Purification and properties of a beta-glycosidase purified from midgut cells of Spodoptera frugiperda (Lepidoptera) larvae

Two beta-glycosidases (BG) (Mr 47,000 and Mr 50,000) were purified from Spodoptera frugiperda (Lepidoptera: Noctuidae) midguts. These two polypeptides associate or dissociate depending on the medium ionic strength. The Mr 47,000 BG probably has two active sites. One of the putative active sites (cellobiase site) hydrolyses p-nitrophenyl beta-D-glucoside (NPbetaGlu) (79% of the total activity in saturated enzyme), cellobiose, amygdalin and probably also cellotriose, cellotetraose and cellopentaose. The cellobiase site has four subsites for glucose residue binding, as can be deduced from cellodextrin cleavage data. The enzymatic activity in this site is abolished after carbodiimide modification at pH 6.0. Since the inactivation is reduced in the presence of cellobiose, the results suggest the presence of a carboxylate as a catalytic group. The other active site of Mr 47,000 BG (galactosidase site) hydrolyses p-nitrophenyl beta-D-galactoside (NPbetaGal) better than NPbetaGlu, cleaves glucosylceramide and lactose and is unable to act on cellobiose, cellodextrins and amygdalin. This active site is not modified by carbodiimide at pH 6.0. The Mr 47,000 BG N-terminal sequence has high identity to plant beta-glycosidases and to mammalian lactase-phlorizin hydrolase, and contains the QIEGA motif, characteristic of the family of glycosyl hydrolases. The putative physiological role of this enzyme is the digestion of glycolipids (galactosidase site) and di- and oligosaccharides (cellobiase site) derived from hemicelluloses, thus resembling mammalian lactase-phlorizin hydrolase.     

Alpha-galactosidases from the larval midgut of Tenebrio molitor (Coleoptera) and Spodoptera frugiperda (Lepidoptera)

There are three midgut alpha-galactosidases (TG1, TG2, TG3) from Tenebrio molitor larvae that are partially resolved by ion-exchange chromatography. The enzymes have approximately the same pH optimum (5.0), pl value (4.6) and Mr value (46000-49000) as determined by gel filtration or native electrophoresis run in polyacrylamide gels with different concentrations. Substrate specificities and functions were proposed for the major T. molitor midgut alpha-galactosidases (TG2 and TG3) based on chromatographic, carbodiimide inactivation, Tris inhibition, and on substrate competition data. Thus, TG2 would hydrolyse alpha-1,6-galactosaccharides, exemplified by raffinose, whereas TG3 would act on melibiose and apparently also on digalactosyldiglyceride, the most important compound in the thylacoid membranes of chloroplasts. Most galactoside digestion should occur in the lumen of the first two thirds of T. molitor larval midguts, since alpha-galactosidase activity predominates there. Spodoptera frugiperda larvae have three midgut alpha-galactosidases (SG1, SG2, SG3) partially resolved by ion-exchange chromatography. The enzymes have similar pH optimum (5.8), pl value (7.2) and Mr value (46000-52000), and at least the major alpha-galactosidase must have an active carboxyl group in the active site. Based on data similar to those described for T. molitor, SG1 and SG3 should hydrolyse melibiose and SG3 should digest raffinose and, perhaps, also digalactosyldiglyceride. The midgut distribution of alpha-galactosidase activity supports the proposal that alpha-galactosidase digestion occurs at the surface of anterior midgut cells in Spodoptera frugiperda larvae.     

Cell-free translation in lysates from Spodoptera frugiperda (Lepidoptera: Noctuidae) cells

1. Conditions for in vitro translation of mRNA in cell-free extracts from cultured Spodoptera frugiperda cells were defined. 2. Incorporation of [35S]methionine into acid-precipitable material increased for approximately 1 hr, and was sensitive to the protein synthesis inhibitors pactamycin and cycloheximide. 3. Micrococcal nuclease-treated lysate, primed with purified rabbit globin mRNA, synthesized a major protein with the size of full length globin, indicating that the lysate supported correct initiation and elongation of polypeptides.     

