紫薯愈伤组织诱导和培养条件优化及绿原酸类化合物的积累

Induction of Ipomoea batatas L. callus and optimization of culture conditions and accumulation of chlorogenic acid compounds

以紫薯叶片为外植体,通过筛选培养基和培养条件建立了紫薯愈伤组织培养体系。研究6-苄基氨基嘌呤(6-BA)及萘乙酸(NAA)不同浓度组合对紫薯愈伤组织诱导的影响,之后分别考察基础培养基MS、1/2MS、B5、WPM、White及NAA、6-BA、2,4-二氯苯氧乙酸(2,4-D)三种激素对紫薯愈伤组织继代生长的影响。确定最适基础培养基后,对激素进行单因素实验后再做正交实验。结果表明1/2MS基本培养基中添加1.5 mg/L 6-BA和0.4 mg/L NAA适合紫薯愈伤组织诱导,诱导率为81.82%。MS为优选基础培养基,通过正交实验优化的激素配比组合为1.0 mg/L NAA,0.4 mg/L 6-BA,1.0 mg/L 2,4-D。最佳继代培养基为MS+1.0 mg/L NAA+0.4 mg/L 6-BA+1.0 mg/L 2,4-D+6 g/L琼脂+30 g/L蔗糖,此优化激素条件下愈伤组织的生物量增长倍数为23.46倍,3-咖啡酰奎宁酸、3,5-二咖啡酰宁酸、4,5-二咖啡酰宁酸三种绿原酸类化合物含量分别为0.30%、1.91%、1.10%,三者总含量3.31%,因此,通过紫薯愈伤组织培养生产绿原酸类化合物是可行的。

With the leaves of purple ipomoea batatas as explants,callus induction was established through selection of medium and conductions of cultivation. The effect of purple potato callus induction with different concentration combinations of 6-BA and NAA was researched. Then five basic culture medium of MS,1/2MS,B5,WPM,White and three hormones of NAA,6-BA,2,4-D were investigated for the purple potato callus transgenerational. After the basic medium was determined,basis on the result of single factor for hormone,the orthogonal experiment was employed. The results showed that 1/2MS medium supplemented with 1.5 mg/L 6-BA plus 0.4 mg/L NAA was suitable for callus formation,the callus induction was 81.82%.MS was the most suitable medium for growth of callus. The optimized combinations with orthogonal test were obtained as follows,1.0 mg/L NAA,0.4 mg/L 6-BA and 1.0 mg/L 2,4-D. The best successive transfer culture medium was MS,1.0 mg/L NAA,0.4 mg/L 6-BA,1.0 mg/L 2,4-D,6 g/L agar,30 g/L sucrose and biomass of callus was increased by 23.46 fold under this condition,the content of 3-caffeoylquinic acid,3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid was 0.30%,1.91% and 1.10% respectively. Total content of the three substance was 3.31% under the optimum conditions. So,it was feasible to produce chlorogenic acid through the callus culture of purple ipomoea batatas.