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1.
BMC Plant Biol ; 21(1): 401, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34461825

RESUMO

BACKGROUND: Timing is everything when it comes to the fitness outcome of a plant's ecological interactions, and accurate timing is particularly relevant for interactions with herbivores or mutualists that are based on ephemeral emissions of volatile organic compounds. Previous studies of the wild tobacco N. attenuata have found associations between the diurnal timing of volatile emissions, and daytime predation of herbivores by their natural enemies. RESULTS: Here, we investigated the role of light in regulating two biosynthetic groups of volatiles, terpenoids and green leaf volatiles (GLVs), which dominate the herbivore-induced bouquet of N. attenuata. Light deprivation strongly suppressed terpenoid emissions while enhancing GLV emissions, albeit with a time lag. Silencing the expression of photoreceptor genes did not alter terpenoid emission rhythms, but silencing expression of the phytochrome gene, NaPhyB1, disordered the emission of the GLV (Z)-3-hexenyl acetate. External abscisic acid (ABA) treatments increased stomatal resistance, but did not truncate the emission of terpenoid volatiles (recovered in the headspace). However, ABA treatment enhanced GLV emissions and leaf internal pools (recovered from tissue), and reduced internal linalool pools. In contrast to the pattern of diurnal terpenoid emissions and nocturnal GLV emissions, transcripts of herbivore-induced plant volatile (HIPV) biosynthetic genes peaked during the day. The promotor regions of these genes were populated with various cis-acting regulatory elements involved in light-, stress-, phytohormone- and circadian regulation. CONCLUSIONS: This research provides insights into the complexity of the mechanisms involved in the regulation of HIPV bouquets, a mechanistic complexity which rivals the functional complexity of HIPVs, which includes repelling herbivores, calling for body guards, and attracting pollinators.


Assuntos
Ritmo Circadiano , Herbivoria/fisiologia , Luz , Tabaco/fisiologia , Compostos Orgânicos Voláteis/metabolismo , Ácido Abscísico/farmacologia , Animais , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/fisiologia , Larva/fisiologia , Mariposas/fisiologia , Fitocromo B/genética , Fitocromo B/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Terpenos/metabolismo
2.
Zool Res ; 42(5): 614-619, 2021 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-34402607

RESUMO

Butterflies are diverse in virtually all aspects of their ontogeny, including morphology, life history, and behavior. However, the developmental regulatory mechanisms underlying the important phenotypic traits of butterflies at different developmental stages remain unknown. Here, we investigated the developmental regulatory profiles of butterflies based on transposase accessible chromatin sequencing (ATAC-seq) at three developmental stages in two representative species ( Papilio xuthus and Kallima inachus). Results indicated that 15%-47% of open chromatin peaks appeared in associated genes located 3 kb upstream (i.e., promoter region) of their transcription start site (TSS). Comparative analysis of the different developmental stages indicated that chromatin accessibility is a dynamic process and associated genes with differentially accessible (DA) peaks show functions corresponding to their phenotypic traits. Interestingly, the black color pattern in P. xuthus 4th instar larvae may be attributed to promoter peak-related genes involved in the melanogenesis pathway. Furthermore, many longevity genes in 5th instar larvae and pupae showed open peaks 3 kb upstream of their TSS, which may contribute to the overwintering diapause observed in K. inachus adults. Combined with RNA-seq analysis, our data demonstrated that several genes enriched in the melanogenesis and longevity pathways also exhibit higher expression, confirming that the expression of genes may be closely related to their phenotypic traits. This study offers new insights into larval cuticle color and adult longevity in butterflies and provides a resource for investigating the developmental regulatory mechanisms underlying butterfly ontogeny.


Assuntos
Borboletas/fisiologia , Cromatina/metabolismo , Pigmentação/genética , Pigmentos Biológicos/metabolismo , Transcriptoma , Animais , Borboletas/anatomia & histologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Tegumento Comum/fisiologia , Larva/anatomia & histologia , Larva/fisiologia , Pigmentação/fisiologia
3.
J Photochem Photobiol B ; 222: 112276, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34358884

