棉铃虫对Bt杀虫剂的抗性遗传方式

随着转基因棉花种植面积的日益增加,棉铃虫Helicoverpa armigera（Hübner)对Bt的抗性已经成为一个不容忽视的问题。发展转多价基因作物是当前缓解害虫对Bt抗性的最有效措施。本研究以经室内多年筛选的、抗性倍数达2 000多倍的Bt杀虫剂（含多种蛋白）抗性品系为材料,通过生物测定和不同的杂交试验,测定棉铃虫对Bt杀虫剂的抗性遗传方式,以期为Bt生物农药的抗性治理提供一定的依据,同时为制定棉铃虫对转多基因作物的抗性治理策略提供一定的参考。对敏感亲本和抗性亲本杂交产生的F1代的研究结果表明,杂交品系的抗性倍数分别为22.2倍和24.6倍;抗性显性度D值均小于0,分别为-0.20和-0.17,抗性为常染色体不完全隐性遗传。对4种回交后代和2种自交后代F2的研究结果表明,实际死亡率与期望死亡率差异较大,说明抗性是由单基因多个位点或多基因控制。     

棉铃虫对转Bt基因棉的抗性筛选及遗传方式的研究

通过室内筛选获得了棉铃虫Helicoverpa armigera (Hübner)对转Bt基因棉的抗性种群,并在此基础上进行了抗性遗传方式的研究。试验结果表明：经16代筛选,棉铃虫对转Bt基因棉的抗性倍数上升到43.3倍。筛选过程中,棉铃虫7日龄幼虫的体重和成虫羽化率变化明显,被选为确定筛选剂量的标准。敏感与抗性棉铃虫杂交,正交、反交的显性度都小于0、杂交过程中雌雄性比基本接近1∶1、假设抗性基因为单基因时的χ²值较低,因此,初步认为棉铃虫对转Bt基因棉的抗性是常染色体单基因控制的不完全隐性遗传。     

棉铃虫对转Bt基因棉的抗性及其治理策略研究进展棉铃虫对转Bt基因棉的抗性及其治理策略研究进展

简述了苏云金杆菌Bacillus thuringiensis（Bt）毒素的作用方式及杀虫机理,分析了Bt棉种植过程中面临的生态风险。综述了昆虫对Bt毒素的抗性机理、监测方法及治理策略方面的研究进展。棉铃虫对Bt棉的抗性可能主要与中肠上皮细胞膜上的特异性结合受体中结合位点的改变有关。在多种抗性治理策略中,庇护所策略被公认为是目前最有效的并已广泛采用的抗性治理措施。应针对Bt棉在我国的种植情况,在棉铃虫还未在田间表现出抗性以前,制定合理的抗性预防、治理措施。     

棉铃虫对Bt生物农药早期抗性及与转Bt基因棉抗虫性的关系

用饲料感染法建立了棉铃虫Helicoverpa rmigera(Hubmer)敏感品系(SUS1)对Bt生物农药的敏感毒力基线和区分剂量,1995年测定了五省六县棉铃虫初孵幼虫对Bt生物农药的敏感性,结果表明：山东阳谷、河北邯郸、河南新乡、安徽萧县及江苏丰县棉铃虫已产生早期抗性,抗性个体百分率为5%～10%,与敏感品系相比,LC₅₀值稍有增加,但斜率b值明显变小;而江苏东台棉铃虫仍属敏感。这是国内外首次诊测到棉铃虫对Bt生物农药抗性。用棉叶喂饲法测定比较了转Bt基因棉花品系对不同种群棉铃虫的抗虫性效果,结果表明：用早期抗性的阳谷和新乡棉铃虫初孵幼虫接虫5d后平均死亡率较敏感品系下降16%～29%,说明棉铃虫对Bt农药与转Bt生物基因棉花品系间存在交互抗性。还讨论了Bt农药的抗性治理对策。     

棉铃虫对Bt生物农药早期抗性及与转Bt基因棉抗虫性的关系

用饲料感染法建立了棉铃虫Helicoverpa rmigera(Hubmer)敏感品系(SUS1)对Bt生物农药的敏感毒力基线和区分剂量,1995年测定了五省六县棉铃虫初孵幼虫对Bt生物农药的敏感性,结果表明：山东阳谷、河北邯郸、河南新乡、安徽萧县及江苏丰县棉铃虫已产生早期抗性,抗性个体百分率为5%～10%,与敏感品系相比,LC₅₀值稍有增加,但斜率b值明显变小;而江苏东台棉铃虫仍属敏感。这是国内外首次诊测到棉铃虫对Bt生物农药抗性。用棉叶喂饲法测定比较了转Bt基因棉花品系对不同种群棉铃虫的抗虫性效果,结果表明：用早期抗性的阳谷和新乡棉铃虫初孵幼虫接虫5d后平均死亡率较敏感品系下降16%～29%,说明棉铃虫对Bt农药与转Bt生物基因棉花品系间存在交互抗性。还讨论了Bt农药的抗性治理对策。     

