Establishing and maintaining colonies of<i>Forcipomyia taiwana</i>in the laboratory

Luo, Yi-Pey

doi:10.1111/jvec.12317

Cited by 7 publications

(7 citation statements)

References 11 publications

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“…All F. taiwana used in this study were collected from Taichung City (GPS 24.171467, 120.751336), where endemism is highest in Taiwan, and then raised in the lab [ 28 ]. For general maintenance, F. taiwana were reared at room temperature in a 20 × 20 × 30 (cm 3 ) transparent acrylic boxes with water and 10% (w/v) honey solution under 10 h light: 14 h dark daily cycles.…”

Section: Methodsmentioning

confidence: 99%

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Transcriptome profiling reveals the developmental regulation of NaCl-treated Forcipomyia taiwana eggs

et al. 2021

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Background The biting midge, Forcipomyia taiwana, is one of the most annoying blood-sucking pests in Taiwan. Current chemical control methods only target the adult, not the immature stages (egg to pupa), of F. taiwana. Discovering new or alternative tactics to enhance or replace existing methods are urgently needed to improve the effectiveness of F. taiwana control. The egg is the least understood life stage in this pest species but may offer a novel point of control as addition of NaCl to the egg environment inhibits development. Thus, the objective of this study was to use RNA profiling to better understand the developmental differences between wild-type melanized (black) and NaCl-induced un-melanized (pink), infertile F. taiwana eggs. Results After de novo assembly with Trinity, 87,415 non-redundant transcripts (Ft-nr) with an N50 of 1099 were obtained. Of these, 26,247 (30%) transcripts were predicted to have long open reading frames (ORFs, defined here as ≥300 nt) and 15,270 (17.5%) transcripts have at least one predicted functional domain. A comparison between two biological replicates each of black and pink egg samples, although limited in sample size, revealed 5898 differentially expressed genes (DEGs; 40.9% of the transcripts with long ORFs) with ≥2-fold difference. Of these, 2030 were annotated to a Gene Ontology biological process and along with gene expression patterns can be separated into 5 clusters. KEGG pathway analysis revealed that 1589 transcripts could be assigned to 18 significantly enriched pathways in 2 main categories (metabolism and environmental information processing). As expected, most (88.32%) of these DEGs were down-regulated in the pink eggs. Surprisingly, the majority of genes associated with the pigmentation GO term were up-regulated in the pink egg samples. However, the two key terminal genes of the melanin synthesis pathway, laccase2 and DCE/yellow, were significantly down-regulated, and further verified by qRT-PCR. Conclusion We have assembled and annotated the first egg transcriptome for F. taiwana, a biting midge. Our results suggest that down-regulation of the laccase2 and DCE/yellow genes might be the mechanism responsible for the NaCl-induced inhibition of melanization of F. taiwana eggs.

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Section: Methodsmentioning

confidence: 99%

“…For the qRT-PCR validation experiments, we changed to feeding the females with an artificial blood substitute, which had been developed in the meantime [ 28 ]. Additionally, knowing that the salt treatment yielded fewer eggs, for each batch the number of adult females for the salt treatment relative to the controls was doubled.…”

Section: Methodsmentioning

confidence: 99%

Transcriptome profiling reveals the developmental regulation of NaCl-treated Forcipomyia taiwana eggs

et al. 2021

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“…All F. taiwana used in this study were collected from Taichung City (GPS 24.171467, 120.751336), where endemism is highest in Taiwan, and then raised in the lab [18]. For general maintenance, F. taiwana were reared at room temperature in a 20*20*30 (cm 3 ) transparent acrylic boxes with water and 10% (w/v) honey solution.…”

Section: Samplesmentioning

confidence: 99%

“…For the qPCR validation experiments, we changed to feeding the females with an arti cial blood substitute, which had been developed in the meantime [18]. Additionally, knowing that the salt treatment yielded fewer eggs, for each batch the number of adult females for the salt treatment relative to the controls was doubled.…”

Section: Samplesmentioning

confidence: 99%

Transcriptome Profiling Reveals The Developmental Regulation of NaCl-Treated Forcipomyia Taiwana Eggs

