GK22、苏抗103及其受体亲本对亚洲玉米螟的抗性比较

通过室内饲养研究转Bt基因抗虫棉GK22、苏抗103对亚洲玉米螟的抗性.用GK22和苏抗103及其受体亲本的不同器官饲喂玉米螟初孵幼虫和3龄幼虫,比较棉花苗期至蕾期、花期和铃期玉米螟幼虫的死亡率,测定死亡率的方差分析差异和平均幼虫存活综合指数.结果表明:GK22和苏抗103与其受体亲本比较对玉米螟都具较高的抗性,且GK22抗性高于苏抗103,GK22和苏抗103不同生育阶段抗性顺序为苗期至蕾期＞花期＞铃期,对玉米螟抗性表现为1龄的抗性高于3龄,     

转cry1Ab基因玉米对粘虫的抗性评价

粘虫Mythimna separata(Walker)是我国玉米上的重要害虫。转Bt基因玉米为防治粘虫提供了新的途径。从室内生测和田间抗性鉴定分别研究了转cry1Ab基因抗虫玉米YieldGard对粘虫的杀虫效果。用4~7叶期转cry1Ab基因玉米心叶室内饲养1~3龄粘虫幼虫3、7和10天的死亡率分别为39.0%~92.0%、96.0%~98.0%和97.0%~100%,第14天全部死亡,而对照的死亡率分别为3.0%~8.0%、6.0%~20.0%和11.0%~28.0%,差异极显著。取食转cry1Ab基因玉米的幼虫不能正常生长发育,存活10天的幼虫其体重仅为取食对照玉米的9.0%。田间玉米4~5叶期,分别接1~6龄粘虫幼虫于转cry1Ab基因和非转基因玉米心叶。转cry1Ab基因玉米上各龄期幼虫平均存活率仅为0.0~6.7%,极显著低于非转基因玉米的8.3%~80.0%。转cry1Ab基因玉米的危害级别0.1~0.7,显著小于非转基因玉米的2.1~7.3。结果说明转cry1Ab基因玉米YieldGard对粘虫各龄幼虫有很高的杀虫作用和田间控制效果,同时能有效保护玉米免受危害。     

小地老虎对几类杀虫剂的毒力反应及其抗药性变化

为了系统地了解小地老虎对几类主要杀虫剂的毒力反应,及其对几种常用杀虫剂的抗性水平,为合理用药和提高现有杀虫剂的杀虫效果提供依据,本文研究了5类、18种杀虫剂对小地老虎五龄幼虫的触杀毒力,以及小地老虎对数种药剂的抗性程度。结果得出,触杀毒力顺序依次为:拟除虫菊酯类>>有机磷剂>六六六和沙蚕毒系杀虫单>>氨基甲酸酯类≥滴滴涕。与1965年的毒力测定资料对比,小地老虎幼虫对敌百虫和滴滴涕的抗性都有明显增加。本试验还发现辛硫磷和敌百虫混配有明显的增效作用,建议在生产上应用合理的混配药剂来提高杀虫效果,减少施药量和次数。     

转Cry1Ab-ma基因玉米CM8101对草地贪夜蛾1龄和2龄幼虫的抗性

为明确转Bt基因玉米对草地贪夜蛾Spodoptera frugiperda的抗性,采用室内玉米离体组织生测法测定取食转Cry1Ab-ma基因抗虫玉米CM8101心叶、花丝和籽粒后草地贪夜蛾1龄和2龄幼虫存活数量、存活幼虫的体重、蛹重、化蛹和羽化等参数,分析转Cry1Ab-ma基因抗虫玉米CM8101三种组织对草地贪夜蛾1龄和2龄幼虫的抗性。结果显示,转Cry1Ab-ma基因玉米CM8101心叶可完全杀死草地贪夜蛾1龄幼虫,2龄幼虫有存活,存活率为10.0%,但不能化蛹与羽化;花丝可杀死大部分草地贪夜蛾1龄幼虫,存活率为3.3%,并阻断其生活史,但对2龄幼虫杀伤力弱,存活率为73.3%,存活幼虫化蛹率和羽化率分别为20.0%和13.3%;籽粒可完全杀死草地贪夜蛾1龄幼虫,2龄幼虫有存活,存活率为13.3%,有少量可化蛹,化蛹率为3.3%、有少量羽化,羽化率为3.3%。表明转Cry1Ab-ma基因玉米CM8101在短期内对草地贪夜蛾1龄和2龄幼虫有良好的控制作用。     

转Bt基因玉米对棉铃虫的抗性评价

棉铃虫Helicoverpa armigem（Htlbner）是常年危害玉米的重要害虫。转Bt基因抗虫玉米为害虫的防治提供了新的途径。采用室内离体生物测定方法测定了孟山都转Bt基因抗虫玉米Yield—Gard不同组织对棉铃虫初孵幼虫的杀虫活性以及田间人工接虫抗性鉴定。Bt玉米花丝、苞叶和幼嫩雌穗对棉铃虫具有较高的杀虫活性，取食Bt玉米花丝和幼嫩雌穗（籽粒和穗轴）的棉铃虫初孵幼虫不能存活和完成幼虫期发育，而对照分别有51．0％和22．0％个体完成幼虫期发育。以Bt玉米雌穗苞叶饲养棉铃虫初孵幼虫，5天后幼虫全部死亡，而非Bt玉米对照仅为16．0％，差异显著。田间花丝期人工接种棉铃虫抗性鉴定试验结果表明，Bt玉米雌穗被害率为3．5％，危害级别为0．007，百株存活幼虫14．3头。而非Bt玉米雌穗被害率达到82．5％，危害级别为2．51，平均百株存活幼虫79．6头，显著高于Bt玉米。这些结果说明转Bt基因玉米穗期对棉铃虫的防治效果明理．     

转Bt基因杨树对杨扇舟蛾的抗性

通过室内饲养和组织切片,研究了表达Cry1A杀虫蛋白的中嘉8号杨树转Bt基因植株对杨扇舟蛾的抗虫效果。结果表明：随着饲养时间的延长,幼虫死亡率和体重抑制率呈上升趋势,处理2d仅分别为10%和6.85%,处理4d分别增加到48.33%和27.93%,处理10d分别达81.67%和63.03%。此外,处理12d取食非转基因杨树幼虫化蛹率100%,而取食转Bt基因杨树幼虫的化蛹率仅为18.18%。组织切片观察表明,取食转Bt基因杨树叶片幼虫的中肠上皮细胞排列松驰,与肠壁基底膜分离,细胞界限较模糊,部分上皮细胞形成空泡。因此,初步认为转Bt基因杨树对杨扇舟蛾幼虫的抗性表现为毒杀作用和抑制其生长发育。     

昆虫病原线虫对小地老虎的致病力测定及防治效果

为探讨应用昆虫病原线虫防治小地老虎技术的可行性,采用培养皿滤纸法比较9个品系的昆虫病原线虫对小地老虎幼虫的致病力,同时通过盆栽试验验证了斯氏线虫属小卷蛾斯氏线虫Steinernema carpocapsae NC116品系、芫菁夜蛾斯氏线虫S.feltiae SF-SN品系和异小杆线虫属嗜菌异小杆线虫Heterorhabditis bacteriphora H06品系的保苗效果和防治效果,并测定了苦参碱与NC116品系混用对小地老虎3龄幼虫的联合作用效果。结果表明,NC116品系对小地老虎3龄幼虫致病力最高,其对3龄、4龄和5龄的LD50分别为2.4、7.5、和31.2条/头,表明随小地老虎幼虫龄期递增,其致病力下降。盆栽玉米苗上分别施用线虫100~200条/头时,对玉米保苗效果及对小地老虎3龄幼虫的控制效果依次为NC116H06SF-SN。0.8 mg/L苦参碱和NC116品系混用后,可使小地老虎3龄幼虫死亡率提高109.89%,二者表现增效作用。     