RESUMO

Despite decades of research, the role of elevated solar ultraviolet-B radiation (UVBR; 280-315 nm) in shaping amphibian populations remains ambiguous. These difficulties stem partly from a poor understanding of which parameters of UVBR exposure - dose, irradiance, and time interval - determine UVBR exposure health risk, and the potentially erroneous assumption that effects are proportional to the dose of exposure, irrespective of the administered regime (Bunsen-Roscoe Law of Reciprocity; BRL). We tested if the BRL holds with respect to UVBR-induced physiological effects in amphibians by acutely exposing tadpoles of the Australian green tree frog (Litoria caerulea) to a combination of different UVBR irradiances and doses in a fully factorial experiment. The BRL was invalid across all metrics assessed, with UVBR irradiance influencing the effects of a given dose on growth, coloration and burst swimming performance of larvae. We demonstrated some of the first empirical evidence for beneficial physiological effects of UVBR exposure in a larval amphibian, with improvements in growth, burst swimming performance and survival at the highest UVBR doses, contrary to hypotheses. Our findings demonstrate the species-specific nature of amphibian responses to UVBR, and the importance of UVBR irradiance in influencing the long-term physiological effects of a given dose of radiation. This work enhances our understanding of which parameters of complex UVBR exposures determine amphibian health risk.


Assuntos
Larva/efeitos da radiação , Raios Ultravioleta , Animais , Anuros/crescimento & desenvolvimento , Austrália , Larva/crescimento & desenvolvimento , Larva/fisiologia , Pigmentação da Pele/efeitos da radiação , Especificidade da Espécie , Natação
4.
BMC Plant Biol ; 21(1): 358, 2021 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-34348650

RESUMO

BACKGROUND: The South America pinworm, Tuta absoluta, is a destructive pest of tomato that causes important losses worldwide. Breeding of resistant/tolerant tomato cultivars could be an effective strategy for T. absoluta management but, despite the economic importance of tomato, very limited information is available about its response to this treat. To elucidate the defense mechanisms to herbivore feeding a comparative analysis was performed between a tolerant and susceptible cultivated tomato at both morphological and transcriptome level to highlight constitutive leaf barriers, molecular and biochemical mechanisms to counter the effect of T. absoluta attack. RESULTS: The tolerant genotype showed an enhanced constitutive barrier possibly as result of the higher density of trichomes and increased inducible reactions upon mild infestation thanks to the activation/repression of key transcription factors regulating genes involved in cuticle formation and cell wall strength as well as of antinutritive enzymes, and genes involved in the production of chemical toxins and bioactive secondary metabolites. CONCLUSIONS: Overall, our findings suggest that tomato resilience to the South America pinworm is achieved by a combined strategy between constitutive and induced defense system. A well-orchestrated modulation of plant transcription regulation could ensure a trade-off between defense needs and fitness costs. Our finding can be further exploited for developing T. absoluta tolerant cultivars, acting as important component of integrated pest management strategy for more sustainable production.


Assuntos
Regulação da Expressão Gênica de Plantas , Lycopersicon esculentum/genética , Doenças das Plantas/genética , Folhas de Planta/genética , Transcriptoma , Animais , Perfilação da Expressão Gênica/métodos , Ontologia Genética , Interações Hospedeiro-Parasita , Larva/fisiologia , Lycopersicon esculentum/metabolismo , Lycopersicon esculentum/parasitologia , Mariposas/fisiologia , Doenças das Plantas/parasitologia , Folhas de Planta/metabolismo , Folhas de Planta/parasitologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , RNA-Seq/métodos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Tricomas/genética , Tricomas/metabolismo , Tricomas/parasitologia
5.
Int J Mol Sci ; 22(13)2021 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-34281244

RESUMO

Olfaction is an important neural system for survival and fundamental behaviors such as predator avoidance, food finding, memory formation, reproduction, and social communication. However, the neural circuits and pathways associated with the olfactory system in various behaviors are not fully understood. Recent advances in optogenetics, high-resolution in vivo imaging, and reconstructions of neuronal circuits have created new opportunities to understand such neural circuits. Here, we generated a transgenic zebrafish to manipulate olfactory signal optically, expressing the Channelrhodopsin (ChR2) under the control of the olfactory specific promoter, omp. We observed light-induced neuronal activity of olfactory system in the transgenic fish by examining c-fos expression, and a calcium indicator suggesting that blue light stimulation caused activation of olfactory neurons in a non-invasive manner. To examine whether the photo-activation of olfactory sensory neurons affect behavior of zebrafish larvae, we devised a behavioral choice paradigm and tested how zebrafish larvae choose between two conflicting sensory cues, an aversive odor or the naturally preferred phototaxis. We found that when the conflicting cues (the preferred light and aversive odor) were presented together simultaneously, zebrafish larvae swam away from the aversive odor. However, the transgenic fish with photo-activation were insensitive to the aversive odor and exhibited olfactory desensitization upon optical stimulation of ChR2. These results show that an aversive olfactory stimulus can override phototaxis, and that olfaction is important in decision making in zebrafish. This new transgenic model will be useful for the analysis of olfaction related behaviors and for the dissection of underlying neural circuits.


