Biotransformation of gamma-terpinene and (-)-alpha-phellandrene by the larvae of common cutworm (Spodoptera litura)

gamma-Terpinene was mixed in artificial diet at a concentration of 1 mg/g of diet, and the diet was fed to the last instar larvae of common cutworm (Spodoptera litura). Metabolites were recovered from frass and analyzed spectroscopically. gamma-Terpinene was transformed mainly to p-mentha-1,4-dien-7-oic acid and p-cymen-7-oic acid (cumic acid). Similarly, (-)-alpha-phellandrene was transformed mainly to (4R)-p-mentha-1,5-dien-7-oic acid and p-cymen-7-oic acid (cumic acid). The C-7 position (allylic methyl group) of gamma-terpinene and (-)-alpha-phellandrene was preferentially oxidized.     

Biotransformation of (R)- and (S)-terpinen-4-ol by the larvae of common cutworm (Spodoptera litura)

(R)-Terpinen-4-ol was mixed in an artificial diet at a concentration of 1 mg/g of diet, and the diet was fed to the last instar larvae of common cutworm (Spodoptera litura). Metabolites were recovered from frass and analyzed spectroscopically. (R)-Terpinen-4-ol was transformed mainly to (R)-p-menth-1-en-4,7-diol. Similarly, (S)-terpinen-4-ol was transformed mainly to (S)-p-menth-1-en-4,7-diol. The C-7 position (allylic methyl group) of (R)- and (S)-terpinen-4-ol was preferentially oxidized.     

Biotransformation of (+)- and (-)-menthol by the larvae of common cutworm (Spodoptera litura).

(+)-Menthol was mixed in an artificial diet at a concentration of 1 mg/g of diet, and the diet was fed to the last instar larvae of common cutworm (Spodoptera litura). Metabolites were recovered from frass and analyzed spectroscopically. (+)-Menthol was transformed mainly to (+)-7-hydroxymenthol. Similarly, (-)-menthol was transformed mainly to (-)-7-hydroxymenthol. The C-7 position of (+)- and (-)-menthol was preferentially oxidized.     

Biotransformation of beta-myrcene by the larvae of common cutworm (Spodoptera litura)

beta-Myrcene was mixed in an artificial diet at a concentration of 1 mg/g of diet, and the diet was fed to the last instar larvae of common cutworm (Spodoptera litura). Metabolites were recovered from frass and analyzed spectroscopically. beta-Myrcene was transformed mainly to myrcene-3(10)-glycol and myrcene-1,2-glycol. Each pair of double bonds of beta-myrcene was converted to the corresponding diol by oxidation, respectively. The 3,10- and 1,2-double bonds of beta-myrcene were respectively oxidized.     

广西斜纹夜蛾核型多角体病毒的分离鉴定

从广西部分地区采集到自然罹病死亡的疑似具有病毒症状的斜纹夜蛾幼虫，对其进行病毒的分离鉴定。用研磨病虫尸体得到的浆液，经分离纯化后感染连续人工饲养至第三代的健康斜纹夜蛾三龄幼虫，部分样品能诱发病毒病症状。经差速离心的提纯物，在相差显微镜下观察到外形不规则的颗粒，进一步在电镜下观察到菱形、五边型、六边形、近似圆形等形状的多角体。用依据SpltNFV DNA的egt基因保守序列设计的引物，对多角体中提取的DNA进行PCR扩增，得到了与预期一致的380bp片段。经测序分析，该PCR片段与Splt NPV egt基因在核苷酸水平上的同源性为99．4％，演绎的氨基酸水平上的同源性为100％。进一步对采自广西16个地域的疑似具有病毒症状的200份斜纹夜蛾幼虫样本进行PCR扩增，其中6个地域的62份样本呈PCR阳性，这表明，广西斜纹夜蛾自然种群中存在着SpltNPV的流行。     

虫害诱导烟草抗性对斜纹夜蛾取食及解毒酶的影响

为探究不同虫害诱导抗性烟草(Nicotiana tabacum L.)对斜纹夜蛾(Spodoptera litura Fabricius)取食和解毒生理的影响,本研究以烤烟品种雄性不育K326为供试材料,观察斜纹夜蛾幼虫对棉铃虫[Helicoverpa armigera(Hübner)]诱导烟草、斜纹夜蛾诱导烟草、烟蚜...     

