冬虫夏草寄主蝠蛾天然饲料珠芽蓼对小菜蛾、斜纹夜蛾生长发育的影响

珠芽蓼Polygonum viviparum L.是冬虫夏草寄主蝠蛾的天然饲料,具有较高的药用、营养与饲用价值。本文分别于小菜蛾Plutella xylostella(L.)与斜纹夜蛾Spodoptera litura Fabricius的人工饲料中添加5%、15%、25%的新鲜珠芽蓼根粉饲喂小菜蛾与斜纹夜蛾,测定了新鲜珠芽蓼根对小菜蛾与斜纹夜蛾生长发育与存活的影响。结果显示,珠芽蓼根对小菜蛾与斜纹夜蛾的1-4龄幼虫具有很强的生长抑制活性及毒杀活性,对斜纹夜蛾5龄幼虫具有明显的毒杀活性。     

植物气味化合物与斜纹夜蛾性信息素的协同作用

为提高现有性信息素对雄蛾的引诱活性, 本研究通过大量的田间试验探索植物气味化合物与斜纹夜蛾Spodoptera litura性信息素（顺9, 反11-十四碳二烯乙酸酯∶顺9, 反12-十四碳二烯乙酸酯=10∶1）的协同作用机制。从斜纹夜蛾寄主植物和花的气味化合物中, 选择9种有代表性的化合物, 并以一定剂量分别加入到斜纹夜蛾性信息素诱芯中, 在田间测试对雄蛾的引诱活性。结果表明: 在测试的9种植源性化合物中, 发现一定剂量(每个诱芯加入0.4 mg)的苯乙醛(PAA), 具有显著提高斜纹夜蛾性信息素的引诱作用, 而高剂量的苯乙醛则强烈抑制性信息素的引诱活性; 此外, 其他各种浓度的测试化合物或混合物对性信息素则没有统计上显著的增效作用。不同剂量的苯乙醛单个化合物及各种植物气味化合物组成的混合物对斜纹夜蛾也有微弱的引诱作用。苯乙醛必须要与性信息素的完整组分(以10∶1比例混合的顺9, 反11-十四碳二烯乙酸酯和顺9反, 12-十四碳二烯乙酸酯)混合才能起作用, 缺少顺9, 反12-十四碳二烯乙酸酯则没有引诱活性。本研究证明, 苯乙醛作为理想的性信息素诱芯增效剂, 可应用于建立更理想的斜纹夜蛾性信息素诱杀技术, 对性诱害虫防治和测报具有应用价值。     

Bacterial compounds,as biocontrol agent against early blight (Alternaria solani) and tobacco cut worm (Spodoptera litura Fab.) of tomato (Lycopersicon esculentum Mill.)

Abstract

Tomato, the important vegetable crop faces yield loses by foliar disease, Early Blight caused by Alternaria solani and infestation by tobacco cut worm, Spodoptera litura. Synthetic pesticides used for disease and pest control resulted in environmental pollution as well as development of resistance. By way of a need to develop a new biopesticide, bacteria were tested for their anti-fungal and insecticidal activity. Volatile compounds and peptides from the bacteria, Bacillus subtilis, Staphylococcus aureus and Pseudomonas aeruginosa were able to inhibit the hyphal growth and melanin production of A. solani. S. aureus showed the highest inhibitory effect as well as reduced the disease severity. B. subtilis exhibited the highest mortality rate of 87% and 83% against I and II instar of S. litura, respectively. The bacteria offered effective biocontrol at 35?°C. Bioactive substances from the bacteria can be used as potential biocontrol agents against the pest and disease of tomato.     

