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 Ca2+ 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.

中文翻译：

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由 AtCPK1 基因的组成型活性 Ca2+ 非依赖性形式转化的 Glycine max 细胞中异黄酮含量增加

钙依赖性蛋白激酶 (CDPK) 代表多基因家族中的一类，在生物和非生物植物胁迫响应中起重要作用，并参与次级代谢物生物合成的调节。我们之前的研究表明，AtCPK1 基因（AtCPK1-Ca）的突变体组成型活性 Ca2+ 独立形式的过表达显着增加了 Rubia cordifolia L. 和 Vitis amurensis Rupr 中蒽醌和芪的生物合成。分别是转基因细胞培养物。在这里，我们建立了大豆植物 Glycine max (L.) Merr 的转基因愈伤组织。表达 AtCPK1-Ca 基因。AtCPK1-Ca 基因的异源表达使异黄酮总产量增加了 5.2 倍，高达 208.09 mg/L，同时异黄酮苷元产量增加了 6.60 mg/L，这比对照培养物大 3 倍。异黄酮的产量显着增加，达到 3.78 mg/L，比对照培养物高 13 倍。4-香豆酸：CoA 连接酶、异黄酮合酶、2-羟基异黄酮脱水酶、异黄酮二甲基烯丙基转移酶和香豆素 4-二甲基烯丙基转移酶基因在转基因细胞培养物中的表达水平显着增加。因此，AtCPK1-Ca 基因的异源表达可用于产生异黄酮的生物工程植物细胞培养物。异黄酮二甲基烯丙基转移酶和香豆雌酚 4-二甲基烯丙基转移酶基因在转基因细胞培养物中显着增加。因此，AtCPK1-Ca 基因的异源表达可用于产生异黄酮的生物工程植物细胞培养物。异黄酮二甲基烯丙基转移酶和香豆雌酚 4-二甲基烯丙基转移酶基因在转基因细胞培养物中显着增加。因此，AtCPK1-Ca 基因的异源表达可用于产生异黄酮的生物工程植物细胞培养物。