Abstract
Increasing leaf chlorophyll content in crops through genetic improvement is a feasible way to increase crop yield. However, the genetic mechanism that regulates the chlorophyll content of rapeseed leaves is not clear, which is not conducive to improving it through genetic pathways. In this study, a population of 189 recombinant inbred lines of Brassica napus was used to identify quantitative trait loci (QTLs) that regulate leaf chlorophyll content. A total of 12 QTLs were identified, of which two QTLs, qCHC.A6-2 and qCHC.C5-1, explained 10.19% and 13.70% of the phenotypic variations in chlorophyll content, respectively, and were considered to be major QTLs. Furthermore, five of the 12 QTLs were inherited from APL01, while seven were inherited from Holly. Comparative transcriptomic analysis revealed that the differential expression of genes associated with the response to light stimulation, chloroplast organization, and glutathione peroxidase activity is the molecular basis for the difference in leaf chlorophyll content between APL01 and Holly. Further combining the variation analysis of the genomic DNA sequence and the functional annotation of homologous genes, four promising candidate genes (BnaA06g37690D, BnaA06g37710D, BnaC05g00280D, and BnaC05g00300D) were obtained, all of which are potentially involved in the response to light stimulation and/or chloroplast organization. These results help to clarify the genetic mechanism of chlorophyll content in B. napus leaves.
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Acknowledgements
This study was funded by Guizhou Provincial Science and Technology Plan Project (Qian Kehe Support [2022]key026); National Natural Science Foundation of China (32060495); Jiangsu Agricultural Science and Technology Innovation Fund (CX(19)3053); Guizhou University Cultivation Project (Guizhou University Cultivation [2019]47); Key Laboratory of Molecular Breeding for Grain and Oil Crops in Guizhou Province (Qiankehezhongyindi (2023) 008); Key Laboratory of Functional Agriculture of Guizhou Provincial Higher Education Institutions (Qianjiaoji (2023) 007). The authors would like to thank Beijing Baimaike Biotechnology Co., Ltd for its great assistance in RNA sequencing. We thank International Science Editing (https://www.internationalscienceediting.com/) for editing this manuscript.
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YL and LC co-wrote the first draft of the manuscript. KY, ZC and XW designed the project and finalized the manuscript. KY and XW acquired funding. YL, LC, YH and TD measured leaf chlorophyll content in the AH population. KY, ET, XW, JZ and YL conducted QTL mapping for leaf chlorophyll content. KY, YH, YL, TD and ET conducted transcriptomics analysis. All authors have reviewed and approved the final version of the manuscript and therefore are equally responsible for the integrity and accuracy of its content.
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Supplementary File 1
The topGO directed acyclic graph of GO terms significantly enriched by up-regulated DEGs in APL01 compared to Holly. (A) The topGO directed acyclic graph of GO terms related to biological processes, (B) cellular components, and (C) molecular functions. In each graph, the square represents the GO terms that were most significantly enriched by DEGs, while the oval represents the GO terms that were less significantly enriched. The description and enrichment significance value (KS) of the GO term are given in each square (or oval). The darker are the color of the squares and ovals, the higher the significance of enrichment. Light yellow represents the range of KS values from 0.5 to 1.0, dark yellow represents the range of KS values from 0.051 to 0.49, orange represents the range of KS values from 0.005 to 0.05, Orange-red represents the range of KS values from 0.0007 to 0.0049, and red represents the range of KS values from 0 to 0.00069. The hierarchical relationship between the GO terms in each diagram was determined based on the functional descriptions described by each GO term, which become increasingly specific from top to bottom. The terms with the most specific functional descriptions located at the extreme ends of each branch are also shown in Figure 2. (PDF 222 kb)
Supplementary Table 1
Epistatic effects between QTLs detected by ICIM-EPI mapping method. (XLS 32 kb)
Supplementary Table 2
Expression levels and functional annotations of genes differentially expressed in APL01 compared to Holly. (XLS 5837 kb)
Supplementary Table 3
TopGO enrichment statistics of up-regulated genes in APL01 compared to Holly. (XLS 4783 kb)
Supplementary Table 4
KEGG enrichment statistics of up-regulated genes in APL01 compared to Holly. (XLS 22 kb)
Supplementary Table 5
DNA sequence variation between APL01 and Holly within the qCHC.A6-2 and qCHC.C5-1 mapping interval detected by genome resequencing. (XLS 156 kb)
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Yuanhong, L., Lei, C., Yuqi, H. et al. Quantitative Trait Locus Mapping Combined with RNA Sequencing Reveals Candidate Genes for Chlorophyll Content in Oilseed Rape Leaves. J Plant Growth Regul 43, 1260–1271 (2024). https://doi.org/10.1007/s00344-023-11181-y
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DOI: https://doi.org/10.1007/s00344-023-11181-y