Efficiency of Different Methods for dsRNA Delivery in Cotton Bollworm （Helicoverpa armigera）

RNAi trigged by dsRNA not only facilitates the development of molecular biology, but also initiates a new way for pest control by silence of fatal genes. However, one of the key limitations in pest control is lack of the convenient and efficient method for dsRNA delivery. In this study, different dsRNA delivery methods at their own optimum conditions were evaluated comparatively for their efficiency with Helicoverpa armigera as test animal. It was found that the popular one- time injection of larvae with dsRNA could reduce the pupation rate by 43.0% and enhance larva mortality by 11.7%. One- time ingestion of dsRNA did not result in any significant effect on phenotype. Continuous ingestion of in vitro synthesized dsRNA by refreshing the bait diet every day caused 40.4% decrease in successful pupation and 10.0% increase in larval mortality, which was similar as one-time injection. The most efficient method was found to be the continuous ingestion of the bacteria containing dsRNA expressed, which reduced the rate of pupation by 68.7% and enhanced the larval mortality by 34.1%. Further analysis found that dsRNA was degraded faster in midgut juice than in hemolymph. However, the cell of bacteria could protect dsRNA and delay the degradation in the midgut juice of H. armigera. These results throw light on the application of dsRNA in pest management with proper ways.     

低剂量60Co-γ辐射对棉铃虫后代的影响

 研究了低剂量γ射线对棉铃虫当代至F6代化蛹率、蛹畸形率、病毒率、成虫羽化率和成虫畸形率的影响。结果显示,辐射后棉铃虫化蛹率和成虫羽化率降低,蛹畸形率、幼虫病毒率和成虫畸形率上升,辐射剂量越大,其影响越明显。其中,对F1代的影响最大,后代有逐渐恢复的趋势。30Gy和20Gy处理的棉铃虫受辐照影响低于50Gy处理,其中30Gy处理可诱导棉铃虫对病毒敏感性增加,导致幼虫死亡率高于其他两个处理,20Gy处理对棉铃虫各生理性状影响低于其他两个处理。     

利用RNA干扰技术沉默稻纵卷叶螟几丁质合成酶A基因

稻纵卷叶螟(Cnaphalocrocis medinalis)是一种主要的水稻害虫,几丁质合成酶是昆虫体内几丁质合成通路中的一个关键酶,在昆虫生长发育中具有重要作用,是控制害虫的理想靶标。本文采用RNA干扰(RNAi)技术,以稻纵卷叶螟几丁质合成酶A基因(CmCHSA)为靶标,设计两个靶位点CmCHSA-I和CmCHSA-II,在体外分别合成其双链RNA(dsRNA),用显微注射法导入3龄幼虫体内,然后通过表型观察和荧光定量PCR(RT-q PCR)检测干扰效应。结果表明,注射两种dsRNA后纵卷叶螟对水稻的取食明显下降,生长发育阻滞,虫体变小变黑和畸形,有的出现死亡。RT-q PCR表明,注射CmCHSA-I和CmCHSA-II两种dsRNA在96 h后,CmCHSA的表达量相比对照分别下降了59%和64%。注射CmCHSA-I和CmCHSA-II两种dsRNA 7 d后,稻纵卷叶螟的死亡率分别为80%和83.33%,与对照组相比,分别增加了66.67%和70%。本研究用注射法RNAi技术成功实现了对稻纵卷叶螟几丁质合成酶A基因的沉默,为利用该技术研究昆虫基因的功能,以及通过转基因RNAi技术防治稻纵卷叶螟奠定了基础。     

Effects of inhibitors on the protease profiles and degradation of activated Cry toxins in larval midgut juices of Cnaphalocrocis medinalis(Lepidoptera: Pyralidae)

