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Fine mapping of the Ca3GT gene controlling anthocyanin biosynthesis in mature unripe fruit of Capsicum annuum L.

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Abstract

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The anthocyanin biosynthesis gene Ca3GT was fine-mapped in a 110.5-kb region through a map-based cloning strategy. Gene expression and promoter analyses confirmed the strong candidate gene Capana10g001978.

Abstract

Pepper (Capsicum annum L.) fruit can be dark green, green, light green, purple, yellow, or ivory at the juvenile stage. Anthocyanins are responsible for fruit color formation in mature unripe pepper fruit, and transient accumulation of anthocyanins is the main problem in breeding pepper plants with mature purple fruit. Only a few genes controlling this trait have been cloned. The present study aimed to map and identify an anthocyanin biosynthesis gene from pepper using an F2 population derived from a cross between line ‘17C3808’ (purple mature unripe fruit) and line ‘17C3807’ (green mature unripe fruit). The trait was mapped on a 110.5-kb interval between markers SSR18213 and SSR18228 on chromosome 10. There were three open reading frames in this region; Capana10g001978 was predicted in this region as markers CAPS-78-708 and InDel146 co-segregated with it. Capana10g001978 is a structural gene encoding the GTB transcription factor involved in the biosynthesis of anthocyanins. Comparing parental sequences, two base mutations were identified in the exon of Capana10g001978, at positions + 528 bp and + 708 bp, which resulted in changes in the 176th and 236th amino acid residues, from glutamine (CAA) to histidine (CAC), causing a nonsense mutation (from CAG to CAA). Additionally, Capana10g001978 was highly expressed in the pericarp of mature, unripe pepper fruit. There were four single nucleotide polymorphisms, three sequence deletions, and one sequence insertion in the promoter region of purple, mature, and unripe pepper fruit, leading to the formation of a W-box and a GT1-motif. Thus, Capana10g001978 is a strong candidate gene of Ca3GT involved in anthocyanin biosynthesis in mature unripe pepper fruit. These results provide important information regarding the isolation and characterization of Ca3GT, and they are the starting point for studying the regulatory pathway responsible for anthocyanin biosynthesis in pepper.

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Acknowledgements

This research was supported by The National Key Research and Development Program of China (2017YFD0101903), the Beijing Fruit Vegetables Innovation Team of Modern Agricultural Industry Technology System (BAIC01-2019), and the Construction of Beijing Science and Technology Innovation and Service Capacity in Top Subjects (CEFF-PXM2019_014207_000032).

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Authors and Affiliations

  1. Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, Department of Vegetable Science, College of Horticulture, China Agricultural University, Beijing, 100193, China

    Jinqiu Liu, Xiyin Ai, Yihao Wang, Qiaohua Lu, Ting Li, Lang Wu, Liang Sun & Huolin Shen

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  1. Jinqiu Liu
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  2. Xiyin Ai
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  3. Yihao Wang
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  4. Qiaohua Lu
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  7. Liang Sun
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  8. Huolin Shen
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Contributions

JQL performed most of the experiments, analyzed the data, and drafted the manuscript. XYA elaborated the figures and revised the manuscript. YHW participated in DNA extraction. TL and LW performed the qRT-PCR analysis. QHL revised the manuscript. HLS designed, and LS directed the entire study. All the authors have read and approved the final manuscript.

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Correspondence to Liang Sun or Huolin Shen.

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Communicated by Amnon Levi.

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Fig. S1 Sequence analysis of

MYBA between 17C3807 and 17C3808. a, CDS sequence of the MYBA between 17C3807 and 17C3808; b, 1071 bp sequence of ATG upstream between 17C3807 and 17C3808 (TIFF 3176 kb)

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Liu, J., Ai, X., Wang, Y. et al. Fine mapping of the Ca3GT gene controlling anthocyanin biosynthesis in mature unripe fruit of Capsicum annuum L.. Theor Appl Genet 133, 2729–2742 (2020). https://doi.org/10.1007/s00122-020-03628-7

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