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1.
梁玉键  张涛  李草  郅军锐 《昆虫学报》2021,64(12):1417-1426
【目的】本研究旨在通过克隆草地贪夜蛾Spodoptera frugiperda的海藻糖合成酶(trehalose-6-phosphate synthase)基因SfTPS,分析其在草地贪夜蛾不同发育阶段、不同组织中的表达水平及不同温度胁迫时5龄幼虫中的相对表达量,为进一步探究TPS在草地贪夜蛾生长发育及抗逆应激反应中的功能奠定基础。【方法】运用RT-PCR技术克隆草地贪夜蛾SfTPS的全长编码区,并进行生物信息学分析。运用RT-qPCR技术检测SfTPS在草地贪夜蛾不同发育阶段(卵、1-6龄幼虫、蛹和成虫)、5龄幼虫不同组织(体壁、中肠和脂肪体)中和经短期(2, 4和8 h)高(35℃)低温(10℃)胁迫后5龄幼虫中的表达变化。【结果】克隆获得2 571 bp的草地贪夜蛾TPS cDNA序列,命名为SfTPS(GenBank登录号: MT920672),全长开放阅读框(ORF)长2 481 bp,编码的826个氨基酸具有TPS和TPP两个保守结构域。同源比对和系统进化分析表明,昆虫TPS蛋白具有较高的保守性,SfTPS与斜纹夜蛾S. litura的TPS亲缘关系最近,序列一致性达到99.15%。SfTPS中α-螺旋、β-折叠和无规则卷曲占比分别为38.14%, 12.23%和48.55%;SfTPS的三级结构为同源二聚体。RT-qPCR结果表明,SfTPS在草地贪夜蛾卵期和1-5龄幼虫期低表达,在6龄幼虫期、蛹期和成虫期高表达,且草地贪夜蛾变态前后SfTPS的表达量变化较大。组织分布结果显示,SfTPS在5龄幼虫脂肪体中表达量最高。草地贪夜蛾5龄幼虫经2~8 h低温(10℃)和高温(35℃)胁迫后,SfTPS的相对表达量显著高于对照(25℃),分别为对照的4.43~9.34和2.50~6.03倍。【结论】SfTPS基因在草地贪夜蛾生长发育过程及抵御高低温度胁迫中可能具有重要作用。  相似文献   
2.
【目的】本研究旨在明确甜菜夜蛾Spodoptera exigua半胱天冬酶(caspase)在细胞凋亡诱导剂诱导和病原微生物胁迫下的表达模式,为丰富鳞翅目昆虫细胞凋亡机制研究奠定基础。【方法】利用RT-PCR技术从甜菜夜蛾3龄幼虫体内扩增两个半胱天冬酶基因(SeCasp-3和SeCasp-4)编码区的全长;利用qPCR技术分别检测两个半胱天冬酶基因在细胞凋亡诱导剂过氧化氢(hydrogen peroxide, H2O2)(100 μmol/L)、放线菌素D(actinomycin D, ActD)(10 μg/mL)和地塞米松(dexamethasone, DEX)(50 μg/mL)诱导后甜菜夜蛾脂肪体细胞中及病原菌苏云金芽孢杆菌Bacillus thuringiensis kurstaki (Btk)(108个细菌/mL)、大肠杆菌TG1菌株Escherichia coli TG1 (E. coli TG1)(108个细菌/mL)、烟芽夜蛾囊泡病毒3h株(Heliothis virescens ascovirus 3h, HvAV-3h)(1.16×1011个基因组拷贝/mL)和苜蓿银纹夜蛾核型多角体病毒(Autographa californica multiple nucleopolyhedrovirus, AcMNPV)(5 000 OBs/μL)感染后甜菜夜蛾3龄幼虫体内的表达模式。【结果】SeCasp-3(GenBank登录号: MW183334)的编码区长942 bp,共编码313个氨基酸;SeCasp-4(GenBank登录号: MW183335)的编码区长843 bp,共编码280个氨基酸。SeCasp-3和SeCasp-4的假定蛋白序列与家蚕Bombyx mori Dronc的氨基酸序列一致性分别为45.54%和58.46%,且SeCasp-3和SeCasp-4之间具有较高的同源性。SeCasp-3和SeCasp-4在不同化学物质诱导下的甜菜夜蛾脂肪体细胞中的表达模式存在明显差异,在100 μmol/L H2O2和10 μg/mL ActD处理后24和48 h,脂肪体细胞中SeCasp-3和SeCasp-4的相对表达量均显著升高;50 μg/mL DEX处理后24和48 h,SeCasp-3的相对表达量未见显著变化,但SeCasp-4的相对表达量升高了数千倍。在不同病原微生物感染后的甜菜夜蛾3龄幼虫体内,SeCasp-3和SeCasp-4的表达模式基本相同。一般线性模型的分析结果表明,Btk和E. coli TG1的感染不造成SeCasp-3和SeCasp-4相对表达量的显著变化,而HvAV-3h和AcMNPV的感染则显著抑制了这两个基因的表达。【结论】本研究鉴定了两个甜菜夜蛾半胱天冬酶基因,并分析了其对细胞凋亡诱导剂和病原微生物感染的表达响应,为进一步探究半胱天冬酶功能和昆虫细胞凋亡过程提供了重要的理论基础。  相似文献   
3.