Chen

Tsai

Huang

et al. 2021

Preprint

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Background: The biting midge, Forcipomyia taiwana, is one of the most annoying blood-sucking pests in Taiwan. Current chemical control only targets the adult, not the immature stages (egg to pupa) of F. taiwana. Discovering new or alternative tactics to enhance or replace existing methods are urgently needed to improve the effectiveness of F. taiwana control. The egg is the least understood life stage in this pest species but may offer a novel point of control as addition of NaCl to the egg environment inhibits development. Thus, the objective of this study was to use RNA profiling to better understand the developmental differences of wild-type melanized and NaCl-induced un-melanized and infertile F. taiwana eggs. Results: After de novo assembly with Trinity, 112,959 non-redundant transcripts (Ft-nr) with an N50 of 1,044 were obtained. Of these, 28,455 (24.8%) transcripts were predicted to have long open reading frames (ORFs, defined here as ≥300 nt) and 17,791 (15.8%) transcripts have at least one predicted functional domain. A comparison between the melanized (control) and un-melanized eggs revealed 15,996 differentially expressed genes (DEGs; 56.2% of the transcripts with long ORFs) with ≥2-fold difference. Of these, 8,865 were annotated to a Gene Ontology biological process and along with gene expression patterns can be separated into 5 clusters. KEGG pathway analysis revealed that 6,888 transcripts could be assigned to 102 significantly enriched pathways in 3 main categories (metabolism, genetic information processing, environmental information processing). As expected, most (78.24%) of these DEGs were down-regulated in un-melanized eggs. Surprisingly, the majority of genes associated with the pigmentation GO term were up-regulated in the pink egg samples; however, two key terminal genes, laccase2 and DCE/Yellow, were significantly down-regulated, and further verified by qPCR. Conclusion: We have assembled and annotated the first egg transcriptome for F. taiwana, a biting midge. We found that down-regulation of laccase2 and DCE/Yellow genes might be the mechanism responsible for the NaCl-induced inhibition of melanization of F. taiwana eggs.

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“…We dissected and examined more than 200 females right after the collection, and none of them had been observed with a developing ovary. The feeding of female midges was performed by using an artificial blood-feeding apparatus described previously [33]. The midges were cultured at 28 • C and relative humidity of 80%.…”

Section: Female Midge Collection Culture and Ovary Dissectionmentioning

confidence: 99%

Oogenesis of Hematophagous Midge Forcipomyia taiwana (Diptera: Ceratopogonidae) and Nuage Localization of Vasa in Germline Cells

Wang

Ching

Krishnaraj

et al. 2020

Insects

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Forcipomyia taiwana is an irritating hematophagous midge that preferentially attacks humans and affects leisure industries in Taiwan. Understanding the female reproductive biology of such pests would facilitate the development of pest control strategies. However, knowledge about oogenesis in the genus Forcipomyia is unavailable. Accordingly, we examined the ovariole structure and features of oogenesis in terms of the oocyte and the nurse cell. After being blood-fed, we observed a high degree of gonotrophic harmony—the synchronization of developing follicles. The follicle of the F. taiwana has only one nurse cell connected to the oocyte, which is distinct among hematophagous midges. In the nurse cell, we identified the perinuclear localization of the germline marker, Vasa. The Vasa localization is reminiscent of the nuclear envelope-associated nuage observed by electron microscopy. To determine whether F. taiwana Vasa (FtVasa) is an authentic nuage component, we produced transgenic flies expressing FtVasa in the female germline and proved that FtVasa was able to be localized to Drosophila nuage. By characterizing the oogenesis and Vasa expression in the germline cells of F. taiwana, this study extends knowledge about the female reproductive biology of hematophagous midges.

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Establishing and maintaining colonies ofForcipomyia taiwanain the laboratory

Cited by 7 publications

References 11 publications

Transcriptome profiling reveals the developmental regulation of NaCl-treated Forcipomyia taiwana eggs

Transcriptome profiling reveals the developmental regulation of NaCl-treated Forcipomyia taiwana eggs

Transcriptome Profiling Reveals The Developmental Regulation of NaCl-Treated Forcipomyia Taiwana Eggs

Oogenesis of Hematophagous Midge Forcipomyia taiwana (Diptera: Ceratopogonidae) and Nuage Localization of Vasa in Germline Cells

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