Anthocyanin production in callus cultures of Cleome rosea: modulation by culture conditions and characterization of pigments by means of HPLC-DAD/ESIMS.
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.
Simões
Claudia
C
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. labplan_uerj@yahoo.com.br
Brasil Bizarri
Carlos Henrique
CH
da Silva Cordeiro
Lívia
L
Carvalho de Castro
Tatiana
T
Machado Coutada
Leonardo César
LC
Ribeiro da Silva
Antônio Jorge
AJ
Albarello
Norma
N
Mansur
Elisabeth
E
D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D000704:Analysis of Variance; D000872:Anthocyanins / Q000032:analysis / Q000096:biosynthesis*; D018533:Biomass; D002851:Chromatography, High Pressure Liquid; D031485:Cleome / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism*; D010848:Picolinic Acids / Q000737:chemistry / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D018547:Plant Stems / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D046509:Tissue Culture Techniques
D000872:Anthocyanins; D010848:Picolinic Acids; D015084:2,4-Dichlorophenoxyacetic Acid; C030614:picolinic acid