亚致死浓度农药对草间钻头蛛生长发育的影响

在室内用两种方法饲喂草间钻头蛛，研究了乐果、溴氰菊酯和氰戊菊酯3种农药对其发育历期和体重的影响。结果表明：以乐果和氰戊菊酯环境染毒法饲喂草间钻头蛛，其雌蛛体重较对照显著下降。以溴氰菊酯毒饵法饲喂草间钻头蛛，其雄蛛体重较对照显著下降。环境染毒法饲喂草间钻头蛛，3种农药对雄蛛体重影响不显著。与对照相比，3种农药饲喂的若蛛在三龄历期上无显著差异，但在四龄和五龄上存在显著差异。总效应是农药延长草间钻头蛛若蛛的发育历期，阻滞个体发育，导致体重下降。     

转基因抗虫棉田棉蚜种群消长关键因素分析

为研究转基因抗虫棉田棉蚜种群动态的主要影响因素,在河北省南皮县采用罩网和开放式观测相结合的方法,以亲本常规棉为对照,对转Bt基因棉田和转Bt CpTI双价基因棉田棉蚜自然种群消长的关键因素进行了分析。与常规棉相比,单、双价抗虫棉对天敌捕食和寄生、病原物、风雨等生物和非生物因素对棉蚜种群的自然控制作用均未产生显著影响;各因素中,除自然死亡及未知因素死亡外,天敌捕食对棉蚜自然种群的控制作用最强,风雨的作用次之,天敌寄生和病原物的作用则较弱。单价抗虫棉田棉蚜的发育历期和产仔量均与亲本常规棉田无显著差异,但双价抗虫棉田棉蚜的发育历期较对照显著延长,产仔量则无显著差异。由此表明,捕食性天敌对单、双价抗虫棉田非靶标害虫棉蚜具有较好的自然控制作用,是影响棉蚜自然种群数量动态的关键因素;单、双价抗虫棉的抗蚜性存在一定差异。     

转基因棉和常规棉叶片对抗Cry1Ac近等基因系棉铃虫生长发育的影响

利用室内饲喂法,以抗Cry1Ac近等基因系棉铃虫为材料,比较转基因棉花33B和SGK321及其对照亲本DP5415和石远321对抗、感棉铃虫生长发育的影响。结果表明,抗性棉铃虫在取食常规棉叶后表现出一定的适合度代价。取食DP5415和石远321两种常规棉花后,抗性品系棉铃虫的幼虫存活率显著低于敏感品系,取食33B和SGK321两种转基因棉花的抗性棉铃虫,不仅其幼虫存活率显著高于敏感品系,而且致死中时间也比敏感品系延长。取食9天后,抗性品系在常规棉花石远321和DP5415上发育到3龄和4龄幼虫的比例显著低于敏感品系,取食33B和SGK321转基因棉花的抗性品系发育到3龄幼虫的比例均显著高于敏感品系。抗性品系在常规棉上的蛹重均显著低于敏感品系,部分取食转基因棉花的抗性品系棉铃虫可以化蛹,而敏感品系不能在转基因棉花上化蛹。     

转cry1Ah基因玉米花粉对龟纹瓢虫生长发育和成虫移动能力的影响

本文利用花粉饲喂法研究了转cry1Ah基因玉米花粉对龟纹瓢虫Propylea japonica生长发育及成虫移动能力的影响。结果显示,与混有适量蚜虫的非转基因亲本对照玉米花粉(花粉与蚜虫重量比为2:1)相比,龟纹瓢虫幼虫取食混有适量蚜虫的转cry1Ah基因玉米花粉(花粉与蚜虫重量比为2:1)后,幼虫总发育历期、蛹期和成虫寿命均无显著差异,但1、3和4龄幼虫的发育历期显著高于对照,2龄幼虫的发育历期显著低于对照;2龄和4龄幼虫的存活率与对照无显著差异,但3龄幼虫的存活率显著增加;幼虫取食Bt花粉对该虫的蛹重、羽化率和雌雄比与对照无显著差异。幼虫期取食Bt玉米花粉而成虫期取食蚜虫对成虫的日产卵量无不利影响,与对照无显著差异。幼虫取食Bt玉米花粉羽化后雄虫和雌虫的步速和翻跃时间与对照均无显著差异。     

