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. 2021 May 10;11(1):9882.
doi: 10.1038/s41598-021-89212-5.

10-hydroxy-2E-decenoic acid (10HDA) does not promote caste differentiation in Melipona scutellaris stingless bees

Affiliations

10-hydroxy-2E-decenoic acid (10HDA) does not promote caste differentiation in Melipona scutellaris stingless bees

Luiza Diniz Ferreira Borges et al. Sci Rep. .

Abstract

In bees from genus Melipona, differential feeding is not enough to fully explain female polyphenism. In these bees, there is a hypothesis that in addition to the environmental component (food), a genetic component is also involved in caste differentiation. This mechanism has not yet been fully elucidated and may involve epigenetic and metabolic regulation. Here, we verified that the genes encoding histone deacetylases HDAC1 and HDAC4 and histone acetyltransferase KAT2A were expressed at all stages of Melipona scutellaris, with fluctuations between developmental stages and castes. In larvae, the HDAC genes showed the same profile of Juvenile Hormone titers-previous reported-whereas the HAT gene exhibited the opposite profile. We also investigated the larvae and larval food metabolomes, but we did not identify the putative queen-fate inducing compounds, geraniol and 10-hydroxy-2E-decenoic acid (10HDA). Finally, we demonstrated that the histone deacetylase inhibitor 10HDA-the major lipid component of royal jelly and hence a putative regulator of honeybee caste differentiation-was unable to promote differentiation in queens in Melipona scutellaris. Our results suggest that epigenetic and hormonal regulations may act synergistically to drive caste differentiation in Melipona and that 10HDA is not a caste-differentiation factor in Melipona scutellaris.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Comparison between expression of HDACs and HAT genes and JH titers in Melipona scutellaris larvae. The graphs show the overlapping of relative expression of hdac1 (a), hdac4 (b), and kat2a (c) quantified by RT-qPCR and JH titers quantified by radioimmunoassay. L2 = larva of the second stage; L3.3 = larva of the third stage in the third instar; LD = defecating larva. The graphs show mean ± SEM (n ≥ 5). Statistical analysis: Kruskal–Wallis test with a post-hoc Dunn’s multiple comparisons test, P < 0.05. JH data are from the work of Cardoso et al..
Figure 2
Figure 2
Expression of HDACs genes in Melipona scutellaris workers and queens. Relative expression of hdac1 (a) and hdac4 (b) quantified by RT-qPCR. Pw = pupa with white body and eyes; Pp = pupa with white body and pink eyes; Pb = pupa with white body and brown eyes; Pbl = pupa with light pigmented body and brown eyes; Pbd = pupa with dark pigmented body and brown eyes; NE = newly emerged adult. The graphs show mean ± SEM (n ≥ 3). Statistical analysis: 2Way-ANOVA with a post-hoc Sidak's multiple comparisons test, P < 0.05.
Figure 3
Figure 3
Metabolomic profiles of Melipona scutellaris larvae. Venn diagram of shared metabolites between larval stages (a) and PCA clustering of samples (b). L2 = larva of the second stage; L3.3 = larva of the third stage in the third instar; LD = defecating larva.
Figure 4
Figure 4
Correlation of metabolomic profiles of each sample of Melipona scutellaris larvae. Correlation heatmap of samples (a) and Hierarchical Clustering Dendrogram of samples (b). L2 = larva of the second stage; L3.3 = larva of the third stage in the third instar; LD = defecating larva.
Figure 5
Figure 5
Effects of treatment of Melipona scutellaris on the third larval instar with 10HDA. Survival rate (a) and Distribution of females in castes, queen and worker (b).

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