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Lucilia cuprina genome unlocks parasitic fly biology to underpin future interventions.

Anstead CA, Korhonen PK, Young ND, Hall RS, Jex AR, Murali SC, Hughes DS, Lee SF, Perry T, Stroehlein AJ, Ansell BR, Breugelmans B, Hofmann A, Qu J, Dugan S, Lee SL, Chao H, Dinh H, Han Y, Doddapaneni HV, Worley KC, Muzny DM, Ioannidis P, Waterhouse RM, Zdobnov EM, James PJ, Bagnall NH, Kotze AC, Gibbs RA, Richards S, Batterham P, Gasser RB - Nat Commun (2015)

Bottom Line: Lucilia cuprina is a parasitic fly of major economic importance worldwide.Here we report the sequence and annotation of the 458-megabase draft genome of Lucilia cuprina.Analyses of this genome and the 14,544 predicted protein-encoding genes provide unique insights into the fly's molecular biology, interactions with the host animal and insecticide resistance.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria 3010, Australia.

ABSTRACT
Lucilia cuprina is a parasitic fly of major economic importance worldwide. Larvae of this fly invade their animal host, feed on tissues and excretions and progressively cause severe skin disease (myiasis). Here we report the sequence and annotation of the 458-megabase draft genome of Lucilia cuprina. Analyses of this genome and the 14,544 predicted protein-encoding genes provide unique insights into the fly's molecular biology, interactions with the host animal and insecticide resistance. These insights have broad implications for designing new methods for the prevention and control of myiasis.

No MeSH data available.


Related in: MedlinePlus

Experimental design for the expression of the L. cuprina nAChR subunit gene (Lcα6) in Drosophila melanogaster and rescue of spinosad resistance in D. melanogaster (Dα6) mutants.(a) Virgin female elav>GAL4;dα6nx flies were crossed with male dα6nx;UAS-Lcα6 flies. The elav-driver produces GAL4 in neuronal cells, and the GAL4 binds to the UAS site to express the Lcα6 subunit that can be assembled into nAChRs. All individuals of the F1 generation have copies of the driver and the construct. First instar larvae were placed in sets of 50 on culture medium containing 0.1, 0.3 and 0.5 p.p.m. of spinosad, and allowed to develop. Mortality (%) was recorded on day 18 and normalized against control mortality. (b) Results from rescue experiments using the D. melanogaster GAL4:UAS system in different Dα6 mutant allele backgrounds: Lcα6–rescue showed no significant mortality in the D. melanogaster line ΦX-86Fb46 (Black) at 0.1, 0.3 and 0.5 p.p.m. of spinosad in a dα6W337* background and low mortality (9.4%±6.8) was seen only at 0.5 p.p.m. but not at the two lower doses in a dα6nx background. The UAS-Dα6 insertion line (Light orange) was susceptible to spinosad at all three doses, whereas the UAS-Lcα6 line (Cyan) was susceptible only at 0.5 p.p.m. (0.1 p.p.m. not tested). The driver line elav>GAL4 expressing Dα6 was highly susceptible at all three doses (Dark orange). The expression cross for the D. melanogaster α6 subunit had high mortality (>90%) on medium containing 0.1 and 0.3 p.p.m. spinosad (0.5 p.p.m. not tested), while the L. cuprina α6 subunit expression cross (Dark blue) showed significant, increasing spinosad susceptibility, with >90% mortality at 0.5 p.p.m. The bars represent 95% confidence intervals (CIs) calculated using a modified Abbott's correction69; five samples at 50 individuals each were tested for each dose. *—Not tested at that dose.
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f3: Experimental design for the expression of the L. cuprina nAChR subunit gene (Lcα6) in Drosophila melanogaster and rescue of spinosad resistance in D. melanogaster (Dα6) mutants.(a) Virgin female elav>GAL4;dα6nx flies were crossed with male dα6nx;UAS-Lcα6 flies. The elav-driver produces GAL4 in neuronal cells, and the GAL4 binds to the UAS site to express the Lcα6 subunit that can be assembled into nAChRs. All individuals of the F1 generation have copies of the driver and the construct. First instar larvae were placed in sets of 50 on culture medium containing 0.1, 0.3 and 0.5 p.p.m. of spinosad, and allowed to develop. Mortality (%) was recorded on day 18 and normalized against control mortality. (b) Results from rescue experiments using the D. melanogaster GAL4:UAS system in different Dα6 mutant allele backgrounds: Lcα6–rescue showed no significant mortality in the D. melanogaster line ΦX-86Fb46 (Black) at 0.1, 0.3 and 0.5 p.p.m. of spinosad in a dα6W337* background and low mortality (9.4%±6.8) was seen only at 0.5 p.p.m. but not at the two lower doses in a dα6nx background. The UAS-Dα6 insertion line (Light orange) was susceptible to spinosad at all three doses, whereas the UAS-Lcα6 line (Cyan) was susceptible only at 0.5 p.p.m. (0.1 p.p.m. not tested). The driver line elav>GAL4 expressing Dα6 was highly susceptible at all three doses (Dark orange). The expression cross for the D. melanogaster α6 subunit had high mortality (>90%) on medium containing 0.1 and 0.3 p.p.m. spinosad (0.5 p.p.m. not tested), while the L. cuprina α6 subunit expression cross (Dark blue) showed significant, increasing spinosad susceptibility, with >90% mortality at 0.5 p.p.m. The bars represent 95% confidence intervals (CIs) calculated using a modified Abbott's correction69; five samples at 50 individuals each were tested for each dose. *—Not tested at that dose.