湖北棉区转Bt基因棉对棉铃虫的控制作用

2000-2001年通过田间系统调查,表明转Bt基因棉（品种为GK19）在湖北江汉平原棉区对棉铃虫抗性稳定。试验设3个处理：转Bt基因棉化防田（使用化学农药控制害虫）、转Bt基因棉自控田（依靠天敌控制害虫）及常规棉对照田（利用综合防治措施控制害虫）。从棉铃虫的第2代到第5代整个发生期内,即使在不进行化学防治的情况下,棉铃虫在Bt棉田的发生量也保持在极低的水平(最高百株虫量为12头)。室内饲养结果表明,转Bt基因棉对棉铃虫的生长发育（幼虫体重、蛹重）有较为明显的影响,使6龄幼虫体重减少25.6%,蛹重减少18.2%。棉铃虫幼虫取食转Bt基因棉组织后,发育迟缓,相对于常规棉喂养的整个发育历期延长17 d,使棉铃虫在田间的危害减少至少一个世代。另外,接虫试验表明,棉铃虫幼虫在常规棉上的取食时间是转Bt基因棉株上的6.1倍,极大地减轻了棉铃虫的危害程度。     

华北地区棉铃虫对转Bt基因抗虫棉抗性适应的模拟模型

通过对华北地区耕作制度和生态系统的了解,在充分考虑种群遗传学、生物学和人为操纵因子等三大因素的基础上,建立了一个预测棉铃虫对转Bt基因抗虫棉抗性适应的模拟模型。在华北地区典型的耕作制度下,如果所有棉田均为Bt棉,则Bt棉的预期寿命为7年;如果只有春播棉为Bt棉(约占棉田总面积的70%),则其寿命为10年。模型的灵敏度分析表明, Bt棉的使用寿命随抗性基因的显性度、初始抗性频率、Bt棉所占比例等因素的增长而迅速缩短。当Bt棉表达的杀虫蛋白量恰好全部杀死敏感基因型(G_SG_S)个体时,Bt棉的预期寿命最短。由于国外采用的“高剂量/庇护所”抗性治理策略不适用于棉铃虫及华北棉区的耕作制度,我国需要加强对其它抗性治理措施(如转双基因抗虫棉)的研究与应用。     

转Bt基因抗虫棉田棉铃虫消长律及危害特点

通过对转Ｂｔ基因抗虫棉田棉铃虫的系统６调查研究表明,（１）转Ｂｔ基因抗虫棉对棉铃虫蛾、卵无抗性表现,田间落卵量与常规棉田无明显差异,相反,二代棉虫百株累计卵量倒比常规棉处理区增加了２１６～２７４粒,增长幅度为３０～３８％,（２）转Ｂｔ基因抗虫棉对二代棉铃虫幼虫的抗性制作强,基本不用进行药剂防治,但对三代棉虫幼虫的抗性控制效果明显处降,田间３龄以上幼虫平均均２０～４０头／百株,多者达６０头以上,仍需     

转Bt基因抗虫棉田棉铃虫消长规律及危害特点

通过对转 Bt基因抗虫棉田棉铃虫的系统调查研究表明 :(1)转 Bt基因抗虫棉对棉铃虫蛾、卵无抗性表现 ,田间落卵量与常规棉田无明显差异 ,相反 ,二代棉铃虫百株累计卵量倒比常规棉处理区增加了 2 16～ 2 74粒 ,增长幅度为 30 %～ 38% ;(2 )转 Bt基因抗虫棉对二代棉铃虫幼虫的抗性及控制作用较强 ,基本不用进行药剂防治 ,但对三代棉虫幼虫的抗性控制效果明显下降 ,田间 3龄以上幼虫平均达2 0～ 4 0头 /百株 ,多者达 6 0头以上 ,仍需进行必要的药剂防治 ;(3)棉铃虫在转 Bt基因抗虫棉株上的危害特点与常规棉株有明显的差异 ,表现在幼虫和危害症状均较隐蔽 ,不易被发现。     

