Abstract
The generation of adventitious roots (ARs) is the key to the success of cuttings. The appropriate environment for AR differentiation in tea plants is acidic. However, the mechanism is unclear. In this study, pH 4.5 was suitable condition for the differentiation of AR in tea plants. At the base of cuttings, the root primordia differentiated ARs more rapidly at pH 4.5 than pH 7.0, and nine AR differentiation-related genes were found to be differentially expressed in 30 days, the result was also validated by qRT-PCR. The promoter regions of these genes contained auxin and brassinosteroid response elements. The expression levels of several genes which were involved in auxin and brassinosteroid synthesis as well as signaling at pH 4.5 compared to pH 7.0 occurred differential expression. Brassinolide (BL) and indole-3-acetic acid (IAA) could affect the differentiation of ARs under pH 4.5 and pH 7.0. By qRT-PCR analysis of genes during ARs generation, BL and IAA inhibited and promoted the expression of CsIAA14 gene, respectively, to regulate auxin signal transduction. Meanwhile, the expression levels of CsKNAT4, CsNAC2, CsNAC100, CsWRKY30 and CsLBD18 genes were up-regulated upon auxin treatment and were positively correlated with ARs differentiation.This study showed that pH 4.5 was the most suitable environment for the root primordia differentiation of AR in tea plant. Proper acidic pH conditions promoted auxin synthesis and signal transduction. The auxin initiated the expression of AR differentiation-related genes, and promoted its differentiated. BL was involved in ARs formation and elongation by regulating auxin signal transduction.
Key message
Acidic environments promote auxin synthesis and regulate root primordia differentiation to form ARs by initiating AR differentiation-related genes expression in Camellia sinensis.
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The dataset used and/or analyzed in this study can be obtained from the author according to reasonable requirements.
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Acknowledgements
We thank the equipment support provided by Institute of Agro-bioengineering and the College of Life Sciences of Guizhou University.
Funding
This work was supported by National Natural Science Foundation of China, "Functional analysis of Brassinosteroid transcription factor CsBZRs in Camellia sinensis under cold stress" (Grant numbers: 32160077). Talent program of the Guizhou Academy of Agricultural Sciences, "Research on functional genomics and high-value utilization technology of important economic plants such as Eucommia ulmoides"[(2022) No.02]. Guizhou Provincial Talent Office "Guizhou Provincial Characteristic Plant Germplasm Resources Utilization and Innovation Talent Base" (RCJD2018-14). Guizhou Provincial High-level Innovative Talent Training Project [QKH Talent (2016) No. 4003].
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11103_2023_1383_MOESM1_ESM.tif
Supplementary file1 (TIF 11661 KB)—Transcriptome sequencing sample location map and auxin standard curve. A The red circle is the sampling site for transcriptome sequencing. B Preparation of auxin standard. The X-axis represents auxin concentration, the Y-axis represents peak area.
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Supplementary file2 (PNG 16 KB)—Number of DEGs. Red represents the DEGs number that is up−regulated, and blue represents the DEGs number that is down−regulated. The x−axis represents the number of corresponding differential genes (DEGs), and the y−axis represents the difference comparison scheme for group.
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Supplementary file3 (TIF 13702 KB)—GO classification of the DEGs. A GO annotation for DEGs. The x−axis is the number of genes in the annotation, and the y−axis represents GO categories of gene function. B Enrichment of the GO annotation of the DEGs.The x−axis represents the percentage of DEGs belonging to the corresponding pathway, and the y−axis representsthe top 20 pathways. The sizes of bubbles represent the number of DEGs in the corresponding pathway, and the colors of the bubbles represent the enrichment P−adjust of the corresponding pathway.
11103_2023_1383_MOESM4_ESM.tif
Supplementary file4 (TIF 18831 KB)—KEGG classification of the DEGs. A and B KEGG annotation for DEGs.The x−axis is represents KEGG categories of gene function, and the y−axis is the number of genes in the annotation. C Enrichment of the KEGG annotation of the DEGs.The x−axis represents the percentage of DEGs belonging to the corresponding pathway, and the y−axis representsthe top 20 pathways. The sizes of bubbles represent the number of DEGs in the corresponding pathway, and the colors of the bubbles represent the enrichment P−adjust of the corresponding pathway.
11103_2023_1383_MOESM5_ESM.tif
Supplementary file5 (TIF 15383 KB)—Enrichment map of KEGG pathway differentially expressed genes in BR and IAA metabolism as well as signal transduction. A KEGG signaling pathway of phytohormone (map04075). B KEGG metabolic pathway of IAA (map00380). C KEGG metabolic pathway of BR (map00905). Red boxes indicate up-regulation of gene expression and blue boxes for down-regulation.
11103_2023_1383_MOESM6_ESM.tif
Supplementary file6 (TIF 1236 KB)—Auxin content at the base of cuttings in the differentiation stage of AR (30 d) under pH 7.0 and pH 4.5. Standard curve: y=37642x -2208.9, R2=0.9999. y: Peak area; x: Concentration. The x-axis represents the pH value and the y-axis represents auxin content. The error bar represents ± SEs, n ≥3. Significant differences at p<0.05 by t-test and is labeled with lower case letters.Different letters showed significant differences (P<0.05), while the same letters showed no significant differences (P>0.05).
11103_2023_1383_MOESM7_ESM.tif
Supplementary file7 (TIF 22343 KB)—The expression trend of DEGs in transcriptome was verified by qRT-PCR. The relative expression amount is calculated as 2−ΔΔCt. The left y-axis is the result of qRT-PCR, and the right y-axis is the result of RNA-seq. The error bar represents ±SEs, n≥3. Different letters showed significant differences (P<0.05), while the same letters showed no significant differences (P>0.05).
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Liu, K., Zhao, Y. & Zhao, DG. Transcriptome analysis reveals the effect of acidic environment on adventitious root differentiation in Camellia sinensis. Plant Mol Biol 113, 205–217 (2023). https://doi.org/10.1007/s11103-023-01383-z
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DOI: https://doi.org/10.1007/s11103-023-01383-z