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利用 Smart-seq2 对 Neopyropia yezoensis 中的无性孢囊和减数分裂相关的合子进行 RNA-seq 分析。

RNA-seq between asexual archeospores and meiosis-related conchospores in Neopyropia yezoensis using Smart-seq2.

机构信息

CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.

Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China.

出版信息

J Phycol. 2021 Oct;57(5):1648-1658. doi: 10.1111/jpy.13197. Epub 2021 Aug 4.

Abstract

In the life cycle of Neopyropia yezoensis, a potential model system for marine macroalgae, both asexual archeospores and meiosis-related conchospores develop into thalli (gametophyte). To understand this special life phenomenon in macroalgae, we picked out the two kinds of spores (10-30 cells in each sample) and conducted RNA-seq using Smart-seq2. Comparative analysis showed that light capture and carbon fixation associated differentially expressed genes (DEGs) were upregulated in archeospores, thus indicating that archeospores are in a state of rapid vegetative growth. In conchospores, protein synthesis and degradation, especially molecular chaperone, associated DEGs were up-regulated, indicating that complex life activities might be occurring in conchospores. There were 68 genes related to DNA replication and repair expressed in conchospores, showing that active DNA replication might occur in conchospores. Moreover, we found that one conchospore specifically expressed DEG (py04595: DNA helicase) only in diploid stages (conchocelis, sporangial filament) and three archeospores specifically expressed DEGs only in haploid stages (thalli). These molecular level results indicated that conchospores were closer to diploid, and might be the meiotic mother cells of N. yezoensis. In addition, we found that the knotted-like homeobox gene (PyKNOX), which might relate to the transition of gametophyte from sporophyte, was only expressed in sporophyte generation but not expressed in conchospores, archeospores and thalli, indicating the morphogenesis of gametophyte sin N. yezoensis might require the inactivation of PyKNOX.

摘要

在 Neopyropia yezoensis 的生命周期中,一种潜在的海洋大型藻类模式系统,无性的原丝体孢子和减数分裂相关的壳孢子都发育成叶状体(配子体)。为了理解这种大型藻类的特殊生命现象,我们挑选了这两种孢子(每个样本 10-30 个细胞),并使用 Smart-seq2 进行了 RNA-seq。比较分析表明,光捕获和碳固定相关差异表达基因(DEGs)在原丝体孢子中上调,表明原丝体孢子处于快速营养生长状态。在壳孢子中,蛋白质合成和降解,特别是分子伴侣,相关的 DEGs 上调,表明壳孢子中可能发生复杂的生命活动。有 68 个与 DNA 复制和修复相关的基因在壳孢子中表达,表明壳孢子中可能发生活跃的 DNA 复制。此外,我们发现一个壳孢子特异性表达的 DEG(py04595:DNA 解旋酶)仅在二倍体阶段(壳状幼体、孢子丝体)表达,三个原丝体孢子特异性表达的 DEG 仅在单倍体阶段(叶状体)表达。这些分子水平的结果表明壳孢子更接近二倍体,可能是 N. yezoensis 的减数分裂母细胞。此外,我们发现与配子体从孢子体过渡相关的结状同源盒基因(PyKNOX)仅在孢子体世代表达,而不在壳孢子、原丝体孢子和叶状体中表达,表明 N. yezoensis 配子体的形态发生可能需要 PyKNOX 的失活。

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