丁烯基多杀菌素高产菌株的巴龙霉素抗性筛选

为了提高须糖多孢菌Saccharopolyspora pogona的丁烯基多杀菌素产量水平,利用核糖体工程技术,对其进行巴龙霉素抗性筛选,在10×MIC巴龙霉素浓度下,分离得到巴龙霉素自发抗性突变株54株。通过对原始菌株和突变株代谢产物变化的初步研究发现,相比于原始菌株,丁烯基多杀菌素组分在抗性突变株中的合成能力有明显差异,有6株抗性突变株的产量明显高于原始菌株,约20%左右的突变株产量提高,约45%左右的突变株产量降低。其中突变株P-7的丁烯基多杀菌素产量提高幅度最大,相比原始菌株提高2.2倍。对巴龙霉素抗性突变株P-7进行DNA序列分析,在编码核糖体S12蛋白的rps L基因保守区域中发现点突变,第314位的C突变为A,由脯氨酸突变为谷氨酰胺;第320位的C突变为T,由丙氨酸突变为缬氨酸。     

MNNG对多杀菌素产生菌的诱变效应

多杀菌素(spinosad)是刺糖多孢菌 Saccharopolyspora spinosa发酵产生的次级代谢产物,有显著杀虫活性.采用N-甲基-N′硝基-N-亚硝基胍(MNNG)作为诱变因子对刺糖多孢菌的孢子进行诱变处理,以高效液相色谱方法检测多杀菌素的发酵产量.研究结果表明:诱变剂量为2mg/ml,诱变40min时多杀菌素产量有较大幅度提高.通过诱变最终得到5株产量分别比出发菌株提高61.1%、80.3%、128.1%、69.9%和77.8%的突变株.     

虫道注射两种生物杀虫剂对桉蝙蝠蛾的防治效果

采用虫道注射方法,研究了吡虫啉和阿维菌素对桉蝙蝠蛾幼虫的防治效果。结果表明,两种杀虫剂对桉蝙蝠蛾幼虫平均虫口密度的防治效果为100%。吡虫啉8个月的持续防治效果稍高于阿维菌素,虫害防治效果达到70.4%,而阿维菌素为69.3%;两种药剂对桉蝙蝠蛾有较强和持续的抑制作用。吡虫啉和阿维菌素每孔注射适宜用药量为5ml(1:200)稀释液。     

二斑叶螨对阿维菌素抗性的CAPS标记

为了实现二斑叶螨Tetranychus urticae Koch对阿维菌素抗药性的快速检测,本研究建立了一种叶螨对阿维菌素抗药性的快速分子检测技术,即酶切扩增多态性序列(cleaved amplified polymorphic sequences,CAPS)标记技术,检测二斑叶螨谷氨酸门控氯离子通道(Glu Cl3)基因片段上G326E的突变频率,并采用琼脂浸叶法测定田间5个种群对阿维菌素的抗药性。结果表明,北京密云、顺义、昌平、浙江宁波和海南吉阳5个种群对阿维菌素的抗性倍数分别达1 155.63、1 317.10、844.19、314.95和1 799.69倍,均为极高抗性水平;CAPS检测发现我国二斑叶螨抗性基因存在G326E位点突变,但在供试的5个不同种群中突变频率变异很大,其中北京昌平种群G326E突变频率高达90.00%,密云和顺义种群均为26.67%,浙江宁波种群为5.00%,而海南吉阳种群不存在该位点的突变。表明该CAPS标记还需与其它抗性机制或检测技术结合来实现叶螨对阿维菌素抗性的早期检测和预警。     

阿维菌素防治水稻中后期二化螟的效果

阿维菌素对早、晚稻后期二化螟有显著的防治效果。试验表明,每667m2用阿维菌素1g(纯药量)防治第2、4代二化螟效果达到89.1%和91.3%,高浓度阿维菌素喷治虫伤株(团)对5龄、6龄幼虫也有较好的杀灭效果。但每667m2用量低于1.0g时,防效显著下降。     