Diet influences rates of carbon and nitrogen mineralization from decomposing grasshopper frass and cadavers

Insect herbivory can produce a pulse of mineral nitrogen (N) in soil from the decomposition of frass and cadavers. In this study, we examined how diet quality affects rates of N and carbon (C) mineralization from grasshopper frass and cadavers. Frass was collected from grasshoppers fed with natural or meridic diets which varied in N content. Frass was also collected from naturally foraging grasshoppers. Nitrogen concentration of frass was directly proportional to diet N, but N content of cadavers was not affected by diet. Incubations of soil plus frass were performed at constant soil moisture and temperature (15°C) for 28 days, after which levels of mineral N (KCl extract) were determined. About 44 % of C and N from the cadavers were mineralized after the 28-day incubation. Carbon mineralization of frass was not affected by diet or frass N but varied considerably among different food treatments: from 15 to 46 % of the C in frass was released as carbon dioxide. Generally, frass with C/N ratio greater than 20 resulted in net immobilization of N. Results suggest that much of the N in grasshopper frass and cadavers is labile and rapidly available for plants, depending on the quality of food consumed by the grasshoppers.     

Secondary metabolites from Glycine soja and their growth inhibitory effect against Spodoptera litura

The wild soybean (Glycine soja Sieb. et Zucc) has been reported to be relatively resistant to insect and pathogenic pests. However, the responsible secondary metabolites in the aerial part of this important plant are largely unknown. From the aerial part of G. soja, 13 compounds were isolated and identified, including seven isoflavonoids (1-7), a cyclitol (8), two sterol derivatives (9 and 10), and three triterpenoids (11-13). Compound 7 is a new isoflavonoid, and compounds 9 and 10 are reported as natural products for the first time. The growth inhibitory activity of 1, 3, 4, and 8 against the larvae of Spodoptera litura was investigated. The most abundant isoflavonoid in the aerial part of G. soja, daidzein (1), which could not be metabolized by S. litura, was found to inhibit the insect larvae growth significantly in 3 days after feeding diets containing the compound. Compounds 3, 4, and 8, which could be partially or completely metabolized, were inactive. Our results suggested that the isoflavonoid daidzein (1) might function as a constitutive defense component in G. soja against insect pests.     

Acute, sublethal, antifeedant, and synergistic effects of monoterpenoid essential oil compounds on the tobacco cutworm, Spodoptera litura (Lep., Noctuidae)

Monoterpenoids (terpenes and biogenically related phenols) commonly found in plant essential oils were tested for acute toxicity via topical application to tobacco cutworms (Spodoptera litura Fab.). The most toxic among 10 such compounds was thymol (LD(50) = 25.4 microg/larva) from garden thyme, Thymus vulgaris. The compounds were then tested for sublethal effects, specifically inhibition of larval growth after topical application of low doses. Among 6 compounds tested, an LD(10) dose reduced growth by 20% on average 3 days after administration. Feeding deterrence was determined using a cabbage leaf disk choice test. The most deterrent compound was thymol, with a DC(50) of 85.6 microg/cm(2) leaf disk area. Because minor constituents in complex essential oils have been suggested to act as synergists, binary mixtures of the compounds were tested for synergy vis à vis acute toxicity and feeding deterrence. trans-Anethole acted synergistically with thymol, citronellal, and alpha-terpineol, in terms of both acute toxicity and feeding deterrence. On the basis of these findings, several complex mixtures were developed and tested as leads for effective control agents. Candidate mixtures demonstrated good synergistic effects. The observed LD(50) of mixture 3 was 40.6 microg/larvae compared to an expected value of 74.6 microg/larvae. The result of this research is a proprietary product suitable for commercial production.     

Evaluation of calcium-alginate gel as an artificial diet medium for bioassays on common cutworms

A calcium-alginate gel diet was developed for Spodoptera litura larvae, and its reliability as a carrier for incorporating antifeedants as well as insecticides was investigated. The alginate gel diet was prepared with a simple protocol, which does not involve any heating process. When tested using this diet, acephate, a Bacillus thuringiensis endotoxin formulation and rotenone reproducibly showed insecticidal activity against the larvae, while neem oil and scabequinone deterred the larval feeding effectively. However, not only the insecticidal activity of acephate but also the antifeedant activity of neem oil was reduced by replacing the alginate component by agar in the diet, suggesting the usefulness of the alginate gel diet as an assay tool for testing a broad range of samples against the larvae.     

