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Induced mutation in ELONGATED HYPOCOTYL5 abolishes anthocyanin accumulation in the hypocotyl of pepper

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Abstract

Key message

The causal gene, CaHY5 of a chemical induced green-hypocotyl mutant was identified by molecular map**. CaHY5 regulates anthocyanin accumulation by directly binding to the promoter of genes in anthocyanin pathway.

Abstract

Morphological markers at seedling stage are useful indicators for F1 hybrid seeds screening. Pepper is a worldwide vegetable with diverse uses, and F1 hybrids are popular in the pepper industry. Hypocotyl color is a useful marker to identify F1 hybrid seeds. However, most pepper accessions have purple hypocotyl caused by anthocyanin accumulation, while green hypocotyl pepper accessions are rare. In this study, we identified a green hypocotyl mutant (e1898) from a pepper ethylmethanesulfonate (EMS) mutant library. By combining bulked segregant RNA-seq (BSR), genome resequencing and recombinant analysis, it was found that CaHY5 is the causal gene of this mutant. Virus-induced gene silencing (VIGS) of CaHY5 resulted in the decrease of anthocyanin accumulation in pepper hypocotyls. RNA-seq data showed that many genes related to anthocyanin biosynthesis and transport decreased significantly in the mutant. Yeast one-hybrid (Y1H) assays showed that CaHY5 can bind to the promoter of CaF3H, CaF3′5’H, CaDFR, CaANS and CaGST, which are important genes in anthocyanin biosynthesis or transport. Our results indicate that CaHY5 directly regulates anthocyanin biosynthesis and transport, thus governing anthocyanin accumulation in pepper hypocotyl. The mutant and gene identified in this work shall be valuable in the purity control of hybrid pepper seeds.

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Data availability

The RNA-seq and resequencing data of this work are available at the NCBI repository (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA842918).

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Acknowledgements

We thank Prof. Jihong Liu for providing the nucleus marker (Mcherry-RFP).

Funding

This study was funded by the National Natural Science Foundation (U21A20230, 31972416, U1906205) and the National Key R&D Program (2018YFD1000800, 2022YFE0100900) of China. Y.T. is grateful to the Tianshan Xuesong Project (2020XS30) of ** Tang, Feng Li, Yongen Lu & Bo Ouyang

Authors
  1. Rong Chen
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  2. Can Yang
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  3. Hu Gao
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  4. Chunmei Shi
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  5. Zhiying Zhang
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  6. Guangyu Lu
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  7. **nyan Shen
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  8. Feng Li
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  9. Yongen Lu
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  10. Bo Ouyang
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Contributions

B.O. and R.C. designed the research and wrote the manuscript; R.C. performed most of the experiments; Y.C. constructed the vectors for Y1H; H.G. and C.S. analyzed data; Z.Z. constructed the pepper EMS mutant library; G.Y., X.S. and Y.T. contributed to the VIGS experiment; F.L. and Y.L. provided valuable suggestions and modified the manuscript.

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Correspondence to Bo Ouyang.

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Chen, R., Yang, C., Gao, H. et al. Induced mutation in ELONGATED HYPOCOTYL5 abolishes anthocyanin accumulation in the hypocotyl of pepper. Theor Appl Genet 135, 3455–3468 (2022). https://doi.org/10.1007/s00122-022-04192-y

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