Assuntos
Channelrhodopsins/metabolismo , Percepção Olfatória/genética , Olfato/genética , Animais , Animais Geneticamente Modificados/genética , Channelrhodopsins/genética , Sinais (Psicologia) , Larva/fisiologia , Luz , Rede Nervosa/metabolismo , Neurônios/metabolismo , Odorantes , Optogenética/métodos , Estimulação Luminosa , Regiões Promotoras Genéticas/genética , Natação , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/metabolismo
6.
J Chem Ecol ; 47(7): 664-679, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34196857

RESUMO

Larval Diaphania indica (Saunders) (Lepidoptera: Crambidae) cause complete defoliation of Trichosanthes anguina L. and reduce crop yield in India. Females lay eggs on the leaf surface, and therefore leaf surface waxes are potentially involved in host selection. Alkanes and free fatty acids are the major constituents of leaf surface waxes, so a study was conducted to determine whether these wax constituents from three T. anguina cultivars (MNSR-1, Baruipur Long, and Polo No.1) could act as short-range attractants and oviposition stimulants in D. indica females. Twenty n-alkanes from n-C14 to n-C36 and 13 free fatty acids from C12:0 to C21:0 were detected in the leaf surface waxes of these cultivars. Heptadecane and stearic acid were predominant among n-alkanes and free fatty acids, respectively, in these cultivars. Females showed attraction towards one leaf equivalent surface wax of each of these cultivars against solvent controls (petroleum ether) in Y-tube olfactometer bioassays. A synthetic blend of heptadecane, eicosane, hexacosane, and stearic acid, a synthetic blend of hexacosane and stearic acid, and a synthetic blend of pentadecane and stearic acid comparable to amounts present in one leaf equivalent surface wax of MNSR-1, Baruipur Long, and Polo No.1, respectively, were short-range attractants and oviposition stimulants in D. indica. Female egg laying responses were similar to each of these blends, providing information that could be used to developing baited traps in integrated pest management (IPM) programs.


Assuntos
Cucurbitaceae/metabolismo , Mariposas/fisiologia , Oviposição/efeitos dos fármacos , Ceras/farmacologia , Alcanos/análise , Alcanos/isolamento & purificação , Alcanos/farmacologia , Animais , Análise Discriminante , Ácidos Graxos não Esterificados/análise , Ácidos Graxos não Esterificados/isolamento & purificação , Ácidos Graxos não Esterificados/farmacologia , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Larva/efeitos dos fármacos , Larva/fisiologia , Mariposas/crescimento & desenvolvimento , Olfatometria , Folhas de Planta/metabolismo , Ceras/química , Ceras/isolamento & purificação
7.
J Chem Ecol ; 47(7): 642-652, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34331170

RESUMO

Finding plant cultivars that are resistant or tolerant against lepidopteran pests, takes time, effort and is costly. We present here a high throughput leaf-disk consumption assay system, to screen plants for resistance or chemicals for their deterrence. A webcam capturing images at regular intervals can follow the feeding activities of 150 larvae placed into individual cages. We developed a computer program running under an open source image analysis program to analyze and measure the surface of each leaf disk over time. We further developed new statistical procedures to analyze the time course of the feeding activities of the larvae and to compare them between treatments. As a test case, we compared how European corn borer larvae respond to a commercial antifeedant containing azadirachtin, and to quinine, which is a bitter alkaloid for many organisms. As expected, increasing doses of azadirachtin reduced and delayed feeding. However, quinine was poorly effective at the range of concentrations tested (10-5 M to 10-2 M). The model cage, the camera holder, the plugins, and the R scripts are freely available, and can be modified according to the users' needs.