4.
The efficacy of different concentrations of the commercial neem-based insecticide, Nimbicidine® and the ecdysone agonist compound Methoxyfenozide (RH-2485), was evaluated against larvae of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae). RH-2485 and Nimbicidine® significantly (p?<?0.001) reduced adult longevity by 2.7 and 1.9 d at the higher concentrations tested, respectively, but no significant differences were observed at low concentration. The tested compounds strongly affected the reproduction of S. littoralis by producing a high percentage of sterility. Fecundity and fertility were significantly affected by both insecticides. No egg laying has been recorded with the higher concentration 0.5% of Nimbicidine® and 0.1?ppm of RH-2485, while no significant (p?>?0.05) difference was noticed on total number of eggs laid by the female when lower concentration 0.0001?ppm of RH-2485 was applied in the same stage as compared to the control. In the second part of this study, the invasion rate of Steinernema feltiae was affected by the addition of both Nimbicidine and RH-2485 to the diet of experimental host. At higher concentration of both compounds, the invasion rate was decreased despite the infection rate, while the percentage of invading nematodes increase to 56.7% with the combination treatment indicating a considerable improvement in the efficacy of S. feltiae nematode applied in combination with the lower concentration of Nimbicidine® over that of nematode alone. On the other hand, the invasion of S. feltiae to the insects that were fed on the diet with the addition of ecdyson compound was strongly decreased with increasing concentration of RH-2485 in host diet.  相似文献   
5.
Laboratory experiments were conducted to assess the efficiency of the new strain CCM 8367 of Isaria fumosorosea on different stages of the Egyptian cotton leafworm, Spodoptera littoralis (Boisd.), including soil treatments with pre-pupal and pupal stages. Treatments of larval instars showed a high susceptibility with 3rd, 5th and last instars to suspension of this fungus with concentration 5?×?107 spores/ml. Larval mortality was over 90%. There were no significant differences (P?=?0.7929, F?=?0.2346) between instar treatments. The commercial fungus, PreFeRal® strain Apopka 97 of I. fumosorosea, which was used in comparison with this new strain caused mortality rates of between 63.33 and 90%. Statistically, differences between the effects of CCM 8367 strain and Apopka 97 were highly significant on the last instar (P?=?0.0064, F?=?6.479) and extremely significant on the 3rd instar (P?<?0.0001, F?=?13.29). No significant differences were recorded between the two strains on the 5th instar (P?=?0.0597, F?=?3.233). Fungal treatments with the late stage insects (end of the last instar or pupal stage) in soil yielded interesting results: the mortality rate on end of final instar larvae was 16.66% when treated with Apopka 97 and 83.33% when treated with CCM 8367. Soil containing pupae of S. littoralis that were inoculated with CCM8367 resulted in a high number of malformed adults, and the mortality rate was 64.52% (32.27% of malformed adults died and 32.25% of pupae fully infected by fungus). Only 3.23% of samples produced morphologically normal adults in this test. The results conclude that the strain of I. fumosorosea CCM 8367 has strong insecticidal effects on S. littoralis and has the potential to be implemented as a novel biocontrol agent.  相似文献   
6.
Egg masses laid by Spodoptera littoralis mated female moths were extracted by petroleum ether (PE), ethanol (E) and Ringer's solution (RS). Egg-wash extracts were evaporated and the weights of crude materials were obtained. Different aqueous concentrations were made. The amount of extracted material increased as the weight of eggs used increased and vice versa. Coating Nerium oleander leaves with aqueous egg-wash extracts prepared from S. littoralis egg-masses deterred the mated conspecific female moths from ovipositing their eggs on treated leaves, as well as causing a decrease in the total number of deposited eggs per female during the moth's life span. The highest deterrent effect on conspecific female moths to oviposit their eggs was obtained after treatment of N. oleander leaves with PE or E egg-wash extract. The deterrent effects of the tested egg-wash extracts was concentration dependant; an increase in the concentration of any extract caused an obvious decrease in the number of deposited egg-masses and the total number of laid eggs on the treated N. oleander leaves.  相似文献   
7.