棉铃虫取食转Bt棉后蛹的抗寒力测定

在棉铃虫HelicoverpaarmigeraH櫣bner人工饲料中分别添加转Bt基因棉叶粉和常规棉叶粉饲喂幼虫,经滞育诱导后,发现2个处理间棉铃虫蛹的滞育率相似,2个处理间滞育蛹的自由水和结合水含量没有差异;但取食含Bt棉叶粉人工饲料的棉铃虫滞育蛹的过冷却点和结冰点显著高于对照组,蛹重及与抗寒性有关的脂肪和糖原含量均显著低于对照组。     

人工释放赤眼蜂对棉铃虫的防治作用及相关生态效应

1998～2000年在河北省南皮县棉区转Bt基因棉田及常规棉田中设置4种不同的组合处理,通过释放螟黄赤眼蜂控制棉铃虫的方法,减少化学农药的使用,达到提升和增强自然生物控制力的生态效应;并以不采取任何防治措施的常规棉田为常规棉对照田。研究表明：（1）转基因棉和常规棉棉田中自然寄生率随棉铃虫世代的增加而逐渐升高,2、3和4代棉铃虫卵被寄生率范围分别为13.3%～14.3%、26.7%～28.2%和60.8%～61.4%。（2）棉铃虫2代期,在常规棉综防田释放赤眼蜂2次,其寄生率为46.4%, 比未释放赤眼蜂的转基因棉棉田和常规棉对照田提高33.1%和32.1%; 3代期,转基因棉棉田释放赤眼蜂1次,其寄生率与常规棉综防田释放4次的效果相当,分别为73.7%和68.1%;但与转基因棉不放蜂田、常规棉化防田及对照田相比,分别提高45.5%、61. 8%和47.0%;4代期,无论放蜂与否,各处理棉田中的寄生率除化防田外（52.1%）均在60%以上。（3）常规棉化防田棉铃虫2代和3代期,分别使用农药2和3次,其自然寄生率分别为5.5%和11.9%,与对照田相比,分别降低8.8%和14.8%;与常规棉综防田相比,分别降低40.9%和56.2%;释放赤眼蜂的效果与施药时间有关,放蜂后1天内施药,寄生率仅为12.5%,施药后2天放蜂,寄生率达45.6%。（4）转基因棉棉田棉铃虫累计数和百株蕾铃被害数比常规棉综防田分别减少74.8%和73.8%,捕食性天敌增加63.0%;放蜂Bt棉田比不放蜂Bt棉田棉铃虫累计数、百株蕾铃被害数分别减少61.8%和33.3%;常规棉综防田棉铃虫累计数、百株蕾铃被害数、农药使用量比化防棉田分别减少29.7%、43.4%和60.0%,捕食性天敌数量增加63.0%。（5）转基因棉田和综防棉田的益害比（捕食性天敌/植食性害虫）分别为0.47∶1和0.30∶1, 而化防棉田为0.24∶1。上述结果表明释放赤眼蜂可弥补抗虫棉后期抗性减弱的不足,增加田间自然天敌的数量,提高对棉田害虫的自然控害功能。     

转CpTI-Bt基因棉和转Bt基因棉对棉铃虫幼虫存活、生长及营养利用的影响

2000年7月中旬和8月中旬, 分别测定了采自田间的转CpTI-Bt基因双价抗虫棉（SGK321, 以下简称CpTI-Bt棉）和转Bt基因抗虫棉（中30,以下简称Bt棉）对棉铃虫Helicoverpa armigera幼虫存活、生长的影响。结果表明：7月中旬两种转基因抗虫棉抗虫效果均较好,尤其是CpTI-Bt棉棉叶和花瓣对4龄幼虫3天内致死率为92%以上;8月中旬两种转基因棉的抗虫活性均明显降低,且Bt棉的杀虫活性显著低于CpTI-Bt棉,其幼虫死亡率与对照受体棉中16的死亡率之间无显著差异,仅显著抑制了幼虫的生长;石远321(SGK321受体品系)的花瓣具有一定的抗虫活性,可显著降低取食幼虫的体重,甚至造成部分幼虫死亡; CpTI-Bt棉中,花瓣和棉叶的抗虫性明显高于蕾和铃心。对5龄幼虫取食棉铃1日后的营养指标测定结果显示： 两种转基因抗虫棉处理的幼虫相对生长率和相对取食量均显著低于石远321,但两者之间无显著差异; CpTI-Bt棉处理的幼虫近似消化率显著低于石远321和Bt棉,但其食物利用率显著高于石远321和Bt棉。     