Assuntos
Bioensaio/métodos , Comportamento Alimentar , Lepidópteros/fisiologia , Animais , Análise por Conglomerados , Comportamento Alimentar/efeitos dos fármacos , Processamento de Imagem Assistida por Computador , Larva/crescimento & desenvolvimento , Larva/fisiologia , Lepidópteros/crescimento & desenvolvimento , Limoninas/farmacologia , Folhas de Planta/química , Folhas de Planta/metabolismo , Quinina/farmacologia , Zea mays/química , Zea mays/metabolismo
8.
J Insect Sci ; 21(3)2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-34113998

RESUMO

Variation in body size has important implications for physical performance and fitness. For insects, adult size and morphology are determined by larval growth and metamorphosis. Female blue orchard bees, Osmia lignaria, (Say) provision a finite quantity of food to their offspring. In this study, we asked how provision-dependent variation in size changes adult morphology. We performed a diet manipulation in which some larvae were starved in the final instar and some were given unlimited food. We examined the consequences on adult morphology in two ways. First, allometric relationships between major body regions (head, thorax, abdomen) and total body mass were measured to determine relative growth of these structures. Second, morphometrics that are critical for flight (wing area, wing loading, and extra flight power index) were quantified. Head and thorax mass had hyperallometric relationships with body size, indicating these parts become disproportionately large in adults when larvae are given copious provisions. However, abdominal mass and wing area increased hypoallometrically with body size. Thus, large adults had disproportionately lighter abdomens and smaller wing areas than smaller adults. Though both males and females followed these general patterns, allometric patterns were affected by sex. For flight metrics, small adults had reduced wing loading and an increased extra flight power index. These results suggest that diet quantity alters development in ways that affect the morphometric trait relationships in adult O. lignaria and may lead to functional differences in performance.


Assuntos
Abelhas , Tamanho Corporal , Aptidão Genética/fisiologia , Asas de Animais , Animais , Abelhas/anatomia & histologia , Abelhas/fisiologia , Tamanho Corporal/fisiologia , Comportamento Alimentar , Feminino , Fertilidade , Himenópteros/anatomia & histologia , Himenópteros/fisiologia , Larva/fisiologia , Masculino , Fatores Sexuais , Asas de Animais/anatomia & histologia , Asas de Animais/fisiologia
9.
Methods Mol Biol ; 2326: 47-54, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34097260

RESUMO

Drosophila melanogaster, the fruit fly, has been widely used in biological investigation as an invertebrate alternative to mammals for its various advantages compared to other model organisms, which include short life cycle, easy handling, high prolificacy, and great availability of substantial genetic information. The behavior of Drosophila melanogaster is closely related to its growth, which can reflect the physiological conditions of Drosophila. We have optimized simple and robust behavioral assays for determining the larvae survival, adult climbing ability (mobility assay), reproductive behavior, and lifespan of Drosophila. In this chapter, we present the step-by-step detailed method for studying Drosophila behavior.


Assuntos
Drosophila melanogaster/fisiologia , Animais , Bioensaio/métodos , Drosophila melanogaster/efeitos dos fármacos , Drosophila melanogaster/crescimento & desenvolvimento , Feminino , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/fisiologia , Locomoção/efeitos dos fármacos , Longevidade/efeitos dos fármacos , Masculino , Reprodução/efeitos dos fármacos , Testes de Toxicidade/métodos
10.
Int J Mol Sci ; 22(10)2021 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-34066037

RESUMO

The multiple functions of the wild type Huntington's disease protein of the sea urchin Hemicentrotus pulcherrimus (Hp-Htt) have been examined using the anti-Hp-Htt antibody (Ab) raised against synthetic oligopeptides. According to immunoblotting, Hp-Htt was detected as a single band at around the 350 kDa region at the swimming blastula stage to the prism larva stage. From the 2-arm pluteus stage (2aPL), however, an additional smaller band at the 165 kDa region appeared. Immunohistochemically, Hp-Htt was detected in the nuclei and the nearby cytoplasm of the ectodermal cells from the swimming blastula stage, and the blastocoelar cells from the mid-gastrula stage. The Ab-positive signal was converged to the ciliary band-associated strand (CBAS). There, it was accompanied by several CBAS-marker proteins in the cytoplasm, such as glutamate decarboxylase. Application of Hp-Htt morpholino (Hp-Htt-MO) has resulted in shortened larval arms, accompanied by decreased 5-bromo-2-deoxyuridin (BrdU) incorporation by the ectodermal cells of the larval arms. Hp-Htt-MO also resulted in lowered ciliary beating activity, accompanied by a disordered swirling pattern formation around the body. These Hp-Htt-MO-induced deficiencies took place after the onset of CBAS system formation at the larval arms. Thus, Hp-Htt is involved in cell proliferation and the ciliary beating pattern regulation signaling system in pluteus larvae.