骆驼蓬种子提取物对斜纹夜蛾的生物活性效应

用甲醇、乙醇、丙酮、氯仿、乙酸乙酯、苯、石油醚等7种溶剂对骆驼蓬种子进行冷浸提取。提取率分别为23．91％。18．68％．17．80％，18．49％，14．31％．15．13％，17．96％。7种提取物中．以种子乙醇提取物(ESPH)对斜纹夜蛾3龄幼虫的非选择性拒食活性最高。24h拒食率为70．15％。采用酸水总生物碱提取法对ESPH进行初步分离。得到氯仿、正丁醇、水及总生物碱萃取物。生物测定结果表明．总生物碱萃取物对斜纹夜蛾幼虫的生长发育抑制作用最为强烈，其次为氯仿萃取物。     

Metabolism of thymol and trans-anethole in larvae of Spodoptera litura and Trichoplusia ni (Lepidoptera: Noctuidae)

Metabolism of the monoterpenoid thymol and the phenylpropanoid trans-anethole, both constituents of essential oils, was investigated following topical and oral administration of the compounds to the tobacco cutworm (Spodoptera litura) and the cabbage looper (Trichoplusia ni). In both species and irrespective of route, administration of thymol resulted in the excretion of its 3-O-beta-glucoside, whereas trans-anethole was hydroxylated on the side chain methyl group. Both metabolites were isolated and their structures elucidated by interpretation of their mass and NMR spectra. Previous experiments indicated that trans-anethole synergized the toxicity of thymol in S. litura, but analyses of feces indicated that metabolism of thymol was not significantly suppressed when the two compounds were orally coadministered to T. ni larvae.     

Antifeedant and growth inhibitory effects of some neo-clerodane diterpenoids isolated from Clerodendron species (Verbenaceae) on Earias vitella and Spodoptera litura

Antifeedant and growth inhibitory effects of various neo-clerodane diterpenoids having a furofuran moiety, isolated from Clerodendron spp., were studied using Earias vitella and Spodoptera litura. The compounds clerodendrin B, 3-epicaryoptin, 15-hydroxyepicaryoptin, and clerodin were effective antifeedants at 10 microg/cm(3) (30 microg/g) of diet against E. vitella and at 10 microg/cm(2) of leaf against S. litura. All of the tested compounds, namely, clerodendrin B, 3-epicaryoptin, clerodendrin C, 15-hydroxyepicaryoptin, clerodendrin B acetate, and clerodin, showed good insect growth inhibitory activity even at lower concentrations.     

6beta-hydroxygedunin from Azadirachta indica. Its potentiation effects with some non-azadirachtin limonoids in neem against lepidopteran larvae

The biological activity of 6beta-hydroxygedunin isolated from Azadirachta indica A. Juss. was assessed using the gram pod borer, Helicoverpa armigera (Hubner), and Asian armyworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), alone and in combination with other limonoids, gedunin, salannin, nimbinene, and azadirachtin. The compound exhibited growth inhibitory activity in artificial diet bioassays, with 24.2 and 21.5 ppm, respectively, inhibiting growth by 50%. This efficacy was higher in comparison to gedunin (EC(50) = 50.8 and 40.4 ppm), salannin (EC(50) = 74.5 and 72.0 ppm), and nimbinene (EC(50) = 391.4 and 404.5 ppm). Azadirachtin, however, remained the most active neem allelochemical against both insect species. Nutritional assays clearly demonstrated that, though relative consumption and growth rates of fourth instar larvae were reduced, gedunin-type compounds induced physiological toxicity, evident by reduced efficiency of conversion of ingested food (ECI) in feeding experiments. Salannin and nimbinene, on the contrary, induced concentration-dependent feeding deterrence only. In feeding experiments, combinations of the compounds revealed that when azadirachtin was present in a mixture, EC(50) values did not deviate from the individual efficacy of azadirachtin (0.26 and 0.21 ppm, respectively) against H. armigera and S. litura larvae. However, a combination without azadirachtin did show a potentiation effect with potent EC(50) values among structurally different molecules, i.e., when salannin or nimbinene was combined with 6beta-hydroxygedunin or gedunin rather than structurally similar salannin + nimbinene or 6beta-hydroxygedunin + gedunin. Obviously, azadirachtin being the most active compound in neem is not synergized or influenced by any other limonoid, but other non-azadirachtin limonoids were more potent in specific combinations vis-à-vis the structural chemistry of the compound. It is obvious from the present study that potentiation among non-azadirachtin limonoids having explicitly two different modes of action, such as feeding deterrence and physiological toxicity, may be playing a significant role in the potentiation effect.     

