Sex-Related Larval Susceptibility of Diamondback Moth,Plutella xylostella （Lepidoptera：Plutellidae） to some Reduced-Risk Insecticides

Five reduced-risk insecticides were compared for their toxicities to male and female third instar larvae of diamondback moth （DBM）, Plutella xylostella L. （Lepidoptera：Plutellidae） obtained from a laboratory colony. Leaf-dip bioassays were used to assess larval mortality at different insecticides rates （ranging from 0.01 mg AI/L to 100 mg A1/L） and exposure times （24, 48 and 72 h after treatment）. Toxicity of the insecticides generally increased with rate and exposure time. At 72 h after treatment, median lethal concentrations （LCs0s） of methoxyfenozide, spinosad, novaluron, indoxacarb and Bacillus thuringiensis （Bt） against the male third instar larvae of P, xylostella were 0.0524, 0.1117, 0.9149, 1.1939 and 4.4983 mg AI/L, respectively, and were 0.1008, 0.2583, 1.0649, 0.2850 and 5.3053 mg AI/L against female third instar larvae, respectively. At 0.1% of the approximate recommended field rates, methoxyfenozide （60%-75%） and spinosad （53%-57%） were the most toxic, while Bt （30%-45% mortality） was the least toxic to male and female P. xylostella larvae. Spinosad was the fastest acting and the only insecticide that caused significant larval mortality （17%） at 24 h after exposure. Significant sexual differences were recorded in the susceptibility of P. xylostella larvae to two of the insecticides： spinosad and indoxacarb. Male larvae were two-fold more susceptible to spinosad but 10-fold more tolerant to indoxacarb than female larvae.     

Separation of Insecticidal Ingredient of Paecilomyces cicadae （Miquel） Samson

In this study, the insecticidal active substance of the fruiting body of Paecilomyces cicadae （Miquel） Samson that can kill the larvae of Plutella xylostella （Lepidoptera： Plutellidae） was isolated and purified by combined separation with biology experiments for the first time. By means of bioactive guided isolation of 40% ethanol-H2O, dichloromethane, ethyl acetate, n-butyl ethanol, macroporous adsorption resin column, high performance liquid chromatography, and sephadex A-20 column, the insecticidal active compound was purified,and a white powder compound was obtained. The compound was active against the third instar larvae ofP. xylostella in 24, 48, and 72 h at the concentration of 10.0 mg mL-1 with a corrective mortality of （95.1 ± 1.8）, （97.7 ± 1.6）, and （99.1 ± 1.7）%, respectively. The compound was strongly sensitive to the larvae. It was stable at acid environment, and was not sensitive to pepsin, and other chracteristics were studied.     

Effect of Rapid Cold Hardening on the Cold Tolerance of the Larvae of the Rice Stem Borer,Chilo suppressalis （Walker）

The effects of rapid cold hardening （RCH） on the cold tolerance of the last instar larvae of Chilo suppressalis （Walker） were evaluated for the first time. The discriminating temperature, induction, detection, duration and extent of RCH of the larvae in the laboratory were tested, and the supercooling points （SCPs） and the contents of water and lipid of the larvae after RCH treatment were determined, respectively. The results showed that the discriminating temperature of the larvae was about -21℃. Mean survival rates of the larvae which exposed to either 0 or 5℃ for 2 and 4 h before exposure to the discriminating temperature for 2 h were significantly higher than those of the control groups （P 〈 0.05）. Moreover, the highest survival rate appeared in the larvae after 0℃ for 4 h treatment. The protection against low temperature gained by RCH at 0℃ for 4 h was rapidly lost on return to 28℃. Mean survival rates of RCH larvae were significantly higher than those of non-acclimated （NACC） larvae and acclimation （ACC） larvae when they were exposed to the discriminating temperature for 2 or 4 h （P〈 0.05）. Moreover, the rates of NACC, ACC, RCH and ACC ＋ RCH larvae from 2 to 6 h to the discriminating temperature resulted in a significant decline. The values of SCPs and the contents of lipid of the larvae which exposed to either 0 or 5℃ for 2 and 4 h showed no significant difference at 0.05 level compared to those of the control groups. But the contents of water in the larvae were obviously decreased. Therefore, it could be concluded that RCH could enhance cold tolerance and affect partly physiological and biochemical components of the larvae of C. suppressalis, but the underlying mechanisms needs to be further explored.     

