Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i>
<i>Lagerstroemia indica</i> is an important woody ornamental plant worldwide. However, the application of many technologies, such as transgenic breeding and genome editing, has been severely hampered due to the lack of efficient calli induction and regeneration technology. Here, we discu...
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MDPI AG
2023-03-01
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| Series: | Forests |
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| Online Access: | https://www.mdpi.com/1999-4907/14/3/517 |
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| author | Shengji Jiang Fenni Lv Lulu Gao Jiaojiao Gu Rutong Yang Sumei Li Ya Li Shaofeng Li Peng Wang |
| author_facet | Shengji Jiang Fenni Lv Lulu Gao Jiaojiao Gu Rutong Yang Sumei Li Ya Li Shaofeng Li Peng Wang |
| author_sort | Shengji Jiang |
| collection | DOAJ |
| description | <i>Lagerstroemia indica</i> is an important woody ornamental plant worldwide. However, the application of many technologies, such as transgenic breeding and genome editing, has been severely hampered due to the lack of efficient calli induction and regeneration technology. Here, we discussed a reliable and efficient calli induction and regeneration protocol using whole-leaf explants. This protocol’s effectiveness for the calli induction and regeneration systems in crape myrtle were up to 70.33% and 44.33%, respectively. Next, an efficient and stable <i>Agrobacterium</i>-mediated genetic transformation system was created from leaf calli, and the green fluorescent protein (GFP) was able to detect up to 90% of its positive frequency. Meanwhile, two positive lines’ transfer DNA insertion sites and directions were identified using whole genome sequencing. LiMYB75, a novel R2R3-MYB transcription factor, was identified and transferred to the <i>L. indica</i> genome to enhance the leaf calli regeneration frequency. Surprisingly, overexpressing LiMYB75 increased the frequency of calli regeneration in the leaf by 1.27 times and the number of regenerated plantlets per callus by 4.00 times compared to the wild type, by regulating the expression levels of genes involved in callus formation, such as <i>SHOOT MERISTEMLESS</i> (<i>STM</i>). Overall, our findings revealed a simple, reliable, and highly efficient transformation approach and identified the desirable candidate gene <i>LiMYB75</i>, which improves <i>L. indica</i>’s calli regeneration efficiency. |
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| institution | Directory Open Access Journal |
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| last_indexed | 2024-03-11T06:32:22Z |
| publishDate | 2023-03-01 |
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| series | Forests |
| spelling | doaj.art-c523530ac50b43d1af0ce3fa3904d74c2023-11-17T11:09:42ZengMDPI AGForests1999-49072023-03-0114351710.3390/f14030517Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i>Shengji Jiang0Fenni Lv1Lulu Gao2Jiaojiao Gu3Rutong Yang4Sumei Li5Ya Li6Shaofeng Li7Peng Wang8Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, ChinaState Key Laboratory of Tree Genetics and Breeding, Experimental Center of Forestry in North China, National Permanent Scientific Research Base for Warm Temperate Zone Forestry of Jiulong Mountain in Beijing, Chinese Academy of Forestry, Beijing 100091, ChinaJiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province & Chinese Academy of Sciences, Nanjing 210014, China<i>Lagerstroemia indica</i> is an important woody ornamental plant worldwide. However, the application of many technologies, such as transgenic breeding and genome editing, has been severely hampered due to the lack of efficient calli induction and regeneration technology. Here, we discussed a reliable and efficient calli induction and regeneration protocol using whole-leaf explants. This protocol’s effectiveness for the calli induction and regeneration systems in crape myrtle were up to 70.33% and 44.33%, respectively. Next, an efficient and stable <i>Agrobacterium</i>-mediated genetic transformation system was created from leaf calli, and the green fluorescent protein (GFP) was able to detect up to 90% of its positive frequency. Meanwhile, two positive lines’ transfer DNA insertion sites and directions were identified using whole genome sequencing. LiMYB75, a novel R2R3-MYB transcription factor, was identified and transferred to the <i>L. indica</i> genome to enhance the leaf calli regeneration frequency. Surprisingly, overexpressing LiMYB75 increased the frequency of calli regeneration in the leaf by 1.27 times and the number of regenerated plantlets per callus by 4.00 times compared to the wild type, by regulating the expression levels of genes involved in callus formation, such as <i>SHOOT MERISTEMLESS</i> (<i>STM</i>). Overall, our findings revealed a simple, reliable, and highly efficient transformation approach and identified the desirable candidate gene <i>LiMYB75</i>, which improves <i>L. indica</i>’s calli regeneration efficiency.https://www.mdpi.com/1999-4907/14/3/517whole-leaf explantscallus inductiongenetic transformationplant hormoneregeneration ability |
| spellingShingle | Shengji Jiang Fenni Lv Lulu Gao Jiaojiao Gu Rutong Yang Sumei Li Ya Li Shaofeng Li Peng Wang Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i> Forests whole-leaf explants callus induction genetic transformation plant hormone regeneration ability |
| title | Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i> |
| title_full | Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i> |
| title_fullStr | Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i> |
| title_full_unstemmed | Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i> |
| title_short | Novel R2R3-MYB Transcription Factor LiMYB75 Enhances Leaf Callus Regeneration Efficiency in <i>Lagerstroemia indica</i> |
| title_sort | novel r2r3 myb transcription factor limyb75 enhances leaf callus regeneration efficiency in i lagerstroemia indica i |
| topic | whole-leaf explants callus induction genetic transformation plant hormone regeneration ability |
| url | https://www.mdpi.com/1999-4907/14/3/517 |
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