Midgut juice plays an important role in food digestion and detoxification in insects. In order to understand the potential of midgut juice of Cnaphalocrocis medinalis(Guenée) to degrade Bt proteins, the enzymatic activity of midgut juice and its degradation of Bt proteins(Cry2 A, Cry1 C, Cry1 Aa, and Cry1 Ac) were evaluated in this study through protease inhibitor treatments. The activities of total protease in midgut juices were significantly inhibited by phenylmethylsulfonyl fluoride(PMSF), tosyl-L-lysine chloromethyl ketone(TLCK), pepstatin A and leupeptin. The enzymatic activity of chymotrypsin was significantly inhibited by PMSF, and enzymatic activity of trypsin was significantly inhibited by ethylenediaminetetraacetic acid(EDTA), PMSF, tosyl phenylalanine chloromethyl ketone(TPCK), TLCK and trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane(E-64). EDTA could significantly inhibit the degradation of Cry2 A by C. medinalis. EDTA, PMSF, TPCK, and TLCK could inhibit the degradation of Cry1 C and Cry1 Aa. EDTA, PMSF, TPCK, TLCK, and E-64 could inhibit the degradation of Cry1 Ac. Our results indicated that some protease inhibitors hindered various enzymatic activities in the larval midgut of C. medinalis, which may reduce the insect's ability to degrade Bt toxins. These findings may aid the application of protease inhibitors in the management of this insect pest in the future.     

棉铃虫鞣化激素β亚基参与脂肪组织免疫的相关功能基因表达模式

【目的】研究棉铃虫鞣化激素β亚基参与脂肪组织免疫的相关功能基因表达模式。【方法】利用多肽激素体外处理与活体RNAi验证,将保存的鞣化激素β亚基同聚体重组蛋白,体外处理棉铃虫幼虫脂肪体组织,分别处理0、0.5、1和3 h,及体内注射鞣化激素β亚基dsRNA,72 h后取样,通过qRT-PCR检测棉铃虫幼虫脂肪体相关免疫基因表达模式。【结果】鞣化激素β亚基同源二聚体蛋白体外处理棉铃虫幼虫脂肪体后,免疫相关基因cecropin1、cecropin2、cecropin3、attacin、cecropin D、gloverin、defensin、moricin和lebocin在 0.5 h转录水平迅速上升,随后在1 h恢复至初始水平,在3 h与初始转录水平亦无显著变化,具有相同的变化趋势;棉铃虫体内注射鞣化激素Burs- β亚基dsRNA,72 h后,moricin转录水平无显著差异, cecropin1、cecropin2、cecropin3、attacin、cecropin D、gloverin、defensin、moricin和lebocin等8个免疫相关基因转录水平的表达均下调。【结论】鞣化激素β亚基同聚体蛋白质能够参与棉铃虫的免疫相关基因转录。     

转基因白桦抗性等级划分及其对幼虫中肠的影响

利用39个转基因白桦无性系和对照值株分别对舞毒蛾幼虫进行了抗虫性测定,结果死亡率75％上的高抗无性系占15．38％,死亡率在41％～75％的中抗无性系占46．15％,死亡率15％～40％的低抗无性系20．51％,死亡率15％以下的无性系占17．95％。通过转基因白桦叶片饲喂舞毒蛾幼虫,取食90h后中肠受到坏,取食量开始明显减少;144h取食量后受到严重影响,中肠内食物与对照相比明显减少,证明转基因白桦叶片的毒素可以影响和破坏舞毒蛾幼虫中肠,降低其取食量,从而达到抑制害虫保护林木的目的。     

