棉铃虫与棉花相互作用研究进展

本文从昆虫对远距离寄主植物或者生境的定向 ,到达寄主植物后对取食或产卵部位的识别和选择 ,以及昆虫对寄主植物的适应及寄主植物抗虫性等 3个方面对棉铃虫与棉花的互作关系进行了综述 ,并展望了其应用前景。     

Cage experiments on Orius tantillus predation of Helicoverpa armigera

The role of Orius tantillus (Motschulsky) (Hemiptera: Anthocoridae) in the predation of H. armigera was studied on both sorghum (Sorghum bicolor L.) and pigeonpea (Cajanus cajan L.), the predominant intercrop of small farmers in Andhra Pradesh, India. Adult Orius tantillus was observed in cages where there were eggs and first-instar larvae of H. armigera on sorghum and pigeonpea. We found that O. tantillus had two diurnal peaks of activity, after sunrise and before sunset. O. tantillus was found preying on first-instar larvae and not on eggs. However, a later immunoassay of O. tantillus collected on sorghum during a peak period of H. armigera oviposition activity showed that predation of eggs did occur under certain conditions. The activity of O. tantillus on sorghum was significantly higher on the flowers than elsewhere but this was not the case on pigeonpea. Therefore O. tantillus was a significantly more efficient predator on sorghum than on pigeonpea. The results indicate that O. tantillus could be an important predator of early stages of an H. armigera infestation, particularly in sorghum.     

Feeding behaviour of Helicoverpa armigera larvae on insect-resistant transgenic cotton and non-transgenic cotton

Abstract: Feeding behaviour of Helicoverpa armigera Hübner (Lep.; Noctuidae) larvae on non‐transgenic Bacillus thuringiensis (Bt) cotton (Gossypium hirsutum L.), Zhong 30, and transgenic cowpea trypsin inhibitor (CpTI)‐Bt cotton, SGK 321, and non‐transgenic cotton, Shiyuan 321, was investigated in both choice tests and no‐choice tests. The results of choice tests suggested that neonates have the ability to detect and avoid transgenic cotton. In the choice tests of neonates with both transgenic and non‐transgenic cotton leaves, a significantly greater proportion of larvae and higher consumption were observed on non‐transgenic cotton than on the transgenic Bt or CpTI‐Bt cotton. In the choice tests with leaves of two transgenic cotton lines, the proportion of neonates on leaf discs of the two lines was not significantly different, but there was significantly higher consumption on CpTI‐Bt transgenic cotton than that on Bt transgenic cotton. In addition, significantly more neonates were found away from the leaf discs, lower consumption and higher mortality were achieved in the choice test with two transgenic cotton leaves than in the choice tests containing non‐transgenic cotton leaves. Leaves and buds were examined in choice tests of fourth instars. It appeared that fourth instars were found in equal numbers on transgenic and non‐transgenic cotton, except when larvae were exposed to leaves for 3 h. However, the total consumption on transgenic cotton was lower than that of the non‐transgenic cotton, so fourth instars may still have the capacity to detect transgenic cotton and reduce feeding on it, although they showed no preference on either transgenic or non‐transgenic cotton. More larvae were found off diet in the treatments with leaves than that of buds, and the number of injured leaf discs by per fourth instar was significantly higher than that of buds in choice tests, suggesting that leaf is a less preferred organ for H. armigera larvae, elicited more larval movements. Similarly, in no‐choice tests of fifth instars, significantly fewer feeding time and more moving time occurred on leaf than that of bud, boll and petal. When cotton line was considered, compared with non‐transgenic cotton, significantly lower feeding time and higher resting time occurred on the two transgenic cottons. Overall, H. armigera larvae have the ability to detect the transgenic Bt and CpTI‐Bt cottons or the less preferred organs and selectively feed more on the non‐transgenic cotton or the preferred organs, especially the neonates, which have a high capacity for avoiding transgenic cotton.     

用性信息素预测棉铃虫发生期的可靠性

利用棉铃虫（Ｈｅｌｉｃｏｖｅｒｉｐａ ａｍｉｇｅｒａ）性信息素诱蛾,预测下一代棉铃虫的卵高峰,幼虫孵化高峰,通过田间调查印证是准确的,它比黑光诱蛾以及田间调查方法简便,省工省时,指导化学农药防治时间准确,经济效益明显。可为棉铃虫发生期预测预报增添一种新的有效方法。     

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.     