Mentions: To examine whether α6-based spinosad resistance might evolve in L. cuprina, we performed heterologous expression of Lcα6 in D. melanogaster (Table 3), and assayed for functional rescue and insecticide susceptibility in transgenic flies. Utilizing the D. melanogaster GAL4:UAS system45, we cloned Lcα6 into either a dα6nx or a dα6W337* spinosad-resistant background (61- and 1,176-fold)44 and expressed Lcα6 in the elav>GAL4 driver line of D. melanogaster (Fig. 3). Rescue experiments showed that Lcα6 restored spinosad susceptibility in D. melanogaster (Fig. 3); no significant mortality in the D. melanogaster line ΦX-86Fb46 was observed using 0.1, 0.3 and 0.5 p.p.m. of spinosad in a dα6W337* background, and low mortality (9.4%±6.8) was seen only at 0.5 p.p.m., but not at the two lower doses in a dα6nx background. The UAS-Dα6 insertion line was susceptible to spinosad at all three doses, whereas the UAS-Lcα6 line was susceptible only at 0.5 p.p.m. (due to ‘leaky expression' at the attP landing site47. The driver line elav>GAL4 expressing Dα6 was highly susceptible at all three doses. Although transgenics with the Lcα6 subunit responded significantly at all doses, mortality at 0.1 p.p.m. was significantly lower than Dα6 in both the backgrounds (dα6nx and dα6W337*) when driven by elav>GAL4, showing that rescue was not as efficient as for Dα6.


Lucilia cuprina genome unlocks parasitic fly biology to underpin future interventions.

Anstead CA, Korhonen PK, Young ND, Hall RS, Jex AR, Murali SC, Hughes DS, Lee SF, Perry T, Stroehlein AJ, Ansell BR, Breugelmans B, Hofmann A, Qu J, Dugan S, Lee SL, Chao H, Dinh H, Han Y, Doddapaneni HV, Worley KC, Muzny DM, Ioannidis P, Waterhouse RM, Zdobnov EM, James PJ, Bagnall NH, Kotze AC, Gibbs RA, Richards S, Batterham P, Gasser RB - Nat Commun (2015)

Experimental design for the expression of the L. cuprina nAChR subunit gene (Lcα6) in Drosophila melanogaster and rescue of spinosad resistance in D. melanogaster (Dα6) mutants.(a) Virgin female elav>GAL4;dα6nx flies were crossed with male dα6nx;UAS-Lcα6 flies. The elav-driver produces GAL4 in neuronal cells, and the GAL4 binds to the UAS site to express the Lcα6 subunit that can be assembled into nAChRs. All individuals of the F1 generation have copies of the driver and the construct. First instar larvae were placed in sets of 50 on culture medium containing 0.1, 0.3 and 0.5 p.p.m. of spinosad, and allowed to develop. Mortality (%) was recorded on day 18 and normalized against control mortality. (b) Results from rescue experiments using the D. melanogaster GAL4:UAS system in different Dα6 mutant allele backgrounds: Lcα6–rescue showed no significant mortality in the D. melanogaster line ΦX-86Fb46 (Black) at 0.1, 0.3 and 0.5 p.p.m. of spinosad in a dα6W337* background and low mortality (9.4%±6.8) was seen only at 0.5 p.p.m. but not at the two lower doses in a dα6nx background. The UAS-Dα6 insertion line (Light orange) was susceptible to spinosad at all three doses, whereas the UAS-Lcα6 line (Cyan) was susceptible only at 0.5 p.p.m. (0.1 p.p.m. not tested). The driver line elav>GAL4 expressing Dα6 was highly susceptible at all three doses (Dark orange). The expression cross for the D. melanogaster α6 subunit had high mortality (>90%) on medium containing 0.1 and 0.3 p.p.m. spinosad (0.5 p.p.m. not tested), while the L. cuprina α6 subunit expression cross (Dark blue) showed significant, increasing spinosad susceptibility, with >90% mortality at 0.5 p.p.m. The bars represent 95% confidence intervals (CIs) calculated using a modified Abbott's correction69; five samples at 50 individuals each were tested for each dose. *—Not tested at that dose.
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Related In: Results  -  Collection

