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Photosynthetic characteristics and genetic mapping of a new yellow leaf mutant crm1 in Brassica napus

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Abstract

Chlorophyll is one of the key factors for photosynthesis and plays an important role in plant growth and development. We previously isolated an EMS mutagenized rapeseed chlorophyll-reduced mutant (crm1), which had yellow leaf, reduced chlorophyll content and fewer thylakoid stacks. Here, we found that crm1 showed attenuated utilization efficiency of both light energy and CO2 but enhanced heat dissipation efficiency and greater tolerance to high-light intensity. BSA-Seq analysis identified a single nucleotide change (C to T) and (G to A) in the third exon of the BnaA01G0094500ZS and BnaC01G0116100ZS, respectively. These two genes encode the magnesium chelatase subunit I 1 (CHLI1) that catalyzes the insertion of magnesium into protoporphyrin IX, a pivotal step in chlorophyll synthesis. The mutation sites resulted in an amino acid substitution P144S and G128E within the AAA+ domain of the CHLI1 protein. Two KASP markers were developed and co-segregated with the yellow leaf phenotype in segregating F2 population. Loss of BnaA01.CHLI1 and BnaC01.CHLI1 by CRISPR/Cas9 gene editing recapitulated the mutant phenotype. BnaA01.CHLI1 and BnaC01.CHLI1 were located in chloroplast and highly expressed in the leaves. Furthermore, RNA-seq analyses revealed the expression of chlorophyll synthesis–related genes were upregulated in the crm1 mutant. These findings provide a new insight into the regulatory mechanism of chlorophyll synthesis in rapeseed and suggest a novel target for improving the photosynthetic efficiency and tolerance to high-light intensity in crops.

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Data availability

The sequencing data have been deposited in the NCBI database under BioProject PRJNA912866, the SRA accession numbers are: SRR12968239, SRR12968240, SRR12968241, and SRR12968242. The data that support the findings of this study are available in the main text and supporting materials of this article.

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Acknowledgements

We thank Dr. Zhongsong Liu for providing L329 (xiangyou 15) seeds, Prof. Wusheng Peng for 2B seeds.

Funding

This work was supported by the National Natural Science Foundation of China (U20A2029), Natural Science Foundation of Changsha city (kq2202150).

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Author notes

  1. These authors contributed equally to this work and share first authorship

Authors and Affiliations

  1. College of Biology, Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation, Hunan Engineering and Technology Research Center of Hybrid Rapeseed, Hunan University, Changsha, 410082, China

    Hui Zhang, Fujiang Xiang, Zhengfeng Zhang, Feifei Duan, Quanyu Zhou, Xin Li, Xinmei Li & Xiaoying Zhao

  2. Crop Research Institute, Hunan Academy of Agricultural Sciences, Changsha, 410125, China

    Hui Zhang, Yiming Guo & Bao Li

  3. College of Agronomy, Hunan Agricultural University, Changsha, 410128, Hunan, China

    Wei Zhang

  4. Hunan Cotton Research Institute, Changde, 415100, Hunan, China

    Tingzhou Chen

  5. Huazhi Biotech Co. Ltd, Changsha, 410125, China

    Miaoquan Fang

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  1. Hui Zhang
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Contributions

HZ, WZ, FX, XinmeiL, BL, and XZ designed the research; HZ, WZ, FX, ZZ, YG, and TC performed the experiments; FD, QZ, XinL, MF, and XinmeiL provided technical assistance to HZ, WZ, and FX; HZ, WZ, FX, XinmeiL, BL, and XZ analyzed the data; HZ, BL, and XZ supervised and completed the writing; HZ, WZ, and FX contributed equally to this work. All authors contributed to the article and approved the submitted version.

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Correspondence to Bao Li or Xiaoying Zhao.

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Zhang, H., Zhang, W., Xiang, F. et al. Photosynthetic characteristics and genetic mapping of a new yellow leaf mutant crm1 in Brassica napus. Mol Breeding 43, 80 (2023). https://doi.org/10.1007/s11032-023-01429-6

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