Study on Active Oxygen Quantum Yield, Insecticidal Activities and Stability of Diphenylthiophene

Active oxygen quantum yield, insecticidal activities and stability of diphenylthiophene were studied for the first time. The results showed that maximum diphenylthiophene absorbency variety was 0.438 after irradiation for 280 min and that of α-terthienyl (α-T) was 0.480 after irradiation for 200 min with UV. LC50 values ofdiphenylthiophene and α-T against 3rd instar larvae of Aedes albopictus were 9.18 × 10-3 and 9.69 × 10-4 μg mL-1 when treated for 24 h, respectively. LC50 values of the two chemicals against the 3rd instar larvae of Plutella xylostella were 267.87 and 222.22 μg mL-1 when treated for 24 h, respectively. The half lives of diphenylthiophene and α-T in methanol were 113.62 and 10.65 h. Difference between quantum yield of diphenylthiophene and α-T was not significant and they all possessed high toxicity to Aedes albopictus and Plutella xylostella, but diphenylthiophene was more stable than α-T. It could be concluded that diphenylthiophene has overcome the deficiency of photoactivated insecticides which degraded quickly in the environment and could not be applied on the field. Diphenylthiophene could kill the insects and accelerate the degradation rate of triazophos. The benefits of diphenylthiophene can be further exploited and applied on the field.     

Antifeedant Activity and Active Ingredients Against Plutella xylostella from Momordica charantia Leaves

With the bioguided fractionation of the ethanol extracts from the leaves of Momordica charantia, we obtained two most active compounds against the feeding of the diamondback moth, Plutella xylostella larvae. The antifeedant activity of momordicine Ⅰ and momordicine Ⅱ against the second and the third instar larvae of Plutella xylostella were tested using leaf discs of cabbage in the laboratory. The results showed that momordicin Ⅰ and momordicin Ⅱ had significant antifeedant activity on the larvae of P. xylostella, and momordicin Ⅱ was more active than momordicin Ⅰ. The concentrations for 50% antifeedant effects （AFC50） of momordicin Ⅱ against the second and the third instar larvae of P. xylostella were 76.69 and 116.24μg mL^-1, whereas that of momordicin I was 144.08 and 168.42μg mL^-1, respectively. In addition, momordicin Ⅰ and momordicin Ⅱ had significant inhibitive effect on the rate of weight gain and survival of P. xylostella larvae.     

Sub-Lethal Effects of Fenvalerate on the Development,Fecundity, and Juvenile Hormone Esterase Activity of Diamondback Moth,Plutella xylostella （L.）

The toxicities of fenvalerate （20% EC） to the 3rd instar larvae of diamondback moth （DBM）, Plutella xylostella （L.）, reared on three host plants viz., radish, oilseed rape, and cabbage were tested. The LC50 values of fenvalerate to the 3rd instar larvae of DBM varied with host plants, however, there wasn＇t any significant difference among them （P〉 0.05）. Similarly, DBM fed on three host plants had same pupal weight, pupal period, pupation rate, adult emergence rate, female ratio, and fecundity. The activity of juvenile hormone esterase （JHE, EC 3.1.1.1） in the 3rd instar larvae of DMB did not significantly vary with host plants, either. These suggested that DBM had similar fitness on the three host plant species. When fed on the host plants pretreated with fenvalerate at the concentrations equivalent to LC20, LC50 and LC50, the pupation rate, pupal weight, adult emergence rate, female ratio, fecundity, and JHE activity of the tested insects were declined as compared with insects in control treatments fed on the same host plant species. Furthermore, the pupal period of the tested insects was extended after fenvalerate treatment. The decrease in JHE activity after fenvalerate treatment in the tested insects could partly explain the changes in the mentioned growth parameters. Whether the role of fenvalerate in the inhibition of JHE activity could serve as a new way to control DBM needs further investigation.     