License
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f3: Experimental design for the expression of the L. cuprina nAChR subunit gene (Lcα6) in Drosophila melanogaster and rescue of spinosad resistance in D. melanogaster (Dα6) mutants.(a) Virgin female elav>GAL4;dα6nx flies were crossed with male dα6nx;UAS-Lcα6 flies. The elav-driver produces GAL4 in neuronal cells, and the GAL4 binds to the UAS site to express the Lcα6 subunit that can be assembled into nAChRs. All individuals of the F1 generation have copies of the driver and the construct. First instar larvae were placed in sets of 50 on culture medium containing 0.1, 0.3 and 0.5 p.p.m. of spinosad, and allowed to develop. Mortality (%) was recorded on day 18 and normalized against control mortality. (b) Results from rescue experiments using the D. melanogaster GAL4:UAS system in different Dα6 mutant allele backgrounds: Lcα6–rescue showed no significant mortality in the D. melanogaster line ΦX-86Fb46 (Black) at 0.1, 0.3 and 0.5 p.p.m. of spinosad in a dα6W337* background and low mortality (9.4%±6.8) was seen only at 0.5 p.p.m. but not at the two lower doses in a dα6nx background. The UAS-Dα6 insertion line (Light orange) was susceptible to spinosad at all three doses, whereas the UAS-Lcα6 line (Cyan) was susceptible only at 0.5 p.p.m. (0.1 p.p.m. not tested). The driver line elav>GAL4 expressing Dα6 was highly susceptible at all three doses (Dark orange). The expression cross for the D. melanogaster α6 subunit had high mortality (>90%) on medium containing 0.1 and 0.3 p.p.m. spinosad (0.5 p.p.m. not tested), while the L. cuprina α6 subunit expression cross (Dark blue) showed significant, increasing spinosad susceptibility, with >90% mortality at 0.5 p.p.m. The bars represent 95% confidence intervals (CIs) calculated using a modified Abbott's correction69; five samples at 50 individuals each were tested for each dose. *—Not tested at that dose.
Mentions: To examine whether α6-based spinosad resistance might evolve in L. cuprina, we performed heterologous expression of Lcα6 in D. melanogaster (Table 3), and assayed for functional rescue and insecticide susceptibility in transgenic flies. Utilizing the D. melanogaster GAL4:UAS system45, we cloned Lcα6 into either a dα6nx or a dα6W337* spinosad-resistant background (61- and 1,176-fold)44 and expressed Lcα6 in the elav>GAL4 driver line of D. melanogaster (Fig. 3). Rescue experiments showed that Lcα6 restored spinosad susceptibility in D. melanogaster (Fig. 3); no significant mortality in the D. melanogaster line ΦX-86Fb46 was observed using 0.1, 0.3 and 0.5 p.p.m. of spinosad in a dα6W337* background, and low mortality (9.4%±6.8) was seen only at 0.5 p.p.m., but not at the two lower doses in a dα6nx background. The UAS-Dα6 insertion line was susceptible to spinosad at all three doses, whereas the UAS-Lcα6 line was susceptible only at 0.5 p.p.m. (due to ‘leaky expression' at the attP landing site47. The driver line elav>GAL4 expressing Dα6 was highly susceptible at all three doses. Although transgenics with the Lcα6 subunit responded significantly at all doses, mortality at 0.1 p.p.m. was significantly lower than Dα6 in both the backgrounds (dα6nx and dα6W337*) when driven by elav>GAL4, showing that rescue was not as efficient as for Dα6.

Bottom Line: Lucilia cuprina is a parasitic fly of major economic importance worldwide.Here we report the sequence and annotation of the 458-megabase draft genome of Lucilia cuprina.Analyses of this genome and the 14,544 predicted protein-encoding genes provide unique insights into the fly's molecular biology, interactions with the host animal and insecticide resistance.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria 3010, Australia.

ABSTRACT
Lucilia cuprina is a parasitic fly of major economic importance worldwide. Larvae of this fly invade their animal host, feed on tissues and excretions and progressively cause severe skin disease (myiasis). Here we report the sequence and annotation of the 458-megabase draft genome of Lucilia cuprina. Analyses of this genome and the 14,544 predicted protein-encoding genes provide unique insights into the fly's molecular biology, interactions with the host animal and insecticide resistance. These insights have broad implications for designing new methods for the prevention and control of myiasis.

No MeSH data available.


Related in: MedlinePlus