Digestive enzymes in midgut cells,endo-and ectoperitrophic contents,and peritrophic membranes of Spodoptera frugiperda (lepidoptera) larvae

In the midgut of Spodoptera frugiperda larvae, subcellular fractionation data suggest that aminopeptidase and part of amylase, carboxypeptidase A, dipeptidase, and trypsin are bound to the microvillar membranes; that major amounts of soluble dipeptidase, cellobiase, and maltase are trapped in the cell glycocalyx; and finally that soluble carboxypeptidase, amylase, and trypsin occur in intracellular vesicles. Most luminal acetylglucosaminidase is soluble and restricted to the ectoperitrophic contents. Aminopeptidase occurs in minor amounts bound to membranes both in the ectoperitrophic contents and incorporated in the peritrophic membrane. Amylase, carboxypeptidase A, and trypsin are found in minor amounts in the ectoperitrophic contents (both soluble and membrane-bound) and in major amounts in the peritrophic membrane with contents. Part of the activities recovered in the last mentioned contents corresponds to enzyme molecules incorporated in the peritrophic membrane. The results suggest that initial digestion is carried out in major amounts by enzymes in the endoperitrophic space and, in minor amounts, by enzymes immobilized in the peritrophic membrane. Intermediate and final digestion occur at the ectoperitrophic space or at the surface of midgut cells. The results also lend support to the hypothesis that amylase and trypsin are derived from membrane-bound forms, are released in soluble form by a microapocrine mechanism, and are partly incorporated into the peritrophic membrane. © 1994 Wiley-Liss, Inc.     

Juvenile hormone biosynthesis in the fall armyworm, Spodoptera frugiperda (Lepidoptera, Noctuidae)

The in vitro production of juvenile hormones (JH) was investigated by using corpora allata (CA) of larvae and corpora cardiaca-corpora allata (CC-CA) complexes of adult females of the fall armyworm Spodoptera frugiperda. In female moths, JH release was high compared to that in 5th and 6th instar larvae. Concentrations of 0.11-0.12 mM methionine, 180-200 mM Na(+), 5.8-8.3 mM K(+), 10-50 mM Ca(2+) and a pH range of 5.7-6.3 yielded optimal incorporation of L-[methyl-(3)H] methionine in vitro by CC-CA complexes. The highest hourly incorporation occurred during a 9-h incubation period following a 1.5-h lag-phase. JH release from CC-CA complexes of adult females was shown to be age-dependent with a peak value on day 2 (approx. 4 pmol h(-1) CA(-1)). By a combination of reversed phase (RP)- and normal phase (NP)-high performance liquid chromatography (HPLC), two major labelled products released by the complex were separated. One compound co-migrated with chemically synthesized JH II diol, the second compound with JH III diol. Only traces of JH II and III could be detected in some samples. Gland extracts also contained both the major radiolabelled products. Double labelling experiments using [3H]methionine and [14C]acetate confirmed their de novo synthesis in CC-CA complexes of female moths. The nature of chemically synthesized reference JH III diol was proved by LC-MS (ESI mass spectrometry) and 1H-NMR (nuclear magnetic resonance spectroscopy).     

Regulation of vitellogenesis in the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae)

Studies were undertaken to investigate vitellogenesis and its regulation in female adults of the fall armyworm, Spodoptera frugiperda. A single female-specific protein, likely to be the S. frugiperda vitellogenin (Vg), appeared approximately 5 h after adult eclosion in the hemolymph of virgin females. The concentration of the protein increased with age as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed. A protein with the same relative molecular mass was also present in egg extracts, but absent from hemolymph samples from male moths. The relative molecular mass of the designated S. frugiperda Vg was determined as 164.5+/-2.5 kDa. Vitellogenic oocytes became visible 36-48 h after emergence and egg deposition began on day 3 of adult life. Vg could not be detected in the hemolymph of females decapitated directly after eclosion. When decapitated virgin females were injected with the JH-mimic methoprene (MP), the level of Vg was comparable to that in non-decapitated moths, indicating that vitellogenesis in S. frugiperda depends on juvenile hormone (JH). However, the number of vitellogenic oocytes was somewhat lower than in non-decapitated virgin females. Injection of 20-hydroxyecdysone (20E) promoted Vg production to a similar extent in decapitated female moths, but in contrast to methoprene injection, treatment with 20E never resulted in the production of vitellogenic oocytes. In vitro cultivated ovaries of adult females dissected directly after eclosion produced lower amounts of ecdysteroids than those isolated on day 1 after emergence. Our results suggest a crucial role for 20E in the induction of vitellogenesis in the noctuid S. frugiperda, while JH seems to be essential for the continued uptake of Vg by developing oocytes and may trigger 20E biosynthesis in the ovary.     