异色瓢虫胁迫对棉铃虫生长发育及压力蛋白基因表达的影响

【目的】明确不同食源异色瓢虫(Harmonia axyridis)胁迫对棉铃虫(Helicoverpa armigera)生长发育和变态发育的影响,探讨棉铃虫是否能感知并分级捕食风险、能否在生长发育和变态发育上体现出权衡效应;明确长时和短时胁迫对棉铃虫压力蛋白基因表达的影响,探讨异色瓢虫胁迫能否引起棉铃虫分子水平上的生理反应。【方法】通过设置7种不同食源的异色瓢虫胁迫处理(饥饿处理、虾卵处理、棉铃虫幼虫处理、棉铃虫卵处理、蚜虫处理、蚜虫对照处理、对照处理),观察记录棉铃虫在胁迫下的生长发育(幼虫历期、蛹历期、雌雄蛾寿命、总寿命)和变态发育(蛹重、化蛹率、羽化失败率、卷翅率)指标;设置长时(1龄幼虫至3日龄成虫)和短时(15 min至6 h)异色瓢虫胁迫处理,利用实时荧光定量PCR(q RT-PCR)技术检测棉铃虫压力蛋白基因(即热激蛋白基因)Hsp70和Hsp90及热激同源蛋白基因Hsc70在胁迫后的表达水平变化。【结果】在捕食性天敌异色瓢虫的胁迫下,棉铃虫幼虫历期、蛹期、雌雄蛾寿命、总寿命均显著性缩短,蛹重和化蛹率显著性下降,成虫卷翅率显著性升高,而羽化失败率无显著性变化。在不同食源的天敌胁迫下,棉铃虫幼虫历期在异色瓢虫取食蚜虫时最短,蛹历期在瓢虫取食棉铃虫卵时最短,总寿命在瓢虫取食虾卵时最短,而雌雄蛾寿命在不同食源天敌胁迫下未表现出显著性差异;棉铃虫成虫卷翅率在瓢虫取食棉铃虫卵时最高,而蛹重、化蛹率、羽化失败率在不同食源天敌胁迫下未表现出显著性差异。压力蛋白基因Hsp70和Hsp90在短时胁迫下(Hsp70:30min至3 h;Hsp90:15 min、1.5 h、2 h、6 h)表达水平显著性上调,热激同源蛋白基因Hsc70在长时胁迫下(5龄幼虫、预蛹、雄蛹、雌蛾阶段)表达水平显著性上调。【结论】在面对捕食性天敌异色瓢虫长时胁迫作用下,棉铃虫各生长发育阶段均出现缩短的现象,即棉铃虫为躲避被捕食风险表现出了发育加速的现象,而快速的生长发育在一定程度上干扰了变态发育,导致蛹重和化蛹率下降,成虫卷翅率提高,符合权衡效应。棉铃虫对不同食源的天敌胁迫具有不同程度的敏感性,即棉铃虫对潜在的捕食风险可能存在一定的分级能力,但这种分级能力没有得到规律性体现。异色瓢虫胁迫能够引起棉铃虫分子层面的生理反应,导致压力蛋白基因的表达上调,其中压力蛋白基因Hsp70和Hsp90受到短时胁迫的反应较为明显,而热激同源蛋白基因Hsc70受到慢性胁迫刺激的反应更为显著。     

Cry1Ac抗、感棉铃虫碱性磷酸酯酶（ALP1）的表达量比较

【目的】通过比较Cry1Ac抗、感棉铃虫昆虫中肠碱性磷酸酶（ALP1）表达量的差异及抗性棉铃虫取食Cry1Ac蛋白对ALP1表达量的影响,分析ALP1表达量变化与抗性的关系,为进一步明确Bt抗性机制、制定抗性治理策略提供理论依据。【方法】利用实时荧光定量PCR技术,比较敏感棉铃虫、Cry1Ac抗性棉铃虫取食含Cry1Ac蛋白的饲料和正常饲料后ALP1表达量的差异。【结果】ALP1在棉铃虫整个发育期都表达,幼虫的表达量最高,蛹期表达量最低,在成虫体内也有较高的表达;ALP1在幼虫中肠表达量最高,其次是后肠、前肠、马氏管,表皮中的表达量最低;与敏感品系相比,对Cry1Ac具有中等水平抗性的棉铃虫ALP1表达量明显增加,尤其是取食含Cry1Ac蛋白饲料的抗性棉铃虫幼虫的ALP1的表达量显著升高,但抗性棉铃虫取食正常饲料后,2、3龄幼虫的ALP1的表达量与敏感棉铃虫差异不显著;所有的抗性品系4龄棉铃虫幼虫中肠ALP1的表达量都显著高于敏感品系,而且随着棉铃虫抗性倍数的升高,ALP1的表达量呈逐渐降低的趋势。【结论】抗性棉铃虫中肠ALP1表达量的改变可能与Cry1Ac抗性、Cry1Ac代谢有一定的关系。     

The Post-exposure Effects of Spinosad and Plant Allelochemicals on Biological Characteristics of Helicoverpa armigera (Lepidoptera:Noctuvidae)

In order to define the combined effects of spinosad and plant allelochemicals on Helicoverpa armigera(Lepidoptera:Noctuvidae), the research fed them on artificial diets with different doses of spinosad, gossypol, quercetin and tannicacid, and the changes of pupal and imaginal biological characteristics were observed and recorded. The results showed that spinosad at sublethal concentrations decreased pupation ratio and pupal weight, prolonged prepupal and pupal periods, and decreased emergence ratio, fecundity and longevity of adults. Combined with plant allelochemicals, spinosad significantly prolonged pupal period and shorten adult longevity. Other life-cycle parameters such as pupation ratio, pupal weights, emergence ratio, eggs laid per female and hatch ratio did not change significantly. The results clearly indicated that plant allelochemicals did not change spinosad efficiency on H. armigera, but the combination of spinosad and plant allelochemicals might affect pest population dynamics significantly by changing its development.     

