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<p><i>ZmAsnS2</i> was identified as a gene with the potential to increase seed protein levels when mis-expressed in maize. <i>ZmAsnS2</i> was cloned into a plant expression vector and transformed into a maize inbred line. Protein immunoblot analysis and activity assays confirmed expression of the protein. Transgenic inbred corn which mis-expressed the gene had significantly higher free asparagine levels in leaf tissue and higher kernel protein levels. These plants also have higher asparagine levels in shank and seed tissue raising the possibility that the additional Asn synthesized in the leaf is being transported to the seed to be used in the synthesis of seed protein. In these transgenic events, asparagine levels were elevated such that asparagine becomes the major amino acid. When these inbred lines were crossed to a tester line, the resulting hybrid plants also exhibited higher leaf kernel protein, albeit to a lesser degree than the inbred maize. Over-expression of an additional asparagine synthetase gene, <i>ZmAsnS3</i>, also led to higher leaf Asn. A hypothesis to explain how the mis-expression of AsnS might increase kernel protein is discussed.</p>

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Overexpression of maize asparagine synthetase increases free asparagine and kernel protein in Zea mays

  • 期刊论文

  • 发表日期：

    2025年1月22日

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  • James H. Crowley,
  • Arick Tuhro,
  • Todd Brown,
  • Anabayan Kessavalou,
  • Bradon J. Fabbri,
  • Stephen M. G. Duff

摘要

ZmAsnS2 was identified as a gene with the potential to increase seed protein levels when mis-expressed in maize. ZmAsnS2 was cloned into a plant expression vector and transformed into a maize inbred line. Protein immunoblot analysis and activity assays confirmed expression of the protein. Transgenic inbred corn which mis-expressed the gene had significantly higher free asparagine levels in leaf tissue and higher kernel protein levels. These plants also have higher asparagine levels in shank and seed tissue raising the possibility that the additional Asn synthesized in the leaf is being transported to the seed to be used in the synthesis of seed protein. In these transgenic events, asparagine levels were elevated such that asparagine becomes the major amino acid. When these inbred lines were crossed to a tester line, the resulting hybrid plants also exhibited higher leaf kernel protein, albeit to a lesser degree than the inbred maize. Over-expression of an additional asparagine synthetase gene, ZmAsnS3, also led to higher leaf Asn. A hypothesis to explain how the mis-expression of AsnS might increase kernel protein is discussed.