Changes in midgut endopeptidase activity of Spodoptera frugiperda (Lepidoptera: Noctuidae) are responsible for adaptation to soybean proteinase inhibitors

The development of transgenic maize plants expressing soybean proteinase inhibitors could reduce the economic damage of one of the major maize pests in Brazil, the fall armyworm, Spodoptera frugiperda (J.E. Smith, 1797). We examined the influence of soybean proteinase inhibitors on digestive enzyme properties and development of S. frugiperda larvae. The inhibition of trypsin and chymotrypsin activities in vitro by soybean proteinase inhibitors suggested that either Kunitz (SBTI) or Bowman-Birk (SBBI) would have a potential antimetabolic effect when ingested by insect larvae. However, chronic ingestion of semipurified soybean inhibitors did not result in a significant reduction of growth and development of fall armyworm. Therefore, digestive serine proteinase activities (trypsin and chymotrypsin) of fall armyworm larvae were characterized. The results suggest that S. frugiperda was able to physiologically adapt to dietary proteinase inhibitors by altering the complement of proteolytic enzymes in the insect midguts.     

Mortality Dynamics of Spodoptera frugiperda (Lepidoptera: Noctuidae) Immatures in Maize

We characterized the dynamics of mortality factors affecting immature developmental stages of the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Multiple decrement life tables for egg and early larval stages of S. frugiperda in maize (Zea mays L.) fields were developed with and without augmentative releases of Telenomus remus Nixon (Hymenoptera: Platygastridae) from 2009 to 2011. Total egg mortality ranged from 73 to 81% and the greatest egg mortality was due to inviability, dislodgement, and predation. Parasitoids did not cause significant mortality in egg or early larval stages and the releases of T. remus did not increase egg mortality. Greater than 95% of early larvae died from predation, drowning, and dislodgment by rainfall. Total mortality due to these factors was largely irreplaceable. Results indicate that a greater effect in reducing generational survival may be achieved by adding mortality to the early larval stage of S. frugiperda.     

草地贪夜蛾的性信息素通讯研究进展

草地贪夜蛾Spodoptera frugiperda原产于美洲,具有长距离迁飞特性。该虫于1988年入侵欧洲,2016年入侵非洲,并引起巨大的粮食损失,2018年入侵亚洲,2019年从缅甸传入中国,并在短时间扩散至全国10多个省。草地贪夜蛾依赖性信息素通讯,实现雌雄间识别、交配和繁衍,为了深入研究草地贪夜蛾的性信息素通讯及其机制并开发更高效的预测预报和防治技术,本文综述了草地贪夜蛾性信息素通讯的研究现状。1967年首次报道草地贪夜蛾性信息素为顺9-十四乙酸酯,随后发现有不同的鉴定结果,1986年确认其性信息素由顺9-十四乙酸酯和顺7-十二乙酸酯两个组分组成,其比例为96.6∶3.4时表现出最强的引诱活性。除此而外,对不同地理种群的研究发现,草地贪夜蛾雌蛾的信息素腺体提取物还存在顺9-十二乙酸酯、顺11-十六乙酸酯和反7-十二乙酸酯,田间实验表明这些物质在性信息素通讯中也可发挥作用。根据分子标记和喜食寄主,草地贪夜蛾可分为玉米品系和水稻品系。两个品系在外部形态上无法区分,性信息素组分也相同,但在组分比例上存在一定的差异。两个品系间存在一定程度的生殖隔离,但室内可以正交和反交并产生可育的F1代,F1代的性信息素比例与母本的比较相似。关于草地贪夜蛾雄性对性信息素的感受机制方面研究较少,仅见有感器形态的鉴定和对性信息素组分的触角电位反应研究的报道。根据目前草地贪夜蛾在我国的发生情况及其性信息素通讯机制的研究现状,我们建议尽快开展以下研究：(1)鉴定草地贪夜蛾我国入侵种群的雌性性信息素,尽快用于其种群动态监测及防治;(2)研究草地贪夜蛾雄性对性信息素的嗅觉编码机制;(3)研究草地贪夜蛾两个品系的行为隔离程度及其演化结局。     

Inga laurina trypsin inhibitor (ILTI) obstructs Spodoptera frugiperda trypsins expressed during adaptive mechanisms against plant protease inhibitors

Plant protease inhibitors (PIs) are elements of a common plant defense mechanism induced in response to herbivores. The fall armyworm, Spodoptera frugiperda, a highly polyphagous lepidopteran pest, responds to various PIs in its diet by expressing genes encoding trypsins. This raises the question of whether the PI‐induced trypsins are also inhibited by other PIs, which we posed as the hypothesis that Inga laurina trypsin inhibitor (ILTI) inhibits PI‐induced trypsins in S. frugiperda. In the process of testing our hypothesis, we compared its properties with those of selected PIs, soybean Kunitz trypsin inhibitor (SKTI), Inga vera trypsin inhibitor (IVTI), Adenanthera pavonina trypsin inhibitor (ApTI), and Entada acaciifolia trypsin inhibitor (EATI). We report that ILTI is more effective in inhibiting the induced S. frugiperda trypsins than SKTI and the other PIs, which supports our hypothesis. ILTI may be more appropriate than SKTI for studies regarding adaptive mechanisms to dietary PIs.     