Potentialities of methoxyfenozide for the integrated management of Helicoverpa armigera (Lepidoptera: Noctuidae) on cotton in Benin,West Africa

Intensive use of chemical insecticides against Helicoverpa armigera has led to the development of resistance to the major chemical families of insecticides. Consequently, management of H. armigera using conventional chemical insecticides is increasingly difficult. Methoxyfenozide is an agonist of the insect moulting hormone 20-hydroxyecdysone that acts faster than chitin synthesis inhibitors. The present work is aimed to assess the effectiveness of methoxyfenozide for use against H. armigera on cotton in Benin, West Africa. Laboratory tests and field experiments have been carried out. For laboratory studies, topical application of methoxyfenozide was done using L2 and L5 instar larva of H. armigera. Tested methoxyfenozide concentrations varied from 24 μl/ml to 144 μl/ml and the control is water only. For field experiments, a complete randomised block design was used with methoxyfenozide (72 μl/ml) and a control (no spraying). Berthoud Ultra Low Volume sprayer was used to spray methoxyfenozide suspension at 60-l per hectare. Two sprays were applied, 7 days apart. Laboratory tests indicated that 24 h after application of methoxyfenozide, about 100% of treated L2 and L5 larva were morbid or dead compared with 0% for the control larva. There are no significant differences between tested concentrations. Morbid larvae died within 2 days. In field experiments, cotton yield harvested on methoxyfenozide treated plants was double of that obtained on untreated plants (9375 kg and 4875 kg per hectare respectively). Thus methoxyfenozide may be used as component of Integrated Pest Control Programme on cotton in Benin, West Africa.     

转Bt棉花对蜘蛛生长发育及捕食行为的影响

室内评价了取食转Bt棉叶的棉铃虫幼虫对草间钻头蛛和八斑鞘腹蛛生长发育的影响.并通过捕食功能反应评价了取食了Bt棉叶的棉铃虫幼虫对成熟草间钻头蛛捕食行为的影响.室内饲养实验结果表明草间钻头蛛、八斑鞘腹蛛取食用转Bt棉叶处理的棉铃虫幼虫与取食普通棉叶处理的棉铃虫幼虫的发育历期、成蛛体重都没有显著差异.捕食功能反应实验结果表明草间钻头蛛对棉铃虫幼虫的捕食功能反应符合HollingⅡ型圆盘方程,两组不同猎物饲养成熟的草间钻头蛛对同种处理的棉铃虫幼虫的捕食行为没有显著差异.综合考虑：转Bt棉对蜘蛛生长发育、捕食能力没有显著的负作用.     

棉铃虫幼虫加单氧酶活性的组织分布

棉铃虫（Ｈｅｌｉｃｏｖｅｒｐａ ａｒｍｉｇｅｒａ）６龄幼虫不同组织的加单氧酶活性的测定结果显示,对－硝基苯甲醚Ｏ－脱甲基酶主要分由于外来物质的入口部位,以中肠和脂肪体的活性较高,在前肠、后肠和马氏管等组织中有相对较低的活性,而在体壁和精巢中未检测到Ｏ－ 甲基作用。体壁表现一定的艾氏剂环氧化作用,但其活性不及中肠的１０％,内源性制剂被证明并非体壁低加单氧酶活性的主要原因。不同组织生物量的差异及其动态     

不同食料植物对棉铃虫生长发育和繁殖的影响

报道了棉花、烟草、番茄和辣椒4种植物对棉铃虫Helicoverpa armigera生长发育和繁殖的影响。棉铃虫成虫喜欢在番茄上产卵,在辣椒上的着卵量最少。初孵幼虫喜选食嫩棉叶,选食辣椒嫩叶的虫数最少。4组幼虫取食嫩叶时的平均相对生长速率都有显著差异,顺序为棉叶组>烟叶组>番茄叶组>辣椒叶组。幼虫存活率以取食棉花时最高,取食番茄时最低。棉花组成虫的产卵量最高,烟草组的产卵量最低。取食棉花的棉铃虫种群增长的速度约为取食番茄时的14倍。6龄幼虫能有效利用和转化棉铃、烟草蒴果、辣椒果实,而对番茄果实的利用和转化效率较低。棉铃虫可分别以这4种植物的不同器官为食完成世代循环。其中,棉花是最适宜的寄主,辣椒和番茄是较不适宜的寄主。     

棉铃虫Polycalin基因的克隆及其表达分析

【目的】通过对棉铃虫Polycalin基因的克隆及其表达谱分析,进一步明确Polycalin基因在抗性中发挥的作用,为棉铃虫Helicoverpa armigera的抗性治理提供理论依据。【方法】通过RACE结合PCR技术克隆获得了棉铃虫Polycalin基因的全长序列,利用实时荧光定量PCR技术测定了在棉铃虫不同发育时期、幼虫肠道不同部位的Polycalin基因表达量,比较了棉铃虫取食含Cry1Ac蛋白的饲料后,Polycalin基因表达量的变化。【结果】该基因全长序列为2 955 bp,开放阅读框为2 781 bp,编码926个氨基酸(Gen Bank登录号为KP100652);预测蛋白的分子量为101.68 ku,等电点为4.57。推导的氨基酸序列中,N末端含有20个氨基酸组成的信号肽,含有8个O-糖基化位点,3个N-糖基化位点,C末端存在2个GPI结合位点。Polycalin在棉铃虫所有发育阶段都可以表达,幼虫期表达量较高,尤其在1~3龄幼虫体内表达量最高,在卵、成虫和蛹中的表达量较低。棉铃虫4龄幼虫取食含活化Cry1Ac蛋白的人工饲料后,Polycalin基因的表达受到抑制。【结论】研究结果可以为进一步揭示Polycalin基因的功能及其在Bt杀虫机制中的作用奠定基础。     

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。     

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。     

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...     

