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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MDPI AG
2025-04-01
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| Series: | Horticulturae |
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| Online Access: | https://www.mdpi.com/2311-7524/11/4/406 |
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| author | Min Su Xinlin Wu Zechen Wang Limei Li Lejun Ouyang |
| author_facet | Min Su Xinlin Wu Zechen Wang Limei Li Lejun Ouyang |
| author_sort | Min Su |
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| description | 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>. |
| format | Article |
| id | doaj-art-c760c5c0d40f4d6ea1e3ca848c1104b6 |
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| issn | 2311-7524 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | MDPI AG |
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| series | Horticulturae |
| spelling | doaj-art-c760c5c0d40f4d6ea1e3ca848c1104b62025-08-20T02:28:23ZengMDPI AGHorticulturae2311-75242025-04-0111440610.3390/horticulturae11040406Construction of an Editing System for Forest Tree Genomes Based on an Efficient Visual Screening Marker in <i>Eucalyptus urophylla</i> × <i>Eucalyptus grandis</i>Min Su0Xinlin Wu1Zechen Wang2Limei Li3Lejun Ouyang4College of Biological and Food Engineering, Guangdong University of Petrochemical Technology, Maoming 525000, ChinaCollege of Biological and Food Engineering, Guangdong University of Petrochemical Technology, Maoming 525000, ChinaCollege of Biological and Food Engineering, Guangdong University of Petrochemical Technology, Maoming 525000, ChinaCollege of Biological and Food Engineering, Guangdong University of Petrochemical Technology, Maoming 525000, ChinaCollege of Biological and Food Engineering, Guangdong University of Petrochemical Technology, Maoming 525000, ChinaHerein, 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>.https://www.mdpi.com/2311-7524/11/4/406<i>Eucalyptus urophylla</i> × <i>Eucalyptus grandis</i>screening markerCRISPR/Cas9C4H |
| spellingShingle | Min Su Xinlin Wu Zechen Wang Limei Li Lejun Ouyang 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> Horticulturae <i>Eucalyptus urophylla</i> × <i>Eucalyptus grandis</i> screening marker CRISPR/Cas9 C4H |
| title | 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> |
| title_full | 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> |
| title_fullStr | 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> |
| title_full_unstemmed | 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> |
| title_short | 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> |
| title_sort | 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 |
| topic | <i>Eucalyptus urophylla</i> × <i>Eucalyptus grandis</i> screening marker CRISPR/Cas9 C4H |
| url | https://www.mdpi.com/2311-7524/11/4/406 |
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