Characterization of a CENP-B homolog in the holocentric Lepidoptera Spodoptera frugiperda

The discovery of an homolog of the human centromeric protein B, CENP-B, in an EST database of the holocentric insect species Spodoptera frugiperda prompted us to further characterize that gene because i) CENP-B has not been described in invertebrates yet ii) it should be a milestone in the molecular characterization of the holocentric centromere of Lepidoptera.Like its human counterpart, the Sf CENP-B protein is related to the transposase of the pogo transposable element (TE) of D. melanogaster. In this paper, we show evidences that the lepidopteran cenpB gene has evolved from domestication of a transposase. Furthermore, the Sf CENP-B nuclear location and its ability to bind to a retrotransposon derived sequence in vivo argue in favor of a functional homology to CENP-B proteins.     

氰氟虫腙对草地贪夜蛾的毒力与田间防效

【目的】本研究旨在明确氰氟虫腙对草地贪夜蛾Spodoptera frugiperda幼虫的毒力水平和田间防治效果,为科学使用氰氟虫腙防治草地贪夜蛾提供参考依据。【方法】采用饲料混毒法在室内测定了氰氟虫腙与4种常用杀虫剂甲维盐、氯虫苯甲酰胺、虱螨脲和茚虫威对草地贪夜蛾3和6龄幼虫的致死中浓度(LC₅₀)及LC₉₀值,以及LC₉₀浓度的这些杀虫剂对3龄幼虫的致死中时(medium lethal time, LT₅₀)值。采用人工喷雾方法测定了玉米田中22%氰氟虫腙悬浮剂(6.6 g/667 m²)、22%氰氟虫腙悬浮剂(17.6 g/667 m²)、5.7%甲维盐水分散剂(1 g/667 m²)和150 g/L茚虫威悬浮剂(2 g/667 m²)对草地贪夜蛾幼虫的防效。【结果】室内生测结果显示,供试的5种杀虫剂中氰氟虫腙对草地贪夜蛾3和6龄幼虫均具有较高的毒力,其LC₅₀值分别为2.64和4.3...     

Host association of Spodoptera frugiperda (Lepidoptera: Noctuidae) corn and rice strains in Argentina, Brazil, and Paraguay

Spodoptera frugiperda (J.E. Smith) is composed of two genetically distinct strains, the so-called corn strain and the rice strain. Whether the two strains differ in their host use is unclear, because laboratory experiments have not been able to show consistent host performance or preference differences between them, and field studies showed high rates of hybridization, as well as some degree asymmetric host use. To determine the distribution of the two strains and their association with host plants, we collected fall armyworm larvae from different crops (corn, rice, alfalfa, and sorghum) and grasses in 15 different localities over 4 yr in Argentina, Brazil, and Paraguay. The strain identity was analyzed using two polymorphisms in the mitochondrial cytochrome oxidase subunit I gene. We identified the corn and rice haplotypes and three types of populations were characterized based on the frequencies of the individuals that belonged to any of these haplotypes: in 44% of populations the corn haplotype predominated, in 44% of populations the rice haplotype was the most frequent, and 11% of populations showed both haplotypes at similar proportions. In total, eight populations (47%) showed the expected pattern, two populations (12%) were polymorphic within the same field, and seven populations (41%) showed the inverse pattern. Taken together, there was no consistent pattern of host association between the two sympatric genotypes and their respective host plants. This investigation supports the need for additional studies to determine which other forces keep the genotypes separate, and what is the degree of genetic differentiation between these populations.     

