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<p>There is emerging interest in tropical C4 forage grasses due to their potential value in changing pasture conditions caused by climate change. In this study, a robust regeneration and transformation system was developed for Rhodes grass (<i>Chloris gayana</i>). From elite commercial cultivar ‘Tolgar’ initial screening for embryogenic callus (EC) responses were generated from cut mesocotyl explants, with 20µM of 2,4-D being most effective at generating EC from cut mesocotyl explants, with an EC induction rate of 3.89% for cv. ‘Tolgar’. From these EC cultures, four isogenic lines were recovered via shoot regeneration and maintained by micropropagation. EC-derived lines were then validated for production of isogenic lines for use in plant transformation, using macerated basal meristem explants from tillers of micropropagated plantlets, with one line (T108) having reliable and efficient regeneration. The most efficient transformation efficiency (2.16%) was obtained using hygromycin selection with isogenic line T108. As determined by ddPCR among all recovered transgenic plants, 12% were single copy and 53% had low transgene copy number (2–4), with 47% of transformed plants expressing the reporter gene (DsRED2). The work here provides an updated transformation protocol for Rhodes grass which will enable future improvement of the species by molecular informed breeding or genome editing.</p>

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Embryogenic callus cultures and genetic transformation of Rhodes grass (Chloris gayana) cv. ‘Tolgar’

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年3月18日

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  • Kellie Maybery-Reupert,
  • Nimal Wijesinghe,
  • Matthew Hayden,
  • Noel Cogan,
  • Daniel Isenegger

摘要

There is emerging interest in tropical C4 forage grasses due to their potential value in changing pasture conditions caused by climate change. In this study, a robust regeneration and transformation system was developed for Rhodes grass (Chloris gayana). From elite commercial cultivar ‘Tolgar’ initial screening for embryogenic callus (EC) responses were generated from cut mesocotyl explants, with 20µM of 2,4-D being most effective at generating EC from cut mesocotyl explants, with an EC induction rate of 3.89% for cv. ‘Tolgar’. From these EC cultures, four isogenic lines were recovered via shoot regeneration and maintained by micropropagation. EC-derived lines were then validated for production of isogenic lines for use in plant transformation, using macerated basal meristem explants from tillers of micropropagated plantlets, with one line (T108) having reliable and efficient regeneration. The most efficient transformation efficiency (2.16%) was obtained using hygromycin selection with isogenic line T108. As determined by ddPCR among all recovered transgenic plants, 12% were single copy and 53% had low transgene copy number (2–4), with 47% of transformed plants expressing the reporter gene (DsRED2). The work here provides an updated transformation protocol for Rhodes grass which will enable future improvement of the species by molecular informed breeding or genome editing.