Selection and Validation of Reference Genes for Gene Expression Analysis in Tuta absoluta Meyrick (Lepidoptera: Gelechiidae).
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ABSTRACT:
SUBMITTER: Yan X
PROVIDER: S-EPMC8305163 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:To reveal the mechanism of temperature preference in Tuta absoluta, one of the top 20 plant pests in the world, we cloned and identified TaTRPA1, TaPain, and TaPyx genes by RACE and bioinformatic analysis, and clarified their expression profiles during different development stages using real-time PCR, and revealed their function in preference temperature by RNAi. The full-length cDNA of TaPain was 3136 bp, with a 2865-bp open reading frame encoding a 259.89-kDa protein; and the partial length cDNA of TaPyx was 2326-bp, with a 2025-bp open reading frame encoding a 193.16-kDa protein. In addition, the expression of TaTRPA1 and TaPyx was significantly lower in larvae than other stages, and it was significantly higher in pupae and newly emerging males for TaPain. After feeding target double-stranded RNA (dsRNA), the preferred temperature decreased 2 °C more than the control group. In conclusion, the results firstly indicated the molecular characterization of TRPA subfamily genes and their key role in temperature perception in T. absoluta, and the study will help us to understand the temperature-sensing mechanism in the pest, and will provide some basis for study of other Lepidoptera insects' temperature preference. Moreover, it is of great significance in enriching the research progress of "thermos TRP".
| S-EPMC8268038 | biostudies-literature
Project description:Endoparasitoids can be killed by host encapsulation, a cellular-mediated host immunological response against parasitism that involves hemocytes aggregation. As a counteracting strategy, many parasitoids can evade this host response through self-superparasitism. The objectives of this study were: 1) to describe the parasitoid Pseudapanteles dignus (Hymenoptera: Braconidae) early immature stages (egg and larva) encapsulation by the host Tuta absoluta (Lepidoptera: Gelechiidae), and 2) to determine the occurrence of self-superparasitism and the rate of escaping to encapsulation of this parasitoid. Knowledge of host-parasitoid immunological interaction is crucial when evaluating the potential of an endoparasitoid as a biological control candidate. Parasitoid-exposed T. absoluta larvae were dissected in vivo under light stereoscope microscope at 24-h intervals, for five days after exposition to detect encapsulation. The preimaginal stages of P. dignus and numbers of healthy and encapsulated immature parasitoids per host were recorded. Samples of parasitoid eggs and larvae were processed for SEM visualization of encapsulation. Necropsies evidenced that only the early first larval instar of P. dignus (up to 96 h-old) was partially or completely encapsulated. A non-melanized capsule, formed by layers of granulocyte-type hemocytes enveloping around the parasitoid body, was recorded. Approximately 50% of the parasitized T. absoluta larvae had significantly only one P. dignus egg, meanwhile supernumerary parasitization yielded up to seven immature parasitoids per host. The proportion of single-early first larval instar of P. dignus reached ? 0.5 and decreased significantly as the number of parasitoid individuals per host increased. P. dignus encapsulation and its ability to overcome with the host immune defense through self-superparasitism indicate that T. absoluta is a semi-permissive host for this parasitoid.
| S-EPMC5061380 | biostudies-literature
Project description:BackgroundTuta absoluta Meyrick 1917 (Lepidoptera: Gelechiidae) is an invasive, pesticide resistant, and a major treat of tomato production in the world. It needs effective management options that naturally infect the insect without causing any identified side effects. Entomopathogenic fungi (EPF) are the most important options. However, geographic origin and climatic condition apparently creates genetic variation among EPF strains that influence on their pathogenicity. Thus, screening of effective EPF strains from the local source is vital to develop environmental friendly pest control tactic for T. absoluta.ResultsIn this study, 27 indigenous Beauveria were isolated from the various types of soil and 12 of the isolates were screened based on their biological efficiency index (BEI). These isolates scored 65.7-95.7% and 68.3-95% of mortality against second and third instar larvae of T. absoluta at concentration of 1 × 107spores·ml-1 in 7 days post inoculation, respectively. Out of these, five (18.5%) isolates scored above 90% mortality on both instar larvae with LT50 value of 3.33 to 5.33 days at the lowest (104 spores·ml-1) and 1.93 to 3.17 days at highest (108 spores·ml-1) spore concentrations and has LC50 value of 1.5 × 103 to 1.1× 105 spores·ml-1. Moreover, isolates exhibited the promising mortality better (1.5 × 106 to 3.5 × 107 spores·ml-1), sporulated over the larval cadavers, well grown at optimal temperature, and produced chitinolytic enzymes. Molecular analysis showed that isolates have nearly monophyletic characters and grouped under species of Beauveria bassiana.ConclusionDifferent types of soil in Ethiopia are an important source of B. bassiana, and these isolates showed promising pathogenicity against T. absoluta, which is crucial for ecofriendly biopesticide development. Although isolates were nearly monophyletic in phylogenetic study, five of them were highly effective in the laboratory bioassays against T. absoluta; however, further field evaluation is required for mass production.
