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不同增殖速度的东海原甲藻细胞周期及转录组比较分析

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为研究不同增殖速度的东海原甲藻(Prorocentrum donghaiense)细胞周期时相分布特点以及内在调控机理,利用流式细胞检测技术比较了两种不同增殖速度的东海原甲藻细胞周期时相差异,并利用RNA-seq技术对两组细胞转录模式进行了比较分析。结果显示,相比增殖速度较快的实验组,增殖速度较慢的实验组S期更短,且部分细胞阻滞在G2/M期。因此,S期的持续时间和G2/M期转换是否被阻滞可以决定东海原甲藻的增殖速率。两组细胞差异表达基因富集程度最大且富集最显著的GO分类均与微管和细胞骨架有关,暗示两实验组细胞周期进程的差异由细胞中的微管动态变化不同造成。注释到49个细胞周期蛋白编码基因、74个细胞周期蛋白依赖性激酶编码基因和26个细胞分裂周期蛋白编码基因。其中,1个CYCB和2个CDK1基因与调节东海原甲藻G2/M期阻滞有关,1个CYCA、1个CYCU、4个CDK2和1个Cdc48基因与藻细胞S期进程调控有关。
Comparative analysis of the cell cycle progression and transcriptome of Prorocentrum donghaiense with different proliferation rates
Prorocentrum donghaiense is one of the main red tide organisms in China's coastal waters. It is characterized by frequent outbreaks,a large area of influence,and serious harm. The cell cycle is an important biological process that regulates cell division. It can be affected by environmental factors,resulting in changes in the growth rate of phytoplankton. Therefore,it is valuable to investigate the differences in the cell cycle progression of P. donghaiense with different proliferation rates to help understanding its proliferation characteristics. The cell cycle pattern of P. donghaiense has been preliminary explored. Genes and proteins related to the cell cycle have been isolated and identified using omics approaches. However,there is a lack of research on the characteristics of the cell cycle with different proliferation rates,as well as the molecular mechanism. Our previous studies demonstrated that the growth rate of P. donghaiense varies under different environmental conditions. To compare the cell cycle progression of P. donghaiense with different proliferation rates and understand its molecular mechanism,we selected two experimental groups with slower (group 3) and faster (group 16) proliferation rates,and analyzed them using flow cytometry and RNA sequencing technology. The results showed that group 3 cells had a shorter S phase and were partially blocked in G2/M phase. A total of 149219 unigenes were obtained,of which 6081 were annotated in the NR,GO,KEGG,eggNOG,Swiss-Prot,and Pfam databases. A total of 114358 unigenes were classified into three terms,namely,cellular components,molecular functions,and bioprocesses,with 57 categories in the GO database. The 30554 unigenes annotated in the eggNOG database were classified into 25 categories. GO classification and pathway enrichment results of differentially expressed genes (DEGs) between the two experimental groups showed that the most important DEGs were related to microtubules and the cytoskeleton. We hypothesized that the differences in cell cycle progression between the two experimental groups were caused by differences in the dynamic changes of microtubules. Among the annotated unigenes,49 encoding cyclins,74 encoding cyclin-dependent kinases (CDKs),and 26 encoding cell division cycle (CDC) proteins were related to cell cycle control. Ten of these comprising one CYCA,one CYCB,one CYCU,one CDK1,four CDK2,and one Cdc48 genes were DEGs. The cyclinA-CDK2 and CDK1-cyclinB complexes participate in the cellular G2/M phase transition. Cells need to retain sufficient levels of CDK activity to maintain the transcription of G2-specific genes and retain their competence to recover from G2 arrest. Since cells in group 3 were partially arrested in G2/M,the transcriptional levels of CDKs,cylinA,and cyclinB were higher in group 3 than in group 16. As Cdc48 is involved in cellular DNA replication,the transcription level of Cdc48 was significantly higher in group 16 than in group 3,which explained why the S phase duration was longer in group 16 than in group 3. In summary,the S phase duration and whether the G2/M phase transition is blocked may decide the rate of P. donghaiense cell proliferation,and one CYCA,one CYCB,one CYCU,two CDK1,four CDK2,and one Cdc48 genes identified from the transcriptome analysis played important roles in cell cycle regulation.

Prorocentrum donghaienseproliferation ratecell cycle phasetranscriptomedifferentially expressed genesred tide

赵鹏珍、王云龙、马欣云、全为民、李楠楠、陈渊戈、范瑞良、张海燕、徐清、欧阳珑玲

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上海海洋大学水产与生命学院,上海 201306

中国水产科学研究院东海水产研究所,农业农村部东海渔业资源开发利用重点实验室,上海 200090

自然资源部宁波海洋中心,浙江 宁波 315012

东海原甲藻 增殖速度 细胞周期时相 转录组 差异表达基因 赤潮

2024

中国水产科学
中国水产科学研究院 中国水产学会

中国水产科学

CSTPCD北大核心
影响因子:1.206
ISSN:1005-8737
年,卷(期):2024.31(9)