Pathogenicity of entomopathogenic fungi to eggs and larvae of Spodoptera litura,the common cutworm

The eggs and second instar larvae of Spodoptera litura were treated with different concentrations of conidial suspensions of six isolates of fungi belonging to five species, Metarhizium anisopliae var. anisopliae (Metschnikov) Sorokin (ARSEF 7487), Lecanicillium muscarium (Petch) Zare & W Gams (ARSEF 7037 and ARSEF 6118), Cordyceps cardinalis (ARSEF 7193), Fusarium lateritium Nees (ARSEF 8291/MTCC 9050) and Aspergillus sp. (ARSEF 8519). In bioassay with unscaled eggs, M. anisopliae, C. cardinalis, F. lateritium and Aspergillus sp. resulted in 100% mortality above 10⁶ conidia/mL. However, with scaled egg masses, the highest hatching rate (56%) was observed with L. muscarium (ARSEF 6118) whereas the lowest hatchings were observed in the case of M. anisopliae followed by L. muscarium (ARSEF 7037), Aspergillus sp., F. lateritium, and C. cardinalis. The larvae were also found susceptible to all isolates in a dose dependent manner. Three promising isolates against larvae, viz. M. anisopliae, F. lateritium and L. muscarium (ARSEF 7037) resulted in average percent mortalities of 88, 89 and 77%, respectively at ≈10⁸ conidia/mL. When both larvae and the leaves (provided as food) were treated with ≈10⁸ conidia/mL, mortality further increased for these isolates. Based on the mortality data and the LC₅₀ values, we suggest that M. anisopliae, F. lateritium and L. muscarium (ARSEF 7037) could be developed as potential biocontrol agents against rice cutworm in IPM programs.     

Laboratory evaluation of flight activity of the common cutworm, Spodoptera litura (Lepidoptera: Noctuidae)

Abstract  The flight activity of Spodoptera litura in tethered conditions is evaluated using a computer-mediated flight-mill in the laboratory. The 3–4-day-old moths fly longer and farther than those of other ages. Male and female moths exhibit similar flight activity. Mating status does not influence the flight duration and distance of 2-day-old females. However, these two flight parameters with a 6-day-old mated female is significantly lower than that of unmated ones. The optimum temperature for flight ranged from 16–24°C, whereas the optimum RH ranged from 60%–100%. During 72-h period, the total flight duration and distance of 1-day-old male and female moths were 19.6 h (± 5.8) and 83.3 km (± 28.4), and 24.0 h (± 7.0) and 105.4 km (± 37.4), respectively. These results indicate that S. litura has a great potential to undertake long-distance migratory flights.     

Effect of plant secondary metabolites on common cutworm,Spodoptera litura (Lepidoptera: Noctuidae)

The effect of various flavonoids, lectins and phenyl β‐_D‐glucoside on larval survival, weights and the activities of digestive (total serine protease and trypsin) and detoxifying (esterase and glutathione‐S‐transferase) enzymes of Spodoptera litura larvae at 7 days after treatment was studied through diet incorporation assay. Flavonoids (rutin, chlorogenic acid, quinic acid, caffeic acid, naringenin, quercitin, kaempferol, myricetin, catechin, and ferulic acid) were incorporated in artificial diet at 100, 500 and 1000 ppm, lectins: groundnut leaf lectin (GLL), concavalin A (ConA) and phenyl β‐_D‐glucoside at 1, 2 and 5 μg/mL. Flavonoids such as rutin, quercitin and kaempferol at 1000 ppm were more toxic to S. litura larvae than quinic acid, caffeic acid, naringenin, myricetin, catechin, and ferulic acid. Larval growth and development were significantly reduced in S. litura larvae fed on a diet with GLL and ConA at 5 μg/mL compared to the larvae fed at 2 and 1 μg/mL concentrations. The larvae fed on flavonoid‐treated diets showed significant reduction in serine protease, trypsin and esterase activities. The flavonoids such as rutin, chlorogenic acid, quinic acid, naringenin, quercitin, kaempferol and myricetin, and lectins, GLL and ConA can be utilized in insect control programs.     

Influence of post-treatment temperature, solvents and emulsifiers on the toxicity of cypermethrin to larvae of Spodoptera litura (Fab.)

