The production of double haploids (“DH”) by microspore-derived embryos is one of the best breeding methods to generate new varieties with better traits and stable genetic background. In the previous study, radish ‘DH’ lines were generated with a different range of glucosinolate (GSL) contents, ranging from low to high levels of GSL. In this study, we performed metabolic and transcriptional profiling to elucidate the regulatory system behind the GSL biosynthesis process using the ‘DH’ radish lines. Four individual ‘DH’ lines were selected, of which two lines named ‘HH’ had high GSL content, while the other two lines, ‘LL’, had low GSL content. HPLC analysis in the vegetative (leaf and root) and reproductive tissue (seed) was detected seven GSL compounds from radish. Among seven GSLs, glucoraphasatin (GRH) occupied the largest proportion of total GSLs in both leaf and root tissues, while glucoraphenin (GRE) was taking a place as the major GSL in radish seeds. Six major genes (RsBCAT4, RsIPMDH1, RsCYP79F1, RsCYP83A1, RsUGT74B1, and RsGRS1) representing the major biosynthesis of aliphatic GSLs were selected for qRT-PCR analysis. Heat map analysis based on Pearson correlation coefficient showed that RsBCAT4, RsIPMDH1, RsCYP79F1, and RsCYP83A1 were positively correlated with the amount of GRH in leaf tissue. Meanwhile, RsGRS1 showed a strong correlation with the amount of GRH and total aliphatic GSLs in both leaf and root tissues. By using the ‘DH’ radish lines, this study revealed that regardless of tissues of radish, accumulation of total GSL is strongly affected by the RsGRS1 and also positively supported by the other biosynthetic genes like RsBCAT4, RsIPMDH1, RsCYP79F1, and RsCYP83A1.

利用小孢子胚产生双单倍体（“DH”）是产生具有更好性状和稳定遗传背景的新品种的最佳育种方法之一。在之前的研究中，萝卜“DH”品系产生了不同范围的芥子油苷（GSL）含量，从低水平到高水平的GSL。在这项研究中，我们进行了代谢和转录分析，以阐明使用“DH”萝卜品系的 GSL 生物合成过程背后的调控系统。选择了四个单独的“DH”品系，其中名为“HH”的两个品系具有高GSL含量，而另外两个品系“LL”具有低GSL含量。通过 HPLC 分析，在萝卜的营养组织（叶和根）和生殖组织（种子）中检测到七种 GSL 化合物。七种GSL中，萝卜苷(GRH)在叶和根组织中所占的总GSL比例最大，而萝卜硫苷(GRE)在萝卜种子中占据主要GSL地位。选择代表脂肪族 GSL 主要生物合成的6 个主要基因（RsBCAT4、RsIPMDH1、RsCYP79F1、RsCYP83A1、RsUGT74B1和RsGRS1 ）进行 qRT-PCR 分析。基于Pearson相关系数的热图分析表明，RsBCAT4、RsIPMDH1、RsCYP79F1和RsCYP83A1与叶组织中GRH量呈正相关。同时，RsGRS1 与叶和根组织中 GRH 和总脂肪族 GSL 的含量呈很强的相关性。通过使用“DH”萝卜品系，这项研究表明，无论萝卜的组织如何，总 GSL 的积累都受到 RsGRS1 的强烈影响，并且还受到其他生物合成基因（如 RsBCAT4、RsIPMDH1、RsCYP79F1 和 RsCYP83A1）的积极支持。