Identification of serine protease,serine protease homolog and prophenoloxidase genes in Spodoptera frugiperda (Lepidoptera: Noctuidae)

In insects, proteolytic cascades medicated by serine proteases (SPs), serine protease homologs (SPHs) and prophenoloxidases (PPOs) control several physiological processes, notably the innate immunity. However, no attempts have been made to identify and characterize these genes in Spodoptera frugiperda, one of the most destructive agricultural pests. In this study, 83 SPs, 26 SPHs and four PPOs were respectively identified in S. frugiperda genome based on homology blast against those of other insects. We then analyzed the domain organization of these proteins and assigned them into different groups by phylogenetic reconstruction. Furthermore, the mRNA levels of clip-domain SPs/SPHs (cSPs/cSPHs) and PPOs were quantified in response to a mixed infection of Micrococcus luteus and Escherichia coli, and obvious accumulations were recorded in immune tissues, including hemocytes and fat body. In the latter study, we profiled the expression patterns of highly expressed cSPs and PPOs in different developmental stages, including egg, larva, pupa, female and male adults. It was shown that most cSPs were abundantly expressed in adults, while PPOs were detected at high levels in both egg and larval stages. These current findings substantially add to our understanding of the roles of S. frugiperda SPs, SPHs and PPOs in immune regulation and further lay a solid foundation for uncovering the interaction mechanisms between insects and pathogens.     

交配和温度对草地贪夜蛾性信息素通讯的影响

【目的】明确交配和温度对草地贪夜蛾Spodopterafrugiperda性信息素通讯的影响,为田间草地贪夜蛾的性信息素防治提供参考。【方法】采用溶剂浸提法提取草地贪夜蛾雌蛾性信息素腺体中的化学组分;利用气相色谱-质谱联用（GC-MS）技术对提取物中的组分进行化学鉴定;利用气相色谱（GC）技术分析雌蛾腺体中各组分的含量;利用触角电位（EAG）技术测定雄蛾对腺体各组分及性信息素类似物的电生理反应。在不同温度（20-35）℃下饲养草地贪夜蛾,以研究温度对其性信息素通讯的影响。【结果】GC-MS分析发现,草地贪夜蛾雌蛾腺体内含有Z7-12:Ac等7种组分。交配实验表明,雌蛾在1个暗期内只交配1次,在羽化后7 d内可交配多次,其中交配2次及以上的占44%（单对放置）和67%(30对放置)。交配显著抑制雌蛾在次日暗期（交配后第1个暗期）的求偶活动,但在交配后第2个暗期基本恢复正常;在交配后第1个暗期,雌蛾腺体中主要组分Z9-14:Ac的含量和比例较对照雌蛾显著升高,其他组分无显著变化。在20-35℃的饲养温度范围内,雌蛾腺体内16:Ac的含量在35℃时最高,在20℃时未能检测出;各测试温度下Z11-16:Ac的含量相差不显著;其余5种组分均在25℃时最高;主要活性组分Z7-12:Ac和Z9-14:Ac在35℃时显著降低。饲养温度对腺体内各组分的相对比例也有较大影响,12:Ac、Z11-14:Ac、Z11-16:Ac和16:Ac所占比例均在30-35℃时较高,而Z9-14:Ac则在20-25℃时较高。不同饲养温度所得雄蛾对腺体主要组分的触角电位反应无显著差异。【结论】交配显著抑制草地贪夜蛾雌蛾在次日的求偶行为;25℃饲养条件最适宜雌蛾腺体内性信息素等组分的合成。     

广东省草地贪夜蛾种群周年动态及发生特征

为了明确广东省草地贪夜蛾Spodoptera frugiperda(J. E. Smith)种群周年消长动态及发生特征,根据2020年和2021年广东省草地贪夜蛾性诱监测及田间调查数据,分析不同生态区草地贪夜蛾成虫及幼虫种群周年发生动态。结果表明：(1)草地贪夜蛾在广东省大部分冬种玉米区可以周年繁殖,仅在清远、韶关、河源等地未发现幼虫为害;不同地区之间草地贪夜蛾越冬虫源基数差异较大,粤西发生为害较重,珠三角及粤东地区发生相对较轻;(2)周年繁殖区草地贪夜蛾成虫和幼虫全年均可发生为害,而季节发生区一般于3-4月才零星始见草地贪夜蛾成虫和幼虫,且诱蛾量、幼虫种群数量及为害程度均相对较低;(3)广东省草地贪夜蛾种群消长动态呈多峰型,发生高峰期主要集中于5-10月,期间出现多个大小不等的高峰,但峰期、蛾量和虫量在不同地区之间差异较大。本研究明确了广东省不同生态区草地贪夜蛾种群的周年消长动态及发生特征,为广东省乃至全国草地贪夜蛾的早期预警和精准防控提供重要参考。     

Partial purification and characterization of Helicoverpa armigera (Lepidoptera: Noctuidae) active aminopeptidase secreted in midgut