Larvae of the tobacco caterpillar, Spodoptera litura (Fab.) were topically bioassayed with cypermethrin emulsion formulation and proprietary EC in combination with different solvents and emulsifiers for their toxicity at 16 ± 2°C and 27 ± 1°C. Among different solvents, the descending order of efficacy (with relative toxicity in parenthesis) against 9 ± 1 day old larvae of S. litura (Fab.) at 16 ± 2°C than 27 ± 1°C was: xylene (7.41), aromex (5.54), cyclohexane (4.93), C-IX (3.93), benzene (2.40) and toluene (2.26). With emulsifier, the order was: Triton X-100 (7.41), SN₄S (4.60), Teepol (2.90), Tween-80 (1.49), Swascofix DP-50 (1.28), SN₄R (1.10) and S₅₂B (1.07). However, the proprietary cypermethrin EC was 2.31 times more toxic at 16 ± 2°C than at 27 ± 1°C.     

Expression of the cry1EC gene in castor (Ricinus communis L.) confers field resistance to tobacco caterpillar (Spodoptera litura Fabr) and castor semilooper (Achoea janata L.)

Castor (cv. DCS-9) has been transformed through Agrobacterium-mediated and particle gun bombardment methods using appropriate vectors containing the Bt chimeric gene cry1EC driven by enhanced 35S promoter. About 81 and 12 putative transformants were regenerated following selection on hygromycin and kanamycin, respectively. Southern analysis of DNA extracted from T₀ plants confirmed integration of the introduced gene in castor genome. The integration and inheritance of the introduced genes was demonstrated up to T₄ generation by PCR and Southern analysis. Southern analysis of two events having single and two copies showed the same pattern of integration in the subsequent generations. Insect feeding experiments conducted in the laboratory by releasing neonate larvae of castor semilooper and S. litura on leaf tissues excised from transgenic and control plants showed varying degrees of larval mortality and slow growth in larvae fed on transgenic leaf tissue. Field bioassays against Spodoptera litura and castor semilooper conducted for eight events in T₁–T₄ generations under net confinement were more informative and events conferring resistance to the two major defoliators were identified.     

Accessory gland proteome of male tobacco cutworm,Spodoptera litura (F.) - An approach to identify proteins influencing reproductive physiology and behaviour

Male-derived accessory glands (MAGs) in insects typically transfer a mixture of proteins and other molecules during mating that have profound effect on female physiology and reproductive behaviour. In the present study, we sought to identify MAG proteins of the serious agricultural pest Spodoptera litura as they have been extensively studied in Drosophila and other insects. A total of 566 distinct proteins across gels of virgin and mated males, as well as 91 differentially expressed proteins in mated contrast to virgin males of S. litura were detected by 2D gel electrophoresis. Here, we successfully identified 20 MAG proteins that appeared in virgin males and depleted in mated males by mass spectrometry, thus predicting the transfer of proteins from males to females during mating. We analyzed the proteins for homology to other proteins with known functions available in Swiss- Prot through Mascot search engine. The proteins were categorized into six diverse functional groups: Metabolism, protein folding, antioxidation, olfaction, DNA binding/replication and cellular activity/response. Most of the MAG proteins fall into the same functional class as in other organisms, although some appear to rapidly evolve. The probable roles of the identified proteins associated with physiology and reproduction have been discussed. Thus, in this study we establish a strong precedent for future research on the potential of MAG proteins to interfere with the pest's own mechanisms resulting in its failure to reproduce successfully.     

Chemical profiling of polyphenols in extracts from bark of Acacia nilotica (Linn.) and their efficacy against Spodoptera litura (Fab.)

Six different crude extracts from bark powder of an important medicinal plant, Acacia nilotica (L.) were investigated for their toxicity against Spodoptera litura (Fab.). Total phenolic content in six crude extracts was measured. Ultra-High Performance Liquid Chromatography (UHPLC) was also performed for qualitative analysis of six crude extracts which revealed the presence of 11 phenolic compounds. Bioassays conducted with all crude extracts revealed maximum adverse effect with acetone extract. Duration of larval period and total development period was found to be prolonged. Various aberrations were observed in pupae and adults in the form of wrinkled pupae, blackened pupae, bulged out head part of pupae, partially emerged adults and adults with crumpled wings. All nutritional indices viz. RGR, RCR, ECI, ECD and AD decreased with treatment. The findings indicated that A. nilotica had considerable potential in the management of this serious insect pest.     