Nicotinamide in relation to dietary nicotinic acid and nine other vitamins and larval development of Ceratitis capitata (Diptera: Tephritidae)

This study was undertaken to understand why Ceratitis capitata larvae reared on a diet fortified with nine vitamins except nicotinic acid had 100% mortality, while those reared on a 10-vitamin-free diet had 66% survival (Chang, C. L.; Li, Q. X. Ann. Entomol. Soc. Am. 2004, 97, 536-540). Our results showed that nicotinamide was detected at a level of 0.07 microg/g in second-instar larvae reared on the 10-vitamin-free diet and 0.30 microg/g in the corresponding spent diet, while it was not detected in either the larvae reared on the diet fortified with 707 microg/g of nine vitamins (nicotinic acid absent) or the corresponding spent diet. Nicotinamide was detected at concentrations of 0.13 and 0.15 microg/g in the larvae fed the diets that were fortified with 707 microg/g of nine other vitamins and 2 and 20 microg/g of nicotinic acid, respectively, but it was not found in the larvae fed the 0.2 microg/g of nicotinic acid diet. Nicotinamide was detected at concentrations of 0.44, 0.52, and 0.55 microg/g in the spent diets that were fortified with the nine vitamins (707 microg/g) and 0.2, 2, and 20 microg/g of nicotinic acid, respectively. Nicotinamide adenine dinucleotide (NAD) was in the live larvae, but not in the dead larvae. These findings indicate that dietary nicotinic acid is converted to nicotinamide, which, in turn, is used to synthesize NAD, and suggest a positive relationship between C. capitata larvae survival rates and concentrations of dietary nicotinic acid and nitcotinamide in the larvae as well as in the spent diets. The result shows that nicotinamide derived from supplemental nicotinic acid is essential for the development and survival of C. capitata larvae. Nicotinamide may be a biomarker for larval survival and development.     

Avocadofurans and their tetrahydrofuran analogues: comparison of growth inhibitory and insecticidal activity

The importance of the double bonds in the furan ring of avocadofurans with relation to their insecticidal activity was examined. The insecticidal activity of two naturally occurring avocadofurans, 2-(pentadecyl)furan and 2-(heptadecyl)furan, was compared to the toxicity of five tetrahydrofurans with alkyl chains at position 2 and varying side chains from 14 to 18 carbons. We found that eliminating the sites of unsaturation in the furan ring of avocadofurans significantly reduced the detrimental effects on the mortality and growth of the generalist insect herbivore Spodoptera exigua. In 7-day bioassays, S. exigua larvae were significantly more affected when fed a diet containing avocadofurans as compared to a larvae fed diet treated with the analogous tetrahydrofurans. Although larvae fed with the tetrahydrofurans showed reduced growth as compared to controls, larval mortality was not significantly increased. We conclude that the double bonds in the furan ring of avocadofurans play an important role in their insecticidal effects.     

Antifeedant activity of some pentacyclic triterpene acids and their fatty acid ester analogues

The 3-O-fatty acid ester derivatives (C(12)-C(18)) of two pentacyclic triterpenic acids, ursolic acid and oleanolic acid, were synthesized under mild esterification conditions in excellent yields (80-85%) and screened for their antifeedant activity, together with the parent acids, against the agricultural pest tobacco caterpillar larvae (Spodoptera litura F) in a no-choice laboratory study. The Urs-12-ene-28-carboxy-3beta-octadecanoate and olean-12-ene-28-carboxy-3beta-hexadecanoate were found to exhibit exceptionally potent antifeedant activities at 50 microg/cm(2) concentration, even after 48 h.     