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。     

苦参碱对棉铃虫幼虫神经细胞钠通道的影响

用全细胞膜片钳技术研究了生物碱类植物杀虫剂苦参碱对棉铃虫Helicoverpa armigera幼虫离体培养中枢神经细胞钠离子通道门控过程的影响。结果表明： 苦参碱对棉铃虫幼虫神经细胞所表达的TTX （tetradotoxin, 河豚毒素)敏感钠通道具有浓度依赖性阻滞作用,1,10和100 μmol/L的苦参碱作用5 min后,分别使钠电流峰值较给药前下降（12.49±1.67）%、（18.79±2.16）%和（43.15±8.17）% (n=8, P<0.05)。苦参碱使钠电流的电流 电压关系曲线上移,但并不改变其激活电压、峰电压和电流电压关系曲线的形状。苦参碱对钠通道的阻滞作用可能是其具有某些毒理效应的离子基础。     

在转Bt基因棉压力下棉铃虫和异色瓢虫的波动性不对称

以取食转Bt基因棉的棉铃虫和取食转Bt基因棉上的蚜虫的异色瓢虫为研究对象,结合各自的适合度参数考察了波动性不对称（fluctuating asymmetry, FA）在不同种群中各对称性状间的表现情况。结果表明： 处理组棉铃虫种群的前、中、后足腿节,前翅肘横脉,前翅副室短径,后翅翅长均表现出FA;与对照种群比较,前足腿节与后翅翅长的FA值较高。 适合度参数测定结果显示,棉铃虫卵孵化率与成虫寿命显著低于对照种群,而成虫产卵量和幼虫发育历期与对照种群差异不显著。异色瓢虫取食了转Bt基因棉上的蚜虫后对其适合度各参数及各虫龄的捕食功能反应均无不良影响,其后翅翅长表现出FA,但与对照种群的FA值无显著差异。FA的大小与种群适合度间存在一定的联系,尚需做进一步的分析。     

Purification and characterization of prophenoloxidase from cotton bollworm,Helicoverpa armigera

Phenoloxidases are oxidative enzymes, which play an important role in both cell mediated and humoral immunity. Purification and biochemical characterization of prophenoloxidase from cotton bollworm, Helicoverpa armigera (Hübner) were carried out to study its biochemical properties. Prophenoloxidase consists of a single polypeptide chain with a relative molecular weight of 85 kDa as determined by SDS–PAGE, MALDI–TOF MS and LC–ESI MS. After the final step, the enzyme showed 71.7 fold of purification with a recovery of 49.2%. Purified prophenoloxidase showed high specific activity and homology with phenoloxidase subunit‐1 of Bombyx mori and the conserved regions of copper binding (B) site of phenoloxidase. Purified prophenoloxidase has pH optima of 6.8 and has high catalytic efficiency towards the dopamine as a substrate in comparison to catechol and L‐Dopa. The PO activity was strongly inhibited by phenylthiourea, thiourea, dithiothreitol and kojic acid.     

Impact of Cheiracanthium pelasgicum (Araneae: Miturgidae) and Chrysoperla carnea (Neuroptera: Chrysopidae) intraguild predation on the potential control of cotton pest Helicoverpa armigera (Lepidoptera: Noctuidae)

Antagonist interactions such as intraguild predation (IGP) or cannibalism among predatory arthropods can reduce the impact of these invertebrates on pest limitation in agroecosystems. Here, the effects of IGP between two major natural enemies of cotton pests, the cursorial spider Cheiracanthium pelasgicum (C.L. Koch) and the common green lacewing Chrysoperla carnea (Stephens), were studied under laboratory conditions. First, a feeding preference test was carried out to determine the degree of C. pelasgicum preference for lacewing larvae, using second-instar Helicoverpa armigera larvae as alternative prey. In a second bioassay, the effects of predator interactions on potential predation of H. armigera larvae were analysed using three treatment combinations (plus a control with no predator): (1) spider alone, (2) lacewing larvae alone, (3) spider + lacewing larvae. Potential predation by C. pelasgicum on lacewing eggs was also studied. C. pelasgicum showed no significant preference for either of the two species, indicating that this spider may impact negatively on the green lacewing population. Findings revealed no additive effects and an antagonist interaction between C. pelasgicum and green lacewing larvae, which adversely affected H. armigera suppression; both predators displayed lower predation rates when kept together than either predator alone. However, presence of lacewing larvae and subsequent unidirectional IGP did not affect the predation capacity of C. pelasgicum. Finally, predation rates of C. pelasgicum on lacewing eggs were very low (mean 2.35 ± 0.71 eggs, 24 h after offering) indicating that the impact of C. pelasgicum on lacewing populations may be limited.     

棉铃虫脑在控制滞育中的作用

脑摘除试验表明: 棉铃虫蛹的初期发育（化蛹后12 h 内）受脑的控制;化蛹12 h后的滞育蛹发育与脑无关。注射活性脑的匀浆液可阻止部分注定滞育的预蛹和化蛹后1～2天的蛹进入滞育状态,说明滞育的个体很有可能从预蛹期开始其脑的活性就已降低。