静电自组装法制备阿维菌素纳米囊悬浮剂及其对黄瓜根结线虫病的防治效果

为提高阿维菌素在土壤中的移动扩散能力,改善其对作物根结线虫病的防治效果,采用静电自组装法制备了3%阿维菌素纳米囊悬浮剂,测定了其贮存稳定性和释放性能,并通过土壤淋溶试验与土壤薄层层析试验评价了其在土壤中的移动性,验证了其对南方根结线虫Meloidogyne incognita 2龄幼虫 (J₂) 的室内毒力和田间防治效果。结果显示:阿维菌素纳米囊为球形,平均粒径 (D₅₀) 为276.81 nm,在释放液中,60 min时纳米囊悬浮剂中阿维菌素的累积释放率为57.59%,而对照乳油制剂中阿维菌素的累积释放率为94.53%。阿维菌素纳米囊悬浮剂在土壤中横向和纵向的移动距离均明显大于阿维菌素乳油,二者对根结线虫J₂在24 h时的LC₅₀值分别为0.257与0.427 mg/L。田间试验中,相同用量下分别采用漫灌法与混土法两种方式施药,药后60 d,3%阿维菌素纳米囊悬浮剂处理比3%阿维菌素乳油处理的防治效果分别高21.82%与4.81%;而药后100 d,前者比后者分别高28.98%与19.79%。表明将阿维菌素制剂纳米化可以增强其在土壤中的移动能力,提高其对黄瓜根结线虫病的防治效果。本研究结果可为开发适合于土壤施用的纳米化农药制剂提供依据。     

不同杀线剂对小麦孢囊线虫病的防治效果

小麦孢囊线虫病目前已在中国13个省(市区)的小麦种植区有发生与分布,其扩散与蔓延将直接威胁我国的粮食生产和经济安全,系统开展小麦孢囊线虫病的防治研究迫在眉睫.本文在大田条件下研究了5％涕灭威颗粒剂(aldicarb)、3％克百威颗粒剂(carbofuran)、5％硫线磷颗粒剂(cadusafos)、10％苯线磷颗粒剂(fenamiphos)和0.5％阿维菌素颗粒剂(abamectin)等5种杀线剂各3个不同剂量在小麦返青期使用对小麦孢囊线虫病的控制效果.结果表明,各药剂处理均对土壤中孢囊线虫的繁殖有明显的抑制作用,但各处理间的校正孢囊减退率差异并不显著;不同药剂处理后的小麦株高、单株根重和单株鲜重均优于对照,且小麦产量与对照相比均有增加.其中0.5％阿维菌素颗粒剂 30 kg/hm2处理后的校正孢囊减退率最大值为55.13％,且增产效果最好,增产率为18.54％;而0.5％阿维菌素颗粒剂60 kg/hm2处理后小麦株高平均值最大,为59.32 cm,促进植物生长的效果明显.由于0.5％阿维菌素颗粒剂能够明显抑制土壤中孢囊线虫的繁殖,同时促进植株生长并减少产量损失,因此,在小麦孢囊线虫重病田于小麦返青期施用0.5％阿维菌素颗粒剂30 kg/hm2进行土壤处理,可在一定程度上减轻小麦孢囊线虫造成的损失.     

8种杀螨剂对马铃薯朱砂叶螨的防治效果

为了筛选在西北旱作区针对马铃薯朱砂叶螨的最佳防治药剂, 选择了8种杀螨剂进行两年的田间药效试验。结果表明, 24%螺螨酯悬浮剂(SC)处理下的马铃薯单株块茎数最高, 螨株率最低, 并且叶片SPAD值及光合参数值最高, 8种药剂中, 24%螺螨酯SC、43%联苯肼酯SC、2%阿维菌素SC、11%乙螨唑SC和73%炔螨特乳油(EC)对马铃薯朱砂叶螨防效高于80%, 可作为防治马铃薯朱砂叶螨的药剂。     

保水剂施用方式对马铃薯产量和水分利用效率的影响

采用盆栽试验,比较深施、浅施、侧施、全施和不施(对照)等保水剂应用方法在充分供水和水分亏缺两种供水条件下,马铃薯(Solanum tuberosum L)生长、产量和水分利用效率(WUE)效应.结果表明,除浅施处理外其它方式施用保水剂处理都能促进马铃薯株高,水分胁迫下能促进马铃薯根系发育.在充分供水和水分胁迫下,深施保水剂使马铃薯产量较对照提高9.4%和16.6%,WUE提高12.82%和54.73%,侧施保水剂使马铃薯产量较对照提高39.4%和21.5%,WUE提高58.79%和59.46%;在水分胁迫下,全施保水剂使马铃薯产量和WUE较对照分别提高15.4%,56.76%.     

添加燃煤脱硫废弃物和专用改良剂对碱化土壤和水稻生长的影响

基于大田试验,通过测定土壤理化性质和水稻成活率、株高和产量等,研究了燃煤脱硫废弃物(简称脱硫废弃物)和3种专业改良剂添加对碱化土壤的改良效果.结果表明:脱硫废弃物和改良剂配合施用显著提高了土壤有机质和养分含量,一定程度上降低了碱化度、pH和全盐含量,使水稻成活率、株高和产量分别提高了30.9%、36.3%和64.8%以...     

