Central Laboratories for Frontier Technology, Kirin Holdings Company, Ltd., Yokohama, Kanagawa, Japan; RIKEN Plant Science Center, Yokohama, Kanagawa, Japan.
Plant Biotechnol J. 2013 Oct;11(8):1017-27. doi: 10.1111/pbi.12095. Epub 2013 Jul 30.
Plants belonging to the Brassicaceae family exhibit species-specific profiles of glucosinolates (GSLs), a class of defence compounds against pathogens and insects. GSLs also exhibit various human health-promoting properties. Among them, glucoraphanin (aliphatic 4-methylsulphinylbutyl GSL) has attracted the most attention because it hydrolyses to form a potent anticancer compound. Increased interest in developing commercial varieties of Brassicaceae crops with desirable GSL profiles has led to attempts to identify genes that are potentially valuable for controlling GSL biosynthesis. However, little attention has been focused on genes of kale (Brassica oleracea var. acephala). In this study, we established full-length kale cDNA libraries containing 59 904 clones, which were used to generate an expressed sequence tag (EST) data set with 119 204 entries. The EST data set clarified genes related to the GSL biosynthesis pathway in kale. We specifically focused on BoMYB29, a homolog of Arabidopsis MYB29/PMG2/HAG3, not only to characterize its function but also to demonstrate its usability as a biological resource. BoMYB29 overexpression in wild-type Arabidopsis enhanced the expression of aliphatic GSL biosynthetic genes and the accumulation of aliphatic GSLs. When expressed in the myb28myb29 mutant, which exhibited no detectable aliphatic GSLs, BoMYB29 restored the expression of biosynthetic genes and aliphatic GSL accumulation. Interestingly, the ratio of methylsulphinyl GSL content, including glucoraphanin, to that of methylthio GSLs was greatly increased, indicating the suitability of BoMYB29 as a regulator for increasing methylsulphinyl GSL content. Our results indicate that these biological resources can facilitate further identification of genes useful for modifications of GSL profiles and accumulation in kale.
芸薹属植物表现出物种特异性的硫代葡萄糖苷(GSL)谱,这是一类防御化合物,可抵御病原体和昆虫。GSL 还具有各种促进人类健康的特性。其中,萝卜硫素(脂肪族 4-甲基亚磺酰基丁基 GSL)最受关注,因为它水解后会形成一种有效的抗癌化合物。人们对开发具有理想 GSL 谱的芸薹属作物商业品种的兴趣日益浓厚,这导致人们试图鉴定出可能对控制 GSL 生物合成有价值的基因。然而,人们对羽衣甘蓝(Brassica oleracea var. acephala)的基因关注甚少。在这项研究中,我们建立了包含 59904 个克隆的全长羽衣甘蓝 cDNA 文库,该文库用于生成包含 119204 个条目表达序列标签(EST)数据集。EST 数据集阐明了羽衣甘蓝中与 GSL 生物合成途径相关的基因。我们特别关注 BoMYB29,它是拟南芥 MYB29/PMG2/HAG3 的同源物,不仅要对其功能进行表征,还要证明其作为生物资源的可用性。BoMYB29 在野生型拟南芥中的过表达增强了脂肪族 GSL 生物合成基因的表达和脂肪族 GSL 的积累。当在没有检测到脂肪族 GSL 的 myb28myb29 突变体中表达时,BoMYB29 恢复了生物合成基因的表达和脂肪族 GSL 的积累。有趣的是,包括萝卜硫素在内的甲基亚磺酰基 GSL 含量与甲基硫代 GSLs 的比率大大增加,表明 BoMYB29 适合作为增加甲基亚磺酰基 GSL 含量的调节剂。我们的结果表明,这些生物资源可以促进进一步鉴定用于修饰羽衣甘蓝 GSL 谱和积累的有用基因。