草间钻头蛛共生菌多样性测定及沃尔巴克氏体感染对宿主发育历期和性比的影响

为了解草间钻头蛛Hylyphantes graminicola体内共生菌的组成及共生菌对宿主的影响,通过高通量测序技术确定其体内的优势共生菌,采用常规PCR法对草间钻头蛛进行优势共生菌的感染检测,并研究了优势共生菌感染对草间钻头蛛子代的发育历期和性比的影响。结果表明:经高通量测序细菌16S r DNA的V3、V4区,确定草间钻头蛛的优势共生菌为沃尔巴克氏体Wolbachia和Cardinium。167头被检测蜘蛛中,Cardinium的感染率为100.00%,沃尔巴克氏体的感染率为38.92%。感染沃尔巴克氏体的草间钻头蛛F1代雌蛛的2龄期、3龄期、4龄期和总历期均极显著低于未感染的草间钻头蛛,感染沃尔巴克氏体的草间钻头蛛F1代雌蛛1龄期显著低于未感染的F1代个体,感染和未感染沃尔巴克氏体的草间钻头蛛F1代雌蛛的卵期和5龄期均无显著差异。感染沃尔巴克氏体后,草间钻头蛛F1代雄蛛的1龄期和总历期极显著缩短,2龄期、3龄期和5龄期显著缩短,而卵期和4龄期与未感染沃尔巴克氏体的F1代雄蛛之间无显著差异。另外,沃尔巴克氏体感染极显著提高了草间钻头蛛的子代性比。     

甜菜夜蛾幼虫对常规棉和转基因棉的取食选择行为

本文研究了甜菜夜蛾Spodoptera exigua幼虫在转基因棉和常规棉上的取食行为反应,以及有/无学习经历对其取食选择的影响及选择行为对蛹重的影响。结果表明,用人工饲料饲养至3龄5、龄的甜菜夜蛾幼虫在常规棉品种33和转基因棉品种33B上、常规棉品种石远321和转基因棉品种SGK321上的取食选择行为除24h内更多的5龄幼虫选择了石远321,在其它时段,幼虫并没有表现出对转基因叶片的规避行为,甚至有些时段更多地选择在转基因叶片上取食;分别用33叶片、33B叶片饲喂至3龄的甜菜夜蛾幼虫进行33和33B的取食选择,以及分别用石远321S、GK321的叶片饲喂至3龄的甜菜夜蛾幼虫进行石远321和SGK321的取食选择均未表现出对转基因棉的规避行为。上述所有处理中,甜菜夜蛾在常规棉上化蛹的蛹重与转基因棉上化蛹的蛹重无显著差异。结果表明,甜菜夜蛾对单价转基因棉33B和双价转基因棉SGK321均未表现出行为抗性。因此,在转基因棉花的害虫治理中要特别关注甜菜夜蛾的防治。     

转cry1Ac基因抗虫棉对土壤三种酶活性的影响

为了明确转cry1Ac基因抗虫棉对土壤酶活性的影响,本研究采集转cry1Ac基因抗虫棉及其亲本棉‘SM 3’种植地的土壤,测定转cry1Ac基因抗虫棉及其亲本棉 ‘SM 3’不同生育期内根区距主根不同距离土壤脱氢酶、脲酶和磷酸酶活性的变化。结果表明,棉花根区土壤脱氢酶活性在整个生育期呈现出先升高后降低再升高的变化趋势,土壤脲酶和磷酸酶活性在整个生育期无明显的起伏变化。转基因棉‘GK 12’盛花期和枯死期根区5 cm的土壤脱氢酶活性显著高于亲本棉‘SM 3’（p<0.05）,转基因棉‘GK 12’盛花期根区15 cm和25 cm的土壤脱氢酶活性也显著高于亲本棉‘SM 3’（p<0.05）;在棉花蕾期,转基因棉‘GK 12’根区5 cm和15 cm的土壤磷酸酶活性显著高于亲本棉‘SM 3’（p<0.05）,而对根区25 cm的土壤磷酸酶活性无显著影响（p>0.05）;转基因棉‘GK 12’在整个生育期对不同距离土壤脲酶活性无显著影响（p>0.05）。上述结果初步看出,在棉花生长的一些生育时期,转cry1Ac基因抗虫棉‘GK 12’对根区土壤酶活性存在明显的影响,但其影响的程度不是很大,后期基本可得到恢复。     

