二斑叶螨田间种群对阿维菌素的抗性及抗性相关基因表达分析

【目的】二斑叶螨Tetranychus urticae Koch是为害多种农作物的世界性重大害螨。本研究旨在明确二斑叶螨不同田间种群对阿维菌素的抗药性、抗性相关基因的突变频率及其表达量变化。【方法】采用药管浸叶法测定了我国二斑叶螨8个地理种群对阿维菌素的抗药性并检测其抗性基因突变频率,结合荧光定量PCR技术分析了高抗种群中抗性相关基因表达量变化。【结果】测试的二斑叶螨8个田间种群均对阿维菌素具有抗药性。北京密云、山东潍坊、海南三亚和湖南长沙种群均对阿维菌素产生了极高水平的抗性,抗性倍数分别为1 526.75,481.00,315.25和160.75倍,而北京通县、北京海淀、山西运城和山东泰安种群对阿维菌素的抗性倍数达54.38~136.38倍,处于高抗性水平。二斑叶螨对阿维菌素抗性相关的谷氨酸氯离子通道基因Glu Cl的突变频率在各个田间种群中存在差异。北京密云种群中Glu Cl的突变频率最高(91.7%),其次是山东潍坊(66.7%)和海南三亚(63.3%)种群;山西运城种群最低(13.3%),且点突变频率与抗性倍数之间呈显著的正相关(P0.05)。与相对敏感种群相比,高抗性二斑叶螨种群中Glu Cl和γ-氨基丁酸受体基因GABAR表达量显著下降。【结论】二斑叶螨田间种群普遍对阿维菌素产生了高水平抗性,抗性相关基因Glu Cl的点突变及其表达量的降低可能与田间抗药性产生相关;田间防治二斑叶螨应避免使用阿维菌素。     

陕西苹果园山楂叶螨抗药性监测

【目的】明确陕西省苹果园的山楂叶螨Tetranychus viennensis Zacher种群对5种药剂的抗性水平。【方法】采用玻片浸渍法,建立了山楂叶螨室内饲养的相对敏感种群对5种杀虫剂的敏感基线,同时从陕西乾县、礼泉、兴平、澄城、安塞、淳化、凤翔和扶风8个不同地区的苹果园采集山楂叶螨,分析这些田间种群的抗药性水平。【结果】山楂叶螨室内相对敏感种群对阿维菌素的敏感性最高,对毒死蜱敏感性最低;各种群对哒螨灵已产生了13.29~69.63倍的抗性;对高效氯氟氰菊酯已经产生了7.99~46.74倍的抗性;除兴平种群对阿维菌素表现为低抗水平外(抗性倍数7.63),其余种群对阿维菌素表现为敏感或者敏感性下降(抗性倍数1.89~3.94);除扶风种群对毒死蜱抗性水平处于敏感性下降的阶段外,其它7个种群对毒死蜱的均处于敏感阶段;各种群对噻虫嗪均处于敏感阶段。【结论】山楂叶螨室内相对敏感种群对5种不同杀虫剂的敏感性不同;各田间种群对哒螨灵和高效氯氟氰菊酯两种药剂已经产生了不同水平的抗药性,除兴平种群对阿维菌素产生低抗水平抗性外,其余田间种群对阿维菌素、毒死蜱和噻虫嗪抗性均表现为敏感或者敏感性下降;田间防治时应该减少哒螨灵和高效氯氟氰菊酯两种药剂的使用,同时注意不同农药的轮换使用,以此延缓山楂叶螨对杀虫剂产生高水平抗药性。     