We report the partial purification to apparent homogeneity of a soluble aminopeptidase (EC 3.4.11.1) from midgut of Helicoverpa armigera larvae, which preferentially degraded Leucine p-nitroanilide (LpNA). After midgut isolation, extraction and precipitation of soluble proteins with acetone, proteins were purified in two consecutive steps including gel filtration and ion-exchange chromatographies. Aminopeptidase activity was increased 8.95 fold after gel filtration chromatography. The purified enzyme appeared as single band with a molecular mass of ～ 112 kDa in SDS-PAGE, with a pH optimum of 7.0. Zymogram analysis revealed two enzymatically active proteinases using LpNA as substrate. The optimal temperature of aminopeptidase activity was 50–60 °C. The enzyme was characterized as metalloprotease as it was strongly inhibited by 1,10 phenanthroline. Strong inhibition was also being observed using the specific aminopeptidase inhibitor bestatin. Heavy metal ions, EDTA and cysteine strongly inhibited the enzyme, while Ca^+ 2, Mn^+ 2 and Mg^+ 2 somewhat stimulated aminopeptidase activity. Besides LpNA, the purified aminopeptidase also cleaved with decreasing activity ApNA, VpNA and BApNA. Study could be helpful to understand the mechanism of action of N-terminal degrading enzymes and also important is to further study the differential interaction of Bacillus thuringiensis cry insecticidal toxin with midgut receptor of insects.     

叉角厉蝽对草地贪夜蛾的捕食功能反应

叉角厉蝽Eocanthecona furcellate(Wolff)是一种极具应用潜力的重要捕食性天敌。为了探明叉角厉蝽3龄若虫对草地贪夜蛾Spodoptera frugiperda(J.E.Smith) 3龄幼虫的控害效果,在室内条件下,观察了叉角厉蝽对草地贪夜蛾幼虫的捕食行为,研究了叉角厉蝽对草地贪夜蛾3龄幼虫的捕食功能反应。结果表明,叉角厉蝽以口针从草地贪夜蛾腹部或者体躯末端插入取食,被取食后的草地贪夜蛾呈干瘪状死亡。叉角厉蝽3龄若虫对草地贪夜蛾3龄幼虫的捕食功能反应模型符合Holling II型方程,为:Na=0.6702 N/(1+0.0133 N),叉角厉蝽若虫对草地贪夜蛾3龄幼虫的日最大捕食量、瞬时攻击率和处理时间分别为50.25头、0.6702 d和0.0199 d。叉角厉蝽的捕食量与猎物密度正相关,寻找效应与猎物密度负相关,叉角厉蝽的捕食作用存在较强的种内干扰作用。试验证实叉角厉蝽对草地贪夜蛾具有较好的控害效果,为其田间应用释放技术提供了理论依据。     

Toxicity and synergism in the feeding deterrence of some coumarins on Spodoptera frugiperda Smith (Lepidoptera: Noctuidae)

The phagodepression activity of five coumarins (= 2H-1-benzopyran-2-ones), 6-hydroxy-7-isoprenyloxycoumarin (1), 6-methoxy-7-isoprenyloxycoumarin (2), 6,7-methylenedioxycoumarin (3), 5-methoxy-6,7-methylenedioxycoumarin (4), and 6-methoxy-7-(2-hydroxyethoxy)coumarin (5), from the Argentine native herb Pterocaulon polystachyum, was tested against Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae. Two analogs, scopoletin (6) and 2-methoxy-2-methyl-3,4,5,6,7,8-hexahydro-3H-chromen-5-one (7), synthesized in our laboratory, were also evaluated for comparison. The compounds were added to an artificial diet at doses ranging from 50 to 200 microg per g of diet. Natural coumarins induced 100% of phagodepression when 200 microg were added per g of diet. Binary equimolar mixtures of the natural coumarins were phagodepressors against S. frugiperda surpassing the expected additive responses, indicating that these compounds can act synergistically against S. frugiperda larvae. Compounds 1 and 3 (non-methoxylated coumarins), and the equimolar mixture of both, displayed the strongest phagodepression. Additionally, 50 microg/g of 1 and 3 incorporated to the larval diet caused 80 and 50% of pupal mortality, respectively, while a 100 microg/g dose of compounds 2, 4, 6, and 7 produced 60, 50, 10, and 80% pupal mortality, respectively. Larval growing rate during the early larval instars was significantly reduced by treatments with the methylenedioxycoumarins 3 and 4. Coincidentally, the larval period duration was significantly increased by the latter compounds.