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<p><i>Gentianella lutescens</i> subsp. <i>carpatica</i> (Beck) Holub, a rich source of xanthones with specific bioactive properties, is limited in nature. To improve shoot proliferation and growth, shoot explants were cultured for five weeks in vitro on Murashige and Skoog medium enriched with individual cytokinins: N<sup>6</sup>-benzyladenine (6-BA), kinetin (KIN), <i>meta</i>-topolin (<i>m</i>T), thidiazuron (TDZ) and forchlorfenuron&#xa0;(CPPU), (concentrations range 0–15&#xa0;µM). The results were in favor of the highest optimal cytokinin concentration 10&#xa0;µM for shoot culture. The efficacy of cytokinins on axillary shoots proliferation and growth was characterized as mT ˃ TDZ ˃ BA = KIN ˃ CPPU. <i>m</i>T and TDZ at 10&#xa0;µM produced the highest number of shoots (6.3 and 5.8, respectively). <i>m</i>T-treated shoots were robust, good quality and displayed the highest fresh mass increase (11.88-fold). Almost all cytokinins at 10&#xa0;µM did not increase the activity of antioxidant enzymes, as well as proline, H<sub>2</sub>O<sub>2</sub> and malondialdehyde content. The photosynthetic pigments were also preserved in all cytokinin-treated shoots compared to control. TDZ showed stronger effect than <i>m</i>T in increasing the content of xanthone glycosides (mangiferin, demethylbellidifolin-8-<i>O</i>-glucoside and bellidifolin-8<i>-O-</i>glucoside), and decreasing the content of their aglycones. Shoot proliferation was enhanced (9.22 shoots/explant) on <i>mT</i> by cultivation in glass jars instead of Erlenmeyer flasks. Compared to TDZ, <i>m</i>T also provided better shoot multiplication and growth under cold conditions (+ 4&#xa0;°C). The results obtained are useful for conservation and further biological and pharmacological studies of G. <i>lutescens</i> subsp. <i>carpatica. m</i>T could be considered to improve the tolerance of <i>Gentianella</i> species to abiotic stressors.</p> Graphical Abstract <p></p>

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The effect of different cytokinins on in vitro shoot culture of an endemic and endangered Gentianella lutescens subsp. carpatica (Beck) Holub

  • 期刊论文

  • 发表日期：

    2025年3月3日

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  • Marija Petrović,
  • Dijana Krstić-Milošević,
  • Mariana Stanišić,
  • Milena Trajković,
  • Nevena Banjac

摘要

Gentianella lutescens subsp. carpatica (Beck) Holub, a rich source of xanthones with specific bioactive properties, is limited in nature. To improve shoot proliferation and growth, shoot explants were cultured for five weeks in vitro on Murashige and Skoog medium enriched with individual cytokinins: N6-benzyladenine (6-BA), kinetin (KIN), meta-topolin (mT), thidiazuron (TDZ) and forchlorfenuron (CPPU), (concentrations range 0–15 µM). The results were in favor of the highest optimal cytokinin concentration 10 µM for shoot culture. The efficacy of cytokinins on axillary shoots proliferation and growth was characterized as mT ˃ TDZ ˃ BA = KIN ˃ CPPU. mT and TDZ at 10 µM produced the highest number of shoots (6.3 and 5.8, respectively). mT-treated shoots were robust, good quality and displayed the highest fresh mass increase (11.88-fold). Almost all cytokinins at 10 µM did not increase the activity of antioxidant enzymes, as well as proline, H2O2 and malondialdehyde content. The photosynthetic pigments were also preserved in all cytokinin-treated shoots compared to control. TDZ showed stronger effect than mT in increasing the content of xanthone glycosides (mangiferin, demethylbellidifolin-8-O-glucoside and bellidifolin-8-O-glucoside), and decreasing the content of their aglycones. Shoot proliferation was enhanced (9.22 shoots/explant) on mT by cultivation in glass jars instead of Erlenmeyer flasks. Compared to TDZ, mT also provided better shoot multiplication and growth under cold conditions (+ 4 °C). The results obtained are useful for conservation and further biological and pharmacological studies of G. lutescens subsp. carpatica. mT could be considered to improve the tolerance of Gentianella species to abiotic stressors.

Graphical Abstract