我国白纹伊蚊对三种拟除虫菊酯类杀虫剂抗性检测诊断剂量的建立

【目的】建立中国白纹伊蚊Aedes albopictus成蚊对溴氰菊酯、氯菊酯和高效氯氟氰菊酯杀虫剂抗性检测的诊断剂量。【方法】应用溴氰菊酯、氯菊酯和高效氯氟氰菊酯原药制作不同浓度的药膜滤纸,接触筒法测定白纹伊蚊实验室敏感品系成蚊对3种拟除虫菊酯类杀虫剂的敏感性,记录1 h的击倒数和24 h的死亡数。应用Excel 2007和SPSS20.0进行数据统计处理,并制作杀虫剂的毒力回归线,计算各自的LC_(50)和LC_(99)值。以2倍LC_(99)值作为区分抗性和敏感种群的诊断剂量,制作药膜滤纸,接触筒法生物测定海口市白纹伊蚊现场种群成蚊对杀虫剂的抗药性。【结果】溴氰菊酯、高效氯氟氰菊酯和氯菊酯杀虫剂对白纹伊蚊实验室敏感品系成蚊的LC_(50)值分别为0.00619%,0.01403%和0.05009%,LC_(99)值分别为0.05175%,0.11859%,和0.53165%,相对应的诊断剂量分别为0.1035%,0.2372%和1.0633%。应用上述溴氰菊酯、氯菊酯和高效氯氟氰菊酯诊断剂量测定的海口市白纹伊蚊现场种群的死亡率分别为22.58%,36.29%和40.83%,表明该种群对这3种菊酯类杀虫剂均已产生了抗性。【结论】本研究建立的白纹伊蚊对3种拟除虫菊酯类杀虫剂的诊断剂量可作为该蚊成蚊抗药性监测的参考。     

湖北四个地区草地贪夜蛾田间种群的杀虫剂敏感性及靶标突变检测

【目的】测定湖北草地贪夜蛾田间种群对4种杀虫剂的敏感性,并对杀虫剂靶标Ace-1,VGSC和RyR的基因型和突变频率进行检测,以明确湖北草地贪夜蛾田间种群药剂敏感性现状,进而指导田间科学用药。【方法】采集湖北黄冈、武穴、咸宁和荆州4个地区玉米田中的草地贪夜蛾幼虫,采用浸叶法测定其对氯虫苯甲酰胺、乙基多杀菌素、茚虫威和甲维盐4种杀虫剂的敏感性;提取4个种群80头个体的单头总RNA合成cDNA,利用特异性引物进行PCR扩增,获得目的基因片段,根据序列比对和测序峰图分析,确定Ace-1,VGSC和RyR的基因型和靶标突变位点的突变频率。【结果】生测结果表明,湖北这4个田间种群的草地贪夜蛾幼虫对氯虫苯甲酰胺、乙基多杀菌素、茚虫威和甲维盐均比较敏感,敏感性从高到低依次为:甲维盐乙基多杀菌素氯虫苯甲酰胺茚虫威。分子检测结果表明,湖北草地贪夜蛾这4个田间种群Ace-1基因在A201S, G227A和F290V位点均存在抗性杂合突变,且在F290V位点检测到抗性纯合突变,而VGSC和RyR两个基因均没有检测到靶标位点突变。【结论】湖北草地贪夜蛾4个田间种群对氯虫苯甲酰胺、乙基多杀菌素、茚虫威和甲维盐这4种杀虫剂均较敏感,但这4个草地贪夜蛾田间种群均携带有对有机磷和氨基甲酸酯类杀虫剂的抗性基因,存在潜在的抗性风险,田间防治建议少用或不用该类杀虫剂,同时需要进一步加强抗性监测工作。     

褐飞虱对五种杀虫剂抗性的快速检测技术

【目的】建立褐飞虱Nilaparvata lugens对常用杀虫剂抗性的快速检测技术,实时掌握田间褐飞虱种群的抗药性水平,以指导褐飞虱防控合理用药。【方法】基于玻璃瓶药膜法,研制褐飞虱3龄若虫对吡虫啉、啶虫脒、醚菊酯、毒死蜱和异丙威5种杀虫剂抗性的快速检测试剂盒;利用试剂盒测定的死亡率与稻苗浸渍法测得的抗性倍数进行相关性分析,并验证利用试剂盒快速测定褐飞虱田间种群对5种杀虫剂 抗性水平的准确性。【结果】处理1 h时吡虫啉、啶虫脒、醚菊酯、毒死蜱和异丙威对褐飞虱室内敏感种群3龄若虫的LD₉₀分别为：30.96, 92.05, 117.24, 514.21和1.24 ng/cm²。在吡虫啉、啶虫脒、醚菊酯、毒死蜱和异丙威相应诊断剂量下,贵州省不同田间种群褐飞虱3龄若虫的校正死亡率分别在23.75%~78.75%, 25.00%~78.75%, 43.75%~88.75%, 36.25%~85.00%和18.75%~67.50%之间。相关性分析表明,上述田间种群褐飞虱3龄若虫的死亡率与稻苗浸渍法测定的抗性倍数呈显著负相关,相关系数在0.8751~0.9754之间。通过快速检测试剂盒获得的死亡率及相关性方程计算得到贵州安龙地区(AL)褐飞虱种群对吡虫啉、啶虫脒、醚菊酯、毒死蜱和异丙威的推测抗性倍数分别为7.23, 3.68, 4.14, 4.12和31.18,采用稻苗浸渍法测得上述5种杀虫剂的实测抗性倍数分别为6.33, 5.24, 3.71, 4.50和26.56,表明推测抗性倍数与实测的抗性倍数结果表现一致。【结论】该快速检测试剂盒可以通过测定褐飞虱田间种群的死亡率,快速评估褐飞虱田间种群对杀虫剂的抗性水平。     

