The black soldier fly(BSF),Hermetia illucens,is a promising insect for miti-gating solid waste problems as its larvae are able to bioconvert organic waste into valuable biomass.We recently reported a high-quality geno...The black soldier fly(BSF),Hermetia illucens,is a promising insect for miti-gating solid waste problems as its larvae are able to bioconvert organic waste into valuable biomass.We recently reported a high-quality genome assembly of the BSF;analysis of this genome sequence will further the understanding of insect biology and identify genes that can be manipulated to improve efficiency of bioconversion.To enable genetic manip-ulation of the BSF,we have established the first transgenic methods for this economically important insect.We cloned and identified the ubiquitous actin5C promoter(Hiactin5C-p3k)and 3 endogenous U6 promoters(HiU6:1,HiU6:2,and HiU6:3).The Hiactin5C pro-moter was used to drive expression of a hyperactive variant of the piggyBac transposase,which exhibited up to 6-fold improvement in transformation rate when compared to the wild-type transposase.Furthermore,we evaluated the 3 HiU6 promoters using this trans-genic system.HiU6:1 and HiU6:2 promoters provided the highest knockdown efficiency with RNAi and are thus promising candidates for future Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)development.Overall,our findings provide valuable genetic engineering toolkits for basic research and genetic manipulation of the BSF.展开更多
Sex-determination pathways are extremely diverse.Understanding the mechanism of sex determination in insects is important for genetic manipulation of the pest population and for breeding of economically valuable insec...Sex-determination pathways are extremely diverse.Understanding the mechanism of sex determination in insects is important for genetic manipulation of the pest population and for breeding of economically valuable insects.Although sex determi-nation has been well characterized in the model species Drosophila melanogaster,little is known about this pathway in Stratiomyidae.In the present study,we first identified the Drosophila intersex(ix)homolog in Hermetia illucens,also known as the black soldier fly,which belongs to the Stratiomyidae family and which is an important insect for the con-version of various organic wastes.Phylogenetic analyses and multiple sequence alignment revealed that Hix is conserved compared with Drosophila.We showed that Hix is highly expressed in internal genitalia.Disruption of the Hix gene using CRISPR/Cas9 resulted in female-specific defects in external genitalia and abnormal and undersized ovaries.Taken together,our study furthers our understanding of sex determination in insects and could facilitatebreedingof H.illucens.展开更多
基金supported by the National Natural Science Foundation of China(32021001,31830093)Strategic Priority Research Program of Chinese Academy of Sciences(Grant no.XDPB16).
文摘The black soldier fly(BSF),Hermetia illucens,is a promising insect for miti-gating solid waste problems as its larvae are able to bioconvert organic waste into valuable biomass.We recently reported a high-quality genome assembly of the BSF;analysis of this genome sequence will further the understanding of insect biology and identify genes that can be manipulated to improve efficiency of bioconversion.To enable genetic manip-ulation of the BSF,we have established the first transgenic methods for this economically important insect.We cloned and identified the ubiquitous actin5C promoter(Hiactin5C-p3k)and 3 endogenous U6 promoters(HiU6:1,HiU6:2,and HiU6:3).The Hiactin5C pro-moter was used to drive expression of a hyperactive variant of the piggyBac transposase,which exhibited up to 6-fold improvement in transformation rate when compared to the wild-type transposase.Furthermore,we evaluated the 3 HiU6 promoters using this trans-genic system.HiU6:1 and HiU6:2 promoters provided the highest knockdown efficiency with RNAi and are thus promising candidates for future Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)development.Overall,our findings provide valuable genetic engineering toolkits for basic research and genetic manipulation of the BSF.
基金funded by the National Key Research and Development Project of China(2021YFD1301000).
文摘Sex-determination pathways are extremely diverse.Understanding the mechanism of sex determination in insects is important for genetic manipulation of the pest population and for breeding of economically valuable insects.Although sex determi-nation has been well characterized in the model species Drosophila melanogaster,little is known about this pathway in Stratiomyidae.In the present study,we first identified the Drosophila intersex(ix)homolog in Hermetia illucens,also known as the black soldier fly,which belongs to the Stratiomyidae family and which is an important insect for the con-version of various organic wastes.Phylogenetic analyses and multiple sequence alignment revealed that Hix is conserved compared with Drosophila.We showed that Hix is highly expressed in internal genitalia.Disruption of the Hix gene using CRISPR/Cas9 resulted in female-specific defects in external genitalia and abnormal and undersized ovaries.Taken together,our study furthers our understanding of sex determination in insects and could facilitatebreedingof H.illucens.
