Constrcting The Hariy Root Bioreactors With High Anthocyanins Content

Anthocyanins are a class of liquid water soluble pigment, existing widely in angiosperms and gymnosperms. Because of its special chemical structure, it can make the lant organs show different colors like pink, blue, purple, etc. in different PH conditions. Anthocyanins are one kind of flavonoids, and have the function of anticancer, anti-inflammatory anti-bacterial, anti aging, reducing blood sugar, protecting liver function. The natural anthocyanin comes from the higher yield of fruit and vegetables (blackwolfberry fruit, blueberry, grape, black currant, purple sweet potato, red cabbage, etc.). Some special Anthocyanin accumulated only in the plant flowers, fruits, tubers. Acquisitions of anthocyanins from more plants were necessary. This study aimed to obtain the high anthocyanin bioreactor through overexpression of MYB and bHLH gene regulation of anthocyanin biosynthesis metabolism in plant hairy root. The experimental results are as follows:1, MYB gene AmROS1 and bHLH gene AmDEL, regulating anthocyanin synthesis metabolism, were construct into two expression vector using Gateway technology. The two expression vectors were electric transferred intoAgrobacterium rhizogenes Ar1193. The leaf of tobacco and the hypocotyl of Petunia were infected with Agrobacterium rhizogenes. The hairy root with high anthocyanin were obtained, and the conversion rate is 40%. The anthocyanin content of the ’Samsun’ tobacco hairy roots was 1.46% of the dry weight.2, The anthocyanins kinds were identified using high performance liquid chromatography-mass spectrometry. The anthocyanin compounds of hairy roots was the same to flowers in tobacco.the peak time of hairy roots and flowers are 11.27min and 11.38min respectively. The molecular weight of anthocyanins was 595.1 Da in the hairy roots, while the molecular weight of anthocyanins is 595Da in flower. Both anthocyanins were cyanidin connected with a coffee acid and a ferulic acid.3, the HBA in the flower of Morning Glory was 1745.5Da, which is composed of a cyanidin,6 glucose, and 3 coffee acid. No HBA was found in the morning glory in hairy roots, and the ferulic acid was detected in hariy root, which didn’t exist in flower.The anthocyanins from hairy root of Morning Glory had 12 peaks and 10 kinds of anthocyanin compound. Their molecular weight were between 773Da and 1111.4Da. The anthocyanin of 773Da was composed of a cyanidin and 3 glucoses, while the 1111.4Da was a peonidin and 5 glucoses.4、Through the excessive expression of MYB gene AmROS and bHLH gene AmDEL regulates anthocyanin biosynthesis metabolism in the hairy root, the hairy root bioreactors with high anthocyanins were obtained. The anthocyanins in hairy root were diverse, which can wide the source of natural anthocyanins.