Assuntos
Cílios/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Proteína Huntingtina/metabolismo , Larva/fisiologia , Ouriços-do-Mar/fisiologia , Natação , Sequência de Aminoácidos , Animais , Proteína Huntingtina/genética , Homologia de Sequência
11.
Int J Mol Sci ; 22(11)2021 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-34070932

RESUMO

The neuronal networks that generate locomotion are well understood in swimming animals such as the lamprey, zebrafish and tadpole. The networks controlling locomotion in tetrapods remain, however, still enigmatic with an intricate motor pattern required for the control of the entire limb during the support, lift off, and flexion phase, and most demandingly when the limb makes contact with ground again. It is clear that the inhibition that occurs between bursts in each step cycle is produced by V2b and V1 interneurons, and that a deletion of these interneurons leads to synchronous flexor-extensor bursting. The ability to generate rhythmic bursting is distributed over all segments comprising part of the central pattern generator network (CPG). It is unclear how the rhythmic bursting is generated; however, Shox2, V2a and HB9 interneurons do contribute. To deduce a possible organization of the locomotor CPG, simulations have been elaborated. The motor pattern has been simulated in considerable detail with a network composed of unit burst generators; one for each group of close synergistic muscle groups at each joint. This unit burst generator model can reproduce the complex burst pattern with a constant flexion phase and a shortened extensor phase as the speed increases. Moreover, the unit burst generator model is versatile and can generate both forward and backward locomotion.


Assuntos
Geradores de Padrão Central/fisiologia , Interneurônios/fisiologia , Locomoção/fisiologia , Atividade Motora/fisiologia , Redes Neurais de Computação , Medula Espinal/fisiologia , Animais , Gatos , Geradores de Padrão Central/citologia , Simulação por Computador , Extremidades/inervação , Extremidades/fisiologia , Humanos , Interneurônios/citologia , Lampreias/fisiologia , Larva/fisiologia , Neurônios Motores/citologia , Neurônios Motores/fisiologia , Músculo Esquelético/inervação , Músculo Esquelético/fisiologia , Roedores/fisiologia , Medula Espinal/citologia , Peixe-Zebra/fisiologia
12.
J Chem Ecol ; 47(7): 680-688, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34101117

RESUMO

Past work shows a significant negative correlation between foliar oregonin concentration and western tent caterpillar (Malacosoma californicum Packard) feeding on red alder (Alnus rubra Bong.). Above an oregonin threshold of 20% leaf dry weight, little feeding by caterpillars is observed. Concentrations of defensive chemicals are influenced by plant genotype, environmental conditions, insect feeding, and the interactions of these factors. Our objective was to measure the effects of nitrogen (N) availability and wounding on foliar oregonin and condensed tannin concentrations in red alder genotypes. One-year-old seedlings from 100 half-sib red alder families were treated with two levels of ammonium nitrate (NH4NO3) for two growing seasons in a common garden. In the second year, leaves from 50 families from the fertilization experiment were used in a bioassay feeding experiment to determine the effects of N fertilization and genotype on WTC damage, and to identify a subset of 20 families with a range of damage to analyze for phytochemical composition. In separate experiments, wound-induction treatments were conducted outdoors and, in a greenhouse using the N treated trees in their third and fourth year, respectively. Foliar condensed tannin, oregonin and N concentrations were measured and ranked among the plant genotypes, and between the two N treatments and two wounding treatments. Results showed that oregonin and condensed tannin concentrations varied among the alder genotypes. Leaf N concentration was negatively correlated with concentration of oregonin. Neither of the measured phenolic compounds responded to wounding. The results suggest that red alder foliar oregonin and condensed tannin are likely constitutive defenses that are largely determined by genotype, and that the negative correlation of defense compounds with plant internal N status holds in this N-fixing tree.


Assuntos
Alnus/química , Diarileptanoides/química , Mariposas/efeitos dos fármacos , Compostos Fitoquímicos/farmacologia , Taninos/análise , Alnus/genética , Alnus/crescimento & desenvolvimento , Animais , Cromatografia Líquida de Alta Pressão , Diarileptanoides/farmacologia , Fertilizantes/análise , Genótipo , Herbivoria/efeitos dos fármacos , Larva/efeitos dos fármacos , Larva/fisiologia , Mariposas/crescimento & desenvolvimento , Mariposas/fisiologia , Nitrogênio/química , Nitrogênio/metabolismo , Compostos Fitoquímicos/química , Folhas de Planta/química , Folhas de Planta/metabolismo , Plântula , Espectrofotometria Ultravioleta , Taninos/farmacologia
13.
J Chem Ecol ; 47(7): 707-718, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34125370

