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<p><i>Hordeum vulgare</i> L. is a diploid monocot cereal crop belonging to the Poaceae grass family. The improvement of the commercially significant crop varieties through molecular breeding depends upon an optimized genetic transformation method without compromising their desired agronomic traits. The current investigation established an efficient regeneration and transformation method for the Indian malting barley cultivar DWRB 160, a genotype previously resistant to in vitro regeneration. The use of mature embryos as a source of explant bypasses the seasonal constraints of immature embryos, thereby improving regeneration efficiency. The selection of Agrobacterium strain AGL1 for genetic transformation at co-cultivation conditions of pH 5.4 and OD 600 = 1.0 achieved a high transient GUS expression frequency of 86.66%, similar to the percentage efficiency documented for the model cultivar Golden Promise. The molecular analysis of the transformed barley lines achieved a stable transformation efficiency of 6%. The quantitative gene expression profiling of the uidA transcript among independent transformed lines showed significant variability in the transcript expression level, suggesting positional and copy number effects. The present study offers an improved genetic transformation protocol with high regeneration potential and stable T-DNA delivery for the effective transformation of elite barley malt germplasm.</p>

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Establishment of an efficient in vitro regeneration and Agrobacterium-mediated transformation system in elite northern plain barley (Hordeum vulgare L.) cv. DWRB 160

  • 期刊论文

  • 发表日期：

    2026年6月27日

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  • Shalu Yadav,
  • Dinesh Kumar,
  • Alka Jangra,
  • Garima Sharma,
  • Santosh Kumar Bishnoi,
  • Vinod Chhokar

摘要

Hordeum vulgare L. is a diploid monocot cereal crop belonging to the Poaceae grass family. The improvement of the commercially significant crop varieties through molecular breeding depends upon an optimized genetic transformation method without compromising their desired agronomic traits. The current investigation established an efficient regeneration and transformation method for the Indian malting barley cultivar DWRB 160, a genotype previously resistant to in vitro regeneration. The use of mature embryos as a source of explant bypasses the seasonal constraints of immature embryos, thereby improving regeneration efficiency. The selection of Agrobacterium strain AGL1 for genetic transformation at co-cultivation conditions of pH 5.4 and OD 600 = 1.0 achieved a high transient GUS expression frequency of 86.66%, similar to the percentage efficiency documented for the model cultivar Golden Promise. The molecular analysis of the transformed barley lines achieved a stable transformation efficiency of 6%. The quantitative gene expression profiling of the uidA transcript among independent transformed lines showed significant variability in the transcript expression level, suggesting positional and copy number effects. The present study offers an improved genetic transformation protocol with high regeneration potential and stable T-DNA delivery for the effective transformation of elite barley malt germplasm.