Cotton bollworm resistance to Bt transgenic cotton: A case analysis

Cotton bollworm (Helicoverpa armigera) is one of the most serious insect pests of cotton. Transgenic cotton expressing Cry toxins derived from a soil bacterium, Bacillus thuringiensis (Bt), has been produced to target this pest. Bt cotton has been widely planted around the world, and this has resulted in efficient control of bollworm populations with reduced use of synthetic insecticides. However, evolution of resistance by this pest threatens the continued success of Bt cotton. To date, no field populations of bollworm have evolved significant levels of resistance; however, several laboratory-selected Cry-resistant strains of H. armigera have been obtained, which suggests that bollworm has the capacity to evolve resistance to Bt. The development of resistance to Bt is of great concern, and there is a vast body of research in this area aimed at ensuring the continued success of Bt cotton. Here, we review studies on the evolution of Bt resistance in H. armigera, focusing on the biochemical and molecular basis of Bt resistance. We also discuss resistance management strategies, and monitoring programs implemented in China, Australia, and India.     

Increasing tolerance to Cry1Ac cotton from cotton bollworm, Helicoverpa armigera, was confirmed in Bt cotton farming area of China

Abstract.  1. Changes in the frequency of Cry1Ac resistance genes and shifts in tolerance of cotton bollworm, Helicoverpa armigera , to the Cry1Ac toxin were assessed using bioassays of F₁ and F₂ offspring of isofemale lines from Anci County of Hebei Province (a multiple-crop system including corn, soybean, peanut, and Bt cotton) and Xiajin County of Shandong Province (an intensive Bt cotton planting area) in Northern China during 2002–2005.
2. A conservative analysis of the overall results indicated that there was a small increase in the frequency of major, non-recessive resistance genes over time.
3. The relative average development ratings [RADR – growth rate of a line on a Bt diet in proportion to the growth rate on a non-Bt (NBT) diet] of the bollworm larvae in F₁ tests increased significantly from year to year, indicating a gradual trend towards higher tolerance to Cry1Ac in the field populations.
4. There were also significant positive correlations between RADR of the lines in the F₁ generation and the RADR of their F₂ offspring, indicating that the tolerance was genetically based.
5. Quantitative genetic simulation analysis showed that resistance of H . armigera to Bt cotton in Xiajin could evolve to a high level in 11–15 years if no effective resistance management measures are carried out.     

Antennal response of cotton bollworm (Heliocoverpa armigera) to volatiles in transgenic Bt cotton

Abstract:  Volatile compositions in transgenic Bt cotton variety 'GK-97' and its parental variety 'Simian No. 3' were analysed, and antennal response of cotton bollworm ( Heliocoverpa armigera ) to the volatiles were investigated. Volatile components in Bt cotton and regular cotton were almost same, except for absence of a sesquiterpene and a minor unknown compound in regular cotton. The results of gas chromatography-electroantennographic detection (GC-EAD) experiments showed that cotton bollworm antennae responded to seven compounds and two minor unknown compounds in Bt volatiles, among which concentrations of α -pinene and β -pinene in Bt cotton were relatively higher than those in regular cotton, and one minor unknown compound was absent in regular cotton.     

南疆三地棉铃虫田间种群对Bt棉的抗性频率监测

为了监测南疆主要Bt棉区棉铃虫田间种群对Bt棉的抗性频率,分别采集库尔勒、阿克苏、泽普三地的棉铃虫单雌系,以Bt毒蛋白作为人工饲料,采用单雌系F1代法进行棉铃虫田间种群抗性个体检测.本文从库尔勒、阿克苏、泽普三地分别筛选了57个、106个、92个棉铃虫单雌系.三地棉铃虫单雌系幼虫在正常饲料和Cry1Ac饲料上的平均发育...     

A review on biological interactions and management of the cotton bollworm,Helicoverpa armigera (Lepidoptera: Noctuidae)

Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is one of the most damaging insect pests globally, causing estimated global economic losses of over 3 billion US dollars annually. Crops most affected include cotton, tomato, soybean, grain crops such as corn and sorghum, chickpea and other pulses. Adults of this species possess strong migratory abilities (>2000 km), high fecundity and rapid reproductive rates; completing 4–6 generations per year in most cropping regions. Furthermore, the larvae are polyphagous, with a wide and diverse host range and possess the ability to enter diapause in order to survive adverse climatic conditions. At present, it is distributed across most of Oceania, Asia, Africa and southern Europe and has recently spread to South America. Various control measures have been trialled or proposed for the treatment of this pest, including synthetic insecticides, phytopesticides, microbial pesticides, macro-biocontrol agents (both parasitoids and predators) and the development of genetically modified crops (e.g. Bt cotton). Successful control necessitates the use of an integrated pest management (IPM) approach, wherein biological, chemical and physical control measures are combined for the greatest control efficacy.