Proteomic profiling of royal jelly produced by Apis mellifera L. exposed to food containing herbicide-based glyphosate

Abstract

Royal jelly (RJ) is rich in protective elements associated with collective immune defenses in the hive of Apis mellifera. Exposure of bees to glyphosate-based herbicides causes ultrastructural changes in the hypopharyngeal glands and a reduction in the production of RJ. However, the effects of glyphosate-based herbicides on the protein composition of RJ and consequences for the hive are unknown. Thus, we performed proteomic profiling of royal jelly produced in hives of A. mellifera exposed to food containing 1,5 μL of Roundup® (2.16 mg. g^-1 of glyphosate). The production of RJ was carried out in six hives, following the method of artificial production of queens. The combs containing 80 grafting cells were introduced into the hives, and the collection of royal jelly was performed after 72 h. Two treatments were determined based on hive feeding and the hive as the experimental unit: Control and "Roundup®". Royal jelly from the Roundup® treatment hives was compared to the Control hives. Proteins with differences in expression were identified by mass spectrometry. Only the proteins present in all three biological replicates were considered in the differential abundance analysis, using Student's t-test (p-value < 0.05, two-tailed). Hives that received food containing Roundup®, analysis showed alterations in protein profile in the RJ produced therein. In total, 24 proteins were identified, and the accumulation of Major royal jelly protein 3 (MRJP3) was downregulated, showing a significant reduction in hives exposed to food containing Roundup® in relation to control hives (t = 0.0017). MRJP3 acts analogously to polyclonal antigen-antibody reactions, performing functions related to immunity in bees of different ages and castes. To the best of our knowledge, this is the first study to demonstrate changes in the proteomic profile of RJ caused by glyphosate-based herbicides, indicating its negative effects on the nutrition and social immunity of bees.

Keywords: MRJP3; Nutrition; Protein composition; Roundup®; Social immunity.