Cry2Ab杀虫蛋白对小地老虎幼虫致死效果及体内酶活性的影响

为治理小地老虎的危害,研究了Bt杀虫蛋白对小地老虎幼虫的控制作用.室内生物测定结果表明,Cry2Ab杀虫蛋白对小地老虎初孵幼虫的致死效果高于Cry1Ac杀虫蛋白.取食含有Cry2Ab杀虫蛋白人工饲料的小地老虎幼虫体内的中肠总蛋白酶和类胰凝乳蛋白酶活性高于对照,当人工饲料中Cry2Ab杀虫蛋白含量达到12μg/g时,小地老虎幼虫中肠氨肽酶活性显著升高;而取食含有Cry1Ac杀虫蛋白人工饲料的小地老虎幼虫中肠蛋白酶活性无显著变化.此外,小地老虎幼虫取食含有Cry2Ab杀虫蛋白的人工饲料后,体内谷胱甘肽-S-转移酶活性显著升高,而取食合有Cry1Ac杀虫蛋白的人工饲料后其变化不明显.     

转Bt基因玉米(瑞丰125、DBN9936、DBN9978)对亚洲玉米螟的抗虫效果研究

评价转Bt基因玉米对靶标生物亚洲玉米螟的杀虫作用是转基因玉米研发的重要一环。本文采用室内生测法对3种转Bt基因抗虫玉米‘瑞丰125’(表达Cry1Ab/Cry2Aj杀虫蛋白),‘DBN9936’‘DBN9978’(表达Cry1Ab杀虫蛋白)对亚洲玉米螟敏感品系ACB-S及抗Cry1Ab品系ACB-AbR、抗Cry1Ac品系ACB-AcR、抗Cry1F品系ACB-FR、抗Cry1Ah品系ACB-AhR、抗Cry1Ie品系ACB-IeR的杀虫活性进行测定,同时采用心叶期和抽丝期人工接虫法进行田间抗虫效果鉴定。结果表明,取食3种Bt玉米的ACB-S幼虫, 3 d死亡率100%,而取食对照常规玉米3 d存活率100%。取食3种Bt玉米的5个抗性品系幼虫除ACB-AbR和ACB-AcR有2%~6%的个体存活4~5 d, 6 d死亡率也达到了100%,其余品系均在3 d全部死亡,而取食对照玉米5~6 d的死亡率仅为4%~14%,差异显著。田间心叶期食叶级别及穗期活虫数、雌穗被害和茎秆被蛀等为害等级说明3种Bt玉米高抗亚洲玉米螟。明确了‘瑞丰125’‘DBN9936’和‘DBN9978’对亚洲玉米螟有很高的杀虫活性和田间防治效果。5个Bt蛋白抗性亚洲玉米螟品系幼虫在常规玉米上显示一定的适合度劣势。     

抗菌肽和几丁质酶基因提高烟草对黑胫病菌和赤星病菌的抗性研究

 通过农杆菌介导法将加信号肽修饰的人工合成杂合抗菌肽CEMA基因(SPCEMA),苜蓿防御素基因(AFP),苦瓜几丁质酶基因(CHI)以及SPCEMA-CHI、AFP-CHI、AFP-SPCEMA双价基因导入本明烟(Nicotiana benthamiana),并对转基因烟草T₀和T₁代进行了抗病性检测,比较了不同转基因植株的抗病效果。研究结果表明,转基因烟草对黑胫病菌(Phytophthora parasitica var.nicotianae)、赤星病菌(Alternaria alternata)的抗性均强于非转基因烟草,病情指数差异达极显著水平,其中转AFP-CHI双价基因烟草具有较强的抗性,与单价转基因烟草的抗性差异达显著水平。但各单价转基因以及双价转基因烟草之间对上述病菌并未表现出显著的抗病性差异。结果表明植物源的抗菌肽基因与几丁质酶基因在抗植物真菌病害中具有协同增效作用。     