棉铃虫磷脂酰乙醇胺结合蛋白的克隆及表达分析

【目的】克隆获得棉铃虫（Helicoverpa armigera）磷脂酰乙醇胺结合蛋白(phosphatidylethanolamine binding protein,PEBP)的cDNA序列,分析HaPEBP在棉铃虫体内的表达规律,检测2-十三烷酮处理条件下棉铃虫体内HaPEBP的变化情况,为进一步明确棉铃虫PEBP的功能和选择该基因作为调控棉铃虫种群数量的分子靶标提供依据。【方法】利用RACE技术从棉铃虫6龄幼虫的中肠组织中扩增得到HaPEBP的cDNA序列,并对其氨基酸序列和蛋白结构进行分析。将HaPEBP的ORF序列连接至pET32a载体并转化大肠杆菌BL21(DE3),IPTG诱导后检测目的蛋白的表达形式,并通过镍柱的亲和层析纯化融合蛋白。最后通过实时荧光定量PCR检测棉铃虫幼虫不同时期和6龄幼虫不同组织中HaPEBP的表达规律,以及2-十三烷酮处理棉铃虫6龄幼虫后中肠内HaPEBP的变化规律。【结果】获得的HaPEBP cDNA序列长760 bp,其中ORF为588 bp,编码195个氨基酸,蛋白质分子量和等电点分别为21.76 kD和5.93。氨基酸序列分析表明HaPEBP无信号肽、跨膜结构域和二硫键,是一个由4个α螺旋和9个β折叠片组成的胞质单体蛋白。将BL21(DE3)-pET32a-HaPEBP重组菌用1 mmol·L^-1的IPTG在37℃条件下诱导4 h,约40 kD的融合蛋白His-HaPEBP能以可溶的形式存在于重组菌中。按照同样的方法诱导1 L的重组菌,将超声处理后的上清液与Ni-NTA柱结合,进行咪唑缓冲液梯度洗涤,200 mmol·L^-1的咪唑洗涤两次后得到约40 kD的单一蛋白,与融合蛋白的大小一致。将此流出液超滤浓缩,获得183.3 ng·μl^-1的融合蛋白,能被抗His-Tag的单克隆抗体识别发生免疫反应。HaPEBP在棉铃虫幼虫的1-6龄期和预蛹期均表达,且6龄幼虫的表达量最高。该基因在6龄幼虫的脂肪体、中肠、头部和体壁中也均有表达,且脂肪体内的表达量最高。不同浓度的2-十三烷酮处理后,棉铃虫6龄幼虫中肠内HaPEBP的表达量均有所降低;低浓度(2.5和5 mg·g^-1)在6 h就能降低HaPEBP的表达量,其mRNA含量随着时间的延长逐渐增加;而高浓度(10和15 mg·g^-1)在12 h才会降低HaPEBP的表达量,其mRNA含量随着时间的延长先下降后上升。【结论】克隆分析了HaPEBP,利用大肠杆菌系统表达出可溶的融合蛋白His-HaPEBP,并通过镍柱-咪唑纯化法得到了高纯度的融合蛋白,时空表达分析显示HaPEBP在棉铃虫6龄幼虫和脂肪体中表达量最高,2-十三烷酮处理6龄幼虫后中肠内HaPEBP的表达量显著降低。     

丙磺舒在阿维菌素作用棉铃虫细胞中的增效机制研究

丙磺舒是ABCC转运蛋白抑制剂,用系列浓度阿维菌素和200μmol/L丙磺舒的食物饲喂棉铃虫,检测中肠组织中阿维菌素的含量。结果表明:与单独使用阿维菌素相比,混合使用丙磺舒后阿维菌素在中肠中的平均驻留时间(MRT)增加了13%,消除半衰期(t1/2z)增加了27%,药物峰值提高了2.8倍。细胞活力测定发现,亚致死剂量阿维菌素与400μmol/L丙磺舒混用后,棉铃虫细胞的活力降低了67.1%。表明抑制ABCC转运蛋白可以提高阿维菌素在棉铃虫中肠的驻留能力,提高药物对棉铃虫细胞的毒力。     

