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Fatemeh Soghra Younesikelaki ¹, Mohammad Hadi Ebrahimzadeh ¹, Mehdi Kiani Desfardi ¹, Mahitha Banala ¹, Rajinikanth Marka ¹, Rama Swamy Nanna ¹

Affiliations
1 Plant Biotechnology Research Laboratory, Department of Biotechnology Kakatiya University, Warangal - 506009, Telangana, IN

Abstract

Objectives: The species Althaea officinalis is belong to Malvaceae family; its common name is Marshmallow. In this study an efficient technique has been optimized for sterilization of seed surface and also in vitro germination of seeds. Methods: Effect of different sterilizing agents with various concentrations and duration times as well as different media with various concentrations of sucrose on MGT (mean of germination time), GP (germination percentage), GR (germination rate), NDFG (number of days to first germination), CP (contamination percentage) and length of ischolar_main and shoot of seedlings were recorded. For each experiment 25 seeds were treated and it was repeated at least three times. Findings: The maximum GP (80.0%) was recorded in sterilized-distilled water for 20 min (control), whereas minimum GP (44.0%) was recorded in experiment using 0.3% mercuric chloride (HgCl2) for 7 min. Between seed surface sterilization techniques used, maximum CP was achieved in control (85.33%) and minimum was found in 4% NaOCl (Sodium Hypochlorite), 10 min (13.33%), 0.3% HgCl2 for 7 min (14.66%) and 4% NaOCl for 5 min (17.33%). We compared the measured seed germination values (NDFG, MGT, GR, GP and CP) and among seed surface sterilization methods tested, NaOCl, 5 min has been found to be the best in A. officinalis. In addition, presence of sucrose in germination medium has shown negative effect on seed germination measured values as well seedling development. The effect of different surface sterilization techniques have no significant effect (p ≤ 0.05) on number of days to first germination (NDFG) but these techniques significantly affected (p ≤ 0.05) the other characteristics of seed sterilization. According to ANOVA results we report for the first time the best sterilization method and medium for in vitro seed germination and seedling development in A. officinalis.

Keywords

Althaea officinalis, Seed Surface Sterilization, Seed Germination.

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Mohammad Hadi Ebrahimzadeh ¹, Fatemeh Soghra Younesikelaki ¹, Mehdi Kiani Desfardi ¹, Mahitha Banala ¹, Rama Swamy Nanna ¹

Affiliations
1 Department of Biotechnology, Kakatiya University, Warangal - 506009, Telangana, IN

Abstract

Objectives: The species Silybum marianum (L.) (Asteraceae) commonly known as Milk thistle is medicinally valuable and due to over exploitation the species may become threatened. Hence, we have made an attempt to optimize the protocol for its direct regeneration through cotyledon explants using in vitro culture technology. Methods/Statistical Analysis: For in vitro multiple-shoot induction, cotyledon explants were excised from surfacesterilized seeds and cultured on MS Basal Medium (MBM) enriched with diverse conc. (10.0–20.0 μM) of BAP/KIN alone and also 10.0 μM BAP/KIN in combination with 1.25–5.0 μM NAA/IAA/IBA. The micro-shoots were cut out and cultured on 1/2 strength MBM enriched with diverse concentrations (5.0-20.0 μM) of NAA/IAA/IBA alone and also coupled with 1.25–5.0 μM BAP/KIN. Findings: In this study, superlative rate of shoot induction, maximal Number of Shoots Per Explant (NSPE) and longest shoot-length were achieved on MBM enriched with 10.0 μM BAP + 2.5 μM NAA. Superlative rate of ischolar_main induction, mean number of ischolar_mains per shoot and mean ischolar_main length were documented on MBM supplemented with 15.0 μM NAA + 2.5 μM BAP. Application/Improvement: Being over exploited, the current technique can be operated for accelerated multiplication and conservation of the species. Further, this protocol can also be operated for transfer of novel genes for enhancement and modulation of bioactive molecules production in S. marianum – a medicinally valuable plant.

Keywords

Cotyledon, In vitro Regeneration, Silybum Marianum.

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