| S-EPMC8390704 | biostudies-literature
Project description:Tomato (Solanum esculentum) is one of the vegetable crops grown by both smallholder and commercial farmers in the Kingdom of Eswatini. Tomato leaf miner, Tuta absoluta M. is a major insect pest of tomatoes resulting in reduced tomato yields throughout the country. The study investigated the virulence of two sub-tropical EPN species on T. absoluta larvae. Steinernema yirgalemense and S. jeffreyense at different concentrations (0, 20, 40, 60 IJs/insect) were screened for efficacy (i.e. mortality rate) against larvae of T. absoluta in laboratory bioassays. Results obtained showed that S. yirgalemense and S. jeffreyense were able to kill T. absoluta larvae without significant differences between the two EPN species in 24-well bioassay trays. Significantly higher (p < 0.05) mortality was observed when 60 IJs/insect was used. The combination of S. yirgalemense at 60 IJs/insect (100%) and S. jeffreyense at 60 IJs/insect (98.3%) resulted in significantly higher (p < 0.05) mortality, compared with the other four combinations of EPN and concentration tested. In the leaf bioassays, S. yirgalemense (58.8%) resulted in significantly higher (p < 0.05) mean mortality compared to S. jeffreyense (46.4%). Steinernema yirgalemense at 60 IJs/insect resulted in significantly higher mean mortality compared to the other EPN and concentration combinations in the leaf bioassay. The results indicated that both EPNs tested were effective against T. absoluta larvae. Steinernema yirgalemense at 60 IJs/insect can effectively find T. absoluta larvae inside leaf mines, but large-scale field trials are recommended to demonstrate the potential use of the biocontrol agent within integrated pest management programs.Tomato (Solanum esculentum) is one of the vegetable crops grown by both smallholder and commercial farmers in the Kingdom of Eswatini. Tomato leaf miner, Tuta absoluta M. is a major insect pest of tomatoes resulting in reduced tomato yields throughout the country. The study investigated the virulence of two sub-tropical EPN species on T. absoluta larvae. Steinernema yirgalemense and S. jeffreyense at different concentrations (0, 20, 40, 60 IJs/insect) were screened for efficacy (i.e. mortality rate) against larvae of T. absoluta in laboratory bioassays. Results obtained showed that S. yirgalemense and S. jeffreyense were able to kill T. absoluta larvae without significant differences between the two EPN species in 24-well bioassay trays. Significantly higher (p < 0.05) mortality was observed when 60 IJs/insect was used. The combination of S. yirgalemense at 60 IJs/insect (100%) and S. jeffreyense at 60 IJs/insect (98.3%) resulted in significantly higher (p < 0.05) mortality, compared with the other four combinations of EPN and concentration tested. In the leaf bioassays, S. yirgalemense (58.8%) resulted in significantly higher (p < 0.05) mean mortality compared to S. jeffreyense (46.4%). Steinernema yirgalemense at 60 IJs/insect resulted in significantly higher mean mortality compared to the other EPN and concentration combinations in the leaf bioassay. The results indicated that both EPNs tested were effective against T. absoluta larvae. Steinernema yirgalemense at 60 IJs/insect can effectively find T. absoluta larvae inside leaf mines, but large-scale field trials are recommended to demonstrate the potential use of the biocontrol agent within integrated pest management programs.
