Koch (Acari: Tetranychidae) to the Acaricides in Beijing Area[J]. Scientia Agricultura Sinica, 2014, 47(15): 2990-2997.

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https://www.chinaagrisci.com/CN/Y2014/V47/I15/2990

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