Cytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analyses
Abstract Background In vitro rooting remains a challenge for many woody species, including Quercus robur. While auxins combined with activated charcoal (AC) have traditionally been used, Q. robur explants exhibit a strong dependence on cytokinins (CKs). This study introduces a novel approach combini...
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BMC
2025-08-01
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| Series: | BMC Plant Biology |
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| Online Access: | https://doi.org/10.1186/s12870-025-07064-x |
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| author | João Paulo Rodrigues Martins Amir Mohammad Mokhtari Mikołaj Krzysztof Wawrzyniak |
| author_facet | João Paulo Rodrigues Martins Amir Mohammad Mokhtari Mikołaj Krzysztof Wawrzyniak |
| author_sort | João Paulo Rodrigues Martins |
| collection | DOAJ |
| description | Abstract Background In vitro rooting remains a challenge for many woody species, including Quercus robur. While auxins combined with activated charcoal (AC) have traditionally been used, Q. robur explants exhibit a strong dependence on cytokinins (CKs). This study introduces a novel approach combining AC with either 6-benzylaminopurine (BAP) or meta-topolin (mT) during the rooting phase to evaluate morphophysiological and hormonal responses. Methods Eight treatments were tested: 0 µM CK + 2 g L−1 AC; 2, 4, and 8 µM BAP + 2 g L−1 AC; 2, 4, and 8 µM mT + 2 g L−1 AC; and to isolate the effect of AC, a treatment without exogenous CK and AC (0 µM CK + 0 g L−1 AC) was also included. Morphophysiological and hormonal analyses were performed after 40 days of cultivation. Results Plantlets grown without AC exhibited a high shoot-to-root biomass ratio, indicating reduced growth balance. Treatments with 0–8 µM BAP combined with 2 g L−1 AC reduced the ratio of vascular cylinder area to total root cross-section by 16.5% compared to the other treatments. Notably, mT-treated plantlets developed root systems with improved anatomical features conducive to transport efficiency. CKs at 8 µM significantly enhanced stem thickening via stimulated cambial activity. Additionally, CK + AC treatments increased chlorophyll (Chl a + b) and carotenoid (Car) contents, while the absence of both CK and AC resulted in higher Car/Chl a + b ratios and an increased photosystem II closure rate (M0), indicating stress. Hormonally, CK-free treatments led to elevated indole-3-acetic acid (IAA) and reduced jasmonic acid (JA) levels, suggesting an imbalance. Conclusion Overall, the combination of AC and 4–8 µM mT during rooting enhanced secondary growth and improved the hormonal balance and morphophysiological traits of Q. robur plantlets. |
| format | Article |
| id | doaj-art-9d49b0422a8a4300a590bc824baa4ddd |
| institution | DOAJ |
| issn | 1471-2229 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | BMC |
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| series | BMC Plant Biology |
| spelling | doaj-art-9d49b0422a8a4300a590bc824baa4ddd2025-08-20T03:04:27ZengBMCBMC Plant Biology1471-22292025-08-0125111410.1186/s12870-025-07064-xCytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analysesJoão Paulo Rodrigues Martins0Amir Mohammad Mokhtari1Mikołaj Krzysztof Wawrzyniak2Wawrzyniak Institute of Dendrology, Polish Academy of SciencesWawrzyniak Institute of Dendrology, Polish Academy of SciencesWawrzyniak Institute of Dendrology, Polish Academy of SciencesAbstract Background In vitro rooting remains a challenge for many woody species, including Quercus robur. While auxins combined with activated charcoal (AC) have traditionally been used, Q. robur explants exhibit a strong dependence on cytokinins (CKs). This study introduces a novel approach combining AC with either 6-benzylaminopurine (BAP) or meta-topolin (mT) during the rooting phase to evaluate morphophysiological and hormonal responses. Methods Eight treatments were tested: 0 µM CK + 2 g L−1 AC; 2, 4, and 8 µM BAP + 2 g L−1 AC; 2, 4, and 8 µM mT + 2 g L−1 AC; and to isolate the effect of AC, a treatment without exogenous CK and AC (0 µM CK + 0 g L−1 AC) was also included. Morphophysiological and hormonal analyses were performed after 40 days of cultivation. Results Plantlets grown without AC exhibited a high shoot-to-root biomass ratio, indicating reduced growth balance. Treatments with 0–8 µM BAP combined with 2 g L−1 AC reduced the ratio of vascular cylinder area to total root cross-section by 16.5% compared to the other treatments. Notably, mT-treated plantlets developed root systems with improved anatomical features conducive to transport efficiency. CKs at 8 µM significantly enhanced stem thickening via stimulated cambial activity. Additionally, CK + AC treatments increased chlorophyll (Chl a + b) and carotenoid (Car) contents, while the absence of both CK and AC resulted in higher Car/Chl a + b ratios and an increased photosystem II closure rate (M0), indicating stress. Hormonally, CK-free treatments led to elevated indole-3-acetic acid (IAA) and reduced jasmonic acid (JA) levels, suggesting an imbalance. Conclusion Overall, the combination of AC and 4–8 µM mT during rooting enhanced secondary growth and improved the hormonal balance and morphophysiological traits of Q. robur plantlets.https://doi.org/10.1186/s12870-025-07064-xChlorophyll a fluorescencePlant anatomyPlant growth regulatorPedunculate oak |
| spellingShingle | João Paulo Rodrigues Martins Amir Mohammad Mokhtari Mikołaj Krzysztof Wawrzyniak Cytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analyses BMC Plant Biology Chlorophyll a fluorescence Plant anatomy Plant growth regulator Pedunculate oak |
| title | Cytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analyses |
| title_full | Cytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analyses |
| title_fullStr | Cytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analyses |
| title_full_unstemmed | Cytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analyses |
| title_short | Cytokinins combined with activated charcoal do not impair in vitro rooting in Quercus robur L.: insights from morphophysiological and hormonal analyses |
| title_sort | cytokinins combined with activated charcoal do not impair in vitro rooting in quercus robur l insights from morphophysiological and hormonal analyses |
| topic | Chlorophyll a fluorescence Plant anatomy Plant growth regulator Pedunculate oak |
| url | https://doi.org/10.1186/s12870-025-07064-x |
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