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<p><i>Arthrocaulon macrostacyhum</i> and <i>Salicornia fruticosa</i> are both halophytes, typical of most Tunisian salt marshes. They are characterized by their tolerance to very high saline conditions, both are potential candidates to remediate salt affected lands and desalinize saline waters. These halophytes are not easy to propagate, since germination and vegetative propagation are highly dependent on abiotic factors. Therefore, in vitro plant tissue culture techniques are alternative ways to improve their handling and production under controlled environmental conditions, and have several advantages over traditional approaches, including as higher multiplication rates, and absence of pathogens. The goal of this study was to develop an efficient mass propagation protocol for <i>S. fructicosa</i> and <i>A. macrostachyum</i> and determine the salinity level for optimal growth and tolerance under in vitro conditions. In order to evaluate the salt tolerance of vitro-plants, cuttings were cultured on MS medium enriched with 0.5 mg l<sup>−1</sup> of 6-benzylaminopurine and supplied with various NaCl concentrations ranging from 200, to 600 mM. Results indicated that the highest number of shoots, the average shoot elongation as well as root number and length were observed on medium containing 450 mM NaCl. Under 450 mM of NaCl, the macro-elements of the vitro-clones like potassium and calcium were not affected compared to control clones, whereas, heavy metals like lead decreased. More interestingly, micronutrients like Strontium, Zinc, Iron, Manganese and Selenium increased in salt treated vitro-halophytes. Following histological and scanning electron microscopy analysis, our results showed that the adaptation of vitro-halophytes clones to salinity was associated with special anatomical features such as succulence, the deposition of salt crystals in well-developed water storage cells, the appearance of dark particles in the epidermis in addition to several prominent closed sunken stomata, sclereids and tracheoidioblasts. This is the first report on salt-tolerance adaptation of Tunisian halophytes conducted under in vitro conditions. The developed protocol and the obtained results will help for selecting clones of these halophytic species with characteristics desirable for utilization and/or restoration in salt affected lands.</p>

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In vitro micropropagation and salt tolerance of Tunisian halophytes: Salicornia fructicosa (L.) A. J. Scott and Arthrocnemum macrostachyum (Moric.) C. Koch: as evidenced by mineral and micromorphological analysis

  • 期刊论文

  • 发表日期：

    2025年8月14日

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  • Badra Bouamama-Gzara,
  • Atrous Ghofrane,
  • Gandoura Samia,
  • Dabebi Afef,
  • Nafati Haythem,
  • Farah Faten,
  • Zemni Hassene,
  • Karim Ben Hamed

摘要

Arthrocaulon macrostacyhum and Salicornia fruticosa are both halophytes, typical of most Tunisian salt marshes. They are characterized by their tolerance to very high saline conditions, both are potential candidates to remediate salt affected lands and desalinize saline waters. These halophytes are not easy to propagate, since germination and vegetative propagation are highly dependent on abiotic factors. Therefore, in vitro plant tissue culture techniques are alternative ways to improve their handling and production under controlled environmental conditions, and have several advantages over traditional approaches, including as higher multiplication rates, and absence of pathogens. The goal of this study was to develop an efficient mass propagation protocol for S. fructicosa and A. macrostachyum and determine the salinity level for optimal growth and tolerance under in vitro conditions. In order to evaluate the salt tolerance of vitro-plants, cuttings were cultured on MS medium enriched with 0.5 mg l−1 of 6-benzylaminopurine and supplied with various NaCl concentrations ranging from 200, to 600 mM. Results indicated that the highest number of shoots, the average shoot elongation as well as root number and length were observed on medium containing 450 mM NaCl. Under 450 mM of NaCl, the macro-elements of the vitro-clones like potassium and calcium were not affected compared to control clones, whereas, heavy metals like lead decreased. More interestingly, micronutrients like Strontium, Zinc, Iron, Manganese and Selenium increased in salt treated vitro-halophytes. Following histological and scanning electron microscopy analysis, our results showed that the adaptation of vitro-halophytes clones to salinity was associated with special anatomical features such as succulence, the deposition of salt crystals in well-developed water storage cells, the appearance of dark particles in the epidermis in addition to several prominent closed sunken stomata, sclereids and tracheoidioblasts. This is the first report on salt-tolerance adaptation of Tunisian halophytes conducted under in vitro conditions. The developed protocol and the obtained results will help for selecting clones of these halophytic species with characteristics desirable for utilization and/or restoration in salt affected lands.