黄皮种子提取物对斜纹夜蛾幼虫的杀虫活性及有效成分鉴定

【目的】测定黄皮Clausena lansium种子提取物对斜纹夜蛾Spodoptera litura幼虫的拒食活性,对有效成分进行分离、鉴定,并进一步测定其拒食活性、生长发育抑制活性及细胞毒性,为筛选斜纹夜蛾生态防治植物源杀虫剂提供依据。【方法】采用叶蝶法测定黄皮种子甲醇提取物及其石油醚、乙酸乙酯、正丁醇和水4种溶剂萃取物对斜纹夜蛾3龄幼虫的非选择性拒食活性和生长发育抑制活性;用硅胶柱层析进行石油醚萃取物和乙酸乙酯萃取物拒食活性成分分离;用核磁共振谱(NMR)进行拒食有效成分结构鉴定;用MTT法测定拒食活性成分对SL细胞毒性。【结果】黄皮种子甲醇提取物对斜纹夜蛾3龄幼虫具有较强的拒食活性,活性成分主要分布于石油醚相和乙酸乙酯相中;石油醚相和乙酸乙酯相经硅胶柱层析分离得10个馏分,以黄色晶体馏分2的拒食活性最强;经分离纯化及核磁共振氢谱和碳谱鉴定,黄皮新肉桂酰胺B(lansiumamide B)为主要的拒食活性成分,对斜纹夜蛾3龄幼虫48 h拒食中浓度为214.95μg/m L,同时具有较强的细胞毒性,对SL细胞48 h抑制中浓度为10.09μg/m L。【结论】黄皮种子提取物对斜纹夜蛾幼虫具有较强的拒食活性及细胞毒性,黄皮新肉桂酰胺B是主要活性成分之一,作为植物源杀虫剂开发具有较好的发展前景。     

Induction of 2-cyanoethyl-isoxazolin-5-one as an antifeedant against the tobacco cutworm (Spodoptera litura) by jasmonic acid in sweet pea leaf

ABSTRACT

Although sweet pea (Lathyrus odoratus) beans contain toxic β-aminopropionitrile, the plant itself is readily attacked by insects and is, therefore, protected through the use of pesticides. Consequently, the induction of L. odoratus resistance to insect attack via exogenous treatment is promising for pest control development. Screening of inducible elicitor effects showed that treatment of sweet pea foliage with jasmonic acid (JA) can induce antifeeding-based resistance to tobacco cutworm (Spodoptera litura) larvae. Spectroscopic analysis identified 2-cyanoethyl-isoxazolin-5-one (2-CEIX) as the antifeedant with a half-maximal effective concentration of 33.6 µmol/g fr. wt., i.e., exogenous JA treatment induced antifeeding activity due to the accumulation of 2-CEIX. Moreover, 2-CEIX-induced mortality of S. litura larvae was evaluated by a dipping test and the half-maximal lethal dose was determined to be 5.9 mg/mL. Therefore, 2-CEIX was concluded to be a suitable induced resistance target for elicitors or a lead compound for insecticide development.     

The effect of dietary nickel on the immune responses of Spodoptera litura Fabricius larvae

By exposing Spodoptera litura Fabricius larvae to nickel (Ni) in artificial diets for successive three generations, we investigated the impacts of the dietary Ni on growth and immune response of the fifth and sixth instar larvae at 24 h intervals. The time of newly moulted fifth instar larvae was labelled as 0 h. After exposure to 5 mg/kg Ni for two generations, Ni exposure significantly improved larval phenoloxidase activity and encapsulation grade in fifth instar larvae when compared to controls, except for encapsulation grade at 72-120 h in the second generation. However, higher concentrations of Ni (≥10 mg/kg) only significantly reduced encapsulation grade at 72-120 h. In the third generation, insects given higher dietary levels of Ni (≥10 mg/kg) showed lower immune responses and retarded relative growth rate (RGR) compared to controls, but those exposed to lower Ni levels (≤5 mg/kg) had a significantly improved encapsulation grade at 24-72 h. Larvae at lower Ni level (≤5 mg/kg) treatments had significantly higher RGR in comparison with that in controls. There was no significant difference in food relative consumption rate (RCR) and RGR among any treatment of the fifth instar larvae in three successive generations. These results indicated that the type and extent of effects on growth and immune responses of S. litura varied with the Ni concentrations and exposure periods.     