Effect of three insect-resistant maizes expressing Cry1Ie,Cry1Ab/Cry2Aj and Cry1Ab on the growth and development of armyworm Mythimna separata(Walker)

Three transgenic maize events(IE09 S034, Shuangkang 12–5 and C0030.3.5) produced Cry1 Ie, Cry1 Ab/Cry2 Aj and G10-EPSPS, Cry1 Ab and EPSPS, respectively, all of which target the Asian corn borer. The oriental armyworm Mythimna separata(Walker) is the secondary target. In this study, the effects of the three Bt maizes on the development and survival of armyworm were studied. The results showed that IE09 S034 had insecticidal activity against 1 st instar larvae, and the survival rate of armyworm fed with Bt maize for 10 days was 46.2%, significantly lower than that of the control. The larvae at 3 rd–6 th instar were more tolerant of the Bt toxin than the early instar larvae. However, Shuangkang 12-5 had good insecticidal activity against 1 st–5 th instar larvae. The mortality was nearly 100% when the larvae were fed with Shuangkang 12-5 before 3 rd instar, and the toxin had quick-acting efficacy. This event significantly inhibited the development of armyworm; that is, the larval duration of the 3 rd and 4 th instar larvae fed with Shuangkang 12-5 was prolonged by 4.5 and 3.0 days, respectively. The pupal weight and egg number were also significantly lower than those of the control. For C0030.3.5, it could control 1 st–5 th instar larvae effectively. The mortality rates were all over 50% if 1 st–3 rd larvae were fed with this event. The pupal weight of 4 th–6 th instar larvae fed with Bt maize were only 53.9, 56.8 and 54.6%, respectively, compared to that of the control. The number of eggs laid was significantly less than the control. The results indicate that all three transgenic maize events exhibit the potential to provide effective control of early instar larvae of armyworm, which can be commercialized in future to control lepidoptera pests such as Asian corn borer and armyworm.     

Comparative Toxicity of Selected Insecticides to Phytoplasma Transmitted Leafhopper Cicadulina bipunctata （Melichar）

Leafhopper Cicadulina bipunctata is represented the main insect as a pathogen for phytoplasma disease occurring by insect-transmitted plant viruses in date palm orchards. Therefore, it is important to investigate the potential effect of some insecticides against such insect. The adults of leafhopper C. bipunctata were collected from date palm orchards in Alhasa, Eastern province, Saudi Arabia. Three insecticides from different classes--beta-cyfluthrin （pyrethroids）, imidacloprid （neonicotinoids） and abamectin （natural compounds）--have been evaluated in vivo against adults C. bipunctata. This stage was exposed to residual film of various concentrations of each insecticide on transparent plastic cups using a Potter precision laboratory spray tower. Bioassay test showed that both beta-cyfluthrin and imidacloprid caused 100% mortality by 500 ppm at 24 h after treatment, whereas abamectin gave the same mortality by 50 ppm at the same time. Toxicity values revealed that abameetin was the most potent insecticide compared with beta-cyfluthrin and imidacloprid, where the lethal concentrations LC50 and LC95 were 24.58 ppm and 116.73 ppm at 3 h after treatment, respectively. Therefore, abamectin can be a possible candidate to be applied on date palm or ground grass by the Ministry of Agriculture after successful field experiments.     

pH influences the profiles of midgut extracts in Cnaphalocrocis medinalis(Guenée) and its degradation of activated Cry toxins

Midgut extracts play crucial roles in food digestion and detoxification. We evaluated the effect of pH on the profiles of the midgut extracts in rice leaffolder, Cnaphalocrocis medinalis and the degradation of activated Bt-toxins by the midgut extracts under different pH conditions. Total protease activity increased slightly with the increase with the simulated pH in the midgut extracts and the maximal protease activity was observed at pH 10.5, while an upward trend was observed as the pH of reaction buffer increased. Activity of chymotrypsin-like enzymes increased with pH, both in the buffer and midgut extracts, while the activity of trypsin-like enzyme was unaffected. Degradation of the activated Cry2 A by the midgut extracts enhanced as the pH increased. Cry2 A was fully degraded into smaller segments at pH 9.0–10.5. Activated Cry1 C protein at pH 9.0–10.5 was partially degraded by the midgut extracts. Activated Cry1 Aa and Cry1 Ac were partially degraded into fragments by the midgut extracts at high pH. These results will facilitate our further understanding of the interactions between C. medinalis and the Cry toxin.     

Antifeedant Activity and Active Ingredients Against Plutella xylostella from Momordica charantia Leaves

With the bioguided fractionation of the ethanol extracts from the leaves of Momordica charantia, we obtained two most active compounds against the feeding of the diamondback moth, Plutella xylostella larvae. The antifeedant activity of momordicine Ⅰ and momordicine Ⅱ against the second and the third instar larvae of Plutella xylostella were tested using leaf discs of cabbage in the laboratory. The results showed that momordicin Ⅰ and momordiein Ⅱ had significant antifeedant activity on the larvae of P. Xylostella, and momordicin Ⅱ was more active than momordicin Ⅰ. The concentrations for 50% antifeedant effects (AFC50) of momordicin Ⅱ against the second and the third instar larvae of P. Xylostella were 76.69 and 116.24 μg mL-1, whereas that of momordicin Ⅰ was 144.08 and 168.42 μg mL-1, respectively. In addition, momordicin Ⅰ and momordicin Ⅱ had significant inhibitive effect on the rate of weight gain and survival of P. Xylostella larvae.     

Effects of Droplet Distribution on Insecticide Toxicity to Asian Corn Borers （ Ostrinia fumaeafis） and Spiders （Xysticus ephippiatus）

Distribution of horizontal boom produced droplets downwards into maize canopies at flowering period and its effects on the efficacies of emamectin benzoate, lambda-cyhalothrin and chlorantraniliprole against the second generation of Asian corn borer （ACB） larvae and their toxicity to spiders were studied. When insecticides were sprayed downwards into the maize canopies, randomly filtering out droplets by upper leaves led to great variations of droplet coverage and density within the canopies. Consequently, the efficacies of lambda-cyhalothrin and emamectin benzoate against ACB larvae were decreased because of randomly filtering out droplets by upper leaves. But field investigation showed that lambda-cyhalothrin was extremely toxic to hunting spiders, Xysticus ephippiatus, and not suitable to IPM programs in regulation of the second generation of ACB. Therefore, randomly filtering out droplets by upper leaves decreased lambda-cyhalothrin＇s efficacy against ACB larvae, but did little to decrease its toxicity to X. ephippiatus. Amamectin benzoate can reduce the populations of X. ephippiatus by 58.1-61.4%, but the populations can recover at the end of the experiment. Chlorantraniliprole was relatively safe to X. ephippiatus. It only reduced the populations of X. ephippiatus by 22.3-33.0%, and the populations can totally recover 9 d after application.     

Toxicity and binding analyses of Bacillus thuringiensis toxin Vip3A in Cry1Ac-resistant and-susceptible strains of Helicoverpa armigera (Hübner)

The Bacillus thuringiensis vegetative insecticidal protein, Vip3 A, represents a new family of Bt toxin and is currently applied to commercial transgenic cotton. To determine whether the Cry1Ac-resistant Helicoverpa armigera is cross-resistant to Vip3 Aa protein, insecticidal activities, proteolytic activations and binding properties of Vip3 Aa toxin were investigated using Cry1Ac-susceptible(96S) and Cry1Ac-resistant H. armigera strain(Cry1Ac-R). The toxicity of Vip3 Aa in Cry1Ac-R slightly reduced compared with 96 S, the resistance ratio was only 1.7-fold. The digestion rate of full-length Vip3 Aa by gut juice extracts from 96 S was little faster than that from Cry1Ac-R. Surface plasmon resonance(SPR) showed there was no significant difference between the binding affinity of Vip3 Aa and BBMVs between 96 S and Cry1Ac-R strains, and there was no significant competitive binding between Vip3 Aa and Cry1 Ac in susceptible or resistant strains. So there had little cross-resistance between Vip3 Aa and Cry1 Ac,Vip3A+Cry proteins maybe the suitable pyramid strategy to control H. armigera in China in the future.     

A Fragment of Cadherin-Like Protein Enhances Bacillus thuringiensis Cry1B and Cry1C Toxicity to Spodoptera exigua（Lepidoptera：Noctuidae）

Cry toxins produced by Bacillus thuringiensis (Bt) are effective biological insecticides against certain insect species.In this study,bioassay results indicated that CrylB and Cry1C were toxic to Spodo...     

Rotate Two Different Mode of Actions of Green Chemicals to Combat Virachola livia （Klug） on Date Palm Fruit,Egypt

Alternation of two different modes of action of green chemicals： spinosad and methoxyfenozide to combat Virachola livia on dates in Dakhla Oasis, New Valley, Egypt, were done in 2010 and 2011. Based on infestation levels in fruit, the three spray programs including rotations with spinosad 24 SC at 20 mL/100 L were not significantly different. Based on minimum effective rotation doses, alternation of spinosad at 20 mL/100 L followed by methoxyfenozide 24 SC at 15 mL/100 L on a three-week interval showed 0.3% and 0.6% infestation, before harvest for 2010 and 2011, respectively, compared to check treatments which showed 33% and 70% infestations. Also, successful penetration and survival of V. livia （% live larvae） into the fruit by following a spinosad and methoxyfenozide rotation program resulted in 1.8% and 1% larvae, before harvest in 2010 and 2011, respectively, compared to fhait infestation levels of 97% and 89% in the untreated in 2010 and 2011, respectively. Fruit were penetrated by first instar larvae upon hatching, in case no chemical treatment. The last data reflect the excellent performance of this rotation. During the same period, reduction of the three rotation programs was 97.8%-100% and 99%-100% for 2010 and 2011, respectively. Both products proved to be working under dry hot temperatures in these areas.     

Comparative Analyses of Proteome Complement Between Worker Bee Larvae of High Royal Jelly Producing Bees （A. m. ligustica） and Carniolian Bees （A. m. camica）

This study is to compare the protein composition of the high royal jelly producing bee （A. m. ligustica） with that of Carniolian bee （A. m. carnica） during their worker larval developmental stage. The experiment was carried out by two- dimensional gel electrophoresis. The results showed that significant higher numbers of total proteins （283） were detected in larvae of high royal jelly producing bees （Jelly bee） than those of Carniolian bees （152） on 2-d-old larvae. Among them, 110 proteins were presented on both strains of bee larvae, whereas 173 proteins were specific to larvae of Jelly bees, and 42 proteins were exclusive to Carniolian larvae. However, on the 4th d, a significant higher number of total proteins （290） were detected in larvae of Jelly bees than those of Carniolian bees （240）, 163 proteins resolved to both bee larvae, and 127 proteins were specific to Jelly bees and 77 proteins to Camiolian bees. Until the 6th d, also a significant higher number of total proteins （236） were detected in larvae of Jelly bees than those of Carniolian bees （180）, 132 proteins were constantly expressed in two bee larvae, whereas 104 and 48 proteins are unique to Jelly bee and Carniolian bee larvae, respectively. We tentatively concluded that the metabolic rate and gene expression of Jelly bees larvae is higher than those of Carniolian bees based proteins detected as total proteins and proteins specific to each stage of two strains of bee larvae. Proteins constantly expressed on 3 stages of larval development with some significant differences between two bee strains, and proteins unique to each stage expressed differences in term of quality and quantity, indicating that larval development needed house keeping and specific proteins to regulate its growth at different development phage, but the expression mold is different between two strains of larval development.     

Effects of Tebufenozide on the Biological Characteristics of Beet Armyworm （Spodoptera exigua Hǖbner） and Its Resistance Selection

In this article, the selection of tebufenozide to beet armyworm （Spodoptera exigua Htibner） was studied by the treatments to alternative generations＇ 3rd-instar larvae with LC50 dose and to continuous generations＇ larvae with LC10 dose; the effects of tebufenozide on the biological characteristics of current and subsequent generations were examined by the treatments to 3rd-instar larvae and egg pods in different concentrations. After treatments with LC50 dose till F11, the toxicity of tebufenozide to beet armyworm had no significant change, whereas the pupation rate, pupal weight, and fecundity were reduced markedly. After treatments with LC10 dose till Fl9, the beet armyworm only developed 3.52-fold resistance, and the main biological characteristics were nearly accordant in each generation. The livability was reduced 72 h later after treatments to 3rd-instar larvae, respectively in 2.5-40 μg mL^-1, and larval duration, pupation rate, and pupal weight changed considerably with the increase in concentrations. The fecundity, larval livability, larval weight and pupal weight of subsequent generations were reduced as the dose increased over 10 μg mL^-1. The hatching rate of egg pods did not differ with that of the controls obviously after treatment in 10-300 μg mL^-1. But the larval livability, larval weight and pupal weight were reduced when eggs were exposed to 50 μg mL^-1 dose or more. The results indicated that tebufenozide had low resistance risk to the current and subsequent generations of beet armyworm even if tebufenozide had significant effects on the biological characteristics of this insect.     

Impacts of Environmental Factors on Degradation of CrylAb Insecticidal Protein in Leaf-Blade Powders of Transgenic Bt Rice

The determination of the environmental fate of Bt insecticidal protein released by Bt rice plants in paddy soils is a key issue in its ecological risk assessment. In this study, the impacts of soil water content, pH, and temperature on the degradation of CrylAb protein expressed in the leaves of Bt rice KMD2 were studied in the laboratory. Three types of paddy soils were used, i.e., blue clayey paddy soil, pale paddy soil on quaternary red soil, and marine-fluvigenic yellow loamy paddy soil. Ground powders of KMD2 leaf blades were mixed with each type of soil, and degradation dynamics of Cry lAb were measured using enzyme-linked immunosorbent assay （ELISA）. The degradation rate of CrylAb was high at the early experimental stage, but slowed down steadily at middle and later stages, which could be described by exponential equations, with the half-life period of degradation determined as 1.8-4.0 d. The soil water content, pH, and temperature could affect the degradation of CrylAb, but the effects of soil pH and temperature were relatively greater. In general, CrylAb degradations were slower under lower soil pH and temperature conditions, especially for marine-fluvigenic yellow loamy paddy soil.     

The Sublethal Effects of Chlorantraniliprole on Helicoverpa armigera (Lepidoptera: Noctuidae)

Sublethal effects of chlorantraniliprole to Helicoverpa armigera (Hübner) larvae were evaluated through exposure of third instar larvae to the insecticide incorporated into an artificial diet. When larvae were fed on the insecticide-containing diet for 7d, the LC10 , LC20 , LC40 , and LC50 values were 3.790, 7.977, 21.577, and 33.121 μg active ingredient L-1, respectively. Chlorantraniliprole at sublethal concentrations significantly reduced the larval body mass, emergence ratio, adult longevity and egg hatching rate in both the parental and offspring generations. The pupation and copulation rate in the parental generation and the pupal mass in the offspring also strongly decreased. Reproduction was seriously disturbed in both the parental and offspring groups even when only one of the partners was exposed to chlorantraniliprole at larval stages. However, at the lowest concentration of exposure (LC10 ), offspring fecundity was strongly reduced only when both partners were exposed. The net reproductive rate (R0 ), intrinsic rate of increase (rm ), and gross reproduction rate (GRR) at LC20 , LC40 concentrations were significantly lower than that of the control. Post-exposure effects also included an extended larval developmental time and increased male proportion in both generations. The doubling time (Dt) at LC20 , LC40 concentrations as well as gross reproduction rate (GRR) at LC10 concentration were also significantly increased. Chlorantraniliprole might have significant effects on H. armigera population dynamics even at sublethal concentrations on both parental and offspring generations.     

Sub-Lethal Effects of Fenvalerate on the Development,Fecundity, and Juvenile Hormone Esterase Activity of Diamondback Moth,Plutella xylostella (L.)

The toxicities of fenvalerate (20% EC) to the 3rd instar larvae of diamondback moth (DBM), Plutella xylostella (L.), reared on three host plants viz., radish, oilseed rape, and cabbage were tested. The LC50 values of fenvalerate to the 3rd instar larvae of DBM varied with host plants, however, there wasn’t any significant difference among them (P 0.05). Similarly, DBM fed on three host plants had same pupal weight, pupal period, pupation rate, adult emergence rate, female ratio, and fecundity. The activity of juvenile hormone esterase (JHE, EC 3.1.1.1) in the 3rd instar larvae of DMB did not significantly vary with host plants, either. These suggested that DBM had similar fitness on the three host plant species. When fed on the host plants pretreated with fenvalerate at the concentrations equivalent to LC20, LC50 and LC80, the pupation rate, pupal weight, adult emergence rate, female ratio, fecundity, and JHE activity of the tested insects were declined as compared with insects in control treatments fed on the same host plant species. Furthermore, the pupal period of the tested insects was extended after fenvalerate treatment. The decrease in JHE activity after fenvalerate treatment in the tested insects could partly explain the changes in the mentioned growth parameters. Whether the role of fenvalerate in the inhibition of JHE activity could serve as a new way to control DBM needs further investigation.