四种Bt毒素蛋白对二点委夜蛾幼虫毒力测定及中肠组织病理学变化

为更好地了解苏云金芽胞杆菌Bacillus thuringiensis毒素蛋白对二点委夜蛾Athetis lepigone的毒力以及作用机理,通过饲喂含有Cry1Ac、Cry1Ab、Cry2Ab和Vip3Aa四种不同Bt毒素蛋白饲料,测定Bt毒素蛋白对二点委夜蛾幼虫的毒力,并观察取食4种毒素蛋白后幼虫中肠组织的病理学变化。结果显示,二点委夜蛾幼虫取食毒素蛋白后72 h,Cry1Ab和Cry1Ac毒素蛋白对二点委夜蛾幼虫的杀虫活性较高,校正死亡率为84.7%和76.4%;Vip3Aa和Cry2Ab毒素蛋白的毒力较弱。二点委夜蛾幼虫取食4种Bt毒素蛋白后,中肠柱状细胞微绒毛脱落,杂乱地分散在肠腔内,杯状细胞变形和腔内微绒毛脱落,线粒体和内质网等变形破裂,细胞核的核膜消失、核质凝聚和形状发生变化,经Cry1Ab和Cry1Ac毒素蛋白处理后中肠细胞的病变症状和速度明显高于Cry2Ab和Vip3Aa毒素蛋白处理。表明Cry1Ab和Cry1Ac毒素蛋白对二点委夜蛾幼虫杀虫活性较高,显著高于Cry2Ab和Vip3Aa毒素蛋白,且对其中肠细胞的破坏作用也较强。     

转Cry1Ac/1Ab基因棉花对棉铃虫的控制效果及对甜菜夜蛾和斜纹夜蛾生长发育的影响

为评估转Cry1Ac/1Ab基因棉花对棉铃虫Helicoverpa armigera的抗性及对非靶标害虫甜菜夜蛾Spodoptera exigua和斜纹夜蛾S.litura生长发育的影响,采用室内生测法测定其对棉铃虫的抗性及对甜菜夜蛾和斜纹夜蛾不同龄期幼虫存活率、营养代谢及中肠酶活性的影响。结果表明,转Cry1Ac/1Ab基因棉花对棉铃虫第2代幼虫的抗性程度最高,幼虫校正死亡率达91.33%,但对甜菜夜蛾和斜纹夜蛾的抗性程度较低,幼虫校正死亡率分别为15.33%和13.33%。甜菜夜蛾和斜纹夜蛾各龄期幼虫取食转Cry1Ac/1Ab基因棉花后,其存活率与取食常规棉花对照无显著差异;甜菜夜蛾对转Cry1Ac/1Ab基因棉花叶片的相对取食量、近似消化率分别为16.68和93.12%,均高于取食常规棉花对照的10.72和92.00%,但差异不显著,而斜纹夜蛾取食转Cry1Ac/1Ab基因棉花后的各项营养指标均低于取食常规棉花对照,差异也不显著。取食转Cry1Ac/1Ab基因棉花后,甜菜夜蛾的过氧化物酶活性为0.02 U/mg prot,显著低于取食常规棉花的0.05 U/mg prot;斜纹夜蛾的酸性磷酸酶活性为0.15 U/mg prot,高于取食常规棉花的0.10 U/mg prot,但差异不显著,其它中肠酶活性均低于对照,亦无显著差异。     