Detrimental effects of endophytic fungus Nigrospora sp. on survival and development of Spodoptera litura

Endophytes have been known to confer resistance to host plants against insect herbivores mediated by fungal alkaloids. In this study we have isolated an endophytic fungus Nigrospora sp. (Ascomycota: Sordariomycetes) from a native plant Tinospora cordifolia. To assess anti-insect potential, we tested to what extent the survival and development of Spodoptera litura (Lepidoptera: Noctuidae), a polyphagous pest, was affected when fed on artificial diet amended with different concentrations of ethyl acetate extract of Nigrospora sp. In feeding assay, the insect suffered significantly higher mortality and showed prolonged development period on amended diet as compared to unamended diet. A significant reduction in relative growth and consumption rate, efficiency of conversion of ingested and digested food and approximate digestibility indicated deterrent as well as toxic effects of the fungal extract. Significant adverse effects were also observed on adult emergence, longevity and reproductive potential of S. litura. This preliminary information on insecticidal properties of Nigrospora sp. may further be used for imparting resistance in plants against insects.     

寄主植物对斜纹夜蛾酯酶和乙酰胆碱酯酶活性的影响

利用生物测定与生物化学的方法,就取食不同寄主植物的2个斜纹夜蛾[Spodoptera litura(Fab.)]品系对丙溴磷和灭多威的敏感性及其体内的酯酶与乙酰胆碱酯酶的活性变化进行了研究。结果表明,取食不同寄主植物的斜纹夜蛾对丙溴磷和灭多威的敏感性不同。在敏感品系中,取食不同食料的斜纹夜蛾后代对灭多威的敏感性顺序为:烟草<棉花<大豆<人工饲料;对丙溴磷而言,敏感性顺序则为:棉花<烟草<大豆<人工饲料。而在田间抗性品系中,对丙溴磷的敏感性顺序为:棉花<烟草<人工饲料<大豆;对灭多威的敏感性顺序为:棉花<烟草<大豆<人工饲料。此外,在田间抗性和敏感品系中,取食不同食料的斜纹夜蛾体内的酯酶活性之间均存在显著差异,对其乙酰胆碱酯酶的活性也有程度不同的诱导效应,但并不会引起乙酰胆碱酯酶的质变。     

Antimicrobial activity of endemic Crataegus tanacetifolia (Lam.) Pers and observation of the inhibition effect on bacterial cells

Up to now an increasing number of antibiotic-resistant bacteria have been reported and thus new natural therapeutic agents are needed in order to eradicate these pathogens. Through the discovery of plants such as Crataegus tanacetifolia (Lam.) Pers that have antimicrobial activity, it will be possible to discover new natural drugs serving as chemotherapeutic agents for the treatment of nosocomial pathogens and take these antibiotic-resistant bacteria under control. The objective of the present study was to determine antimicrobial activity and the activity mechanism of C. tanacetifolia plant extract. The leaves of C. tanacetifolia, which is an endemic plant, were extracted using methanol and tested against 10 bacterial and 4 yeast strains by using a drop method. It was observed that the plant extract had antibacterial effects on Bacillus subtilis, Shigella, Staphylococcus aureus, and Listeria monocytogenes among the microorganisms that were tested. Minimum inhibitory concentration (MIC) results obtained at the end of an incubation of 24 h were found to be > or =6.16 mg ml(-1) for B. subtilis, < 394 mg ml(-1) for Shigella, and > or =3.08 mg ml(-1) for L. monocytogenes and S. aureus and minimum bactericidal concentration (MBC) were found as > or =24.63 mg ml(-1) for B. subtilis, > or =394 mg ml(-1) for Shigella, > or =6.16 mg ml(-1) for L. monocytogenes, and > or =98.5 mg ml(-1) for S. aureus. According to the MBC results, it was found that the plant extract had bactericidal effects and in order to explain the activity mechanism and cell deformation of bacterial strains treated with plant extract, the scanning electron microscopy (SEM) was used. The results of SEM showed that the treated cells appeared shrunken and there was degradation of the cell walls. This study, in which the antibacterial effect of C. tanacetifolia was demonstrated, will be a base for further investigations on advanced purification and effect mechanism of action of its active compounds.     

