Núcleo de Biotecnologia Vegetal, Universidade do Estado do Rio de Janeiro (UERJ), Rua São Francisco Xavier, 524 - PHLC, sala 509, Maracanã, Rio de Janeiro, RJ, Brazil.
Plant Physiol Biochem. 2009 Oct;47(10):895-903. doi: 10.1016/j.plaphy.2009.06.005. Epub 2009 Jun 18.
Leaf and stem explants of Cleome rosea formed calluses when cultured on MS medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or 4-amino-3,5,6-trichloropicolinic acid (PIC). The highest biomass accumulation was obtained in the callus cultures initiated from stem explants on medium supplemented with 0.90 microM 2,4-D. Reddish-pink regions were observed on callus surface after 6-7 months in culture and these pigments were identified as anthocyanins. Anthocyanins production was enhanced by reducing temperature and increasing light irradiation. Pigmented calluses transferred to MS1/2 with a 1:4 ratio NH(4)(+)/NO(3)(-), 70 g L(-1) sucrose and supplementation with 0.90 microM 2,4-D maintained a high biomass accumulation and showed an increase of 150% on anthocyanin production as compared with the initial culture conditions. Qualitative analysis of calluses was performed by high performance liquid chromatography coupled to diode array detector and electrospray ionization mass spectrometry (HPLC-DAD/ESIMS). Eleven anthocyanins were characterized and the majority of them were identified as acylated cyanidins, although two peonidins were also detected. The major peak was composed by two anthocyanins, whose proposed identity were cyanidin 3-(p-coumaroyl) diglucoside-5-glucoside and cyanidin 3-(feruloyl) diglucoside-5-glucoside.
长春花叶片和茎段外植体在添加不同浓度 2,4-二氯苯氧乙酸（2,4-D）或 4-氨基-3,5,6-三氯吡啶羧酸（PIC）的 MS 培养基上培养时形成愈伤组织。从茎外植体起始的愈伤组织在添加 0.90 μM 2,4-D 的培养基上获得最高的生物量积累。在培养 6-7 个月后，在愈伤组织表面观察到红粉色区域，这些色素被鉴定为花青素。通过降低温度和增加光照来增强花青素的产生。将有色愈伤组织转移到 MS1/2，其中 NH(4)(+) / NO(3)(-) 的比例为 1:4，70 g L(-1)蔗糖，并添加 0.90 μM 2,4-D，与初始培养条件相比，保持了较高的生物量积累，并使花青素产量增加了 150%。通过高效液相色谱-二极管阵列检测器-电喷雾电离质谱联用（HPLC-DAD/ESIMS）对愈伤组织进行定性分析。鉴定了 11 种花青素，其中大部分为酰化氰定，尽管也检测到两种芍药苷。主要峰由两种花青素组成，其拟议的身份为氰定 3-（对香豆酰）二葡萄糖苷-5-葡萄糖苷和氰定 3-（阿魏酰）二葡萄糖苷-5-葡萄糖苷。