松毛虫赤眼蜂对三种农田常用杀虫剂的敏感性

为明确常用杀虫剂对松毛虫赤眼蜂Trichogramma dendrolimi的安全性,室内评估了高效氯氰菊酯、阿维菌素和氯虫苯甲酰胺3种药剂对其成蜂的急性毒力以及田间推荐使用剂量对赤眼蜂各发育阶段的间接影响,并模拟自然情况,测定了不同残留期的3种药剂对赤眼蜂存活和寄生能力的影响.结果表明:以LC₅₀进行评估,对松毛虫赤眼蜂毒力最高的为阿维菌素,其次是高效氯氰菊酯,氯虫苯甲酰胺毒力最小;在田间推荐使用剂量下,阿维菌素、高效氯氰菊酯和氯虫苯甲酰胺的安全系数分别为0.017、0.066和69.175;在推荐使用剂量下,氯虫苯甲酰胺处理含有不同发育阶段赤眼蜂的米蛾Corcyra cephalonica卵后,平均羽化率最高为90.43%,其次是高效氯氰菊酯为56.46%,阿维菌素仅为9.84%;接触0~7 d残留期阿维菌素的玉米叶片24 h,松毛虫赤眼蜂全部死亡,无米蛾卵被寄生,但接触3~7 d残留期氯虫苯甲酰胺的玉米叶片,赤眼蜂的死亡率和寄生米蛾卵数量与清水对照无明显差异.表明在实施释放赤眼蜂的害虫综合治理方案中,应尽量避免使用阿维菌素,而氯虫苯甲酰胺对赤眼蜂风险性极低,具有很好的相容性,可以推广使用.     

Transcription and induction profiles of two esterase genes in susceptible and acaricide-resistant Tetranychus cinnabarinus

The carmine spider mite Tetranychus cinnabarinus is the most serious of crop mite pests in China. Their ability to rapidly develop resistance to acaricides has caused difficulty in controlling this mite. In this study, the molecular mechanism of acaricide resistance associated with esterase genes TCE1 and TCE2 was investigated in susceptible and acaricide-resistant strains of T. cinnabarinus. The quantitative real-time PCR (qrtPCR) method was adopted to compare the expression level of two esterase genes TCE1 and TCE2 among four different strains (abamectin-resistant, AbR; fenpropathrin-resistant, FeR; omethoate-resistant, OmR and susceptible strains, S) of T. cinnabarinus. The relative expression level of TCE2 was 1.39-2.47 fold in the three resistant strains compared with the S strain. And after inducing with abamectin, fenpropathrin, and omethoate the highest expression level of TCE2 in the S was 1.64-, 2.92- and 2.24-fold compared with the control, respectively, and this difference was found to be significant. However, there was no obvious difference of the mRNA relative expression levels of TCE1 genes among the four strains, and those of TCE1 were not higher than the control throughout the study. Furthermore, the expression modes of TCE1 and TCE2 in AbR and FeR were similar with that in the S after being treated with abamectin and fenpropathrin, respectively. These results indicated that the enhanced expression of esterase gene TCE2 was associated with acaricide-resistance in T.cinnabarinus.     

Resistance selection and biochemical mechanism of resistance to two Acaricides in Tetranychus cinnabarinus (Boiduval)

A Tetranychus cinnabarinus strain was collected from Chongqing, China. After 42 generations of selection with abamectin and 20 generations of selection with fenpropathrin in the laboratory, this T. cinnabarinus strain developed 8.7- and 28.7-fold resistance, respectively. Resistance to abamectin in AbR (abamectin resistant strain) and to fenpropathrin in FeR (fenpropathrin resistant strain) was partially suppressed by piperonyl butoxide (PBO), diethyl maleate (DEM) and triphenyl phosphate (TPP), inhibitors of mixed function oxidase (MFO), glutathione S-transferases (GST), and hydrolases, respectively, suggesting that these three enzyme families are important in conferring abamectin and fenpropathrin resistance in T. cinnabarinus. The major resistant mechanism to abamectin was the increasing activities of carboxylesterases (CarE), glutathione-S-transferase (GST) and mixed function oxidase (MFO), and the activity in resistant strain developed 2.7-, 3.4- and 1.4-fold contrasted to that in susceptible strain, respectively. The activity of glutathione-S-transferase (GST) in the FeR strain developed 2.8-fold when compared with the susceptible strain, which meant the resistance to fenpropathrin was related with the activity increase of glutathione-S-transferase (GST) in T. cinnabarinus. The result of the kinetic mensuration of carboxylesterases (CarE) showed that the structure of CarE in the AbR has been changed.     

