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.

中文翻译：

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抑制消减杂交技术筛选Nosema ceranae感染蜜蜂的差异表达微孢子虫基因

微小孢子虫（Nosema ceranae）是欧洲蜜蜂（Apis mellifera）的高寄生虫。这种寄生虫正在世界范围内传播到其新颖的宿主中。深入研究了猪笼草感染蜜蜂的发育过程以及一些机理。然而，在感染过程中，对于中华猪笼草的基因表达机制尚未进行过研究。因此，我们进行了抑制性消减杂交（SSH）方法来研究中华猪笼草的候选基因。在其感染过程中。将感染的（正向）和未感染的（反向）文库的所有96个克隆浸入膜上进行杂交。总共112个差异表达的序列标签（EST）已测序。对于宿主反应，来自正向文库的20％EST（13个EST，10个基因和1个非编码RNA）和来自逆向文库的93.6％的EST（44 EST，28个基因）被鉴定为差异表达基因（主机）。参与催化活性和代谢过程的DEGs的高百分比表明，在猪笼草感染后宿主基因表达的变化可能导致生理机制的失衡。在前向文库的EST中，有75.4％的EST（49个EST属于24个基因）被鉴定为cer.ceranae基因。在24个cer。ceranae基因中，对9个DEG进行了实时定量逆转录PCR（real-time qRT-PCR）的验证。结果表明，这些基因在cerenae ceranae感染期间高表达。间9个N. ceranae基因，一个N. ceranae基因（AAJ76_1600052943）显示感染后的最高表达水平。这些从SSH鉴定的差异表达基因可以提供关于猪笼螺的病理效应的信息。九种上调的猪笼草基因的验证揭示了该领域检测早期鼻涕病的高潜力，并为进一步的应用提供了见识。