江西省柑橘木虱对五种常规药剂的抗药性

【目的】针对江西省柑橘木虱Diaphorina citri对常规杀虫剂抗药性水平的研究相对匮乏,研究不同地区柑橘木虱种群对常用杀虫剂的抗性水平,为生产实践合理化用药提供依据。【方法】采用浸叶法测定毒死蜱、联苯菊酯、噻虫嗪、烯啶虫胺和虫螨腈5种杀虫剂对上高县（SG）、宜黄（YH）、南丰（NF）、宁都（ND）、大余（DY）和寻乌县（XW）6个不同地区的田间柑橘木虱的毒力,结合毒力参考基线,得到抗性倍数。【结果】上高、宜黄和南丰县种群对毒死蜱的抗性倍数在中等水平（9.01-15.01）,而宁都、大余和寻乌县则是高水平抗性（128.36-308.82）。大余种群对联苯菊酯的抗性水平为133.71倍,其他5个地区均在中等水平（17.08-88.53）。不同地区柑橘木虱种群对噻虫嗪的抗性倍数在13.62-25.92,仅为中等偏低水平;而它们对烯啶虫胺的抗性倍数为77.37-257.71,已经达到极高水平。与联苯菊酯类似,除了大余种群对虫螨腈为高水平抗性外,其余5个地区种群均为中等水平。【结论】赣南脐橙产区的宁都县、大余县和寻乌县的柑橘木虱种群对5种常用杀虫剂均表现出高水平抗性,显著高于非赣南脐橙产区种群（上高县、宜黄县和南丰县）。推荐在高抗地区使用抗性水平较低的药剂,并注意轮换用药;在中等抗性地区应有限制地使用药剂,以延缓抗性增长。     

我国草地贪夜蛾田间种群有机磷和氨基甲酸酯类杀虫剂靶标基因ace-1的基因型和突变频率

【目的】对入侵我国的草地贪夜蛾Spodoptera frugiperda有机磷和氨基甲酸酯类杀虫剂靶标基因ace-1的基因型进行分子检测,明确抗性基因频率,进而指导田间科学用药。【方法】采集中国12省份的草地贪夜蛾田间种群幼虫样本,提取单头样本的基因组DNA,利用特异性引物进行PCR扩增,获得ace-1基因片段。根据碱基、氨基酸序列比对和测序峰图分析,明确与有机磷和氨基甲酸酯类杀虫剂抗性相关的3个氨基酸突变位点A201S, G227A和F290V的基因型和抗性基因频率。【结果】通过DNA检测分析中国12省份草地贪夜蛾田间种群589头个体ace-1基因的基因型和突变频率发现,在A201S位点检测到137头个体为抗性杂合基因型,抗性基因频率为11.6%,未发现抗性纯合基因型个体;G227A位点589头个体均为敏感纯合基因型;F290V位点的抗性基因频率最高,达到57.1%,携带抗性基因的个体数量达到523头(占样本总数的88.8%)。【结论】结果表明入侵我国的草地贪夜蛾种群携带高频率的对有机磷和氨基甲酸酯类杀虫剂抗性基因。田间防治建议不用或少用有机磷和氨基甲酸酯类杀虫剂,同时进一步加强田间抗性监测工作。     