寄生不同龄期草地贪夜蛾的斑痣悬茧蜂生长发育表现

草地贪夜蛾Spodoptera frugiperda(J. E. Smith),又称秋黏虫,是新入侵我国的外来物种。为探究本土天敌斑痣悬茧蜂寄生不同龄期草地贪夜蛾幼虫后的生长发育表现,本研究检测了斑痣悬茧蜂寄生2龄、3龄和4龄草地贪夜蛾幼虫后子代蜂幼虫发育历期、茧历期、羽化率、茧重、成蜂个体大小和成蜂寿命。结果表明,寄主龄期对子代蜂的幼虫发育历期、茧历期和成蜂寿命存在显著影响。与寄生4龄幼虫的子代蜂相比,寄生2龄和3龄幼虫的子代蜂幼虫发育历期分别平均延长了2.08 d(z=5.42,P0.05)和1.64 d(z=4.23,P0.05);寄生2龄幼虫的子代蜂茧历期平均延长了0.46 d(z=3.51,P0.05),但寄生4龄幼虫的子代蜂茧历期与寄生3龄幼虫的子代蜂茧历期无显著差异。4龄幼虫体内羽化的子代蜂成蜂寿命比在2龄和3龄幼虫体内羽化的子代蜂成蜂寿命分别平均延长了1.46 d(z=-2.60,P0.05)和1.68 d(z=-2.98,P0.05)。寄主龄期对子代蜂羽化率、茧重和成蜂个体大小无显著影响。随寄主龄期增大,子代蜂的发育速度加快,但寿命缩短。研究结果将为揭示斑痣悬茧蜂与寄主草地贪夜蛾的互作关系提供参考。     

Cry1Ac和Cry2Ab蛋白对大草蛉生长发育及酶活力的影响

为明确转Cry1Ac和Cry2Ab基因棉花对大草蛉的影响,运用Bt蛋白与人工饲料混合的方法,以大于转基因棉花叶片中蛋白含量20倍的剂量饲喂大草蛉初孵幼虫,初步研究了Cry1Ac和Cry2Ab对大草蛉生物学参数和消化酶、解毒酶活性的影响。结果表明：取食含Bt蛋白饲料的大草蛉幼虫的发育历期、体重、蛹重、成虫体重、羽化率等生物学参数与对照相比均没有显著差异;在大草蛉幼虫体内可以检测到含量较高的Cry1Ac和Cry2Ab蛋白,分别为974.92~1 282.39 ng/g鲜重和5 592.62~6 082.92 ng/g鲜重,而在大草蛉成虫体内检测到的Cry1Ac和Cry2Ab蛋白含量非常低,分别为0.29~0.39 ng/g鲜重和50.34~56.71 ng/g鲜重;取食含Bt蛋白的饲料对大草蛉幼虫的类胰蛋白酶、类胰凝乳蛋白酶、氨肽酶和谷胱甘肽-S-转移酶活性有一定的影响,但对大草蛉成虫影响与对照差异不显著。表明大草蛉取食含Bt蛋白的人工饲料后,虽然体内可以检测到一定含量的Bt蛋白,但对大草蛉的生长发育并没有显著的直接不利影响。     

草间小黑蛛人工饲料研究初报

草间小黑蛛Erigonidium graminicolum Sundevall是我国农田蜘蛛中种群数量最大的一种,由于它分布广,抗药力强,发育历期短,捕食范围广,很有利用前途。关于蜘蛛的人工饲料,国内外报导较少。我们自1978年以来对草间小黑蛛的人工饲料进行了研究,利用正交试验法从30多个配方中筛选出H_1和L_3两种配方。用此两种配方配制的饲料饲养草间小黑蛛,获得较好的效果。其中L_3配方已连续饲养五个世代。用H_1配方饲料饲养食虫瘤胸蛛Oedothorax insecticeps,粽管巢蛛Clubiona japonicola,叉斑巨齿蛛Enoplognatha japonica,均能取食、蜕皮、交配、产卵传代。用L_3配方饲料     

