Screening of Differentially Expressed Microsporidia Genes from Nosema ceranae Infected Honey Bees by Suppression Subtractive Hybridization
| Autor: | Chong Yu Ko, Ming Ren Yen, Yu-Shin Nai, Zih Ting Chang, Yue Wen Chen |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
cDNA subtraction
Genetics 0303 health sciences Candidate gene Expressed sequence tag biology 030306 microbiology food and beverages Honey bee Nosema ceranae biology.organism_classification 03 medical and health sciences CDNA Subtraction Suppression subtractive hybridization Insect Science Microsporidia Gene expression honey bee lcsh:Q lcsh:Science Gene 030304 developmental biology |
| Zdroj: | Insects Volume 11 Issue 3 Insects, Vol 11, Iss 3, p 199 (2020) |
| ISSN: | 2075-4450 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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