水稻基腐病菌毒素对烟草活性氧代谢及细胞超微结构的影响

 用水稻基腐病菌毒素注射烟草叶片后,烟草细胞中O₂^-_·和NO大量增加,H₂O₂含量明显降低,SOD、CAT和POD等酶的活性发生变化。毒素引起烟草细胞死亡,但没有细胞程序性死亡(PCD)的DNA梯特征。病菌毒素处理烟草叶片后,电导率增大,细胞膜透性增加。电镜观察烟草细胞的超微结构表明,毒素处理8h后,叶绿体变形,叶绿体基质片层大部分消解,基粒结构消失,叶绿体外膜和内膜剥离,质壁分离和细胞膜内陷,细胞器消解;处理48h后,细胞内含物完全消解,细胞膜消失,细胞壁皱缩变形。本研究表明,毒素引起烟草细胞死亡与植物抗病反应HR中细胞坏死的生理生化机制不同,但可能存在相似的信号传导途径。     

暗黑鳃金龟幼虫取食Bt后中肠细胞形态变化的初步研究

本研究利用透射电镜技术观察了暗黑鳃金龟幼虫取食Bt后不同时间其中肠细胞形态变化过程。结果表明：幼虫取食Bt后1~7 d中肠不同的组织结构逐渐发生变化,第1天就出现了细胞膜溶解的现象,在第7天细胞膜完全溶解消失;其他器官出现如细胞核核膜溶解,细胞核消失,内质网消失等现象。经过中肠细胞一系列的病变,最终导致幼虫死亡。     

草地贪夜蛾对玉米和烟草的偏好性研究

自2019年初首先在云南省发现草地贪夜蛾入侵中国以来,该虫已对国家粮食安全构成严重威胁。草地贪夜蛾寄主范围广,主要喜食禾本科植物,同时也能为害多种双子叶植物。本研究以玉米和烟草为测试对象,比较研究了草地贪夜蛾在两种寄主上的产卵量及两种寄主植物对草地贪夜蛾生长发育的影响。结果表明,每个养虫笼中放入2对雌雄成虫,玉米植株上的卵块数量(17±2.65)块/笼显著多于烟草叶片上的卵块数量(3±1.73)块/笼,玉米上的幼虫数量(834.67±275.16)头/笼显著多于烟草上的幼虫数量(92±55.83)头/笼,表明草地贪夜蛾更喜欢选择玉米进行产卵和取食。烟草上草地贪夜蛾幼虫死亡率达92.53%±1.46%,但其能在烟草上产卵和存活。取食玉米籽粒的草地贪夜蛾幼虫发育历期、死亡率、体重、蛹重分别为(17.22±1.90)d、24.35%±8.13%、(0.261±0.112)g/头、(0.187±0.030)g/头,取食烟草叶片的草地贪夜蛾对应的参数分别为(26.80±1.89)d、85.49%±4.16%、(0.034±0.028)g/头、(0.131±0.028)g/头,表明取食烟草叶片的草地贪夜蛾幼虫发育历期显著延长,幼虫死亡率显著升高,幼虫体重和蛹重显著下降。与取食玉米籽粒的草地贪夜蛾相比,取食烟草叶片的个体适合度显著降低,但成虫配对后能够产卵并孵化出幼虫。与非嗜好寄主烟草相比,草地贪夜蛾取食嗜好寄主玉米表现出更高的适合度,其偏好选择玉米进行产卵和取食,但在烟草上也能完成生活史,种群密度大时存在为害烟草的潜在风险。     

Optical brightener M2R destroys the peritrophic membrane of Spodoptera exigua (Lepidoptera: Noctuidae) larvae

Observations under an environmental scanning electron microscope showed that the peritrophic membrane (PM) of Spodoptera exigua (Hübner) was smooth without pores, but there were many pores and slits in the PM of larvae fed with 10 g L(-1) optical brightener Calcofluor white M2R. After incubation of S. exigua PM in vitro with 10 g L(-1) M2R, a significant amount of proteins was released from the PM structure. M2R disrupted the structure of larval PM, resulting in greatly increased permeability and increased larval susceptibility to Syngrapha falcifera multiple nucleopolyhedrovirus infection. Continuous feeding of larvae on a diet treated with a 10 g L(-1) M2R solution significantly retarded larval development and resulted in high mortality. The destructive effect of M2R on PM was transient and reversible, depending on the presence of M2R within the midgut.     

