Federal Scientific Center of the East Asia Terrestrial Biodiversity of the Far East Branch of the Russian Academy of Sciences, Vladivostok, 690022, Russia.
Federal Scientific Center of the East Asia Terrestrial Biodiversity of the Far East Branch of the Russian Academy of Sciences, Vladivostok, 690022, Russia; Institute of Marine Biology of the Far East Branch of the Russian Academy of Sciences, Vladivostok, 690041, Russia.
Phytochemistry. 2019 Jan;157:111-120. doi: 10.1016/j.phytochem.2018.10.023. Epub 2018 Nov 3.
Calcium-dependent protein kinases (CDPKs) represent a class within a multigene family that plays an important role in biotic and abiotic plant stress responses and is involved in the regulation of secondary metabolite biosynthesis. Our previous study showed that overexpression of the mutant constitutively active Ca independent form of the AtCPK1 gene (AtCPK1-Ca) significantly increased the biosynthesis of anthraquinones and stilbenes in Rubia cordifolia L. and Vitis amurensis Rupr. transgenic cell cultures, respectively. Here, we have established transgenic calli of soybean plants Glycine max (L.) Merr. that express the AtCPK1-Ca gene. Heterologous expression of the AtCPK1-Ca gene provoked a 5.2-fold increase in total isoflavone production up to 208.09 mg/L, along with an increase in isoflavone aglycones production up to 6.60 mg/L, which is 3-fold greater than that of the control culture. The production of prenylated isoflavones significantly increased, reaching 3.78 mg/L, 13-fold higher than in the control culture. The expression levels of 4-coumarate:CoA ligases, isoflavone synthases, 2-hydroxyisoflavanone dehydratase, isoflavone dimethylallyltransferase, and coumestrol 4-dimethylallyltransferase genes in transgenic cell cultures significantly increased. Thus, heterologous expression of the AtCPK1-Ca gene can be used to bioengineer plant cell cultures that produce isoflavonoids.
钙依赖型蛋白激酶（CDPKs）是一个多基因家族中的一类，在植物的生物和非生物胁迫反应中发挥着重要作用，并参与次生代谢物生物合成的调控。我们之前的研究表明，过表达突变的、组成型激活的拟南芥钙不依赖形式的 AtCPK1 基因（AtCPK1-Ca），分别显著增加了小酸模 Rubia cordifolia L. 和山葡萄 Vitis amurensis Rupr. 转基因细胞培养物中蒽醌和芪类化合物的生物合成。在这里，我们建立了转 AtCPK1-Ca 基因的大豆 Glycine max（L.）Merr. 愈伤组织。AtCPK1-Ca 基因的异源表达导致总异黄酮产量增加了 5.2 倍，达到 208.09mg/L，同时异黄酮苷元产量增加了 6.60mg/L，是对照培养物的 3 倍。烯丙基异黄酮的产量显著增加，达到 3.78mg/L，是对照培养物的 13 倍。转基因细胞培养物中 4-香豆酸：CoA 连接酶、异黄酮合酶、2-羟基异黄酮脱水酶、异黄酮二甲烯丙基转移酶和雌马酚 4-二甲基烯丙基转移酶基因的表达水平显著升高。因此，异源表达 AtCPK1-Ca 基因可用于生物工程生产异黄酮的植物细胞培养物。