Biochemical mechanisms conferring cross-resistance between tebufenozide and abamectin in Plutella xylostella

Experiments have been carried out to confirm the cross-resistance between abamectin and tebufenozide in Plutella xylostella and demonstrate its mechanism. The results showed that the resistant strain of P. xylostella selected by tebufenozide (RF 99.38) really showed high cross-resistance to abamectin (RF 29.25). When this strain was subjected to resistance decaying treatment, breeding without contacting any insecticides, and abamectin resistance selection for 20 generations, the former resulted in decrease of its resistance to both tebufenozide and abamectin to about one third of the original (RF 35.03 and 11.67, respectively), and the later enhanced its resistance to abamectin dramatically (RF 303.77), but not to tebufenozide(RF 50.04). PBO showed high synergism to abamectin (SR 2.11-12.23), and the synergism ratio positively related to the resistance level among different strains. Enzyme analysis also proved that the activity of cytochrome P450 monooxygenase (MFO) was notable enhanced in the strains resistant to both tebufenozide and abamectin (1.71- to 3.01-fold). Based on discussion, it was concluded that tebufenozide selection could resulted in significant cross-resistance of P. xylostella to abamectin. The major mechanism for the cross-resistance should be the enhancement of MFO activity. For resistance management, tebufenozide and abamectin would not recommend for rotational use.     

Selection and characterization of spinosad resistance in Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae)

The cross-resistance and biochemical mechanism of the beet armyworm, Spodoptera exigua (Hübner), to spinosad was studied in the laboratory. S. exigua population were collected from Shanghai suburb. After five generations of selection, the resistance of S. exigua to spinosad increased 345.4 times compared with the susceptible strain. There was no cross-resistance between spinosad and fenvalerate, phoxim, methomyl, abamectin, and cyfluthrin. When the inhibitors, PBO, TPP, DEF, and DEM were used as synergist in the susceptible strain and resistant strain, the synergistic ratio was 0.7-, 0.5-, 1.0-, and 0.6- fold for the susceptible strain, and 9.8-, 1.5-, 2.6-, and 1.5-fold for the resistant strain, respectively. The results revealed that PBO had significant synergistic effect on the resistant strain. The activity in vitro of microsomal-O-demethylase and glutathione S-transferase in the resistant strain was 5.2- and 1.0-fold of the susceptible strain, respectively. The results implied that microsomal-O-demethylase might be important in conferring spinosad resistance in the S. exigua population.     

二斑叶螨对阿维菌素的抗药性及抗性基因的PASA检测技术

为了明确二斑叶螨Tetranychus urticae Koch对阿维菌素的抗性水平,采用玻片浸渍法测定了北京4个地区二斑叶螨雌成螨对阿维菌素的抗药性,建立了特异性等位基因PCR（PASA）方法,并检测了二斑叶螨谷氨酸门控氯离子通道（GluCl）基因片段上G323D的突变频率。结果显示,北京昌平、海淀、密云和怀柔4个田间二斑叶螨种群对阿维菌素均达极高抗性水平,其中昌平种群抗性最高,LC₅₀值为448.93 mg/L,抗性倍数为4 988.11倍。室内敏感种群未见抗性个体,昌平和密云种群G323D等位基因突变频率为100.00%,怀柔和海淀种群分别为86.25%和90.00%。北京4个地区二斑叶螨种群对阿维菌素达极高抗性水平,抗性基因的突变频率也很高,表明阿维菌素不适宜用来防治这些地区的二斑叶螨种群。     

八种杀虫剂对黑粪蚊的防治效果及残留分析

选择8种杀虫剂,分别在实验室和菇房条件下研究其对黑粪蚊的防治效果和对平菇菌丝生长的影响,并分析施药后不同时间平菇中的农药残留.结果表明,40%辛硫磷EC、40%氧乐果EC、48%毒死蜱EC对黑粪蚊具有很好的防治效果,但对平菇菌丝的抑制率明显高于其它药剂;10%吡虫啉WP、4.5%高效氯氰菊酯EC不仅对黑粪蚊成、幼虫具有良好的防治效果,而且使用后对平菇菌丝生长的影响较小;2.5%高效氯氟氰菊酯EW、4.5%高效氯氰菊酯EC、1.8%阿维菌素EC等使用5天后均未检测到农药残留,5%氟虫腈SC和10%吡虫啉WP药后15天仍能检测到残留.     

