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白菜( Brassica campestris L. ssp. pekinensis )叶片和叶柄发育相关的 ceRNA 调控网络的全转录组分析与构建。

Whole transcriptome analysis and construction of a ceRNA regulatory network related to leaf and petiole development in Chinese cabbage (Brassica campestris L. ssp. pekinensis).

机构信息

Department of Horticulture, Shenyang Agricultural University, 120 Dongling Road, Shenhe District, Shenyang, 110866, China.

Vegetable Research Institute of Liaoning Academy of Agricultural Sciences, Shenyang, 110161, China.

出版信息

BMC Genomics. 2023 Mar 24;24(1):144. doi: 10.1186/s12864-023-09239-y.

Abstract

BACKGROUND

The growth and development of leaves and petioles have a significant effect on photosynthesis. Understanding the molecular mechanisms underlying leaf and petiole development is necessary for improving photosynthetic efficiency, cultivating varieties with high photosynthetic efficiency, and improving the yield of crops of which the leaves are foodstuffs. This study aimed to identify the mRNAs, long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and circular RNAs (circRNAs) related to leaf and petiole development in Chinese cabbage (Brassica campestris L. ssp. pekinensis). The data were used to construct a competitive endogenous RNA (ceRNA) network to obtain insights into the mechanisms underlying leaf and petiole development.

RESULTS

The leaves and petioles of the 'PHL' inbred line of Chinese cabbage were used as research materials for whole transcriptome sequencing. A total of 10,646 differentially expressed (DE) mRNAs, 303 DElncRNAs, 7 DEcircRNAs, and 195 DEmiRNAs were identified between leaves and petioles. Transcription factors and proteins that play important roles in leaf and petiole development were identified, including xyloglucan endotransglucosylase/hydrolase, expansion proteins and their precursors, transcription factors TCP15 and bHLH, lateral organ boundary domain protein, cellulose synthase, MOR1-like protein, and proteins related to plant hormone biosynthesis. A ceRNA regulatory network related to leaf and petiole development was constructed, and 85 pairs of ceRNA relationships were identified, including 71 DEmiRNA-DEmRNA, 12 DEmiRNA-DElncRNA, and 2 DEmiRNA-DEcircRNA pairs. Three LSH genes (BrLSH1, BrLSH2 and BrLSH3) with significant differential expression between leaves and petioles were screened from transcriptome data, and their functions were explored through subcellular localization analysis and transgenic overexpression verification. BrLSH1, BrLSH2 and BrLSH3 were nuclear proteins, and BrLSH2 inhibited the growth and development of Arabidopsis thaliana.

CONCLUSIONS

This study identifies mRNAs and non-coding RNAs that may be involved in the development of leaves and petioles in Chinese cabbage, and establishes a ceRNA regulatory network related to development of the leaves and petioles, providing valuable genomic resources for further research on the molecular mechanisms underlying leaf and petiole development in this crop species.

摘要

背景

叶片和叶柄的生长发育对光合作用有重要影响。了解叶片和叶柄发育的分子机制对于提高光合作用效率、培育具有高光效的品种以及提高以叶片为食的作物产量是必要的。本研究旨在鉴定与白菜( Brassica campestris L. ssp. pekinensis )叶片和叶柄发育相关的 mRNA 、长非编码 RNA ( lncRNA )、 microRNA ( miRNA )和 circular RNA ( circRNA )。利用这些数据构建了一个竞争性内源 RNA ( ceRNA )网络,以深入了解叶片和叶柄发育的机制。

结果

以白菜自交系“PHL”的叶片和叶柄为研究材料进行全转录组测序。共鉴定出叶片和叶柄之间差异表达的 10646 个 mRNA 、 303 个 DElncRNA 、 7 个 DEcircRNA 和 195 个 DEmiRNA 。鉴定出在叶片和叶柄发育中起重要作用的转录因子和蛋白,包括木葡聚糖内转糖苷酶/水解酶、扩张蛋白及其前体、转录因子 TCP15 和 bHLH 、侧生器官边界域蛋白、纤维素合酶、 MOR1 样蛋白和与植物激素生物合成相关的蛋白。构建了一个与叶片和叶柄发育相关的 ceRNA 调控网络,共鉴定出 85 对 ceRNA 关系,包括 71 个 DEmiRNA-DEmRNA 、 12 个 DEmiRNA-DElncRNA 和 2 个 DEmiRNA-DEcircRNA 对。从转录组数据中筛选出叶片和叶柄间差异表达显著的 3 个 LSH 基因( BrLSH1 、 BrLSH2 和 BrLSH3 ),并通过亚细胞定位分析和转基因过表达验证探索其功能。 BrLSH1 、 BrLSH2 和 BrLSH3 均为核蛋白, BrLSH2 抑制拟南芥的生长发育。

结论

本研究鉴定了可能参与白菜叶片和叶柄发育的 mRNAs 和非编码 RNA ,并建立了一个与叶片和叶柄发育相关的 ceRNA 调控网络,为进一步研究该作物叶片和叶柄发育的分子机制提供了有价值的基因组资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34b5/10039531/a59e84bb8f21/12864_2023_9239_Fig1_HTML.jpg

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