RESUMO

Flooding is a major plant abiotic stress factor that is frequently experienced by plants simultaneously with other biotic stresses, including herbivory. How plant volatile emissions, which mediate interactions with a wide range of organisms, are influenced by flooding and by multiple co-occurring stress factors remains largely unexplored. Using Spodoptera frugiperda (Lepidoptera: Noctuidae) (fall armyworm) as the insect pest and two maize (Zea mays, L. Poaceae) hybrids differentially marketed for conventional and organic production, we assessed the effects of flooding, herbivory, and both stress factors on the composition of blends of emitted volatiles. Headspace volatiles were collected from all treatment combinations seven days after flooding. We documented metrics indicative of biomass allocation to determine the effects of individual and combined stressors on plant growth. We also evaluated relationships between volatile emissions and indicators of soil chemical characteristics as influenced by treatment factors. Flooding and herbivory induced the emission of volatile organic compounds (VOCs) in similar ways on both maize hybrids, but the interaction of both stress factors produced significantly larger quantities of emitted volatiles. Thirty-eight volatile compounds were identified, including green leaf volatiles, monoterpenes, an aldehyde, a benzoate ester, sesquiterpenes, a diterpene alcohol, and alkane hydrocarbons. The hybrid marketed for organic production was a stronger VOC emitter. As expected, plant biomass was detrimentally affected by flooding. Soil chemical properties were less responsive to the treatment factors. Taken together, the results suggest that flooding stress and the interactions of flooding and insect attack can shape the emission of plant volatiles and further influence insect-plant interactions.


Assuntos
Inundações , Spodoptera/fisiologia , Compostos Orgânicos Voláteis/química , Zea mays/química , Animais , Quimera , Cromatografia Gasosa-Espectrometria de Massas , Herbivoria , Larva/fisiologia , Folhas de Planta/química , Folhas de Planta/metabolismo , Solo/química , Spodoptera/crescimento & desenvolvimento , Compostos Orgânicos Voláteis/análise , Zea mays/metabolismo
14.
J Chem Ecol ; 47(7): 689-706, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34056671

RESUMO

A large percentage of crop loss is due to insect damage, especially caterpillar damage. Plant chitinases are considered excellent candidates to combat these insects since they can degrade chitin in peritrophic matrix (PM), an important protective structure in caterpillar midgut. Compared to chemical insecticides, chitinases could improve host plant resistance and be both economically and environmentally advantageous. The focus of this research was to find chitinase candidates that could improve plant resistance by effectively limiting caterpillar damage. Five classes of endochitinase (I-V) genes were characterized in the maize genome, and we isolated and cloned four chitinase genes (chitinase A, chitinase B, chitinase I, and PRm3) present in two maize (Zea mays L.) inbred lines Mp708 and Tx601, with different levels of resistance to caterpillar pests. We also investigated the expression of these maize chitinases in response to fall armyworm (Spodoptera frugiperda, FAW) attack. The results indicated that both chitinase transcript abundance and enzymatic activity increased in response to FAW feeding and mechanical wounding. Furthermore, chitinases retained activity inside the caterpillar midgut and enzymatic activity was detected in the food bolus and frass. When examined under scanning electron microscopy, PMs from Tx601-fed caterpillars showed structural damage when compared to diet controls. Analysis of chitinase transcript abundance after caterpillar feeding and proteomic analysis of maize leaf trichomes in the two inbreds implicated chitinase PRm3 found in Tx601 as a potential insecticidal protein.


Assuntos
Quitinases/farmacologia , Proteínas de Plantas/farmacologia , Spodoptera/efeitos dos fármacos , Zea mays/metabolismo , Sequência de Aminoácidos , Animais , Quitinases/classificação , Quitinases/genética , Quitinases/metabolismo , Clonagem Molecular , DNA de Plantas/química , DNA de Plantas/genética , DNA de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Herbivoria/efeitos dos fármacos , Larva/efeitos dos fármacos , Larva/fisiologia , Filogenia , Folhas de Planta/metabolismo , Proteínas de Plantas/classificação , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Análise de Sequência de DNA , Spodoptera/crescimento & desenvolvimento , Spodoptera/fisiologia
15.
Nat Methods ; 18(5): 557-563, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33963344

RESUMO

Visualizing dynamic processes over large, three-dimensional fields of view at high speed is essential for many applications in the life sciences. Light-field microscopy (LFM) has emerged as a tool for fast volumetric image acquisition, but its effective throughput and widespread use in biology has been hampered by a computationally demanding and artifact-prone image reconstruction process. Here, we present a framework for artificial intelligence-enhanced microscopy, integrating a hybrid light-field light-sheet microscope and deep learning-based volume reconstruction. In our approach, concomitantly acquired, high-resolution two-dimensional light-sheet images continuously serve as training data and validation for the convolutional neural network reconstructing the raw LFM data during extended volumetric time-lapse imaging experiments. Our network delivers high-quality three-dimensional reconstructions at video-rate throughput, which can be further refined based on the high-resolution light-sheet images. We demonstrate the capabilities of our approach by imaging medaka heart dynamics and zebrafish neural activity with volumetric imaging rates up to 100 Hz.


