Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis
Abstract Background Hairy root (HR) transformation assays mediated by Agrobacterium rhizogenes, both in vitro and ex vitro, are essential tools in plant biotechnology and functional genomics. These assays can be significantly influenced by various factors, which ultimately can enhance the efficiency...
Saved in:
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-07-01
|
| Series: | BMC Biotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12896-025-00990-6 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849769234532401152 |
|---|---|
| author | Ladan Ajdanian Davoud Torkamaneh |
| author_facet | Ladan Ajdanian Davoud Torkamaneh |
| author_sort | Ladan Ajdanian |
| collection | DOAJ |
| description | Abstract Background Hairy root (HR) transformation assays mediated by Agrobacterium rhizogenes, both in vitro and ex vitro, are essential tools in plant biotechnology and functional genomics. These assays can be significantly influenced by various factors, which ultimately can enhance the efficiency. In this study, we optimized a two-step ex vitro HR transformation method using the actual mother plant combined with the RUBY system and compared with existing methods. Results The two-step ex vitro method proved more efficient than both the one-step ex vitro and in vitro methods, with the highest transformation efficiency of 90% observed in the actual plant. This technique also demonstrated a faster and less complicated approach, reducing time to achieve massive transgenic HR formation by 9–29 days compared to other methods. Conclusions A novel, quicker, less complicated, and more efficient two-step transformation method for cannabis has been established, presenting a significantly lower risk of contamination. This protocol is particularly interesting to produce secondary metabolites using the CRISPR/Cas system in cannabis. We anticipate that this method will facilitate substantial time savings by rapidly producing hundreds of transformed samples. |
| format | Article |
| id | doaj-art-de529bc1fcb042aaa8f007dba824235a |
| institution | DOAJ |
| issn | 1472-6750 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj-art-de529bc1fcb042aaa8f007dba824235a2025-08-20T03:03:28ZengBMCBMC Biotechnology1472-67502025-07-012511710.1186/s12896-025-00990-6Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for CannabisLadan Ajdanian0Davoud Torkamaneh1Département de Phytologie, Université LavalDépartement de Phytologie, Université LavalAbstract Background Hairy root (HR) transformation assays mediated by Agrobacterium rhizogenes, both in vitro and ex vitro, are essential tools in plant biotechnology and functional genomics. These assays can be significantly influenced by various factors, which ultimately can enhance the efficiency. In this study, we optimized a two-step ex vitro HR transformation method using the actual mother plant combined with the RUBY system and compared with existing methods. Results The two-step ex vitro method proved more efficient than both the one-step ex vitro and in vitro methods, with the highest transformation efficiency of 90% observed in the actual plant. This technique also demonstrated a faster and less complicated approach, reducing time to achieve massive transgenic HR formation by 9–29 days compared to other methods. Conclusions A novel, quicker, less complicated, and more efficient two-step transformation method for cannabis has been established, presenting a significantly lower risk of contamination. This protocol is particularly interesting to produce secondary metabolites using the CRISPR/Cas system in cannabis. We anticipate that this method will facilitate substantial time savings by rapidly producing hundreds of transformed samples.https://doi.org/10.1186/s12896-025-00990-6Agrobacterium rhizogenesCRISPRin-vitroMother plantSecondary metabolite |
| spellingShingle | Ladan Ajdanian Davoud Torkamaneh Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis BMC Biotechnology Agrobacterium rhizogenes CRISPR in-vitro Mother plant Secondary metabolite |
| title | Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis |
| title_full | Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis |
| title_fullStr | Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis |
| title_full_unstemmed | Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis |
| title_short | Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis |
| title_sort | mother transformer a high throughput cost effective in planta hairy root transformation method for cannabis |
| topic | Agrobacterium rhizogenes CRISPR in-vitro Mother plant Secondary metabolite |
| url | https://doi.org/10.1186/s12896-025-00990-6 |
| work_keys_str_mv | AT ladanajdanian mothertransformerahighthroughputcosteffectiveinplantahairyroottransformationmethodforcannabis AT davoudtorkamaneh mothertransformerahighthroughputcosteffectiveinplantahairyroottransformationmethodforcannabis |