我国韭菜主产区韭菜迟眼蕈蚊田间种群的抗药性监测

【目的】建立韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang对10种常用药剂的敏感基线,并对4省7个主要韭菜产区的田间种群进行6种常用杀虫剂的抗药性水平监测。【方法】采用胃毒触杀联合毒力法对韭菜迟眼蕈蚊3龄幼虫进行室内生物测定。【结果】建立了敏感品系对新烟碱类、有机磷类、菊酯类、昆虫生长调节剂类、吡咯类药剂的敏感基线。对7个地区的田间韭菜迟眼蕈蚊种群监测结果表明:其对有机磷类药剂均产生了抗药性,其中河南郑州种群对毒死蜱和辛硫磷产生了极高水平抗性;河南郑州种群对高效氯氰菊酯产生了中等水平抗性,其他各地区均处于敏感状态;大部分种群对新烟碱类药剂处于低等或中等水平抗性,但山东李坡种群对噻虫嗪产生了高水平抗性。【结论】本文建立的韭菜迟眼蕈蚊对10种杀虫剂的敏感基线及抗药性监测数据为抗性治理提供一定参考。     

湖南宁乡烟区烟蚜的抗性监测及其相关酶的活性测定

为明确湖南宁乡烟区烟蚜对杀虫剂的抗性现状,测定烟蚜田间种群对吡虫啉、啶虫脒、氯氟氰菊酯、辛硫磷及灭多威5种杀虫剂的敏感性。结果表明,烟蚜田间种群对烟碱类的啶虫脒和吡虫啉表现出中等抗性,与敏感品系相比,抗性倍数分别为25. 20和21. 03;对菊酯类的氯氟氰菊酯表现为低抗水平,抗性倍数为5. 19;对辛硫磷和灭多威均表现为敏感性降低,抗性倍数分别为2. 38和2. 86。本研究又测定了烟蚜田间与室内敏感种群相关酶活力的变化,结果表明,田间种群体内解毒酶、保护酶与消化酶活力均显著高于室内敏感种群。试验结果可为延缓田间烟蚜抗性及烟蚜的综合治理提供理论依据。     

不同葫芦科作物对二斑叶螨的抗性评价

【目的】二斑叶螨Tetranychus urticae Koch是世界性重大农业害螨,葫芦科作物是其重要寄主植物。为明确不同葫芦科作物对二斑叶螨生长发育的抗性/适合性,开展了二斑叶螨在葫芦科作物上的产卵和取食选择性研究。【方法】选择苦瓜、葫芦、南瓜、丝瓜、西瓜、甜瓜和黄瓜作为供试寄主植物,室内采用离体叶片和室外采用活体植株比较了二斑叶螨对7种作物的取食选择性及其种群生存与繁殖动态。【结果】供试7种葫芦科作物中,二斑叶螨对黄瓜的选择率最高,为35.49%,其次为甜瓜和西瓜;二斑叶螨在南瓜和黄瓜叶碟上的生存率和繁殖率高,12 h单雌产卵数分别为9.72粒和7.83粒;在葫芦和苦瓜叶碟上,二斑叶螨48 h螨口减退率达47.5%和32.5%,表现为较低的取食和生存适合度,同时葫芦和苦瓜活体植株上二斑叶螨繁殖率显著低于其他5种葫芦科作物。【结论】二斑叶螨在黄瓜、西瓜和甜瓜上的种群发育适合度高,在葫芦和苦瓜上适合度低。这一结果可为田间防控二斑叶螨生态措施的制定及开展葫芦科作物抗螨性机理和抗螨育种提供科学依据。     

Mechanisms of drug resistance and reversal of the resistance

The mechanisms for the resistance to anticancer agents have been vigorously studied and many factors that are involved in the resistance were found. Among the members of ABC transporter superfamily, P-glycoprotein, MRP1-5 and BCRP are involved in the drug resistance. LRP, identified as the major vault protein, is also related to drug resistance.     

