Construction of an Editing System for Forest Tree Genomes Based on an Efficient Visual Screening Marker in <i>Eucalyptus urophylla</i> × <i>Eucalyptus grandis</i>
Herein, the clustered regularly interspaced short palindromic repeats (CRISPRs)/CRISPRs-associated protein 9 (Cas9) technology for genome editing was used to develop an efficient gene editing system for <i>Eucalyptus urophylla</i> × <i>Eucalyptus grandis</i> and generate a ne...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-04-01
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| Series: | Horticulturae |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2311-7524/11/4/406 |
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| Summary: | Herein, the clustered regularly interspaced short palindromic repeats (CRISPRs)/CRISPRs-associated protein 9 (Cas9) technology for genome editing was used to develop an efficient gene editing system for <i>Eucalyptus urophylla</i> × <i>Eucalyptus grandis</i> and generate a new <i>Eucalyptus</i> germplasm with reduced lignin content in the pulp for environmental sustainability in papermaking. By targeting the cinnamate-4-hydroxylase (<i>C4H</i>) gene in <i>E. urophylla</i> × <i>E. grandis</i>, the recombinant plasmid pHEE401E-<i>35S-RUBY-EuC4H</i> was constructed through homologous recombination. This plasmid was then transformed into <i>E. urophylla</i> × <i>E. grandis</i> callus tissue. Using the <i>RUBY</i> gene as a marker, positive transformants were screened based on the callus tissue phenotype. Subsequent PCR and sequencing confirmed the successful creation of mutants with a significantly edited <i>EuC4H</i> gene. This method offers a valuable framework and guidance for genetically improving and establishing an efficient gene editing system in <i>Eucalyptus</i>. |
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| ISSN: | 2311-7524 |