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Integrative analysis in WGBS and transcriptome profiles of an temperature sensitive etiolation tea plant cultivar ‘Zhonghuang 2’

ABSTRACT: The green tea made from the etiolation shoots and leaves have fresh taste and strong aroma. ‘Zhonghuang 2 (ZH2) is a natural etiolation mutant and its shoots are etiolation in spring and turn green in summer. Two conditions including the relative high temperature (LT, 26 / 22oC) and the relative low temperature (LT, 18 / 14oC) were to find out the etiolated reason of ZH2. We confirmed ZH2 was a low-temperature sensitive tea plant cultivar, and the reason of shoots etiolation at LT was the reduction of stacked grana, which could cause the lack of photoreaction. We identified 1279 differentially expressed genes (DEGs), most of them were related to cytochrome synthesis and chloroplast development and function by RNA-seq. Through the whole-genome bisulfite sequencing (WGBS) assay, we found that the dynamic of DNA methylation level in three contexts (CG, CHG and CHH) was all decreased at LT, and the change was most obvious in the CHH context. According the result of cross analysis, 56 genes were identified with significant changes in both expression and DNA methylation levels. A large number of them were related to chloroplast development and chlorophyll synthesis. This study demonstrated that the DNA methylation was involved in the low-temperature regulated etiolation process of young shoots in ZH2. Change of the DNA methylation level was associated with the gene expression level, thus affecting the structure and function of chlorophyll and chloroplast, even reduced photosynthesis, which might have a phenotypic impact on the color of shoots and leaves. And it laid the foundation for studying the etiolation process from an epigenetic perspective.

ORGANISM(S): Camellia sinensis

PROVIDER: GSE234849 | GEO | 2023/10/19

REPOSITORIES: GEO

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Project description:Background: Low temperature (LT) often occurs at the seedling stage in the early rice-growing season, especially for direct seeded early-season indica rice, and using flooding irrigation can mitigate LT damage in rice seedlings. The molecular mechanism by which flooding mitigates the damage induced by LT stress has not been fully elucidated. Thus, LT stress at 8C, LT accompanied by flooding (LTF) and CK (control) treatments were established for three days to determine the transcriptomic, proteomic and physiological response in direct seeded rice seedlings at the seedling stage. Results: LT damaged chloroplasts, and thylakoid lamellae, and increased osmiophilic bodies and starch grains compared to CK, but LTF alleviated the damage to chloroplast structure caused by LT. The physiological characteristics of treated plants showed that compared with LT, LTF significantly increased the contents of rubisco, chlorophyll, PEPCK, ATP and GA3 but significantly decreased soluble protein, MDA and ABA contents. 4D-label-free quantitative proteomic profiling showed that photosynthesis-responsive proteins, such as phytochrome, as well as chlorophyll and the tricarboxylic acid cycle were significantly downregulated in LT/CK and LTF/CK comparison groups. However, compared with LT, phytochrome, chlorophyllide oxygenase activity and the glucan branching enzyme in LTF were significantly upregulated in rice leaves. Transcriptomic and proteomic studies identified 72818 transcripts and 5639 proteins, and 4983 genes that were identified at both the transcriptome and proteome levels. Differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were significantly enriched in glycine, serine and threonine metabolism, biosynthesis of secondary metabolites, glycolysis/gluconeogenesis and metabolic pathways. Conclusion: Through transcriptomic, proteomic and physiological analyses, we determined that a variety of metabolic pathway changes were induced by LT and LTF. GO and KEGG enrichment analyses demonstrated that DEGs and DEPs were associated with photosynthesis pathways, antioxidant enzymes and energy metabolism pathway-related proteins. Our study provided new insights for efforts to reduce the damage to direct seeded rice caused by low-temperature stress and provided a breeding target for low temperature flooding-resistant cultivars. Further analysis of translational regulation and metabolites may help to elucidate the molecular mechanisms by which flooding mitigates low-temperature stress in direct seeded early indica rice at the seedling stage.

Project description:Purpose: Many studies have demonstrated that ZFPs have important functions in many biological processes, including plant growth and development, stress response, and phytohormone response. RanBP2-type zinc finger transcription factors have been characterized in animals and humans. However, their functions remain largely unknown in plants. Therefore, the functions of RanBP ZFPs in plants need further study to shed new insights into the importance of these transcription factors in developmental processes. Methods: The transcriptomes of between overexpression of SlRBZ transgenic and wild-type tomato by RNA-seq analysis were evaluated using the Illumina HiSeqâ¢ 2000 sequencing platform. Raw sequences were filtered and the resulting sets of clean reads were used for the following analysis by Tophat and DEGseq software. qRTâPCR validation was performed using SYBR Green assays. Results: In this study, we identified a RanBP2-type zinc finger protein gene (SlRBZ) in tomato. SlRBZ was constitutively expressed in roots, stems, leaves, flowers and fruits. Overexpression of SlRBZ resulted in etiolated and dwarf phenotypes in tomato. Determination of physiological index showed that chlorophyll, carotenoid, and GAs contents were evidently decreased in transgenic plants. Furthermore, the qRT-PCR and RNA-Seq analyses demonstrated that the transcription of the genes involved in these biosynthesis pathways obviously decreased in SlRBZ-OE plants. In addition, ultrastructural observation by transmission electron microscopy indicated that plastids could not develop into mature chloroplasts with normal chloroplast membrane and thylakoid membrane system in SlRBZ-OE plants. Conclusions: The results suggest that overexpression of SlRBZ may impair the biosynthesis of chlorophyll, carotenoid, and gibberellin through blocking chloroplast development, resulting in etiolation and dwarfism in tomato. The transcriptomes of between overexpression of SlRBZ transgenic and wild-type tomato by RNA-seq analysis were evaluated, in duplicate, using the Illumina sequencing platform.

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Integrative analysis in WGBS and transcriptome profiles of an temperature sensitive etiolation tea plant cultivar ‘Zhonghuang 2’

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