The biological activities of two species of bacteria isolated from soil of cotton fields identified as Bacillus subtilis strain NRC313 (BS NRC313) and Bacillus thuringiensis strain NRC335 (BT NRC335) were evaluated against the third larval instar of the cotton leafworm, Spodoptera littoralis (Boisd.). The different entomopathogenic bacteria of BS NRC313and BT NRC335 contained 10 × 108 cell/ml, and caused mortality of 100 and 97.3% for the above mentioned strains, respectively. Concentrations of 2.5 × 108 to 10 × 108 cell/ml of strains BS NRC313 and BT NRC335 were applied to the larvae: LC50 were 3.3 × 108 and 3.9 × 108 cell/ml respectively. The influence of exposure to toxin concentrations manifested in terms of decreasing the adult emergence and prolongation of the generation period. The percentage of larvae that survived and succeeded to pupate increased by decreasing the concentration. The longevity of adult emergence that resulted from larvae treated with Bacillus subtilis were 6.0 ± 0.51 and 9.0 ± 0.63 days at 5 × 108 and 2.5 × 108 cell/ml, respectively compared with 9.8 ± 0.47 in control. The results indicated that Bacillus subtilis was more potent than Bacillus thuringiensis. Field applications of B. thuringiensis, B. subtilis and Reldan achieved 55.6, 67.4 and 89.4% reduction of the cotton leafworm larvae Spodoptera littoralis in clover plants under field conditions.  相似文献   
8.
为探讨淡足侧沟茧蜂Microplitis pallidipes Szepligeti调控寄主的生理机制,测定了淡足侧沟茧蜂寄生后,甜菜夜蛾Spodoptera exigua(Hübner)幼虫血淋巴总糖、蛋白质及虫体脂类含量的变化。研究结果表明,寄生后连续5 d的观察时间内,甜菜夜蛾幼虫血淋巴总糖含量从寄生后第1天开始就高于未寄生寄主幼虫,且在寄生后第2至5天达到显著水平;除寄生后第3天外,被寄生寄主幼虫血淋巴总蛋白质含量始终低于未寄生幼虫,且在寄生后第1、4、5天达到显著水平;甜菜夜蛾被寄生后,虫体脂质含量始终高于未寄生幼虫,且从寄生后第2天开始达到显著水平。本研究揭示,淡足侧沟茧蜂寄生刺激了寄主甜菜夜蛾幼虫血淋巴总糖合成,但抑制了其蛋白质合成,同时也刺激了甜菜夜蛾虫体脂质合成。  相似文献   
9.
Series of experiments were undertaken to investigate whether application of Precocene II (PII) to parasitized S. littoralis larvae can influence their THCs and DHCs. PII was applied in single dose treatments (70μg/5 μl/dose). Haemolymph samples were taken at daily intervals from day 3 to day 8 post‐treatments. Two distinct alterations were noted; first, PII‐treatments caused significant decrease in THCs. Once the parasitoid larvae emerge from either treated or untreated hosts, a sharp decline in THCs was observed. The second effect was changes in DHCs of S. littoralis larvae. In controls, haemocyte types could be arranged in descending order due to their relative occurrence into plasmatocytes (PLs), prohaemocytes (PRs), spherule cells (SPs), granular cells (GRs) and oenocytoids (OEs). This order changed in treated hosts to PLs, GRs, PRs, SPs and OEs, i.e., GRs that were less prevalent in the haemolymph of control larvae increased significantly in PII‐treated hosts. In the latters, sharp rise in GRs levels followed parasitoid emergence. OEs cells were scarce in both treated and untreated hosts. No significant difference was observed between the overall means of PLs in both types of host larvae. But GRs‐PRs was markedly altered in treated larvae.  相似文献   
10.
The biochemical effects of the chlorpyrifos organophosphate insecticide and camphor plant oil in addition to their combination were studied and compared against the fourth larval instars of the cotton leaf worm Spodoptera littoralis under laboratory condition. Biochemical analysis showed that the total protein content of the cotton leaf worm larval instars was decreased at 31, 26 and 13.5% by using a camphor extract, chlorpyrifos and combination, respectively. In addition, the activity of acid phosphatase, α-esterase was significantly declined. Biochemical analysis also showed that the alkaline phosphatase activity was increased comparing with control in another side. Acetylcholinesterase enzyme amount and activity were increased with the treatment of camphor extract followed by chlorpyrifos, even though the mixture of them nearly showed the same amount in control trail. In contrast, chitinase enzyme showed a negative effect of both camphor and chlorpyrifos with nearly about the same lowering in the amount and activity of chitinase, while the mixture of them revealed a high positive increasing in the amount and activity of chitinase. On the opposite trend, phenoloxidase enzyme of treated larvae increased by action of chlorpyrifos, followed by camphor, while the mixture of them showed a negative decrease when comparing with the control.  相似文献   
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