棉铃虫对久效磷抗性和敏感性品系的生物适合度

通过构建种群生命表，观察比较了棉铃虫[Helicoverpa armigera (Huebner)]对久效磷抗性品系和敏感性品系的一系列生长发育和繁殖特征。结果表明，两品系的适合度差异显著。相对于敏感品系，抗性品系生长发育上表现为幼虫期和蛹期延长，蛹重减轻，化蛹率和羽化率降低；繁殖上表现为交本配率、产卵量和卵孵化率降低。用种群数量趋势指数（I）来确定两个品系的相对适合度，抗性品系对敏感品系的相对适合度仅为0．14。认为这是生产上棉铃虫对有磷的抗性发展较慢的原因之一。     

Comparative Analysis of Bacillus thuringiensis Vip3Aa57 and Vip3Aa62 Insecticidal Activities

Bacillus thuringiensis (Bt) exhibits strong insecticidal activity and is harmless to non-target organisms such as human and animals. Bt becomes the most commonly used environment-friendly insecticidal microorganism. However, the insecticidal activities of different Bt strains variy significantly. Therefore, it is particularly important to compare the insecticidal activities of different strains and explore their insecticidal effector mechanisms to expand Bt insecticidal spectrum and enrich transgenic resources. Here, the insecticidal activities of Vip3Aa57 and Vip3Aa62 strains, carrying vegetative insecticidal protein-encoding genes that were inserted into the expression vector pET-21b and transformed into Escherichia coli Rosetta (DE3) strain, expressing 88 ku protein, were compared. Vip3Aa57 protein reportedly displayed body weight inhibition effect on Spodoptera exigua without affecting Heliothis armigera while Vip3Aa62 protein was known to have strong insecticidal activity against S. exigua (LC50=5.124 ng ? mg-1). A low H. armigera activity (LC50=870.1 ng ? mg-1) was observed. The paraffin sectioning results showed that Vip3Aa57 protein affected S. exigua midgut cell morphology. The laser confocal microscopic imaging results showed that Vip3Aa57 bound to receptors in the midgut without damaging the midgut tissue morphology. This study would be conducive for making full use of Bt strains in the soil to compare the insecticidal activities of different Vip insecticidal genes. It could thus provide significant help in revealing the underlying insecticidal mechanisms of Vip3Aa insecticidal genes, developing new insecticidal proteins and delaying pest resistance problems.     

Feeding effects of dsNPF interference in Ostrinia furnacalis

The corn borer is a world-wide agricultural pest. In this study, a high-efficiency RNAi method was explored to knock down the neuropeptide F(NPF) to determine if NPF regulates larval feeding in the Asian corn borer, Ostrinia furnacalis. Results showed that the expression peaks of npf mRNA in both midgut and fat body are at 28 h of early 5th instar larvae. When NPF was knocked down either by feeding dsNPF-containing artificial diet or direct dsNPF injection into the 5th instar larvae, npf expression was effectively inhibited in the midgut. Larval feeding, body weight and development time were significantly impacted. In contrast, the expression of the npf receptors npfr1 and npfr2 in fat body of 5th instar larvae was significantly increased by dsNPF injection. These results indicate that NPF can be effectively knocked down in O. furnacalis, and dsNPF by injection is an effective and fast way to silence npf expression. This study provides a critical basis for further exploration in mechanism of feeding regulation in O. furnacalis.     

菜喜和集琦虫螨克对小菜蛾生产发育、存活和生殖的影响

在室内测定了抗生素杀虫剂菜喜和集琦虫螨克对小菜蛾卵及幼早生长发育，成虫存活及生殖的影响，结果表明，这两种药剂对小菜峨卵的孵化率没有影响，但能大大降低初孵幼虫的存活率，对幼虫的生物活性，均随虫龄的增加面下降，且集琦虫螨克的活性远比菜喜高，菜喜对小菜蛾4龄幼虫化蛹率有显著降低作用，但对肾的羽化率，雌成虫寿命及产卵量却没有明显影响；而集琦虫螨克则对4龄幼虫的化蛹率、蛹羽化率及羽化后成虫的产卵量均有显著的降低作用，用菜喜和集琦虫螨克的田间推荐浓度药液饲喂小菜蛾成虫，均能显著减少雌成虫的产卵量，但对雌雄成虫的寿命影响不大。     