Assuntos
Aprendizado Profundo , Coração/fisiologia , Processamento de Imagem Assistida por Computador/métodos , Microscopia/métodos , Animais , Fenômenos Biomecânicos , Cálcio/química , Larva/fisiologia , Oryzias/fisiologia , Reprodutibilidade dos Testes , Peixe-Zebra/fisiologia
16.
Ecotoxicol Environ Saf ; 220: 112324, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34015630

RESUMO

Insecticides are extensively used worldwide to kill insect pests, yet organisms are most often exposed to insecticides at sublethal concentrations. Our understanding of sublethal effects on life histories is needed to predict the impact of insecticides on population dynamics and improve insecticide use and pest control. Sublethal concentrations can impact life histories directly and indirectly through changes in the intraspecific competition. Yet, few studies have evaluated the sublethal effects on intraspecific competition and these do not disentangle the insecticide effects on interference competition versus exploitative competition. As such, sublethal effects on the relative contribution of each pathways in shaping life histories are largely unknown, despite the fact that this can impact population dynamics. In this study, we focused on the neurotoxic insecticide spinosad and investigated its sublethal effects on interference among the aggressive larvae of the tortrix moth Adoxophyes honmai and the consequences for life histories. We conducted a set of paired experiments to disentangle the insecticide effects on interference from the ones on exploitation. Spinosad was found to amplify interference with most effects on mortality which lets us suggest that the insecticide likely increases the level of aggressive interactions resulting in more conspecific killings (e.g. cannibalism). Spinosad exposure was found to impair movement ability. Less movements may increase susceptibility to conspecific attacks and or increase aggresivity for better defence, two plausible mechanisms that could explain the increase in interference with insecticide. This study shows that insecticide at sublethal concentration can impact life histories by altering the strength of interference competition. Many organisms (pest and non-target species) compete through interference and theory predicts that a change in interference can substantially change dynamics. Our finding therefore reveals the importance of assessing the effect of insecticides on the mechanisms of competition when predicting their impact on populations.


Assuntos
Controle de Insetos , Inseticidas/farmacologia , Macrolídeos/farmacologia , Mariposas/efeitos dos fármacos , Agressão/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/fisiologia , Mariposas/crescimento & desenvolvimento , Mariposas/fisiologia , Movimento/efeitos dos fármacos , Dinâmica Populacional
17.
Nat Commun ; 12(1): 2943, 2021 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-34011945

RESUMO

Typical patterned movements in animals are achieved through combinations of contraction and delayed relaxation of groups of muscles. However, how intersegmentally coordinated patterns of muscular relaxation are regulated by the neural circuits remains poorly understood. Here, we identify Canon, a class of higher-order premotor interneurons, that regulates muscular relaxation during backward locomotion of Drosophila larvae. Canon neurons are cholinergic interneurons present in each abdominal neuromere and show wave-like activity during fictive backward locomotion. Optogenetic activation of Canon neurons induces relaxation of body wall muscles, whereas inhibition of these neurons disrupts timely muscle relaxation. Canon neurons provide excitatory outputs to inhibitory premotor interneurons. Canon neurons also connect with each other to form an intersegmental circuit and regulate their own wave-like activities. Thus, our results demonstrate how coordinated muscle relaxation can be realized by an intersegmental circuit that regulates its own patterned activity and sequentially terminates motor activities along the anterior-posterior axis.


