Additional file 3: of An IMD-like pathway mediates both endosymbiont control and host immunity in the cereal weevil Sitophilus spp.

List of primers used for dsRNA synthesis and RT-qPCR. (XLSX 12 kb

Average amounts of lipid, glycogen, and sugar in infected larvae (red) and non-infected larvae (blue) of <i>Ceratitis capitata</i>.

Error bars indicate the standard error. (*) indicates a significant difference between columns (P<0.05).</p

Digestive glycosyl hydrolase activities after fungal inoculation to <i>Ceratitis capitata</i> (24 to 144 hours post-inoculation).

Blue and red bars chart represent glycosyl hydrolase activities mg protein for non-infected and infected flies respectively. (A) β –galactosidase activities; (B) α-glucosidase activities; (C) α-galactosidase activities and (D) β–glucosidase activities. Substrates used: PNPaglu, PNPbglu, PNPagal, PNPbgal. Error bars indicate the standard error. (* p<0.05, ** p<0.01, *** p<0.001).</p

Digestive proteinase activities after fungal inoculation to <i>Ceratitis capitata</i> (24 to 144 hours post-inoculation).

(A) Blue bar charts represent azocaseinolytic activities per mg of protein for non-infected flies; red bar charts represent azocaseinolytic activities per mg of protein for infected flies. (B) Blue bar charts represent hemoglobinasic activities per mg of protein for non-infected flies; red bar charts represent hemoglobinasic activities per mg of protein for infected flies. Error bars indicate the standard error. (* p<0.05, ** p<0.01, *** p<0.001).</p

Expression of 7 immune genes in <i>Ceratitis capitata</i> inoculated with <i>P</i>. <i>lilacinum</i> for the times analyzed (0h, 24h, 72h, and 144h).

The bottom and top “whiskers” represent minimum and maximum values, respectively. The thick line bisecting the box represents the median. The bottom and top of the box represent the 25th and 75th percentiles, respectively. Asterisks indicate a significant difference (* pS1 Data.</p

Variance Analysis tables of the complete linear models for the 7 genes of the study.

Variance Analysis tables of the complete linear models for the 7 genes of the study.</p

Survival analysis of <i>Ceratitis capitata</i> males and females, infected and non-infected (controls) with <i>P</i>. <i>lilacinum</i>.

Survival analysis of Ceratitis capitata males and females, infected and non-infected (controls) with P. lilacinum.</p

Primers sequences used in q-PCR.

The medfly Ceratitis capitata is one of the most damaging fruit pests with quarantine significance due to its extremely wide host range. The use of entomopathogenic fungi constitutes a promising approach with potential applications in integrated pest management. Furthermore, developing insect control methods can involve the use of fungal machinery to cause metabolic disruption, which may increase its effectiveness by impairing insect development. Insect species, including C. capitata, relies on reproduction potential, nutrient reserves, metabolic activities, and immune response for survival. Accordingly, the purpose of this study was to investigate the impacts of the entomopathogenic fungus Purpureocillium lilacinum on C. capitata pre-mortality. The medfly V8 strain was subjected to laboratory bioassays, which consisted on determining the virulence of P. lilacinum on the medfly. Purpureocillium lilacinum was applied on abdominal topical of 5-day-old males and females. Following the fungal inoculation, we have confirmed (i) a significant increase in tissue sugar content, (ii) a significant decrease in carbohydrase activities, digestive glycosyl hydrolase, and proteinase activities in whole midguts of treated flies, (iii) the antimicrobial peptides (AMPs) genes expression profile was significantly influenced by fly gender, fly status (virgin, mature, and mated), and time after infection, but infection itself had no discernible impact on the AMPs for the genes that were examined. This study provides the first insight into how P. lilacinum could affect C. capitata physiological mechanisms and provides the foundation for considering P. lilacinum as a novel, promising biocontrol agent.</div

Effect of <i>P</i>. <i>lilacinum</i> on fecundity and fertility of <i>Ceratits capitata</i>.

(A) Bar chart illustrating the mean number of eggs laid by non-infected and infected females of C. capitata; (B) Bar chart showing the mean proportion of eggs that hatched from eggs laid by non-infected and infected females of C. capitata. Error bars indicate the standard error. (*) indicates a significant difference between infected adults and non-infected (p<0.05).</p

Digestive disaccharide and polysaccharide activities after fungal inoculation to <i>Ceratitis capitata</i> (24 to 144 hours post-inoculation).

Blue and red bars chart illustrating enzymatic activities per mg protein for non-infected and infected flies respectively. (A) represent enzymatic activities on starch; (B) represent enzymatic activities on maltose (C) represent enzymatic activities on sucrose; (D) represent enzymatic activities on pectin. Error bars indicate the standard error. (* p<0.05, ** p<0.01, *** p<0.001).</p