| S-EPMC7265902 | biostudies-literature
Project description:In this study, we conducted tests on the isolation, identification, characterization, and extraction of chemical molecules from Beauveria bassiana against Tuta absoluta larvae. The enzyme responses of T. absoluta to the crude extract were examined 24 h after treatment, and the number of dead larvae was calculated 24 and 48 h after treatment. Molecular docking studies were conducted to assess the interaction of important molecules with the acetylcholinesterase enzyme. The larvicidal activity of crude chemicals from fungi was high 24 h after treatment, with LC50 and LC90 values of 25.937 and 33.559 μg/mL, respectively. For a period of 48 h, the LC50 and LC90 values were 52.254 and 60.450 μg/mL, respectively. The levels of acetylcholinesterase, α-carboxylesterase, and β-carboxylesterase enzymes were lower in the treatment group after 24 h compared to the control group. The GC-MS test revealed that the crude extract consisted mainly of 9,10-octadecadienoic acid, which was the primary compound. Docking results indicated that 9,10-octadecadienoic acid showed a strong interaction with acetylcholinesterase (AChE). Our findings suggest that the chemical molecule 9,10-octadecadienoic acid derived from the entomopathogenic fungus B. bassiana is more toxic to T. absoluta larvae. We plan to conduct studies to test its effectiveness in semi-field conditions and to evaluate its stability in field conditions. We believe that this 9,10-octadecadienoic acid molecule could be used to control T. absoluta larvae in the near future without causing environmental pollution.
| S-EPMC10901010 | biostudies-literature
Project description:(1) Background: The tomato leafminer Tuta absoluta (Lepidoptera: Gelechiidae) is a key tomato insect pest. At present, it is considered to be a serious threat in various countries in Europe, North Africa, and Middle East. The extensive use and the developed resistance of T. absoluta to spinosad causes some concern, which leads to the need for alternative products. (2) Materials and Methods: Several laboratory experiments were conducted to investigate the ovicidal properties of a zeolite particle film on T. absoluta. The toxicity of three different zeolites and six zeolite formulations to T. absoluta eggs and larvae was determined using different exposure methods. (3) Results: In general, the formulated zeolites yielded higher egg and larvae mortality values, especially when the zeolite particle film was residually applied. Notable differences in mortality rates from exposure to zeolites compared to other products, such as kaolin, its formulated product Surround, and the insecticide spinosad, were observed. Kaolin and Surround exhibited little or no effect for both application methods, while the hatch rate was reduced by 95% when spinosad was applied topically. Spinosad yielded egg and larvae mortality rates of 100% for both application methods. Additionally, increased oviposition activity was observed in adults exposed to the wettable powder (WP) formulations. These WP formulations increased egg deposition, while Surround and spinosad elicited a negative oviposition response. (4) Conclusions: It can be derived that the tested products, zeolites BEA (Beta polymorph A), FAU (Faujasite), LTA (Linde type A), and their formulations, had no real insecticidal activity against the eggs of T. absoluta. Nevertheless, egg exposure to zeolites seemed to affect the development process by weakening the first instar larvae and increasing their mortality. Subsequently, based on the choice test, no significant difference was observed between the number of eggs laid on the treated leaves and control leaves.
| S-EPMC5198220 | biostudies-other
Project description:Entomopathogenic nematodes (EPN) are excellent biocontrol agents against various insect pests. Novel biotechnological approaches can enhance their utility against insects above-ground, opening a new venue for selecting superior EPN against certain insects. We hypothesize that different populations of the same species but from different origins (habitat, ecoregion) will differ in their virulence. This study aimed to evaluate the virulence of various EPN populations against two pests of worldwide incidence and damage to high value crops: Frankliniella occidentalis (Thysanoptera: Thripidae) and Tuta absoluta (Lepidoptera: Gelechiidae). We tested 10 EPN populations belonging to three EPN species: Heterorhabditis bacteriophora (Koppert, MG-618b, AM-203, RM-102), Steinernema feltiae (Koppert, RS-5, AM-25, RM-107), and Steinernema carpocapsae (Koppert, MG-596a). Each EPN population was tested at two concentrations. Frankliniella occidentalis was tested at 160 and 80 IJs/cm2 and T. absoluta at 21 and 4 IJs/cm2. Control treatments followed the same experimental procedure but only adding distilled water. Overall, whenever different, higher IJs concentration resulted in lower adult emergence, higher larval mortality, and shorter time to kill the insects. Considering the low concentration, S. feltiae provided the best results for both insects and instars investigated, while H. bacteriophora and S. carpocapsae required a high concentration to reach similar or slightly better results. Differences among populations of each of the species were detected, but only the native populations of H. bacteriophora populations showed consistently higher control values against both insects/instar compared with the commercial one. Differences among S. feltiae and S. carpocapsae populations depended on the IJs concentration, insect, and instar. We consider S. feltiae a very promising species for their application against F. occidentalis and T. absoluta, with the Koppert population as the most consistent among the populations tested. Specific EPN-populations of S. carpocapsae and H. bacteriophora were good candidates against certain instar/insects at high concentrations. This study emphasized the importance of intraspecific variability for EPN virulence.
