继代周期和接种量对葡萄细胞培养的影响

在每种不同的继代周期和接种量条件下,葡萄细胞在连续10次继代培养过程中的生物量、花青素含量、胞内糖、胞内蛋白及胞内总磷均表现出不同程度的波动。不同接种量对培养不稳定性的影响比不同继代周期大;在所考察的条件中,7d继代周期与1.60g接种量组合的继代条件下花青素合成相对稳定;花青素合成与胞内蔗糖或胞内总磷水平呈负相关。

Impact of subculture cycles and inoculum sizes on suspension cultures of Vitis vinifera

The commercial application of plant cell cultures is often hindered by the instability of secondary metabolite biosynthesis,where the metabolite yield fluctuates and decline dramatically over subcultures. This study proposed that such instability is due to the fluctuations of culture variables. To validate this hypothesis,the effects of the fluctuations of two culture variables (subculture cycle and inoculum size) on the biomass,anthocyanin biosynthesis,intracellular carbon,nitrogen and phosphate during continuous 10 subculture cycles were investigated. The subculture cycle was fluctuated for 12h in a 7 day cycle (6.5,7 and 7.5d),and the inoculum size was fluctuated by 20% on basis of 2.00g (1.60,2.00 and 2.40g). It was found that all the measured culture parameters fluctuated over the 10 subculture cycles. The fluctuation in terms of inoculum sizes had a greater effect on the stability of anthocyanin biosynthesis in suspension cultures of V. vinifera. Among all the subculture conditions investigated,7d-subculture cycle and 1.60 g-inoculum size was the best one to hold the relatively stable anthocyanin production. The anthocyanin yield presented a negative correlation with intracellular sucrose content or intracellular total phosphate content.