Abstract
Key message
This study demonstrated high expression and accumulation of human α-lactalbumin in transgenic maize, and significant improvement of lysine content in maize endosperm.
Abstract
As a high-yield crop, lack of lysine in endosperm storage protein is a major defect of maize (Zea mays L.). Specifically expression of foreign proteins is a potential way to improve lysine content in maize endosperm. Human α-lactalbumin is such a protein with high lysine content and high nutritional value. In this study, the codon-optimized human lactalbumin alpha (LALBA) gene was driven by maize endosperm-specific 27 kD γ-zein promoter, and transformed into maize. Five independent transgenic lines were obtained, and LALBA was highly expressed in endosperm in all these lines. Protein assay indicated that human α-lactalbumin was highly accumulated in maize endosperm. Immuno-localization assay indicated that human α-lactalbumin was mainly deposited into the protein body (PB). Protein interaction assay showed that human α-lactalbumin interacted with 16 kD γ-zein, which might lead to its deposition to the PBs. Amino acid analysis of two independent transgenic lines showed significant increase of lysine contents in transgenic endosperm, with 47.26% and 45.15% increase to their non-transgenic seeds, respectively. We obtained transgenic maize with endosperm-specific accumulation of human α-lactalbumin at high level and increased the lysine content in maize endosperm. This study demonstrated an effective way to improve the nutritional value of maize seeds.
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Funding
This work was supported by the Research Program of Sanya Yazhou Bay Science and Technology City (SYND-2021-22 to R.S.), the Hainan Yazhou Bay Seed Lab (B21HJ8105 to R.S.), and the National Key Research and Development Program of China (grant 2016YFD0100503 to W.Q.).
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RTS and WWQ conceived and designed research. CWLL and LFJ performed experiments. CWLL and WM collected and analyzed the data. CWLL, WWQ and RTS wrote the manuscript. All authors read and approved the final manuscript.
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299_2022_2906_MOESM1_ESM.pdf
Supplementary file1 Figure S1. The sequence of 27 kD γ-zein promoter, modified LALBA (opening reading frame) and 27 kD γ-zein terminator. Figure S2. RT-qPCR analysis of LALBA expression in 18 DAP seeds with Ubiquitin as reference gene. Figure S3. Protein interaction between human α-lactalbumin and zeins. Figure S4. Schematic structure of chimeric constructs used in transient expression. Figure S5. Phenotypic features of human α-lactalbumin transgenic line #1. Table S1. Primers used in this research. Table S2. Amino acid content in #1 endosperm. Table S3. Amino acid content in #4 endosperm. Table S4. Other transgenic maize for increasing lysine content. Table S5. The contents of five amino acids in human α-lactalbumin and α-zeins (PDF 807 KB)
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Li, C., Ma, W., Jin, L. et al. Endosperm-specific accumulation of human α-lactalbumin increases seed lysine content in maize. Plant Cell Rep 41, 2023–2035 (2022). https://doi.org/10.1007/s00299-022-02906-6
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DOI: https://doi.org/10.1007/s00299-022-02906-6