Instituto de Conservación y Mejora de la Agrodiversidad Valenciana, Universitat Politècnica de València, Valencia, Spain.
Methods Mol Biol. 2021;2264:197-206. doi: 10.1007/978-1-0716-1201-9_14.
The use of antimitotic agents such as colchicine has been common to obtain polyploid organisms. However, this approach entails certain problems, from its toxicity to the operators for being carcinogenic compounds to the instability of the individuals obtained, and the consequent reversion to its original ploidy because the individuals obtained in most cases are chimeric. In vitro culture allows taking advantage of the full potential offered by the cellular totipotence of plant organisms. Based on this, we present a new in vitro culture protocol to obtain polyploid organisms using zeatin riboside (ZR) and eggplant as a model organism. Flow cytometry is used to identify tetraploid regenerants. The regeneration of whole plants from the appropriate tissues using ZR allowed developing polyploid individuals in eggplant, a crop that tends to be recalcitrant to in vitro organogenesis. Thanks to the use of the polysomatic pattern of the explants, we have been able to develop a methodology that allows to obtain stable non-chimeric polyploid individuals from organogenic processes.
秋水仙素等抗有丝分裂剂的使用已被普遍用于获得多倍体生物。然而,这种方法存在一些问题,包括其对操作人员的毒性(因为这些化合物是致癌物质)、获得的个体的不稳定性,以及由于大多数情况下获得的个体是嵌合体,因此会恢复到其原始的倍性。体外培养可以充分利用植物生物的细胞全能性所提供的潜力。基于这一点,我们提出了一种新的体外培养方案,使用玉米素核苷(ZR)和茄子作为模型生物来获得多倍体生物。利用流式细胞术鉴定四倍体再生体。使用 ZR 从适当的组织再生出完整的植株,使得茄子中能够产生多倍体个体,而茄子在体外器官发生方面往往比较难处理。由于利用了外植体的多体细胞模式,我们已经能够开发出一种从器官发生过程中获得稳定非嵌合体多倍体个体的方法。