Effect of Bt cotton expressing Cry1Ac and Cry2Ab, non-Bt cotton and starvation on survival and development of Trichoplusia ni (Lepidoptera: Noctuidae)

Effects of Bollgard II cotton containing two Bacillus thuringensis var. kurstaki Berliner (Bt) toxin proteins (Cry1Ac and Cry2Ab), non-Bt cotton (DPL 491) and starvation on survival and development of cabbage looper, Trichoplusia ni (Hübner), were determined in the laboratory. Larvae of the first four larval instars died when they fed on the terminal leaves of Bt cotton plants at 50 days after planting (DAP). However, 51.3% of fifth instars that fed on 50 DAP Bt cotton leaves pupated, and 87.1% of the pupae successfully developed into adults. Of the unfed fifth instars (starved), 55.6% pupated and 88.1% of the pupae emerged. Pupae that developed from larvae fed on Bt cotton leaves and unfed were significantly smaller, being 89.7 and 73.2% of the weight of the pupae that developed from larvae fed on non-Bt cotton leaves. Leaves of 120 DAP Bt cotton were less toxic to T. ni larvae. When the first instars continuously fed on 120 DAP Bt cotton leaves, 75.9, 60.6, 56.4 and 38.4% of larvae survived to second, third, fourth and fifth instars respectively, and 20.9% pupated and 17.9% successfully became adults. However, it took the surviving first instars 37.1 days to become adults, which was 7.2 and 8.9 days longer than those fed on 50 and 120 DAP non-Bt cottons respectively. Pupae that developed from larvae that fed on 120 DAP Bt cotton leaves were only 50.9 and 52.6% of the weight of those developed from larvae that fed on 50 and 120 DAP non-Bt cotton respectively. Non-Bt cotton, both 50 and 120 DAP, did not exhibit significant effects on larval survival and development, except that the pupae in the 50 DAP non-Bt cotton treatments developed over a significantly longer time than those in the 120 DAP non-Bt cotton treatment.     

转cry1Ac玉米对亚洲玉米螟的抗性评价

目标性状的有效性评价是转Bt基因抗虫玉米研发和安全性评价研究的重要内容之一。采用酶联免疫吸附剂测定（ELISA）、室内生测和田间人工接虫鉴定研究了转cry1Ac玉米纯合群体BT-X和分离群体BT-38、BT-181、BT-43、BT-105等转化事件Cry1Ac蛋白的表达量以及对亚洲玉米螟Ostrinia furnacalis（Guenée）的室内杀虫和田间抗虫效果。转cry1Ac玉米鲜组织中Cry1Ac蛋白含量在44.07～438.00 ng/g之间,不同转化事件及不同组织之间差异显著,其中BT-X鲜花丝中Bt蛋白表达量显著低于未展开心叶。室内生测试验中除BT-43与对照没有显著差异外,取食其它转基因玉米心叶的亚洲玉米螟初孵幼虫6或7天的存活率为0～15.3%,显著低于非转基因对照;取食夏播BT-X鲜花丝6天的幼虫存活率为62.5%,显著低于Bt11及非转基因对照。田间心叶期接虫鉴定显示BT-X高抗亚洲玉米螟。说明转cry1Ac玉米BT-X、BT-181、BT-38和BT-105对亚洲玉米螟有很高的杀虫作用,BT-X田间抗虫效果良好。     