Assuntos
Drosophila melanogaster/fisiologia , Interneurônios/fisiologia , Relaxamento Muscular/fisiologia , Animais , Animais Geneticamente Modificados , Neurônios Colinérgicos/citologia , Neurônios Colinérgicos/fisiologia , Drosophila melanogaster/anatomia & histologia , Interneurônios/citologia , Larva/anatomia & histologia , Larva/fisiologia , Locomoção/fisiologia , Modelos Neurológicos , Neurônios Motores/citologia , Neurônios Motores/fisiologia , Rede Nervosa/anatomia & histologia , Rede Nervosa/fisiologia , Optogenética
18.
PLoS One ; 16(4): e0246215, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33831004

RESUMO

Vale do Rio Juruá in western Acre, Brazil, is a persistent malaria transmission hotspot partly due to fish farming development that was encouraged to improve local standards of living. Fish ponds can be productive breeding sites for Amazonian malaria vector species, including Nyssorhynchus darlingi, which, combined with high human density and mobility, add to the local malaria burden.This study reports entomological profile of immature and adult Ny. darlingi at three sites in Mâncio Lima, Acre, during the rainy and dry season (February to September, 2017). From 63 fishponds, 10,859 larvae were collected, including 5,512 first-instar Anophelinae larvae and 4,927 second, third and fourth-instars, of which 8.5% (n = 420) were Ny. darlingi. This species was most abundant in not-abandoned fishponds and in the presence of emerging aquatic vegetation. Seasonal analysis of immatures in urban landscapes found no significant difference in the numbers of Ny. darlingi, corresponding to equivalent population density during the rainy to dry transition period. However, in the rural landscape, significantly higher numbers of Ny. darlingi larvae were collected in August (IRR = 5.80, p = 0.037) and September (IRR = 6.62, p = 0.023) (dry season), compared to February (rainy season), suggesting important role of fishponds for vector population maintenance during the seasonal transition in this landscape type. Adult sampling detected mainly Ny. darlingi (~93%), with similar outdoor feeding behavior, but different abundance according to landscape profile: urban site 1 showed higher peaks of human biting rate in May (46 bites/person/hour), than February (4) and September (15), while rural site 3 shows similar HBR during the same sampling period (22, 24 and 21, respectively). This study contributes to a better understanding of the larvae biology of the main malaria vector in the Vale do Rio Juruá region and, ultimately will support vector control efforts.


Assuntos
Anopheles/fisiologia , Aquicultura , Malária , Mosquitos Vetores/fisiologia , Lagoas , Estações do Ano , Animais , Brasil , Larva/fisiologia , Dinâmica Populacional
19.
PLoS One ; 16(4): e0249422, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33852607

RESUMO

Response to human impacts on the environment are typically initiated too late to remediate negative consequences. We present the novel use of stable isotope analysis (SIA) of blow flies to determine human influences on vertebrate communities in a range of human-inhabited environments, from a pristine national park to a dense metropolitan area. The refrain "you are what you eat" applies to the dietary isotope record of all living organisms, and for carrion-breeding blow flies, this translates to the type of carcasses present in an environment. Specifically, we show that carnivore carcasses make up a large proportion of the adult fly's prior larval diet, which contrasts to what has been reportedly previously for the wild adult fly diet (which consists of mostly herbivore resources). Additionally, we reveal the potential impact of human food on carcasses that were fed on by blow flies, underscoring the human influences on wild animal populations. Our results demonstrate that using SIA in conjunction with other methods (e.g., DNA analysis of flies) can reveal a comprehensive snapshot of the vertebrate community in a terrestrial ecosystem.


Assuntos
Calliphoridae/fisiologia , Dieta , Cadeia Alimentar , Animais , Calliphoridae/crescimento & desenvolvimento , Marcação por Isótopo , Larva/fisiologia
20.
Elife ; 102021 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-33875133

RESUMO

Insect herbivores use different cues to locate host plants. The importance of CO2 in this context is not well understood. We manipulated CO2 perception in western corn rootworm (WCR) larvae through RNAi and studied how CO2 perception impacts their interaction with their host plant. The expression of a carbon dioxide receptor, DvvGr2, is specifically required for dose-dependent larval responses to CO2. Silencing CO2 perception or scrubbing plant-associated CO2 has no effect on the ability of WCR larvae to locate host plants at short distances (<9 cm), but impairs host location at greater distances. WCR larvae preferentially orient and prefer plants that grow in well-fertilized soils compared to plants that grow in nutrient-poor soils, a behaviour that has direct consequences for larval growth and depends on the ability of the larvae to perceive root-emitted CO2. This study unravels how CO2 can mediate plant-herbivore interactions by serving as a distance-dependent host location cue.


Assuntos
Dióxido de Carbono/metabolismo , Herbivoria , Mariposas/fisiologia , Zea mays/metabolismo , Animais , Cadeia Alimentar , Larva/crescimento & desenvolvimento , Larva/fisiologia , Mariposas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo
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