Anthocyanin production in callus cultures of Cleome rosea: modulation by culture conditions and characterization of pigments by means of HPLC-DAD/ESIMS.
Anthocyanin production in callus cultures of Cleome rosea: modulation by culture conditions and characterization of pigments by means of HPLC-DAD/ESIMS.
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DOI：
10.1016/j.plaphy.2009.06.005
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发表时间：
2009-10
期刊：
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通讯作者：
C. Simões;Carlos Henrique Brasil Bizarri;Lívia da Silva Cordeiro;Tatiana Carvalho de Castro;Leonardo César Machado Coutada;A. J. Ribeiro da Silva;N. Albarello;E. Mansur
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文献类型：
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作者：
C. Simões;Carlos Henrique Brasil Bizarri;Lívia da Silva Cordeiro;Tatiana Carvalho de Castro;Leonardo César Machado Coutada;A. J. Ribeiro da Silva;N. Albarello;E. Mansur
玫瑰花叶、茎外植体在附加不同浓度2，4-二氯苯氧乙酸(2，4-D)或4-氨基-3，5，6-三氯烟酸(PIC)的MS培养基上均可形成愈伤组织。茎段外植体在附加0.90μM2，4-D的愈伤组织培养中，生物量积累最高。培养6-7个月后，在愈伤组织表面观察到红粉色的区域，这些色素被鉴定为花青素。降低温度和增加光照可提高花色苷的产量。有色愈伤组织以1：4的NH_4~+/NO_3−、70 mg的L−1蔗糖和0.90%的μM_2，4-D转化至MS_1/2，4-D，保持了较高的生物量积累，花色苷产量较初始培养条件提高150%。采用高效液相色谱-二极管阵列检测器-电喷雾电离质谱仪对愈伤组织进行定性分析。鉴定了11个花青素，其中大部分为酰化花青素，但也检测到两个牡丹素。主峰由两个花色苷组成，分别为花青素3-(p-香豆酰基)二葡糖苷-5-葡萄糖苷和花青素-3-(阿魏酰基)二葡萄糖苷-5-葡萄糖苷。
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 μM 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 NH4+/NO3−, 70 g L−1sucrose and supplementation with 0.90 μM 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.