Hydraulic resistance of two sorghums varying in drought resistance

Genotypes of sorghum [Sorghum bicolor (L.) Moench] vary in drought resistance. Yet it is not known if their hydraulic resistances vary. The objective of this study was to determine if the hydraulic resistance of a drought-resistant sorghum was the same as that of a drought-sensitive sorghum. Leaf water and osmotic potentials were measured daily, during a 14-d period, in leaves of a drought-resistant (‘KS9’) and a drought-sensitive (‘IA25’) sorghum, which had the roots in pots with a commercial potting soil that was either well watered or allowed to dry. Soil water potential, adaxial stomatal resistance, and transpiration rate were determined daily. Hydraulic resistance of the plants was calculated from the slope of the line relating soil water potential minus leaf water potential versus transpiration rate. When the soil was not watered, the drought-sensitive sorghum had a water potential that averaged −0.50 MPa lower and an osmotic potential that averaged −0.57 MPa lower, but a similar adaxial stomatal resistance (1.19 s mm⁻¹), compared with the drought-resistant sorghum. Seven days after the beginning of the experiment, the water potential of the soil with the drought-sensitive sorghum was −0.25 MPa lower than that of the soil with the drought-resistant sorghum. With the water-limited conditions, the drought-sensitive sorghum depleted the soil-water reserve more quickly and died 2 d before the drought-resistant sorghum. Under well watered conditions, the two sorghums had similar water potentials (−1.64 MPa), osmotic potentials (−2.83 MPa), and adaxial stomatal resistances (0.78 s mm⁻¹). The calculated hydraulic resistance of the two sorghums did not differ and averaged 3.4 × 10⁷ MPa s m⁻¹. The results suggested that the variation in susceptibility to drought between the two genotypes was due to differences in rate of soil-water extraction. Contribution No. 86-249-J from the Kansas Agricultural Experiment Station. The paper is dedicated to the memory of Dr Dan M Rodgers.     

Components of partial resistance as criteria for identifying resistance

The components of partial resistance, incubation period, lesion area, latent period and sporulation were recorded on plants of six winter and two spring wheat cultivars which had been artificially inoculated with Septoria nodorum spores. Incubation period gave a guide as to how the cultivars would respond in the field to Septoria nodorum but statistical analysis showed that it could not be used alone to predict accurately the resistance of each cultivar to the pathogen. Average sporulation, however, could be used with more confidence for predicting the field resistance of the cultivars. From a regression analysis of NIAB rating versus incubation period, lesion area, latent period and sporulation, an equation was devised to obtain resistance indices for each cultivar. These resistance indices clearly reflected the NIAB ratings for the cultivars. It would therefore appear that resistance indices could be used as a pre-field evaluation method for identifying resistance to Septoria nodorum and thus be a valuable technique in breeding programmes.     

Costs of resistance

Studies of the reduction of fitness in plants expressing resistance characteristics have always been popular. New techniques for manipulating defense expression have recently resulted in a greater understanding of the mechanisms through which different types of resistance strategies produce costs, especially those costs associated with inducible defenses.     

山西省黍稷种质资源抗倒性鉴定及相关形态特征研究

黍稷种质的倒伏是造成黍稷减产的一大要素,如何防止黍稷的倒伏,筛选和培育抗倒的种质是解决黍稷倒伏最根本和有效的方法。通过对山西省的1192份黍稷种质资源进行抗倒性鉴定,筛选出71份高抗倒的种质。同时对高抗倒种质和不抗倒种质茎、根的形态特征进行了比较研究,证明黍稷种质的抗倒性与茎、根的形态特征有密切关系,高抗倒种质在茎、根形态特征上所占的优势是形成高抗倒的重要原因。     

The evolution of resistance genes in multi-protein plant resistance systems

The genomic perspective aids in integrating the analysis of single resistance (R-) genes into a higher order model of complex plant resistance systems. The majority of R-genes encode a class of proteins with nucleotide binding (NB) and leucine-rich repeat (LRR) domains. Several R-proteins act in multi-protein R-complexes that mediate interaction with pathogen effectors to induce resistance signaling. The complexity of these systems seems to have resulted from multiple rounds of plant-pathogen co-evolution. R-gene evolution is thought to be facilitated by the formation of R-gene clusters, which permit sequence exchanges via recombinatorial mispairing and generate high haplotypic diversity. This pattern of evolution may also generate diversity at other loci that contribute to the R-complex. The rate of recombination at R-clusters is not necessarily homogeneous or consistent over evolutionary time: recent evidence suggests that recombination at R-clusters is increased following pathogen infection, suggesting a mechanism that induces temporary genome instability in response to extreme stress. DNA methylation and chromatin modifications may allow this instability to be conditionally regulated and targeted to specific genome regions. Knowledge of natural R-gene evolution may contribute to strategies for artificial evolution of novel resistance specificities.