An efficient <i>in vitro</i> propagation, antioxidant and antimicrobial activities of <i>Aphyllorchis montana</i> Rchb.f.
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An <i>in vitro</i> plant regeneration protocol was successfully established in <i>Aphyllorchis montana</i>, a saprophytic achlorophyllous orchid by culturing immature seeds. Among the six basal media evaluated for seed germination, BM-1-Terrestrial Orchid medium was found to be the best followed by Knudson C medium. Half-strength BM-1-Terrestrial Orchid medium was supplemented with different growth regulators either individually or in combinations for multiplication of shoots induction. Of the five cytokinins tested, thidiazuron at 6.8 μM was found to be most effective for multiple shoot induction yielding 17.24 ± 0.27 shoots after 10 weeks of culture. Addition of low concentration of α-Naphthaleneacetic acid (1.3 μM) in MS medium supplemented with the cytokinin thidiazuron (6.8 μM) favored shoot multiplication. The addition of organic additives to the medium containing thidiazuron enhanced the number of regenerated shoots. The plantlets were acclimatized, and the survival rate was 100%. Screening of the antioxidant and antimicrobial activities and estimation of total phenolics and flavonoid content of methanolic extracts of micropropagated plants were also carried out and compared with that of the wild-grown plants. In all the tests, methanolic extract from wild-grown plants showed higher antioxidant, antimicrobial activity, total phenolics and flavonoid content than <i>in vitro</i> propagated plants.
本研究以腐生无叶绿素兰科植物无叶兰（Aphyllorchis montana）为材料，通过培养未成熟种子，成功建立了该物种的离体（in vitro）植株再生体系。在所评估的6种用于种子萌发的基础培养基中，BM-1陆生兰培养基（BM-1-Terrestrial Orchid medium）的效果最佳，其次为Knudson C培养基。以减半浓度的BM-1陆生兰培养基为基础，分别单独或复配添加不同生长调节剂，用于诱导丛生芽增殖。在所测试的5种细胞分裂素（cytokinins）中，浓度为6.8 μM的噻苯隆（thidiazuron）对丛生芽诱导效果最优，培养10周后可获得17.24 ± 0.27个不定芽。在添加6.8 μM噻苯隆的MS培养基（MS medium）中，加入1.3 μM的低浓度α-萘乙酸（α-Naphthaleneacetic acid）可促进芽的增殖。向含有噻苯隆的培养基中添加有机添加物，可提升再生芽的数量。对获得的组培苗进行炼苗驯化，其成活率达100%。此外，本研究还对微繁殖植株（micropropagated plants）的甲醇提取物进行了抗氧化与抗菌活性筛选，并测定了其中总酚类物质与总黄酮含量，同时与野生植株的对应指标进行了对比。所有测试结果均显示，野生植株的甲醇提取物在抗氧化活性、抗菌活性、总酚类物质含量及总黄酮含量上，均优于离体繁殖植株。
