Lu Yin, Dai Shuangyan, Gu Aixia, Liu Mengyang, Wang Yanhua, Luo Shuangxia, Zhao Yujing, Wang Shan, Xuan Shuxin, Chen Xueping, Li Xiaofeng, Bonnema Guusje, Zhao Jianjun, Shen Shuxing
Key Laboratory of Vegetable Germplasm and Utilization of Hebei, Collaborative Innovation Center of Vegetable Industry in Hebei, College of Horticulture, Agricultural University of Hebei Baoding, China.
Plant Breeding, Wageningen University & Research Wageningen, Netherlands.
Front Plant Sci. 2016 Nov 28;7:1780. doi: 10.3389/fpls.2016.01780. eCollection 2016.
Chinese cabbage buds were soaked with Ethyl methanesulfonate (EMS) to induce mutagenesis. The influence of different EMS concentrations and treatment durations on microspore development, embryo production rate and seedling rate were evaluated in five Chinese cabbage genotypes. Mutations in four color-related genes were identified using high resolution melting (HRM) curves of their PCR products. The greatest embryo production and seedling rates were observed when buds were treated with 0.03 to 0.1% EMS for 5 to 10 min, while EMS concentrations greater than 0.1% were lethal to the microspores. In total, 142 mutants with distinct variations in leaf shape, leaf color, corolla size, flower color, bolting time and downy mildew resistance were identified from 475 microspore culture derived Doubled Haploids. Our results demonstrate that microspore derived Doubled Haploids from EMS soaked buds represents an efficient approach to rapidly generate homozygous Chinese cabbage mutants.
用甲基磺酸乙酯（EMS）浸泡大白菜花蕾以诱导诱变。评估了不同EMS浓度和处理时间对5种大白菜基因型小孢子发育、胚发生率和成苗率的影响。利用4个与颜色相关基因的PCR产物的高分辨率熔解（HRM）曲线鉴定突变。当花蕾用0.03%至0.1%的EMS处理5至10分钟时，观察到最高的胚发生率和成苗率，而EMS浓度大于0.1%对小孢子具有致死性。从475个小孢子培养获得的双单倍体中，共鉴定出142个在叶形、叶色、花冠大小、花色、抽薹时间和霜霉病抗性方面有明显变异的突变体。我们的结果表明，EMS浸泡花蕾获得的小孢子双单倍体是快速产生纯合大白菜突变体的有效方法。
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