Activity of abamectin against larval stages of Spodoptera littoralis boisduval and Heliothis armigera hübner (Lepidoptera: Noctuidae) and possible mechanisms determining differential toxicity

Marked changes in the relative toxicity of topically-applied abamectin were found between larval instars of Spodoptera littoralis, toxicity decreasing up to the fifth instar but increasing over 500-fold (at LD₅₀ level) in the sixth instar. By contrast, the toxicity of abamectin remained constant from fifth to sixth instar Heliothis armigera and there was an increase in the toxicity of two chemically unrelated insecticides, malathion (4-fold) and lambda-cyhalothrin (2.5-fold), from fifth to sixth instar S. littoralis. Prior topical application or injection of the microsomal oxidase inhibitor, piperonyl butoxide (PB) increased the toxicity of abamectin (6–8 and 16-fold respectively) against fifth instar S. littoralis, while topically-applied PB increased the proportion of radioactivity present as abamectin in the ventral nerve cord of this instar following topical application with [³H]abamectin. Topically-applied PB also enhanced the toxicity of abamectin against third (4-fold) and fourth instar (5-fold) S. littoralis but had no significant effect on sixth instar S. littoralis, fifth instar H. armigera, or on the toxicity of malathion and lambda-cyhalothrin against fifth instar S. littoralis. Topical application of the esterase inhibitor, S,S,S-tributyl phosphorothioate (DEF) significantly increased the toxicity of abamectin at the LD₅₀ level (3-fold) against fifth instar S. littoralis. The toxicity of injected abamectin against fifth instar S. littoralis was greater (20-fold) than with topical application but injected abamectin was less toxic (2-fold) against sixth instar S. littoralis and had no significant effect on fifth instar H. armigera. It is suggested that differential toxicity of abamectin is due in part to greater metabolism and reduced penetration in fifth instar S. littoralis than in sixth instar S. littoralis or fifth instar H. armigera.     

常用杀虫剂对异色瓢虫的毒力及其保护酶的影响

为明确杀虫剂对异色瓢虫Harmonia axyridis(Pallas)幼虫、成虫的毒力及其体内保护酶的影响,采用喷雾法测定了7种杀虫剂对异色瓢虫3龄幼虫和成虫的毒力,同时测定了3种杀虫剂对3龄幼虫和成虫体内超氧化物歧化酶(superoxide dismutase)、过氧化物酶(peroxidase)和过氧化氢酶(catalase)活性的影响。结果显示:7种杀虫剂对3龄幼虫的毒力大小为阿维菌素、啶虫脒吡虫啉、虫酰肼哒螨灵噻嗪酮吡蚜酮,对成虫的毒力大小为阿维菌素、啶虫脒虫酰肼、吡虫啉哒螨灵噻嗪酮吡蚜酮,吡蚜酮对异色瓢虫最安全。亚致死浓度LC10药剂可促进保护酶活性的提高,但随着浓度提高和处理时间增长,3种酶的活性总体呈逐渐下降趋势,表明低浓度的杀虫剂对异色瓢虫体内酶活性存在一定的不利影响。     

溴氰虫酰胺和氯虫苯甲酰胺对三种鳞翅目害虫的毒力作用比较

为比较溴氰虫酰胺和氯虫苯甲酰胺对甜菜夜蛾、玉米螟和小地老虎的作用差异,采用浸卵法、浸叶法、饲喂毒营养液法等测定了溴氰虫酰胺和氯虫苯甲酰胺对3种害虫的卵、3龄幼虫和成虫的毒力,以及对其3龄幼虫的拒食活性。结果表明,溴氰虫酰胺和氯虫苯甲酰胺均无杀卵作用,但能降低初孵幼虫存活率。溴氰虫酰胺对甜菜夜蛾、玉米螟和小地老虎3龄幼虫的LC₅₀值分别为0.11、0.05和0.13 mg/L,相对毒力分别是氯虫苯甲酰胺的2.8、2.7和5.1倍。溴氰虫酰胺对3种害虫成虫的LC₅₀值分别为0.11、0.09和0.22 mg/L,相对毒力分别是氯虫苯甲酰胺的3.9、3.8和2.7倍。溴氰虫酰胺在亚致死浓度0.10、0.05和0.14 mg/L的剂量下分别对甜菜夜蛾、玉米螟和小地老虎3龄幼虫表现出较强的拒食作用,处理48 h后达到最高,拒食率分别为85.58%、81.91%和86.11%。药剂处理试虫虫体皱缩,体节缩短,体重增加率明显低于对照处理。表明溴氰虫酰胺对3种鳞翅目害虫的毒力作用比氯虫苯甲酰胺好,可作为氯虫苯甲酰胺的替代药品。