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<p>The current study demonstrates the production of synthetic seeds, short-term cold storage, and genetic uniformity analysis of synthetic seed-derived plantlets of <i>Coleus forskohlii</i> (Willd) Briq., using Inter Simple Sequence Repeat (ISSR) molecular markers. <i>C. forskohlii</i> is a highly valuable aromatic medicinal herb of the family Lamiaceae<i>.</i> To optimize synthetic seed production, non-embryogenic explants (nodal segments with axillary buds), of 12&#xa0;week old in vitro grown plantlets of <i>C. forskohlii</i> were encapsulated using 2%, 3% and 4% Sodium alginate (SA) with 75&#xa0;mM and 100&#xa0;mM Calcium chloride (CaCl<sub>2</sub>·2H<sub>2</sub>O). The encapsulated beads were short term cold stored at different time periods under refrigeration conditions (4&#xa0;°C) and another at culture room conditions (25&#xa0;°C) in dark. The synthetic seeds produced using 3% SA and 75&#xa0;mM CaCl<sub>2</sub>·2H<sub>2</sub>O showed the optimal encapsulation matrix formation favoured superior regeneration ability (63.33%) on the pre-optimized regeneration medium (½ MS medium with 0.5 mgL<sup>−1</sup> BAP + 0.1 mgL<sup>−1</sup> IAA)<b>.</b> The synthetic seeds-derived shoots exhibited the highest mean number of multiple shoot (17.21 ± 0.14) proliferation tendencies till 3rd subculture, wherein adequate in vitro rooting (94.33 ± 0.33%) was recorded on ½ MS medium contained 0.5 mgL<sup>−1</sup> IBA. The plantlets were hardened in the greenhouse and successfully transplanted to the field conditions with 90% success rate. The banding patterns of ISSR molecular marker analysis between the synthetic seed derived plants and mother plant was noticed to be monomorphic. This is the first report on synthetic seed preparation and cold storage of <i>C. forskohlii,</i> demonstrating that synthetic seed-derived plantlets maintained true-to-type characteristics. This approach offers a viable alternative for the micropropagation of this species and facilitates the transfer and exchange of germplasm over distant places.</p>

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Production of non-embryogenic synthetic seeds for short-term cold storage and ISSR marker-assisted genetic fidelity analysis of Coleus forskohlii (Willd) Briq. plants

  • 期刊论文

  • 发表日期：

    2025年1月31日

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  • Mahesh Kumar Badhepuri,
  • Mahender Aileni,
  • Prabhakar Rao Beeravelli,
  • Sathish Kumar Durgam,
  • Dulam Sandhya,
  • Mahipal S. Shekhawat,
  • Nageswara Rao Singisala

摘要

The current study demonstrates the production of synthetic seeds, short-term cold storage, and genetic uniformity analysis of synthetic seed-derived plantlets of Coleus forskohlii (Willd) Briq., using Inter Simple Sequence Repeat (ISSR) molecular markers. C. forskohlii is a highly valuable aromatic medicinal herb of the family Lamiaceae. To optimize synthetic seed production, non-embryogenic explants (nodal segments with axillary buds), of 12 week old in vitro grown plantlets of C. forskohlii were encapsulated using 2%, 3% and 4% Sodium alginate (SA) with 75 mM and 100 mM Calcium chloride (CaCl2·2H2O). The encapsulated beads were short term cold stored at different time periods under refrigeration conditions (4 °C) and another at culture room conditions (25 °C) in dark. The synthetic seeds produced using 3% SA and 75 mM CaCl2·2H2O showed the optimal encapsulation matrix formation favoured superior regeneration ability (63.33%) on the pre-optimized regeneration medium (½ MS medium with 0.5 mgL−1 BAP + 0.1 mgL−1 IAA). The synthetic seeds-derived shoots exhibited the highest mean number of multiple shoot (17.21 ± 0.14) proliferation tendencies till 3rd subculture, wherein adequate in vitro rooting (94.33 ± 0.33%) was recorded on ½ MS medium contained 0.5 mgL−1 IBA. The plantlets were hardened in the greenhouse and successfully transplanted to the field conditions with 90% success rate. The banding patterns of ISSR molecular marker analysis between the synthetic seed derived plants and mother plant was noticed to be monomorphic. This is the first report on synthetic seed preparation and cold storage of C. forskohlii, demonstrating that synthetic seed-derived plantlets maintained true-to-type characteristics. This approach offers a viable alternative for the micropropagation of this species and facilitates the transfer and exchange of germplasm over distant places.