Unan Rasim, Deligoz Ilyas, Al-Khatib Kassim, Mennan Husrev
Black Sea Agricultural Research Institute, Samsun, Turkey.
University of California, Davis, CA, USA.
Open Res Eur. 2022 Feb 14;1:19. doi: 10.12688/openreseurope.13317.3. eCollection 2021.
Non-transgenic chemical mutagen application, particularly ethyl methanesulfonate (EMS), is an important tool to create mutations and gain a new genetic makeup for plants. It is useful to obtain a sufficient number of mutant plants instead of working with a severe mutation in a few plants. EMS dose and exposure period have been previously studied in several crops; however, EMS used to create point mutations in presoaked rice seeds has not been sufficiently studied and there is no standard protocol for such treatment. The aim of this study is to establish a standard protocol for EMS mutagenesis application in rice. Two studies were conducted to evaluate the effect of four durations of rice seed presoaking (0, 6, 12, and 24 hours), four EMS concentration doses (0.0%, 0.5%, 1.0%, and 2.0%), and four EMS exposure periods (6, 12, 24, and 48 hours). Germination rate, plumula and radicle length, seedling survival, LD (Lethal Dose) determination, shoot length, root length and fresh seedling weight were evaluated. Results showed that a 12-hour presoaking duration, 0.5% EMS dose, and six hours of EMS exposure were the best practices for the optimum number of mutant plants. In light of both this study and the literature, a standard application protocol was established. This application protocol, detailed in this article, contains the following guidelines: (1) Presoaking: 12 hours, (2) EMS application: 0.5% dose EMS and six hours, (3) Final washing: six hours, (4) Drying: 72 hours at 38°C. A user-friendly protocol has been presented for utilization by researchers.
非转基因化学诱变剂的应用，特别是甲磺酸乙酯（EMS），是为植物创造突变并获得新遗传组成的重要工具。获得足够数量的突变植株而不是仅研究少数植株中的严重突变是很有用的。此前已在几种作物中研究过EMS剂量和处理时间；然而，用于在预浸水稻种子中产生点突变的EMS尚未得到充分研究，且尚无此类处理的标准方案。本研究的目的是建立水稻EMS诱变应用的标准方案。进行了两项研究，以评估水稻种子四种预浸时间（0、6、12和24小时）、四种EMS浓度剂量（0.0%、0.5%、1.0%和2.0%）以及四种EMS处理时间（6、12、24和48小时）的效果。评估了发芽率、胚芽和胚根长度、幼苗存活率、致死剂量测定、地上部分长度、根长度和幼苗鲜重。结果表明，12小时的预浸时间、0.5%的EMS剂量和6小时的EMS处理时间是获得最佳数量突变植株的最佳做法。结合本研究和文献，建立了标准应用方案。本文详细介绍的该应用方案包含以下指南：（1）预浸：12小时，（2）EMS处理：0.5%剂量的EMS处理6小时，（3）最终冲洗：6小时，（4）干燥：在38°C下干燥72小时。已提出一个便于用户使用的方案供研究人员使用。
