Liu, H and Hu, D and Du, P and Wang, L and Liang, X and Li, H and Lu, Q and Li, S and Liu, H and Chen, X and Varshney, R K and Hong, Y (2021) Single-cell RNA-seq describes the transcriptome landscape and identifies critical transcription factors in the leaf blade of the allotetraploid peanut (Arachis hypogaea L.). Plant Biotechnology Journal (TSI), 19 (11). pp. 2261-2276. ISSN 1467-7644
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Abstract
Single-cell RNA-seq (scRNA-seq) has been highlighted as a powerful tool for the description of human cell transcriptome, but the technology has not been broadly applied in plant cells. Herein, we describe the successful development of a robust protoplast cell isolation system in the peanut leaf. A total of 6,815 single cells were divided into eight cell clusters based on reported marker genes by applying scRNA-seq. Further, a pseudo-time analysis was used to describe the developmental trajectory and interaction network of transcription factors (TFs) of distinct cell types during leaf growth. The trajectory enabled re-investigation of the primordium-driven development processes of the mesophyll and epidermis. These results suggest that palisade cells likely differentiate into spongy cells, while the epidermal cells originated earlier than the primordium. Subsequently, the developed method integrated multiple technologies to efficiently validate the scRNA-seq result in a homogenous cell population. The expression levels of several TFs were strongly correlated with epidermal ontogeny in accordance with obtained scRNA-seq values. Additionally, peanut AHL23 (AT-HOOK MOTIF NUCLEAR LOCALIZED PROTEIN 23), which is localized in nucleus, promoted leaf growth when ectopically expressed in Arabidopsis by modulating the phytohormone pathway. Together, our study displays that application of scRNA-seq can provide new hypotheses regarding cell differentiation in the leaf blade of Arachis hypogaea. We believe that this approach will enable significant advances in the functional study of leaf blade cells in the allotetraploid peanut and other plant species.
| Item Type: | Article |
|---|---|
| Divisions: | Center of Excellence in Genomics and Systems Biology |
| CRP: | UNSPECIFIED |
| Uncontrolled Keywords: | scRNA-seq, plant singlecell, leaf cell, leaf development, peanut improvement |
| Subjects: | Others > Crop Improvement Mandate crops > Groundnut |
| Depositing User: | Mr Nagaraju T |
| Date Deposited: | 06 Feb 2025 05:03 |
| Last Modified: | 06 Feb 2025 05:03 |
| URI: | http://oar.icrisat.org/id/eprint/12939 |
| Official URL: | https://onlinelibrary.wiley.com/doi/full/10.1111/p... |
| Projects: | UNSPECIFIED |
| Funders: | UNSPECIFIED |
| Acknowledgement: | This study was supported by Key-Area Research and Development Program of the Guangdong Province (2020B020219003), National Natural Science Foundation of China (32001442), Guangdong Basic and Applied Basic Research Foundation (2020A1515010021), China Agriculture Research System of MOF and MARA (CARS-13), Technology Special Fund of Guangdong Province Agriculture and Rural Affairs Department (2019KJ136-02), Agricultural Competitive Industry Discipline Team Building Project of Guangdong Academy of Agricultural Sciences (202104TD), Special Fund for Scientific Innovation Strategy-Construction of High Level Academy of Agriculture Science (R2016PY-JX001, R2017YJ-YB2001, R2018PY-JX001, R2018QD-049, R2018QD-050, R2019PY-QF006). R.K.V. is thankful to Science & Engineering Research Board (SERB) of Department of Science & Technology (DST), Government of India for providing the J C Bose National Fellowship (SB/S9/Z-13/2019). |
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