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Key message <p>The targeted mutation of the strigolactone biosynthetic gene, <i>CCD8,</i> through genome-editing CRISPR–Cas9 induces dwarfism and enhanced branching in <i>Eustoma grandiflorum.</i></p> Abstract <p>In ornamental plants, modifying shoot architecture is a major breeding objective for diverse applications; however, achieving this goal using conventional techniques remains challenging. Strigolactones are plant hormones that inhibit shoot branching. We aimed to generate a strigolactone-deficient mutant of <i>Eustoma grandiflorum</i> using CRISPR–Cas9 genome editing. Four guide RNAs targeting <i>CAROTENOID CLEAVAGE DIOXYGENASE 8</i> (<i>EgCCD8</i>) were designed, and transgenic <i>E. grandiflorum</i> plants were produced. Three independent lines carried deletions or insertions in all <i>EgCCD8</i> alleles, indicating high editing efficiency. Two lines exhibited increased branching and pronounced dwarfism. In one of these lines, plant height in T<sub>1</sub> null segregants was reduced to 47% of wild-type (WT) levels. Branch number increased 5.0-fold compared with the WT, and flower bud formation increased 1.7-fold alongside enhanced branching. These findings demonstrate that the targeted mutation of <i>CCD8</i> through genome editing induces dwarfism and enhanced branching without compromising other ornamental traits.</p>

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Engineering plant architecture in the ornamental species Eustoma grandiflorum by knockout of strigolactone biosynthesis

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年6月2日

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  • Kana Yamaguchi,
  • Eri Tomizawa,
  • Noriko Kanematsu,
  • Kimitoshi Sakaguchi,
  • Takashi Kasai,
  • Ryo Ebisawa,
  • Akihiro Hasumi,
  • Kyoko Kawakatsu,
  • Yuriko Osakabe,
  • Keishi Osakabe,
  • Takashi Nakatsuka

摘要

Key message

The targeted mutation of the strigolactone biosynthetic gene, CCD8, through genome-editing CRISPR–Cas9 induces dwarfism and enhanced branching in Eustoma grandiflorum.

Abstract

In ornamental plants, modifying shoot architecture is a major breeding objective for diverse applications; however, achieving this goal using conventional techniques remains challenging. Strigolactones are plant hormones that inhibit shoot branching. We aimed to generate a strigolactone-deficient mutant of Eustoma grandiflorum using CRISPR–Cas9 genome editing. Four guide RNAs targeting CAROTENOID CLEAVAGE DIOXYGENASE 8 (EgCCD8) were designed, and transgenic E. grandiflorum plants were produced. Three independent lines carried deletions or insertions in all EgCCD8 alleles, indicating high editing efficiency. Two lines exhibited increased branching and pronounced dwarfism. In one of these lines, plant height in T1 null segregants was reduced to 47% of wild-type (WT) levels. Branch number increased 5.0-fold compared with the WT, and flower bud formation increased 1.7-fold alongside enhanced branching. These findings demonstrate that the targeted mutation of CCD8 through genome editing induces dwarfism and enhanced branching without compromising other ornamental traits.