不同寄主植物与斜纹夜蛾喜食程度、生长发育及存活率的关系研究

研究了不同寄主植物与斜纹夜蛾喜食程度、生长发育及存活率的关系结果表明 ,斜纹夜蛾喜食植物依次为槟榔芋 >莲藕 >甘蓝 >青菜 >水蕹菜 >黄牙白 >棉花 >大豆 >萝卜 >豇豆 >木耳菜 ;较喜食植物依次为花生 >芝麻 >酸模叶蓼 >四叶萍 >甘薯 >绿豆 >黄瓜 ;其余为次要寄主植物。斜纹夜蛾幼虫取食不同寄主植物 ,其发育历期、蛹重、蛹羽化出成虫比率均有显著差异 ,取食芋头、水蕹菜、青菜、甘蓝、豇豆和莲藕等作物叶片的幼虫发育历期较短 ,蛹较重 ,蛹羽化率较高 ,而取食棉花、大豆和向日葵叶片的幼虫历期相对较长 ,蛹较小 ,蛹羽化率相对较低。     

黑水虻虫粪对水稻生长及土壤理化性质的影响

  【目的】  黑水虻虫粪为黑水虻幼虫生物转化畜禽粪便所得,其有机质、养分含量高,可作为有机肥施用。研究黑水虻虫粪施用对水稻生长和土壤理化性质的影响,为其安全、合理使用提供科学依据。  【方法】  采用盆栽试验,设置施用盆栽土壤重量的0%、2%、4%、6%和8%虫粪肥5个处理 (依次记为CK、T1、T2、T3、T4),一次施肥后连续两年种植水稻,研究不同施肥处理对水稻株高、叶片酶活性、稻米产量、稻米氮磷钾含量、土壤养分指标、土壤酶活性的影响。  【结果】  施肥后第一年 (2018年),T2处理的水稻株高、产量最高,分别较CK提高了5.8%、43.7%,而T4处理的株高、产量分别较CK降低了8.0%、20.0%。在第二年 (2019年) 水稻株高和产量随着虫粪施用量的增加而提高,施虫粪肥处理提高了11.5%～33.5%的株高和33.9%～195.5%的产量。虫粪肥的施用在两年中都提高了水稻叶片的SOD、POD、CAT活性和稻米中的氮磷钾含量,且随着虫粪用量的增加而提高。虫粪肥的施用使土壤pH两年中分别由6.46、6.51提升到7.13、7.14。有机质含量提高了7.1%～32.5%,EC值提高了13.6%～99.9%,铵态氮含量提高了25.2%～195.4%,有效磷含量提高了45.7%～312.0%,速效钾含量提高了1.7～20.1倍。虫粪肥提高了13.1%～109.5%的脲酶活性,提高了14.5%～45.4%的磷酸酶活性,提高了25.2%～286.5%的蔗糖酶活性。  【结论】  黑水虻虫粪的施用可以提高土壤养分含量和酶活性,适宜的施用量能促进当季和后茬水稻生长,提高水稻产量,因此黑水虻虫粪作为一种新型有机肥料,具有广阔的应用潜力。     

Expression of the Truncated vip3A Gene at the 3'-terminus from Bacillus thuringiensis in Escherichia coli

将苏云金芽孢杆菌(Bacillus thuringiensis)缺失C端154个氨基酸编码区的vip3A基因(vip3T)插入原核表达载体pQE30，构建了重组表达载体pQEvip3T，并转化大肠杆菌(Escherichia coli ) M15进行IPTG诱导表达，比较了完整的Vip3A蛋白和C端缺失的蛋白Vip3T的可溶性和杀虫活性。与Vip3A不同，融合蛋白Vip3T以不可溶的包含体形式存在，诱导表达的菌液中没有检测到可溶性Vip3T蛋白。生物测定结果表明，M15(pOTP)诱导表达的Vip3A蛋白对初孵斜纹夜蛾(Spodoptera litua)和甜菜夜蛾(S. exigua)幼虫具有较高的杀虫活性，其提纯的包含体无毒，但包含体的碱性裂解液却又恢复了对夜蛾科害虫的活性；M15(pQEvip3T)菌液、包含体及其碱性裂解液对这两种昆虫幼虫则完全无毒，说明在大肠杆菌中，Vip3A蛋白C端氨基酸可能对Vip3A蛋白的可溶性和杀虫活性具有重要的影响。