Alterations in Histone Methylation States Increased Profusion of Lethal(2)-Essential-for-Life-Like (l(2)elf), Trithorax and Polycomb Genes in Apis mellifera under Heat Stress.

Histone post-translational modifications (PTMs) represent a key mechanism in the thermal adaptation of the honeybee Apis mellifera. In this study, a chromatin immunoprecipitation assay and qPCR were employed to explore the changes in the methylation states of H3K4m2, H3K4m3, H3K27m2 and H3K27m3 associated with l2efl (ID: 72474, 724405, 724488), histone methyltransferases (HMTs) ((trx) and PR-set7) and Polycomb (Pc) and (Su(z)12) genes in A. m. jemenitica (tolerant subspecies) and A. m. carnica (susceptible subspecies) in response to heat treatment (42 °C for 1 h). The results revealed significant enrichment fold changes in the methylation/demethylation of most H3K4 and H3K27 marks at all targeted genes. These changes increased the profusion of l2efl (ID: 72474, 724405, 724488), histone methyltransferases (HMTs) (trx) and Polycomb (Pc) and Su(z)12 and decreased the profusion of HMT (PR-set7) in both honeybee subspecies. The changes in the methylation enrichment folds of histone methyltransferases (HMTs) ((trx), PR-set) and Polycomb (Pc), Su(z)12 genes demonstrate the well-harmonized coordination of epigenetic gene regulation in response to heat treatment. Compared to the control, the changes in the methylation enrichment folds of H3K4m3 at Polycomb Su(z)12 were about 30× and 100× higher in treated A. m. jemenitica and A.m. carnica, respectively. Similarly, changes in the methylation/demethylation enrichment folds of HMT (trx) and Polycomb (Pc) and Su(z)12 were 2-3× higher in A. m. carnica than in A. m. jemenitica after treatment (42 °C). It is evident that post-translational chromatin modification in both honeybee subspecies can diminish heat stress impact by (I) increasing the transcriptional provision of l2efl associated with survival and (II) increasing the silencing of genes associated with general cellular activities.

Linking Histone Methylation States and hsp Transcriptional Regulation in Thermo-Tolerant and Thermo-Susceptible A. mellifera L. Subspecies in Response to Heat Stress.

Genetic and epigenetic responses to environmental cues of worker honeybees mediate hsp synthesis, a key mechanism to tolerate high ambient temperatures in Apis mellifera. In this study, the chromatin immunoprecipitation assay followed by qPCR were used to determine alterations in histone methylation states (H3K27me2, H3K27me3, H3K4me2, and H3K4me3) associated with hsp/hsc/trx in A. m. jemenetica (thermo-tolerant subspecies) and A. m. carnica (thermo-susceptible subspecies) after heat treatment. The results revealed significant changes in enrichment folds of histone methylation states associated with hsp/hsc/trx. Indeed, the enrichment of H3K27me2 decreased strongly in response to heat stress. Changes in histone methylation states were significantly higher in A. m. carnica samples compared to A. m. jemenitica samples. Our study provides a new perception on linking histone post-translational methylation as an epigenetic mechanism of gene regulation with hsp/hsc/trx in A. mellifera subspecies exposed to heat stress.

Expression Levels of Heat-Shock Proteins in Apis mellifera jemenetica and Apis mellifera carnica Foragers in the Desert Climate of Saudi Arabia.

A. m. jemenetica is the indigenous honeybee of the Arabian Peninsula. It is highly adapted to extreme temperatures exceeding 40 °C, yet important molecular aspects of its adaptation are not well documented. In this study we quantify relative expression levels of small- and large-molecular-weight heat-shock proteins (hsp10, hsp28, hsp70, hsp83, hsp90 and hsc70 (mRNAs)) in the thermos-tolerant A. m. jemenetica and thermosusceptible A. m. carnica forager honeybee subspecies under desert (Riyadh) and semi-arid (Baha) summer conditions. The results showed significant day-long higher expression levels of hsp mRNAs in A. m. jemenetica compared to A. m. carnica under the same conditions. In Baha, the expression levels were very modest in both subspecies compared those in Riyadh though the expression levels were higher in A. m. jemenetica. The results also revealed a significant interaction between subspecies, which indicated milder stress conditions in Baha. In conclusion, the higher expression levels of hsp10, hsp28, hsp70ab, hsp83 and hsp90 mRNAs in A. m. jemenetica are key elements in the adaptive nature of A. m. jemenetica to local conditions that enhance its survival and fitness in high summer temperatures.