Departamento de Biologia, Laboratório de Genética e Biotecnologia, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Juiz de Fora, Brazil.
Methods Mol Biol. 2024;2827:267-278. doi: 10.1007/978-1-0716-3954-2_18.
In ornamental plants, artificial polyploidization has enabled the creation of new cultivars. Due to their high commercial value in the international flower market and their ornamental characteristics, such as the shape, size, color, and durability of their flower, orchids have received great attention in studies of artificial polyploidization. Here we described the protocol used for polyploid induction in Oncidium crispum, an epiphyte species native of southeastern Brazil, of great ornamental interest and widely sold in flower shops. The species stands out for having inflorescence with large flowers, brown in color with yellow spots. In addition, O. crispum has great potential for use in genetic improvement programs since the species is widely used in interspecific crosses. Closed capsules containing mature O. crispum seeds were subjected to running sterilized water for 10 min and then to a 1.5% sodium hypochlorite solution for 10 min. Small portions of seeds were introduced into 50 mL of water-soluble fertilizer with macro- and micronutrients (B&G) plus 0.7% agar. Explants originating from seeds previously in vitro germinated were submitted to 0.05% and 0.1% of colchicine for 4 days and 8 days. Flow cytometry and chromosome counts confirmed that the protocol successfully produced synthetic polyploid plants.
在观赏植物中，人工多倍体化已经创造了许多新的品种。由于其在国际花卉市场上具有很高的商业价值，以及其观赏特性，如花的形状、大小、颜色和耐久性，兰花在人工多倍体化的研究中受到了极大的关注。在这里，我们描述了在原产于巴西东南部的附生植物硬叶兜兰（Oncidium crispum）中诱导多倍体的方法，硬叶兜兰具有很大的观赏价值，在花店中广泛销售。该物种的花序具有大花朵，颜色为棕色，带有黄色斑点，非常引人注目。此外，硬叶兜兰具有很大的遗传改良潜力，因为该物种广泛用于种间杂交。将含有成熟硬叶兜兰种子的封闭蒴果在流动的无菌水中处理 10 分钟，然后在 1.5%次氯酸钠溶液中处理 10 分钟。将小部分种子接种到含有大量和微量元素（B&G）的 50 毫升水溶性肥料和 0.7%琼脂中。先前在体外萌发的种子起源的外植体分别在 0.05%和 0.1%的秋水仙素中处理 4 天和 8 天。流式细胞术和染色体计数证实，该方案成功地产生了合成的多倍体植物。
