Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseases
Abstract The citrus industry is confronted with significant challenges posed by viral diseases. However, commercially favored seedless cultivars cannot be propagated through seeds to generate virus-free plants. Obtaining virus-free plants from seedless cultivars has been an important goal for resear...
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2024-12-01
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| Series: | Horticulture Advances |
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| Online Access: | https://doi.org/10.1007/s44281-024-00054-3 |
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| author | Zhehui Zhang Yongjun Hu Ming Wang Ruihan Nie Manli Sun Zongzhou Xie Lijun Chai Junli Ye Xiuxin Deng |
| author_facet | Zhehui Zhang Yongjun Hu Ming Wang Ruihan Nie Manli Sun Zongzhou Xie Lijun Chai Junli Ye Xiuxin Deng |
| author_sort | Zhehui Zhang |
| collection | DOAJ |
| description | Abstract The citrus industry is confronted with significant challenges posed by viral diseases. However, commercially favored seedless cultivars cannot be propagated through seeds to generate virus-free plants. Obtaining virus-free plants from seedless cultivars has been an important goal for research and practical purposes. In this study, virus-free plants were successfully regenerated from undeveloped ovules of seedless citrus cultivars through somatic embryogenesis. Following rigorous sterilization, the undeveloped ovules, collected from fruits harvested at 210 DAF (days after flowering), were cultured in darkness on MGS medium (MS medium containing 0.5 g L−1 malt extract, 1 mg L−1 GA3, 40 mg L−1 SAD [adenine hemisulfate]) to induce embryoid formation. The embryoids were then transferred to a 16-h photoperiod to promote bud regeneration. Thereafter, the regenerated buds were grafted onto in vitro rootstocks to recover entire plants. Four viruses, including Citrus exocortis viroid (CEVd), Citrus yellow vein clearing virus (CYVCV), Citrus tristeza virus (CTV), and Citrus tatter leaf virus (CTLV), were confirmed to be successfully eradicated in the regenerated plants. Both whole-genome resequencing and single-gene sequence alignment analyses confirmed that the regenerated plants retained genetic fidelity to the maternal plants. Furthermore, we established that the regeneration protocol from undeveloped ovules is an effective and practical method for virus elimination across diverse citrus varieties. These findings provide an alternative strategy for combating citrus viral diseases and achieving germplasm conservation. |
| format | Article |
| id | doaj-art-a34acf8acf3b48bcb18c1b67bafe19a8 |
| institution | DOAJ |
| issn | 2948-1104 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Springer |
| record_format | Article |
| series | Horticulture Advances |
| spelling | doaj-art-a34acf8acf3b48bcb18c1b67bafe19a82025-08-20T02:43:33ZengSpringerHorticulture Advances2948-11042024-12-012111210.1007/s44281-024-00054-3Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseasesZhehui Zhang0Yongjun Hu1Ming Wang2Ruihan Nie3Manli Sun4Zongzhou Xie5Lijun Chai6Junli Ye7Xiuxin Deng8National Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityNational Key Laboratory for Germplasm Lnnovation & Utilization of Horticultural Crops, Huazhong Agricultural UniversityAbstract The citrus industry is confronted with significant challenges posed by viral diseases. However, commercially favored seedless cultivars cannot be propagated through seeds to generate virus-free plants. Obtaining virus-free plants from seedless cultivars has been an important goal for research and practical purposes. In this study, virus-free plants were successfully regenerated from undeveloped ovules of seedless citrus cultivars through somatic embryogenesis. Following rigorous sterilization, the undeveloped ovules, collected from fruits harvested at 210 DAF (days after flowering), were cultured in darkness on MGS medium (MS medium containing 0.5 g L−1 malt extract, 1 mg L−1 GA3, 40 mg L−1 SAD [adenine hemisulfate]) to induce embryoid formation. The embryoids were then transferred to a 16-h photoperiod to promote bud regeneration. Thereafter, the regenerated buds were grafted onto in vitro rootstocks to recover entire plants. Four viruses, including Citrus exocortis viroid (CEVd), Citrus yellow vein clearing virus (CYVCV), Citrus tristeza virus (CTV), and Citrus tatter leaf virus (CTLV), were confirmed to be successfully eradicated in the regenerated plants. Both whole-genome resequencing and single-gene sequence alignment analyses confirmed that the regenerated plants retained genetic fidelity to the maternal plants. Furthermore, we established that the regeneration protocol from undeveloped ovules is an effective and practical method for virus elimination across diverse citrus varieties. These findings provide an alternative strategy for combating citrus viral diseases and achieving germplasm conservation.https://doi.org/10.1007/s44281-024-00054-3Seedless citrusVirus eliminationUndeveloped ovulesPlant regeneration |
| spellingShingle | Zhehui Zhang Yongjun Hu Ming Wang Ruihan Nie Manli Sun Zongzhou Xie Lijun Chai Junli Ye Xiuxin Deng Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseases Horticulture Advances Seedless citrus Virus elimination Undeveloped ovules Plant regeneration |
| title | Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseases |
| title_full | Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseases |
| title_fullStr | Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseases |
| title_full_unstemmed | Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseases |
| title_short | Regeneration of virus-free plants from undeveloped ovules of seedless citrus cultivars: a practical approach for combating citrus viral diseases |
| title_sort | regeneration of virus free plants from undeveloped ovules of seedless citrus cultivars a practical approach for combating citrus viral diseases |
| topic | Seedless citrus Virus elimination Undeveloped ovules Plant regeneration |
| url | https://doi.org/10.1007/s44281-024-00054-3 |
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