杆状病毒介导RNAi抑制棉铃虫细胞色素P450 CYP9A14基因的转录

采用实时荧光定量PCR,测定棉铃虫Helicoverpa armigera(Hübner)细胞色素P450 CYP9A14基因在氯氰菊酯抗性棉铃虫中肠里的表达量,结果表明抗性棉铃虫是敏感棉铃虫中肠表达量的17倍。将棉铃虫中肠P450 CYP9A14基因的一个490bp反向重复片段以双链RNA干扰(double-stranded RNA interference,dsRNAi)的方法重组到棉铃虫核型多角体病毒(helicoverpa nuclear polyhedrosis virus,HaNPV)中,结果表明以该重组病毒注射氯氰菊酯抗性棉铃虫体内后,幼虫中肠CYP9A14基因的转录水平显著下降。     

嗜线虫致病杆菌杀虫毒素对棉铃虫的中肠组织病理学研究

 嗜线虫致病杆菌HB310菌株与小卷蛾斯氏线虫HB310共生，对多种昆虫均具有较高生物活性。本实验通过盐析和非变性凝胶电泳等方法从嗜线虫致病杆菌HB310菌株的细胞内分离纯化出具胃毒活性的蛋白毒素Ⅱ。经毒素Ⅱ处理的棉铃虫幼虫，中肠组织的病理学变化类似于发光杆菌Tca毒素和Btδ-内毒素。棉铃虫4龄幼虫口服毒素Ⅱ 6 h后中肠组织开始发生变化：首先围食膜前端破碎，中肠柱状细胞伸长；随着时间推移，12 h后整个围食膜被破坏消失，中肠组织病变加重，肠壁细胞排列疏松混乱；随着毒素作用的逐渐减弱，处理72 h后棉铃虫中肠围食膜又重新出现。毒素Ⅱ离体处理棉铃虫围食膜，同样可以导致围食膜破碎。     

牛角瓜(Calotropis procera Ait.)提取物对非洲瓜瓢虫(Henosepilachna elaterii Rossi)的杀虫潜能评价

室内评价了牛角瓜(Calotropis procera Ait.)提取物对非洲瓜瓢虫(Henosepilachna elaterii Rossi)的生物活性及提取方法.结果表明,浸泡提取法的提取效率显著比电动振荡提取法高;植物的不同部分提取效率存在显著差异,以叶最高,花次之,根最低;叶提取物的生物活性高于花和根;初孵的幼虫不取食牛角瓜叶、花和根提取物处理过的南瓜叶;用提取物处理过的南瓜叶饲养4龄幼虫,幼虫不能化蛹就死亡;4龄幼虫前胸背板点滴牛角瓜叶或花提取物试验表明,点滴处理后幼虫的死亡率、随后的蛹死亡率和羽化的畸形成虫比例增加.但4龄幼虫先用牛角瓜叶、花和根提取物处理过的南瓜叶饲养2天,随后用未经提取物处理的南瓜叶饲养,则这些幼虫仍能完成幼虫和蛹的发育,而且育出的成虫寿命和生殖力不受影响.     

晋中盆地夏玉米田棉铃虫发生危害特点的研究

通过对晋中盆地夏玉米田棉铃虫发生危害的调查研究 ,结果表明 ,第 3代棉铃虫幼虫是夏玉米生长后期的重要害虫之一 ,该虫主要危害玉米的雄穗和雌穗 ,其危害高峰期在 8月中旬～ 9月上旬 ,通常年份蛀穗率可达 5 9.3%以上。     

Bt杀虫蛋白对甜菜夜蛾生长发育的影响

 实验室内研究了Bt杀虫蛋白对甜菜夜蛾生长发育的影响。结果表明：甜菜夜蛾二龄幼虫取食含Bt杀虫蛋白浓度为0.5μg/g,1.0μg/g,2.0μg/g,4.0μg/g,8.0μg/g的人工饲料后,随着饲料中杀虫蛋白浓度增加,幼虫死亡率增加、幼虫发育历期延长、蛹的发育历期缩短、蛹重变轻、化蛹率降低。甜菜夜蛾四龄幼虫取食含Bt杀虫蛋白浓度为2.0μg/g,4.0μg/g,8.0μg/g,16.0μg/g的人工饲料后,幼虫死亡率、化蛹率、蛹的羽化率均无明显差异;而幼虫发育历期延长、蛹的发育历期缩短,蛹重减轻。杀虫蛋白浓度为8.0μg/g,16.0μg/g时,幼虫发育历期明显长于对照。杀虫蛋白浓度为16.0μg/g时, 蛹的发育历期和蛹重与对照差异显著