Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion Fruit
Passion fruit (<i>Passiflora edulis</i> Sims), belonging to the Passifloraceae family, is an economically important plant in tropical and subtropical regions. The advances in functional genomics research of passion fruit have been significantly hindered by its recalcitrance to regenerati...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-07-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2223-7747/14/15/2312 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849239793229103104 |
|---|---|
| author | Jiayi Pan Yiping Zheng Tiancai Wang Pengpeng Xiong Kaibo Cui Lihui Zeng Ting Fang |
| author_facet | Jiayi Pan Yiping Zheng Tiancai Wang Pengpeng Xiong Kaibo Cui Lihui Zeng Ting Fang |
| author_sort | Jiayi Pan |
| collection | DOAJ |
| description | Passion fruit (<i>Passiflora edulis</i> Sims), belonging to the Passifloraceae family, is an economically important plant in tropical and subtropical regions. The advances in functional genomics research of passion fruit have been significantly hindered by its recalcitrance to regeneration and stable transformation. This study establishes the first efficient <i>Agrobacterium rhizogenes</i>-mediated hairy root transformation system for passion fruit. Utilizing the <i>eGFP</i> marker gene, transformation efficiencies of 11.3% were initially achieved with strains K599, MSU440, and C58C1, with K599 proving most effective. Key transformation parameters were systematically optimized to achieve the following: OD<sub>600</sub> = 0.6, infection duration 30 min, acetosyringone concentration 100 μM, and a dark co-cultivation period of 2 days. The system’s utility was further enhanced by incorporating the red visual marker RUBY, enabling direct, instrument-free identification of transgenic roots via betaxanthin accumulation. Finally, this system was applied for functional analysis using <i>PeMYB123</i>, which may be involved in proanthocyanidin accumulation. Overexpression of <i>PeMYB123</i> produced a higher content of proanthocyanidin in hairy roots. Additionally, the <i>PeANR</i> gene involved in the proanthocyanidin pathway was strongly activated in the transgenic hairy roots. This rapid and efficient visually simplified hairy root transformation system provides a powerful tool for functional gene studies in passion fruit. |
| format | Article |
| id | doaj-art-2268fe8c4b434e3ca1482dfad7d633da |
| institution | Kabale University |
| issn | 2223-7747 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj-art-2268fe8c4b434e3ca1482dfad7d633da2025-08-20T04:00:50ZengMDPI AGPlants2223-77472025-07-011415231210.3390/plants14152312Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion FruitJiayi Pan0Yiping Zheng1Tiancai Wang2Pengpeng Xiong3Kaibo Cui4Lihui Zeng5Ting Fang6College of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaCollege of Horticulture, Institute of Genetics and Breeding in Horticultural Plants, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaPassion fruit (<i>Passiflora edulis</i> Sims), belonging to the Passifloraceae family, is an economically important plant in tropical and subtropical regions. The advances in functional genomics research of passion fruit have been significantly hindered by its recalcitrance to regeneration and stable transformation. This study establishes the first efficient <i>Agrobacterium rhizogenes</i>-mediated hairy root transformation system for passion fruit. Utilizing the <i>eGFP</i> marker gene, transformation efficiencies of 11.3% were initially achieved with strains K599, MSU440, and C58C1, with K599 proving most effective. Key transformation parameters were systematically optimized to achieve the following: OD<sub>600</sub> = 0.6, infection duration 30 min, acetosyringone concentration 100 μM, and a dark co-cultivation period of 2 days. The system’s utility was further enhanced by incorporating the red visual marker RUBY, enabling direct, instrument-free identification of transgenic roots via betaxanthin accumulation. Finally, this system was applied for functional analysis using <i>PeMYB123</i>, which may be involved in proanthocyanidin accumulation. Overexpression of <i>PeMYB123</i> produced a higher content of proanthocyanidin in hairy roots. Additionally, the <i>PeANR</i> gene involved in the proanthocyanidin pathway was strongly activated in the transgenic hairy roots. This rapid and efficient visually simplified hairy root transformation system provides a powerful tool for functional gene studies in passion fruit.https://www.mdpi.com/2223-7747/14/15/2312passion fruit<i>Agrobacterium rhizogenes</i>hairy root<i>PeMYB123</i>proanthocyanidin |
| spellingShingle | Jiayi Pan Yiping Zheng Tiancai Wang Pengpeng Xiong Kaibo Cui Lihui Zeng Ting Fang Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion Fruit Plants passion fruit <i>Agrobacterium rhizogenes</i> hairy root <i>PeMYB123</i> proanthocyanidin |
| title | Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion Fruit |
| title_full | Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion Fruit |
| title_fullStr | Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion Fruit |
| title_full_unstemmed | Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion Fruit |
| title_short | Establishment of an Efficient <i>Agrobacterium rhizogenes</i>-Mediated Hairy Root Transformation System for Functional Analysis in Passion Fruit |
| title_sort | establishment of an efficient i agrobacterium rhizogenes i mediated hairy root transformation system for functional analysis in passion fruit |
| topic | passion fruit <i>Agrobacterium rhizogenes</i> hairy root <i>PeMYB123</i> proanthocyanidin |
| url | https://www.mdpi.com/2223-7747/14/15/2312 |
| work_keys_str_mv | AT jiayipan establishmentofanefficientiagrobacteriumrhizogenesimediatedhairyroottransformationsystemforfunctionalanalysisinpassionfruit AT yipingzheng establishmentofanefficientiagrobacteriumrhizogenesimediatedhairyroottransformationsystemforfunctionalanalysisinpassionfruit AT tiancaiwang establishmentofanefficientiagrobacteriumrhizogenesimediatedhairyroottransformationsystemforfunctionalanalysisinpassionfruit AT pengpengxiong establishmentofanefficientiagrobacteriumrhizogenesimediatedhairyroottransformationsystemforfunctionalanalysisinpassionfruit AT kaibocui establishmentofanefficientiagrobacteriumrhizogenesimediatedhairyroottransformationsystemforfunctionalanalysisinpassionfruit AT lihuizeng establishmentofanefficientiagrobacteriumrhizogenesimediatedhairyroottransformationsystemforfunctionalanalysisinpassionfruit AT tingfang establishmentofanefficientiagrobacteriumrhizogenesimediatedhairyroottransformationsystemforfunctionalanalysisinpassionfruit |