Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico.
Consejo Nacional de Humanidades, Ciencias y Tecnologías, Laboratorio Nacional de Investigación y Servicio Agroalimentario y Forestal, Universidad Autónoma de Chapingo, Texcoco, Estado de México, Mexico.
PeerJ. 2024 Nov 22;12:e18545. doi: 10.7717/peerj.18545. eCollection 2024.
Linton & Epling subsp. (Lamiaceae) is an aromatic medicinal plant, characterized by a high concentration of tilianin, a flavonoid with therapeutic potential in cardiovascular diseases. In this study, we have explored the use of colchicine to obtain autotetraploid lines of and analyze their morphological characteristics. In addition, we aimed to identify polyploid plants with a high content of tilianin.
seedlings at the stage of cotyledon emergence were dipped in colchicine solution at 0.0%, 0.1%, 0.3%, and 0.5% (w/v) for 6, 12, and 24 h. Seedlings were cultured on half-strength basal Murashige and Skoog medium supplemented with 20 g/L sucrose. After 2 months, the shoots from surviving seedlings were excised and grown individually in the same medium to obtain plantlets. The ploidy level of all materials was verified through flow cytometry and chromosome counting before acclimatization and transfer to the greenhouse. The investigated characteristics included length, density and stomatal index, leaf area, chlorophyll content, flower size and color, and tilianin content measured by high-performance liquid chromatography.
The most efficient production of tetraploid in terms of percentage was achieved with 0.1% colchicine for 6 h resulting in no generation of mixoploids. Tetraploid plants had twice the number of chromosomes (2 = 4x = 36) and nearly twice the total DNA content (2.660 ± 0.236 pg) of diploids. Most tetraploid plants showed variations in flower and leaf characteristics compared to the diploid controls. High-performance liquid chromatography analysis showed that tetraploid plants with small leaves produced the greatest amount of tilianin; up to 32.964 ± 0.004 mg/g dry weight (DW), compared to diploid plants with 6.388 ± 0.005 mg/g DW.
polyploidization using colchicine demonstrates potential for enhancing bioactive constituents of . This approach has proven effective in generating elite tetraploid lines with increased tilianin production.
Linton & Epling 亚种（唇形科）是一种芳香药用植物，其特征是含有高浓度的Tilianin，这是一种具有治疗心血管疾病潜力的类黄酮。在这项研究中，我们探索了使用秋水仙素获得 和分析其形态特征的同源四倍体系。此外，我们旨在鉴定Tilianin 含量高的多倍体植物。
在子叶出现阶段将幼苗浸泡在 0.0%、0.1%、0.3%和 0.5%（w/v）的秋水仙素溶液中 6、12 和 24 小时。幼苗在半强度 Murashige 和 Skoog 培养基上培养，补充 20 g/L 蔗糖。2 个月后，从存活幼苗中取出芽并在相同培养基中单独生长以获得植物。在适应和转移到温室之前，通过流式细胞术和染色体计数验证所有材料的倍性水平。调查的特征包括长度、密度和气孔指数、叶面积、叶绿素含量、花大小和颜色以及高效液相色谱法测量的Tilianin 含量。
以 0.1%秋水仙素处理 6 小时，获得了最高的四倍体生成效率，没有产生混倍体。四倍体植物的染色体数增加了一倍（2n=4x=36），总 DNA 含量增加了近一倍（2.660±0.236 pg）。与二倍体对照相比，大多数四倍体 植物的花和叶特征发生了变化。高效液相色谱分析表明，与二倍体植物的 6.388±0.005 mg/g DW 相比，小叶片的四倍体植物产生的 Tilianin 最多；达到 32.964±0.004 mg/g DW。
秋水仙素的多倍体化显示出增强 的生物活性成分的潜力。这种方法已被证明可有效生成Tilianin 产量增加的优良四倍体系。