Developmental studies of transgenic maize expressing Cry1Ab on the African stem borer, Busseola fusca; effects on midgut cellular structure

BACKGROUND: Busseola fusca is a major pest of maize in Africa but unfortunately is difficult to control using chemical insecticides. Insect‐resistant transgenic crops may provide an alternative viable strategy to control this pest. RESULTS: Recombinant Cry1Ab (1%) reduced larval weight by 60% over the trial period, while larval weight in the control group increased by 25%; no effects on mortality were observed. Insect survival, developmental rate and pupal and adult weight were significantly reduced (P < 0.05) on maize expressing Cry1Ab (MON810) compared with the non‐transformed parental line. These differences were more pronounced with second‐instar larvae than with third‐instar larvae. Leaf area consumed by Bacillus thuringiensis (Bt)‐fed larvae was significantly lower (0.5 cm² larva^?1 day^?1) compared with the area consumed by control‐fed insects (3.3 cm² larva^?1 day^?1). EM studies revealed that consumption of Bt maize deleteriously affected gut integrity. Effects were observed in columnar cells of the midgut epithelium, with the cytoplasm becoming highly vacuolated; the microvilli were disorganised, the mitochondria were abnormal and there was an increase in the number of lysosomal bodies. The rough endoplasmic reticulum had also become dilated. CONCLUSION: This study confirms the potential for Bt maize, when used as part of an IPM programme, for control of B. fusca. Copyright © 2011 Society of Chemical Industry     

转基因抗虫棉花对棉铃虫生长发育影响的系统评价

以我国研制的转基因抗虫棉(Bt棉)品系CK595-1为材料,系统研究了对棉铃虫1～5龄幼虫存活及生长发育的影响。结果表明,Bt棉对棉铃虫的杀虫活性和体重抑制率有明显的器官和幼虫龄期间差异,其活性顺序为叶>蕾>铃>花,其作用随着幼虫龄期的增大而降低。棉铃虫1～4龄幼虫连续取食Bt棉叶片和棉蕾不能正常化蛹,5龄幼虫可部分化蛹及羽化;1～4龄幼虫取食Bt棉的花及2～5龄幼虫连续取食Bt棉铃后均可部分化蛹及羽化,化蛹率随龄期增大而增加。     

Acquisition of Tomato spotted wilt virus by Adults of Two Thrips Species

ABSTRACT Only larval thrips that acquire Tomato spotted wilt virus (TSWV), or adults derived from such larvae, transmit the virus. Nonviruliferous adults can ingest virus particles while feeding on TSWV-infected plants, but such adult thrips have not been shown to transmit TSWV. Immunofluorescence microscopy was used to show that thrips 1, 5, 10, and 20 days after adult emergence (DAE) fed on TSWV-infected plants acquired TSWV with virus replication and accumulation occurring in both epithelial and muscle cells of Frankliniella fusca (tobacco thrips [TT]) and F. occidentalis (western flower thrips [WFT]), as indicated by immunodetection of the nonstructural (NSs) protein encoded by the small RNA and the nucleocapsid (N) protein, respectively. Adult WFT acquired TSWV more efficiently than TT. There was no significant effect of insect age on TSWV acquisition by TT. In contrast, acquisition by adult WFT at 1 and 5 DAE was higher than acquisition at 10 and 20 DAE. Subsequent transmission competence of adult cohorts was studied by vector transmission assays. All adult thrips tested that had an acquisition access period as an adult were unable to transmit the virus. These results indicate the susceptibility of adult TT and WFT to infection of midgut cells by TSWV and subsequent virus replication and confirm earlier studies that adult thrips that feed on virus-infected plants do not transmit the virus. The role of a tissue barrier in TSWV movement and infection from midgut muscle cells to the salivary glands is discussed.