Reference no: EM132775249

The presented temporal change in gene expression subdivided lengthier stages (e.g., third instar, post-feeding and pupae) into smaller developmental units (i.e. third instar, early post-feeding, late post-feeding, early pupae, mid pupae and late pupae) which were of great significance to improve the precision of insects age estimation. Thus, the pattern of the gene expression is useful for determining age, but single gene should not be used to estimate age within developmental stages. Potential of gene expression as an aging tool comes by using combination of genes. During present study three reference (RP49, GPDH and AlphaTub84B) and five potential target genes (LSP-1α, Tango 5, Cytp450, SDCB and JHEBP) were analyzed by using RT-qPCR to age larvae and pupae. Seventy-two samples ranging from third instar to late puparial stage were analyzed with RT-PCR. Only one of these samples did not yield profiles for the three references genes rp49, Gpdh and AlphaTub84B, and was therefore excluded from analyses. The stability measure (M) was less than 1.0 for all the reference genes, indicating that the selected genes are stable and reliable for normalizing target genes expression. When analyzed with ANOVA, all genes of interest were found to have significant differences in gene expression between developmental time points. Tukey HSD was performed as a post-hoc analysis. The significant threshold was set to P < 0.05. Box and whisker plots with Tukey HSD significance were made to visualize gene expression. Different letters above each box indicates that expression is significantly different, whereas shared letters indicate that expression is not significantly different at the 0.05 level. According to Tukey HSD post-hoc analyses, Larval serum protein -1α expression were not significantly different from each other in larval stage, but found to be significantly higher than the early, mid and late puparial stages. Transport and Golgi organization 5 (Tango5) was found to have significantly high expression levels in feeding third instar and early pupae stage than another stages Cytochrome p450 (Cytp450) expression decreases as the larvae and pupae develop stages. Succinate dehydrogenase cytochrome B (SDCB) and Juvenile hormone esterase binding protein (JHEBP) were highly expressed in late pupal stage and showed significant difference in expression from other immature stages.

Why Transport golgi organisation gene was used to study an expression in immature development stages of Chrysomya rufifacies?