Reproducibility and desensitization of the power frequency magnetic field effect on movement of the common cutworm Spodoptera litura

All creatures on Earth, including human beings, can be influenced by the power frequency electromagnetic field (EMF), even though the consequence and degree of the effect may vary due to regional context, species, etc. Most of the outstanding scientific achievements about the EMF effect on life have come from behavioral studies. In such studies, in contrast to the geomagnetic field or static magnetic field (MF), the oscillating MF has attracted far less attention so far. Following a previous report, to attain deep basic knowledge about the effect of an extremely low frequency (ELF) MF on animal behavior, we characterized the 60‐Hz MF‐responsive movement activity of common cutworm larvae using sophisticated experimental schemes. The MF‐exposed third instar larvae showed significantly reduced locomotive activity compared to the matching sham‐exposed larvae. Moreover, repeated MF exposure to the same larvae up to three times also showed similar behavioral responsiveness even though the extent of movement decrease was attenuated by the repetition time. These results suggest that sinusoidal power frequency MF could disrupt the normal locomotory activity of insect larvae, and the insects may show adaptive desensitization to the same MF.     

Effectiveness of plant extracts to control purple blotch and Stemphylium blight diseases of onion (Allium cepa L.) in Assiut,Egypt

The present investigation aimed to isolate the causative agents of onion purple blotch and Stemphylium blight diseases and evaluate the efficacy of certain plant extracts against the two identified pathogens, in vitro and under greenhouse condition. Fourteen isolates of S. vesicarium and two isolates of Alternaria porri were tested for pathogenicity. The results indicated that all isolates were able to produce the symptoms of Stemphylium blight and onion purple blotch diseases with different degrees of severity ranging from 10.42 to 81.25%. A. porri No. 6022 caused the highest disease severity (81.25%), while S. vesicarium No. 6003 was the best one out of the tested 14 isolates (37.5%). Antifungal activity of some aqueous plant extracts (Azadirachta indica, Cydonia oblonga, Datura stramonium, Eucalyptus globulus, Foeniculum vulgare, Ocimum basilicum, Rosmarinus officinalis and Salix mucronata) was assayed in vitro by dry weight technique. The data indicated that there were significant differences between these extracts in their effect on fungal growth of A. porri and S. vesicarium and the best were A. indica and D. stramonium. Under greenhouse conditions, application of the aqueous extract of A. indica either before or after 48?h A. porri inoculation produced the highest reduction in disease severity comprising 70 and 74.7%, respectively. On the other hand, the highest percentage of disease reduction before and after 48?h S. vesicarium inoculation was produced by Ridomil gold plus reached to 84.4 and 95.8% respectively, followed by the aqueous extract of A. indica (74.1 and 89.7, respectively). According to our results, it can be concluded that plant extracts of A. indica and D. stramonium can be used for the biocontrol of purple blotch and Stemphylium blight diseases instead of fungicides to minimise the risks and hazards of using toxic fungicides.     

Role of leaf extracts of some medicinal plants in the management of fungal diseases of okra (Abelmoschus esculentus (L.) Moench)

In order to manage the fungal pathogens in okra, seeds of variety Arka Anamika were subjected to soaking treatment with the aqueous leaf extracts of Coleus aromaticus, Adathoda vesica, Vitex negundo, Solanum nigrum, Leucas aspera, Ocimum sanctum and Catharanthus roseus. Among the extracts used, C. aromaticus, V. negundo extracts were found superior in reducing the incidence of mycoflora. These leaf extract treatments also resulted in increased seed germination and vigour of the seedlings. Both in green-house and field conditions also these extracts were proved their efficient in the enhancement of biomass, number of leaves, number of seeds per fruit, seed density and ascorbic acid content in the raw fruits.