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<p>Currently, the use of CRISPR/Cas9 gene editing technologies in plants is growing. Since Hu et al. created the CRISPR/Cas9 system in bananas in 2017, researchers have employed CRISPR/Cas9 gene editing technology to change the banana <i>MaACO1</i> and <i>MaGA20ox2</i> genes, resulting in novel banana germplasm banana. However, there are no reports on base editing in banana until now. In this study, adenine base editor (ABE) was implemented in banana with banana <i>PDS</i> (<i>MaPDS</i>) as the target gene, and a mutant was successfully generated. In the mutant, the leaves showed chlorosis and lesion spots. Deep amplicon sequencing analysis revealed that the fifth and sixth bases of the target sequence exhibited the A·T to G·C mutation, and the sixth base conversion mutation caused the amino acid to change from isoleucine to valine. Moreover, the mutation efficiency of the sixth base in the leaf sample with lesion spots was much greater than that in the adjacent normal leaf samples from the same leaf. In addition, the structure of the leaf samples was examined using paraffin section analysis. The results showed that the number of cells in the upper cuticle, upper epidermis, lower epidermis and lower cuticle of the chlorosis spot regions in the leaves of the mutant line #1 was significantly lower, by 16.89, 91.67, 14.38 and 30.69%, respectively, than that in the wild type. Taken together, these results indicated that ABE works well in banana and could provide a basis for the genetic improvement of important agronomic traits via precise base editing.</p>

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Precise A·T-to-G·C Base Editing in Banana

  • 期刊论文

  • 发表日期：

    2025年7月21日

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  • Shaoping Wu,
  • Chunhua Hu,
  • Haocheng Zhu,
  • Jinxing Liu,
  • Tongxin Dou,
  • Fei Qin,
  • Binhui Yin,
  • Fangcheng Bi,
  • Qiaosong Yang,
  • Ganjun Yi

摘要

Currently, the use of CRISPR/Cas9 gene editing technologies in plants is growing. Since Hu et al. created the CRISPR/Cas9 system in bananas in 2017, researchers have employed CRISPR/Cas9 gene editing technology to change the banana MaACO1 and MaGA20ox2 genes, resulting in novel banana germplasm banana. However, there are no reports on base editing in banana until now. In this study, adenine base editor (ABE) was implemented in banana with banana PDS (MaPDS) as the target gene, and a mutant was successfully generated. In the mutant, the leaves showed chlorosis and lesion spots. Deep amplicon sequencing analysis revealed that the fifth and sixth bases of the target sequence exhibited the A·T to G·C mutation, and the sixth base conversion mutation caused the amino acid to change from isoleucine to valine. Moreover, the mutation efficiency of the sixth base in the leaf sample with lesion spots was much greater than that in the adjacent normal leaf samples from the same leaf. In addition, the structure of the leaf samples was examined using paraffin section analysis. The results showed that the number of cells in the upper cuticle, upper epidermis, lower epidermis and lower cuticle of the chlorosis spot regions in the leaves of the mutant line #1 was significantly lower, by 16.89, 91.67, 14.38 and 30.69%, respectively, than that in the wild type. Taken together, these results indicated that ABE works well in banana and could provide a basis for the genetic improvement of important agronomic traits via precise base editing.