利用单细胞测序技术发掘小立碗藓发育过程中的差异基因

	利用单细胞测序技术发掘小立碗藓发育过程中的差异基因; Transcriptomic profiling of Physcomitrium patens development by single cell sequencing
	周士钊
导师	吴建强
摘要	Bryophyta is the very early land plant in the evolution from aquatic to terrestrial. During the landing process, it displayed a unique morphological change. It will help us to have a deeper understanding of the evolution of terrestrial plants to study the mechanism of morphological change. The transformation from 2-D protonema to 3-D gametophyte is an important stage in the development of Bryophyta. Compared with 2-D protonema, the 3-D leaf-like gametophyte enables P.patens to capture more light and carbon dioxide from air , which potentially increased its photosynthesis and thus enhancing its adaptabiliy to the terrestrial environment. However, due to technical limitations, only a few genes involves in this transformation process have been identified. In recent years, the rise of single-cell sequencing technology has provided us a new opportunity to analyze plant development at single-cell resolution. In this study, microfluidic single-cell transcriptome was used to analyze the transcriptional changes in transformation process,trying to identify specific genes and key genes that involved in transformation process of Physcomitrium patens(P.patens). Transgenic methods were used to verify the functions of specific candidate genes. The main research results are as follows: 1. Protoplasts were successfully prepared with diameters less than 40 μm and cell activity more than 85%. After that, three replicates were performed for single-cell transcriptome sequencing using approximately 8000 cells per sample. After filtering out the cells with expressed-genes detected less than 200(which may be impurity or dead cells), the qualified cell number was about 7000. 2. In order to find out the differential expressed genes during the development of wild-type P.patens, cells were clustered to 9 clusters, which were identified and named as gametophyte (cluster 0), transformation (cluster 3, 5), protonema (cluster 1, 2, 4), chloronema (cluster 3, 5, 7) and caulonema (cluster 6). 3. Further analysis of the transcription factors in different genes showed that the cell-specific transcription factors were enriched in cluster3 and cluster5, mainly including WRKY, WOX, NAC, MYB, LBD, GRAS and AP2 / ERF. This finding was consistent with the increased number and type of transcription factors in the evolution from aquatic plants to terrestrial plant species, which suggested that cluster 3 and 5 may be closely related to the transition period from 2-D growth into a 3-D growth. Thus these two clusters were named “transformation cells”. Notably, AP2/ERF transcription factors, that had been reported to play an important role in transition, also were found in cluster 3 and cluster 5, indicating that our analytical method is reliable and accurate. 4. Pseudo-time analysis of cluster 3 and cluster 0 cells showed that the presumed developmental trajectory of the cells is from cluster 3 to cluster 0, which was consistent with the developmental state of the cells. Moreover, the express
	2021-05
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74561
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	周士钊. 利用单细胞测序技术发掘小立碗藓发育过程中的差异基因, Transcriptomic profiling of Physcomitrium patens development by single cell sequencing[D],2021.

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