Induction of Ipomoea batatas L. callus and optimization of culture conditions and accumulation of chlorogenic acid compounds
-
摘要: 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。Abstract: 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.
-
Keywords:
- purple ipomoea batatas /
- callus /
- chlorogenic acid compound /
- hormone /
- medium
-
[1] 陈炼红,伍红,李明珠.紫薯花青素提取工艺优化研究[J].西南民族大学学报·自然科学版,2012,38(3):396-400. [2] Ishida H,Suzuno H,Sugiyama N,et al.Nutritive evaluation on chemical components of leaves,stalks and stems of sweet potatoes(Ipomoea batatas poir)[J].Food Chemistry,2000,68(3):359-367.
[3] Einbond L S,Reynertson K A,Luo X D,et al.Anthocyanin antioxidants from edible fruits[J].Food Chemistry,2004,84(1):23-28.
[4] Nems'A,P誰ksa A,Kucharska A Z,et al.Anthocyanin and antioxidant activity of snacks with coloured potato[J].Food Chemistry,2015,172:175-182. [5] Szymanowska U,Z覥otek U,Karas'M,et al.Anti-inflammatory and antioxidative activity of anthocyanins from purple basil leaves induced by selected abiotic elicitors[J].Food Chemistry,2015,172:71-77. [6] Kruger M J,Davies N,Myburgh K H,et al.Proanthocyanidins,anthocyanins and cardiovascular disease[J].Food Research International,2014,59:42-52.
[7] Speciale A,Cimino F,Saija A,et al.Bioavailability and molecular activities of anthocyanins as modulators of endothelial function[J].Genes&nutrition,2014,9(4):1-19.
[8] Terahara N,Konczak-Islam I,Nakatani M,et al.Anthocyanins in callus induced from purple storage root of Ipomoea batatas L[J].Phytochemistry,2000,54(8):919-922.
[9] Yoshimoto M,Yahara S,Okuno S,et al.Antimutagenicity of mono-,di-,and tricaffeoylquinic acid derivatives isolated from sweetpotato(Ipomoea batatas L.)leaf[J].Bioscience,biotechnology,and biochemistry,2002,66(11):2336-2341.
[10] Kim D W,Yokozawa T,Hattori M,et al.Inhibitory effects of an aqueous extract of Apocyum venetum leaves and its constituents on Cu2+induced oxidative modification of low density lipoprotein[J].Phytother Res,2000,14(7):501-504.
[11] 陈绍华,王亚琴,立新.天然产物绿原酸的研究进展[J].食品科技,2008,33(2):195-199. [12] Jiang Y,Kusama K,Satoh K,et al.Induction of cytotoxicity by chlorogenic acid in human oral tumor cell lines[J].Phytomedicine,2000,7(6):483-491.
[13] Rodriguez de Sotillo D V,Hadley M.Chlorogenic acid modifies plasma and liver concentrations of:Cholesterol,and minerals in(fa/fa)Zucker rats[J].Nutr Biochem,2002(13):717-726.
[14] 席利莎,木泰华,孙红男.绿原酸类物质的国内外研究进展[J].核农学报,2014,28(2):0292-0301. [15] Wang J L,Liao X Y,Zhang H M,et al.Accumulation of chlorogenic acid in cell suspension culutures of Eucommia Ulmoides[J].Plant cell,Tissue and organ culture,2003(74):193-195.
[16] 张志斌,颜日明,邱晓芳,等.杜仲愈伤组织与悬浮细胞中黄酮和绿原酸积累研究[J].中国中药杂志,2009,34(13):1636-1639. [17] 邓倩,王芳,刘小锐,等.响应曲面法优化罗布麻愈伤组织中绿原酸的提取工艺[J].中国农学通报,2012,28(36):107-112. [18] 李琰.杜仲愈伤组织培养及次生代谢产物含量的研究[D].西安:西北农林科技大学,2003. [19] Zhou Y,Hirotani M,Takafumi,et al.Flavonoids and phenylethanoids from hairy root cultures of Scutellaria baicalensis Georgi[J].Phytochemistry,1997,44(1):83-87.
[20] 闫晓慧.狭叶松果菊组织培养、毛状根诱导及松果菊苷和绿原酸积累的研究[D].重庆:西南大学,2007. [21] 李品,岳彩鹏,王新建,等.植物细胞培养生产次生代谢物的影响因素[J].河南林业科技,2007,27(1):21-23. [22] 上官新晨,郭春兰,蒋艳,等.培养基和植物激素对青钱柳茎段和叶片愈伤组织诱导的研究[J].江西农业大学学报,2006,28(5):678-682. [23] 刘小锐,朱国丽,贾红丽,等.罗布麻愈伤组织中绿原酸的提取与测定[J].药物分析杂志,2009,29(12):2116-2119. [24] 张志斌,颜日明,邱晓芳,等.高效液相色谱法测定杜仲愈伤组织和悬浮细胞中绿原酸[J].时珍国医国药,2008,19(9):2082-2083. [25] 张延红,杜弢,王惠珍,等.激素和外植体对黄芩短枝生根和愈伤诱导的影响[J].甘肃中医学院学报,2010,27(1):57-59. [26] 孙爱花,华玉伟,黄天带,等.不同激素配比对橡胶树叶片愈伤组织诱导的影响[J].中国农学通报,2012,28(4):24-27. [27] 李宝健,曾庆平.植物生物技术原理与方法[M].长沙:湖南科学技术出版社,1990:285. [28] 郭紫娟,上官新晨,蒋艳,等.白背三七愈伤组织诱导及培养条件优化[J].江西农业大学学报,2013,35(1):74-79. [29] 谢从华,柳俊.植物细胞工程[M].北京:高等教育出版社,2004.
计量
- 文章访问数: 173
- HTML全文浏览量: 27
- PDF下载量: 145
下载:
