Screening of Differentially Expressed Microsporidia Genes from Nosema ceranae Infected Honey Bees by Suppression Subtractive Hybridization

Abstract

The microsporidium Nosema ceranae is a high prevalent parasite of the European honey bee (Apis mellifera). This parasite is spreading across the world into its novel host. The developmental process, and some mechanisms of N. ceranae-infected honey bees, has been studied thoroughly; however, few studies have been carried out in the mechanism of gene expression in N. ceranae during the infection process. We therefore performed the suppressive subtractive hybridization (SSH) approach to investigate the candidate genes of N. ceranae during its infection process. All 96 clones of infected (forward) and non-infected (reverse) library were dipped onto the membrane for hybridization. A total of 112 differentially expressed sequence tags (ESTs) had been sequenced. For the host responses, 20% of ESTs (13 ESTs, 10 genes, and 1 non-coding RNA) from the forward library and 93.6% of ESTs (44 ESTs, 28 genes) from the reverse library were identified as differentially expressed genes (DEGs) of the hosts. A high percentage of DEGs involved in catalytic activity and metabolic processes revealed that the host gene expression change after N. ceranae infection might lead to an unbalance of physiological mechanism. Among the ESTs from the forward library, 75.4% ESTs (49 ESTs belonged to 24 genes) were identified as N. ceranae genes. Out of 24 N. ceranae genes, nine DEGs were subject to real-time quantitative reverse transcription PCR (real-time qRT-PCR) for validation. The results indicated that these genes were highly expressed during N. ceranae infection. Among nine N. ceranae genes, one N. ceranae gene (AAJ76_1600052943) showed the highest expression level after infection. These identified differentially expressed genes from this SSH could provide information about the pathological effects of N. ceranae. Validation of nine up-regulated N. ceranae genes reveal high potential for the detection of early nosemosis in the field and provide insight for further applications.

Transcriptome of Nosema ceranae and Upregulated Microsporidia Genes during Its Infection of Western Honey Bee (Apis mellifera)

“…Therefore, the upregulation of AAJ76_1600052943 and AAJ76_2000141845 might also play roles similar to those mentioned above to facilitate spore germination and mature spore development. The highly expressed SPW was consistent with our previous study on N. ceranae gene expression in A. mellifera [ 33 ] and the research on the N. ceranae gene expression in A. cerana, suggesting a significant role for SPW during N. ceranae infection [ 53 ].…”

“…Moreover, the genome of N. ceranae has been studied and reported for a better understanding of the architecture, regulation, and evolution of this pathogen [ 31 , 32 ]. From our previous study, the identification of N. ceranae -specific genes during its infection of A. mellifera by suppressive subtractive hybridization (SSH) has been performed [ 33 ], but comprehensive data on the gene expression profile during the infection process are still unavailable. In this study, we attempted to reveal the transcriptomic profiling of N. ceranae during its infection of A. mellifera to complete the information on the infection cycle of this parasite.…”

Fungi of entomopathogenic potential in Chytridiomycota and Blastocladiomycota, and in fungal allies of the Oomycota and Microsporidia

“…Microsporidial infection can also alter host metabolism and induce both local and systemic innate immune responses (Pan et al 2018). For example, N. ceranae infection is believed to suppress immune defence mechanisms in honey bees; studies have indicated that infection downregulates some immune-related genes, including abaecin, apidecin, defensin, hymenoptaecin, glucose dehydrogenase (GLD) and vitellogenin (Vg), and upregulates Jun-related antigen Jra and pi3k (Chang et al 2020). Additionally, transcription of the inhibitor of apoptosis protein (iap-2) gene was found to be upregulated in Nosema-tolerant honeybees (Kurze et al 2015); it is possible that inhibition of apoptosis might help the parasites optimize their environment and extend the period during which they can grow and differentiate within host cells (Higes et al 2013).…”

Monitoring the Season–Prevalence Relationship of Vairimorpha ceranae in Honey Bees (Apis mellifera) over One Year and the Primary Assessment of Probiotic Treatment in Taichung, Taiwan

“…Infection by V. ceranae weakens A. mellifera , and severe cases can result in bee mortality, which is often correlated with significant colony losses. These infections are also associated with various physiological impairments, contributing to reduced honey production, malnutrition, shortened lifespans, and increased mortality among adult honey bees [ 2 , 22 ].…”