Bt棉种植时期和作物布局方式对龟纹瓢虫生长发育和繁殖的影响

采集多年连作Bt棉区、Bt棉与春玉米等作物混栽区(过渡区)、多年连作果园(非Bt区)的龟纹瓢虫幼虫,分别用Bt棉上的棉蚜、Bt棉和非Bt植物上的蚜虫、非Bt植物上的蚜虫饲养,模拟研究Bt棉不同推广种植时期和作物布局方式对龟纹瓢虫发育、繁殖生物学特性的影响。结果显示,采自不同种植区域的龟纹瓢虫,雌雄成虫的体型由小到大依次为:Bt棉区<过渡区、非Bt区;体重依次为:Bt棉区、过渡区<非Bt区;幼虫历期依次为:Bt棉区>过渡区>非Bt区;新羽化的雌雄成虫体重均依次为:Bt棉区<过渡区<非Bt区;雌成虫比例依次为:Bt棉区<过渡区、非Bt区;羽化成虫的单雌产卵量依次为:Bt棉区、非Bt区<过渡区。表明采自不同种植区域的龟纹瓢虫,体型、体重、发育和繁殖特性均有较大差异,与多年连作果园和过渡区相比,多年连作的Bt棉区中,龟纹瓢虫成虫的体型和体重降低、幼虫发育历期延长、新羽化成虫的体重减轻、雌虫比例降低、单雌产卵量下降。     

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。     

The selective feeding of cotton bollworms (Helicoverpa armigera) on transgenic and non-transgenic cotton leaves from consecutive cultivation fields

Abstract

The aim of this study was to determine whether target-insects selectively feed on transgenic or non-transgenic crops after consecutive cultivation in fields. We sampled leaves from insect-resistant single-transgenic cotton (expressing Cry1Ac gene), pyramided-transgenic (expressing Cry1Ac and CPTI genes) and conventional cotton that were consecutively cultivated for three years in a field. Cotton bollworm (Helicoverpa armigera) collected from the cotton field was reared and its next-generation larvae were fed the three types of cotton leaves in a laboratory. The leaf area damaged by the cotton bollworm larvae was not significantly different among single-transgenic, pyramided transgenic and conventional cotton leaves. There was no significant difference found in the leaf damage among the combination groups of the three cotton types. The survival rate of the bollworms was comparable among the combination groups. The results demonstrated that the cotton bollworms did not selectively feed on transgenic or non-transgenic cotton leaves after consecutive cultivation, which may be due to the resistance evolution of the target-insects and decreased Bt content in transgenic cotton leaves.     

陷阱法调查转Bt棉对棉田地面蜘蛛群落的影响

为明确转Bt棉对非靶标节肢动物棉田蜘蛛群落的影响,在棉花全生长期利用陷阱法对转Bt棉(中棉29)和传统棉田(湘杂3号)蜘蛛的群落多样性及数量动态变化进行了系统调查。结果表明:共采集棉田蜘蛛5 020头,隶属13科33种,其中转Bt棉田2 548头,隶属12科27种,传统棉田2 472头,隶属12科27种;转Bt棉田和传统棉田均以狼蛛科Lycosidae、平腹蛛科Gnaphosidae和隐石蛛科Titanoecidae为优势科,优势科的优势种也均为沟渠豹蛛Pardosa laura、蛞平腹蛛Gnaphosa kompirensis和白斑隐石蛛Nurscia albofasciata,2种棉田在地面蜘蛛群落组成上除少部分稀有物种存在差异外,优势科及优势科的优势种均无显著差异;且3种优势种蜘蛛数量、Shannon-Wiener多样性指数以及物种丰富度均无显著差异;2种棉田地面蜘蛛数量动态变化也无显著差异,均有2个高峰期,分别在苗期和铃期。表明转Bt棉对棉田地面蜘蛛群落无显著影响。     

The effects of diet on the detection of resistance to Cry1Ac toxin in Australian Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

The effects of raw or heat-denatured soybean flour in an artificial diet on the detection of Cry1Ac resistance in Helicoverpa armigera were examined. Resistant neonate larvae reared on denatured soybean flour diet showed resistance factors of 7980 and 16,901 at the LC₅₀ and LC_99.9 levels, respectively. By comparison, resistance could not be detected in neonate larvae reared on raw flour diet. Third instar larvae reared on denatured flour diet showed resistance factors of 322 and 21,190 at the LC₅₀ and LC_99.9 levels. Resistance was not detected in third instar larvae reared on raw flour diet. There was 68% survival of resistant neonate larvae on Bollgard II cotton leaf feeding assays, compared to 100% mortality in a susceptible strain. We conclude that detection of CRY1Ac resistance in H. armigera from Australia can be masked, if an artificial diet gives chronic exposure to potent, protease inhibitors present in raw soy flour.