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Fine mapping and identification of a novel albino gene OsAL50 that is required for chlorophyll biosynthesis and chloroplast development in rice (Oryza sativa L.)

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Abstract

Leaf color is a highly important agronomic trait, and mutants with altered leaf coloration can serve as excellent models for studies on chloroplast development and chlorophyll biosynthesis, enabling the cloning of genes involved in these processes in rice (Oryza sativa L.). In this study, we isolated a stable genetic rice mutant, oryza sativa albino leaf 50 (osal50), from a breeding population of the japonica cultivar GP50. This mutant exhibited a distinctive albino phenotype, with white-striped leaves in seedlings and white panicles at the heading stage. Compared with wild-type GP50, the osal50 mutant showed lower chlorophyll and carotenoid accumulation, together with abnormal chloroplast ultrastructure. Genetic analysis demonstrated that a recessive nuclear gene was responsible for the albino phenotype of osal50, and a map-based cloning strategy delimited OsAL50 to a 160-kb physical interval on chromosome 1, flanked by two single nucleotide polymorphism (SNP) markers, CAPS-08 and CAPS-37, that included 26 putative open reading frames. Sequence and expression analyses revealed LOC_Os01g20110 as the candidate OsAL50 gene, which was confirmed by knockout using CRISPR/Cas9. Subcellular localization and protein sequence analyses suggested that OsAL50 likely encodes an endoribonuclease E-like protein localized to the chloroplasts. Further investigation indicated that OsAL50 plays a vital role in the regulation of photosynthetic pigment metabolism, photosynthesis, and chloroplast biogenesis. In summary, we identified a novel albino mutant that will serve as useful genetic material for studies of chlorophyll biosynthesis and chloroplast development in rice.

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Acknowledgements

This study was supported by the China Agriculture Research System of MOF and MARA (CARS-01), the Fujian Provincial Natural Science Foundation (2021J01535, 2021J01536), and the Sanming Municipal Science and Technology Project (2022-N-7).

Funding

Funding was supported by Agriculture Research System of China,CARS-01 ,Natural Science Foundation of Fujian Province,2021J01536, 2021J01535, Sanming Municipal Science and Technology Project,2022-N-7.

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  1. Yuehui Zeng and Xinyu Wei have contributed equally to this work.

Authors and Affiliations

  1. Biotechnology Research Institute, Sanming Academy of Agricultural Sciences, Sanming, 365500, China

    Yuehui Zeng, Xinyu Wei, Changchun Xiao, Rui Zhang & Jianhong Huang

  2. Fujian Key Laboratory of Crop Genetic Improvement and Innovative Utilization for Mountain Area, Sanming, 365500, China

    Yuehui Zeng, Xinyu Wei, Changchun Xiao, Rui Zhang, Jianhong Huang & Xuming Xu

  3. Rice Research Institute, Sanming Academy of Agricultural Sciences, Sanming, 365500, China

    Xuming Xu

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  1. Yuehui Zeng
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Contributions

YZ and XW contributed equally to this work. YZ carried out molecular-marker development, genetic analysis, molecular mapping, candidate gene analysis, physical-map and phylogenetic-tree construction, and wrote the manuscript. XW carried out molecular-marker development, genetic analysis, molecular mapping and expression analysis. CX and RZ participated in genetic analysis, molecular mapping, and physical-map construction. JH carried out mapping-population construction and participated in genetic analysis and molecular mapping. XX designed the research and wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Xuming Xu.

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Communicated by Dawei Xue.

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Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 554 KB)—Sequences of PCR primers used for gene mapping

Supplementary file2 (PDF 554 KB)—Primers used for expression analysis of candidate genes

Supplementary file3 (PDF 554 KB)—Primers used for sequencing of candidate genes

10725_2023_1116_MOESM4_ESM.pdf

Supplementary file4 (PDF 554 KB)—Primers used for expression analysis of genes associated with chloroplast development, chlorophyll biosynthesis, and photosynthesis

10725_2023_1116_MOESM5_ESM.pdf

Supplementary file5 (PDF 554 KB)—Functional annotation of 26 candidate genes for OsAL50 based on the RGAP http://rice.plantbiology.msu.edu/

10725_2023_1116_MOESM6_ESM.pdf

Supplementary file6 (PDF 457 KB)—Mutation analysis within the promoter region of LOC_Os01g20110 in 20 rice varieties, where “--”indicates base deletion

Supplementary file7 (PDF 551 KB)—Analysis of cis-acting regulatory elements in the promoter region of OsAL50

10725_2023_1116_MOESM8_ESM.pdf

Supplementary file8 (PDF 404 KB)—Subcellular localization of OsAL50 in rice protoplasts. a Observation of free green fluorescent protein (GFP) signals in rice protoplasts. b Observation of OsAL50-GFP fluorescent signals in rice protoplasts. Bars: 10 μm

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Zeng, Y., Wei, X., Xiao, C. et al. Fine mapping and identification of a novel albino gene OsAL50 that is required for chlorophyll biosynthesis and chloroplast development in rice (Oryza sativa L.). Plant Growth Regul (2024). https://doi.org/10.1007/s10725-023-01116-8

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