| S-EPMC8672422 | biostudies-literature
Project description:Tuta absoluta (the tomato pinworm) is an invasive insect pest with a highly damaging effect on tomatoes causing between 80 and 100% yield losses if left uncontrolled. Resistance to chemical pesticides have been reported in some T. absoluta populations. Insect microbiome plays an important role in the behavior, physiology, and survivability of their host. In a bid to explore and develop an alternative control method, the associated microbiome of this insect was studied. In this study, we unraveled the bacterial biota of T. absoluta larvae and adults by sequencing and analyzing the 16S rRNA V3-V4 gene regions using Illumina NovaSeq PE250. Out of 2,092,015 amplicon sequence variants (ASVs) recovered from 30 samples (15 larvae and 15 adults), 1,268,810 and 823,205 ASVs were obtained from the larvae and adults, respectively. A total of 433 bacterial genera were shared between the adults and larval samples while 264 and 139 genera were unique to the larvae and adults, respectively. Amplicon metagenomic analyses of the sequences showed the dominance of the phylum Proteobacteria in the adult samples while Firmicutes and Proteobacteria dominated in the larval samples. Linear discriminant analysis effect size (LEfSe) comparison revealed the genera Pseudomonas, Delftia and Ralstonia to be differentially enriched in the adult samples while Enterococcus, Enterobacter, Lactococcus, Klebsiella and Wiessella were differentially abundant in the larvae. The diversity indices showed that the bacterial communities were not different between the insect samples collected from different geographical regions. However, the bacterial communities significantly differed based on the sample type between larvae and adults. A co-occurrence network of significantly correlated taxa revealed a strong interaction between the microbial communities. The functional analysis of the microbiome using FAPROTAX showed that denitrification, arsenite oxidation, methylotrophy and methanotrophy as the active functional groups of the adult and larvae microbiomes. Our results have revealed the core taxonomic, functional, and interacting microbiota of T. absoluta and these indicate that the larvae and adults harbor a similar but transitory set of bacteria. The results provide a novel insight and a basis for exploring microbiome-based biocontrol strategy for this invasive insect pest as well as the ecological significance of some of the identified microbiota is discussed.
| S-EPMC11003966 | biostudies-literature
Project description:Microorganisms in the guts of insects enhance the adaptability of their hosts with different lifestyles, or those that live in different habitats. Tuta absoluta is an invasive pest that is a serious threat to tomato production in China. It has quickly spread and colonized Xinjiang, Yunnan and other provinces and regions. We used Illumina HiSeq next generation sequencing of the 16S rRNA gene to study and analyze the composition and diversity of the gut microbiota of three geographical populations of T. absoluta. At the phylum level, the most common bacteria in T. absoluta across all three geographical populations were Proteobacteria and Firmicutes. An uncultured bacterium in the Enterobacteriaceae was the dominant bacterial genus in the T. absoluta gut microbiotas. There were no significant differences in alpha diversity metrics among the Spanish, Yunnan and Xinjiang populations. The structures of the gut microbiota of the three populations were similar based on PCoA and NMDS results. The results confirmed that the microbial structures of T. absoluta from different regions were similar.
| S-EPMC8951508 | biostudies-literature
Project description:Phthorimaea absoluta is an invasive solanaceous plant pest with highly devastating effects on tomato plant. Heavy reliance on insecticide use to tackle the pest has been linked to insecticide resistance selection in P. absoluta populations. To underline insights on P. absoluta insecticide resistance mechanisms to diamides and avermectins, we evaluated the transcriptomic profile of parental (field-collected) and F8 (lab-reared) populations. Furthermore, to screen for the presence of organophosphate and pyrethroid resistance, we assessed the gene expression levels of acetylcholinesterase (ace1) and para-type voltage-gated sodium channel (VGSG) genes in the F1 to F8 lab-reared progeny of diamide and avermectin exposed P. absoluta field-collected populations. The VGSG gene showed up-regulation in 12.5% and down-regulation in 87.5% of the screened populations, while ace1 gene showed up-regulation in 37.5% and down-regulation in 62.5% of the screened populations. Gene ontology of the differentially expressed genes from both parental and eighth generations of diamide-sprayed P. absoluta populations revealed three genes involved in the metabolic detoxification of diamides in P. absoluta. Therefore, our study showed that the detoxification enzymes found could be responsible for P. absoluta diamide-based resistance, while behavioural resistance, which is stimulus-dependent, could be attributed to P. absoluta avermectin resistance.
| S-EPMC10995152 | biostudies-literature