RESUMEN
Cytochrome P450 (CYP) is a group of important detoxification enzymes found in insects related to their resistance to insecticides. To elucidate the CYP6 family genes of P450, which are potentially related to imidacloprid resistance in Aphis glycines, the CYP6 cDNA sequences of A. glycines were studied. The transcriptome of A. glycines was constructed, and the CYP6 cDNA sequences of A. glycines were screened. Their relative expression levels in response to imidacloprid induction were examined through qRT-PCR, and the CYP6s with higher expression levels were used to study the detoxification of imidacloprid through RNA interference and a bioassay. Twelve CYP6s were obtained from the A. glycines transcriptome. These samples were named by the International P450 Nomenclature Committee and registered in GenBank. After 3, 6, 12, 24 and 48 h of induction with LC50 concentrations of imidacloprid, the relative expression levels of these CYP6s increased; the expression level of CYP6CY7 experienced the highest increase, being more than 3-fold higher than that of those of the non-imidacloprid-induced CYP6s. After RNA interference for CYP6CY7, the relative expression level of CYP6CY7 significantly decreased after 3, 6 and 12 h, while the corresponding P450 enzyme activity decreased after 12 and 24 h. The mortality of A. glycines due to imidacloprid treatment increased by 14.71% at 24 h. CYP6CY7 might detoxify imidacloprid in A. glycines. This study provides a theoretical basis for the further study of the mechanism of action of CYP6s and potential new methods for improving insecticidal efficacy.
RESUMEN
Little is known about the complex molecular and cellular events occurring during implantation, which represents a critical step for pregnancy. The conventional 2D culture could not support postimplantation embryos' normal development, and 3D conditions shed light into the "black box". 3D printing technology has been widely used in recapitulating the structure and function of native tissues in vitro. Here, we 3D printed anisotropic microporous scaffolds to culture embryos by manipulating the advancing angle between printed layers, which affected embryo development. The 30° and 60° scaffolds promote embryo development with moderate embryo-scaffold attachments. T-positive cells and FOXA2-positive cells were observed to appear in the posterior region of the embryo and migrated to the anterior region of the embryo on day 7. These findings demonstrate a 3D printed stand that supports embryonic development in vitro and the critical role of 3D architecture for embryo implantation, in which additive manufacturing is a versatile tool.
