Male germline specification is a crucial step in double fertilization in flowering plants. Determining which genes are activated in the male germline, and how those genes are regulated, is essential to understand double fertilization. However, transcription activities in the male germline may be not easy to detect due to low-level expression of some genes and technical difficulties of isolating male germline cells. Here, through a series of gene reporter assays in Arabidopsis, we showed a weak male germline expression pattern of the SOLO DANCERS (SDS) gene, which was confirmed by RT-PCR. Compared to directly fusing the SDS sequence with GFP, adding the coding sequences of other genes such as CDKA1 can greatly enhance the detection of the male germline expression pattern of SDS. We found that SDS expression in the male germline is activated by a novel pathway that differs from the well-known DUO1 regulon. We also developed an SDS-based fluorescence reporter to analyze posttranscriptional regulation in the male germline. Our data suggest that stable gene products of CDKA1 and others may enhance the sensitivity of gene reporters, and the male germline may use diverse pathways to activate gene expression.

中文翻译：

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使用周期蛋白依赖性激酶 A1 编码序列检测雄性生殖系中 SOLO DANCERS 的弱表达

雄性种系规范是开花植物双重受精的关键步骤。确定哪些基因在雄性生殖系中被激活，以及这些基因是如何被调控的，对于理解双受精至关重要。然而，由于某些基因的低水平表达和分离雄性生殖细胞的技术困难，雄性生殖细胞中的转录活性可能不易检测。在这里，通过拟南芥中的一系列基因报告分析，我们显示了 SOLO DANCERS (SDS) 基因的弱雄性种系表达模式，这通过 RT-PCR 得到证实。与直接将SDS序列与GFP融合相比，加入CDKA1等其他基因的编码序列可以大大增强对SDS雄性生殖系表达模式的检测。我们发现雄性生殖系中的 SDS 表达被一种不同于众所周知的 DUO1 调节子的新途径激活。我们还开发了一种基于 SDS 的荧光报告基因来分析雄性生殖系的转录后调控。我们的数据表明，CDKA1 和其他稳定基因产物可能会增强基因报告基因的敏感性，而雄性种系可能会使用多种途径来激活基因表达。