Synergistic effect of morphactin on cytokinin-induced production of isoflavonoids in cell cultures of Pueraria tuberosa (Roxb. ex. Willd.) DC

Abstract

Isoflavonoids, the functional molecules of Fabaceae, are under clinical trials against cancer, osteoporosis and cardiovascular diseases. In this study, the efficacy of different plant growth regulators was evaluated for optimizing the production of isoflavonoids in Pueraria tuberosa. The cultures were maintained in Murashige and Skoog's medium containing 0.1 mg l -1 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and 0.1 mg l -1 kinetin. The addition of 5.0 mg l -1 N 6 -(2-Isopentenyl) adenine (2iP) resulted in about *32-folds increase in production of isoflavonoids, while about *23-folds increase was recorded in the absence of kinetin in the maintenance medium. A maximum yield of isoflavonoids (*80 mg l -1 ; 82-folds increase) was obtained in cultures grown at 0.1 mg l -1 morphactin and 5.0 mg l -1 of 2iP. However, 2,4,5-T in combination with 2iP was ineffective for their production. Among different plant growth regulators tested, maximum yields of puerarin, genistin, daidzein and genistein were 17.4, 15.9, 69.0 and 0.04 mg l -1 , respectively. The study suggested that the presence of two cytokinins or 2iP with morphactin in the culture medium markedly enhanced the production of isoflavonoids in P. tuberosa.

Plant growth regulators affect biosynthesis and accumulation profile of isoflavone phytoestrogens in high-productive in vitro cultures of Genista tinctoria

“…As in the case of auxins, the observed effects depended largely on the type of PGR used. In general, the experiment confirmed that the simultaneous use of auxin and cytokinin often stimulates the production of secondary metabolites (Goyal and Ramawat 2008). Of all the tested cytokinins, KIN at 0.5 mg l -1 exerted the strongest stimulatory effect on isoflavone production (Figs.…”

“…Apart from the phytohormones commonly applied to cell cultures of legume plants, such as BAP, 2,4-D, IAA, KIN, NAA and ZEA (Fedoreyev et al 2000;Luo et al 2003;Matkowski 2004;Vaishnav et al 2006;Tůmová et al 2007;Shinde et al 2010;Pu et al 2013), the study included PGRs which so far have not been widely examined with respect to their influence on isoflavone metabolism. Considering that the biosynthesis of isoflavonoids under in vitro conditions depends not only on PGR type, but also on its amount in the growth medium (Fedoreyev et al 2000;Vaishnav et al 2006;Tůmová et al 2007;Goyal and Ramawat 2008;Shinde et al 2010), the particular auxins and cytokinins were supplemented in two extreme concentrations of 0.5 and 5.0 mg l -1…”

“…Goyal and Ramawat (2008) reported that 2iP stimulated isoflavone production in P. tuberosa callus when applied alone, together with 2,4,5-T or jointly with 2,4,5-T ? KIN (Goyal and Ramawat 2008).…”

“…Goyal and Ramawat (2008) reported that 2iP stimulated isoflavone production in P. tuberosa callus when applied alone, together with 2,4,5-T or jointly with 2,4,5-T ? KIN (Goyal and Ramawat 2008). Similarly, Udomsuk et al (2009Udomsuk et al ( , 2012 described the stimulatory effect of TDZ, used as the sole PGR or conjointly with NAA ?…”

“…(Łuczkiewicz and Głód 2003;Tůmová and Tůma 2011), Pueraria sp. (Matkowski 2004;Goyal and Ramawat 2008;Udomsuk et al 2009Udomsuk et al , 2012, Psoralea corylifolia (Shinde et al 2010), Maackia amurensis (Fedoreyev et al 2000(Fedoreyev et al , 2008Luo et al 2003Luo et al , 2004Luo et al , 2007, Trifolium pratense (Engelmann et al 2009;Kašparová et al 2012), Medicago truncatula (Lei et al 2010) and Sophora japonica (Pu et al 2013). Moreover, attempts are being made to improve isoflavone productivity of the obtained systems, either by means of genetic and molecular engineering (Du et al 2010) or classical experimental techniques including clone selection and medium optimization (Luczkiewicz 2008).…”