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. 2021 Nov 5:12:762265.
doi: 10.3389/fpls.2021.762265. eCollection 2021.

TbMYC4A Is a Candidate Gene Controlling the Blue Aleurone Trait in a Wheat- Triticum boeoticum Substitution Line

Xin Liu  1 , Minghu Zhang  1 , Xiaomei Jiang  1 , Hui Li  1 , Zhenjiao Jia  1 , Ming Hao  1 , Bo Jiang  1 , Lin Huang  1   2 , Shunzong Ning  1 , Zhongwei Yuan  1 , Xuejiao Chen  1 , Xue Chen  1 , Dengcai Liu  1   2 , Baolong Liu  3   4 , Lianquan Zhang  1   2
Affiliations

TbMYC4A Is a Candidate Gene Controlling the Blue Aleurone Trait in a Wheat- Triticum boeoticum Substitution Line

Xin Liu et al. Front Plant Sci. .

Abstract

Triticum boeoticum Boiss (AbAb, 2n = 2x = 14) is one of the sources of the blue grain trait controlled by blue aleurone layer 2 (Ba2). However, the underlying genes have not been cloned. In this study, a transcriptomic comparison between a blue-grained wheat-T. boeoticum substitution line and its wheat parent identified 41 unigenes related to anthocyanin biosynthesis and 29 unigenes related to transport. The bHLH transcription factor gene TbMYC4A showed a higher expression level in the blue-grained substitution line. TbMYC4A contained the three characteristic bHLH transcription factor domains (bHLH-MYC_N, HLH and ACT-like) and clustered with genes identified from other wheat lines with the blue grain trait derived from other Triticeae species. TbMYC4A overexpression confirmed that it was a functional bHLH transcription factor. The analysis of a TbMYC4A-specific marker showed that the gene was also present in T. boeoticum and T. monococcum with blue aleurone but absent in other Triticeae materials with white aleurone. These results indicate that TbMYC4A is a candidate gene of Ba2 controlling the blue aleurone trait. The isolation of TbMYC4A is helpful for further clarifying the genetic mechanism of the blue aleurone trait and is of great significance for breeding blue-grained wheat varieties.

Keywords: TbMYC4A; Triticum boeoticum; anthocyanin biosynthesis; blue aleurone; transcriptome analysis.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Expression profiles of DEGs [|log2(fold change)| ≥ 1, FDR < 0.05] related to the anthocyanin biosynthesis and transport processes in blue and white aleurone. The color scale from green (low) to purple (high) represents the log2FPKM values measured in white and blue aleurones.
FIGURE 2
FIGURE 2
Phylogenetic tree of bHLH proteins regulating anthocyanin biosynthesis. The accession numbers of these proteins (or translated products) in the GenBank database are as follows: Nicotiana tabacum/NtAN2:ACO52472.1; Oryza/OsbHLH14:XP_015649359.1; Arabidopsis thaliana/bHLH025:Q9T072.2; Arabidopsis thaliana/bHLH018:Q1PF17.1; Brassica oleracea var.botrytis/BoTT8: ADP76654.1; Gentiana triflora/GTbHLH1:BAH03387.1; Lilium hybrid division I/LHbHLH2:BAE20058.1; Diospyros kaki/MYC1:AEC03343.1; Vitis vinifera/MYC1: ACC68685.1; Malus domestica/MdbHLH3:ADL36597.1; Malus domestica/MdbHLH33:AAC49219.1; Arabidopsis thaliana/MYC1:NP_191957; Arabidopsis thaliana/bHLH12:AAL55719.1; Gerbera hybrid cultivar/GMYC1: CAA07615.1; Perilla frutescens/MYC-GP:BAA75514.1; Petunia x hybrida/JAF13: AAC39455.1; Oryza/Ra:AAC49219; Maize/RS:NP_001106073; Zea mays/ZmR: NP_001105339.2; Hordeum_vulgare/MYC2:MF679157.1; Thinopyrum ponticum/MYC4E:KX914905.1; Aegilops tauschii/R-S:KD566857.1; Triticum urartu/RS: KD032825.1; and Triticum aestivum/MYC1:KY499900.1.
FIGURE 3
FIGURE 3
Comparison of the amino acid sequence of TbMYC4A with those of ThNYC4E, HvMYC2, and bHLH transcription factors known to regulate anthocyanin biosynthesis from Thinopyrum ponticum and Hordeum vulgare, with black lines representing bHLH-MYC_N, HLH and ACT-like domains. The accession numbers of these proteins (or translated product) in the GenBank database are as follows: Thinopyrum ponticum/MYC4E:KX914905.1 and Hordeum vulgare/MYC2:MF679157.1.
FIGURE 4
FIGURE 4
Wheat coleoptiles 16 h after bombardment with different plasmids. TaMYB7D, TbMYC4A, and TaMYC1 represent the constructed vectors pBRACT214:TaMYB7D, pLGY-02: TbMYC4A, and pBRACT214:TaMYC1, respectively.
FIGURE 5
FIGURE 5
Distribution of TbMYC4A in Z18-1244, Crocus, G52, and 14 examples selected from 7 Triticeae species. (A) lanes 1 and 2: Crocus; lanes 3 and 4: T. boeoticum G52; lanes 5 and 6: Z18-1244; (B) T. monococcum accessions PI 10474T (1) and PI 168805 (2); T. boeoticum accessions PI 401416 (3) and PI 427506 (4); T. timopheevii accessions AS270 (5) and AS272 (6), T. zhukovskyi accessions TRI 7270-1 (7) and TRI 7270-3 (8); Ae. tauschii accessions AS60 (9) and AS62 (10); T. urartu accessions PI428224 (11) and PI428274 (12); and T. turgidum accessions PI525355 (13) and AS2239 (14).
FIGURE 6
FIGURE 6
Relative expression of TbMYC4A in the seeds of Z18-1244 and Crocus at different developmental stages.
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