前体饲喂、诱导子和光照联合使用对葡萄细胞培养合成花青素的影响

Significant Improved Anthocyanins Biosynthesis in Suspension Cultures of Vitis vinifera by Process Intensification

苯丙氨酸前体饲喂分别和环糊精、葡聚糖、茉莉酸甲酯、黑曲霉和直喙镰孢茵提取液五种诱导子联合作用，其中以与茉莉酸甲酯的联合作用时葡萄细胞培养生产花青素的影响最大，可使单位鲜细胞花青素含量提高2．7倍，花青素产量提高3．4倍，实验证明两者在培养后第4天加入效果最好。在30μmol／L苯丙氨酸、218μmol／L茉莉酸甲酯和3000-4000lx光照条件下，不同花青素产量的细胞株都能显著提高花青素产量，但低产株VV06比高产株VV05具有更大的产率提高潜力。该条件下VV05和VV06花青素产量分别达到2975和4090CV／L，是对照组的2．5倍和5.2倍。

The low-production is a ubiquitous problem and has prevented the commercialization of secondary metabolite production in plant cell culture. In order to examine the effective approaches to improvement of secondary metabolite production in plant cell culture, the investigation of anthocyanins accumulation in suspension cultures of Vitis vinifera, as a model system, had been initiated in our laboratory. In this present research, various elicitors and the precursor of phenylalanine were used in combination to enhance the anthocyanins production in suspension cultures of Vitis vinifera. And an integrated process with the combination of elicitation, precursor feeding and light irradiation was reported for rational bioprocess design. Among the combination treatment of phenylalanine feeding and several elicitors （methyl-β-cyclodextrin, dextran T-40, methyl jasmonate, extracts of Aspergillus niger and Fusarium orthoceras ）, the combination with methyl jasmonate gave the highest anthocyanins production in suspension cultures of Vitis vinifera. When compared to the controls, the anthocyanins content （CV/g, FCW） and production （CV/L） increased by 2.7-fold and 3.4-fold, respectively. The optimum time for the addition of phenylalanine and methyl jasmonate was 4 days after inoculation. Two cell lines with different anthocyanins-producing capacity responded differently to the optimum combination treatment of 30μmol/L phenylalanine feeding, 218μmol/L methyl jasmonate elicitation and 3000 -4000lx light illumination. The high-and low-anthocyanins-producing cell lines of VV05 and VV06 produced the maximum of 2975 and 4090 CV/L of anthocyanins that were 2.5- and 5.2-fold of the controls, respectively.