外源激素IBA和真菌诱导子对八角莲离体根生长及其鬼臼毒素合成的影响
|
摘要
该实验以White为基本培养基,研究了不同浓度外源激素IBA和真菌诱导子对八角莲离体根生长和次生代谢产物鬼臼毒素合成的影响。结果表明,经过3周的培养,生长激素IBA和内生真菌Zasmidium syzygii诱导子均能提高八角莲离体根中鬼臼毒素的含量;在添加3 mg·L~(-1) IBA的White培养基中,离体根中鬼臼毒素的质量分数最高为1 830.86μg·g~(-1),是对照组的2.07倍;其次是添加1.5 mg·L~(-1) IBA,Z.syzygii真菌诱导子,1,0.5,4.5 mg·L~(-1) IBA,鬼臼毒素的含量分别是对照组的1.82,1.71,1.63,1.43,1.1倍。离体根生长与IBA浓度的变化呈一定的正相关性。上述结果初步说明,最适于八角莲离体根代谢产生鬼臼毒素的IBA质量浓度为3 mg·L~(-1);Z.syzygii真菌诱导子的促进作用效果介于1.5 mg·L~(-1)和1 mg·L~(-1)的IBA之间,是将来生产中诱导八角莲中鬼臼毒素积累的潜在天然诱导子。
Using the White as basic medium, the effects of the exogenous IBA and endophytic fungal elicitor on the growth of in vitro roots cultures of Dysosma versipellis and production of podophyllotoxin were investigated in this study. The results showed that the IBA and the endophytic fungus Zasmidium syzygii elicitor could increase the content of podophyllotoxin of in vitro roots of D. versipellis after 3 weeks. The White medium added with 3 mg·L~(-1) IBA induced the highest increase of podophyllotoxin(1 830.86 μg·g~(-1)), which was 2.07 folds greater than the control, and followed by 1.5 mg·L~(-1) IBA, fungal elicitor, 1 mg·L~(-1) IBA, 0.5 mg·L~(-1) IBA and 4.5 mg·L~(-1) IBA, which was 1.82, 1.71, 1.63, 1.43 and 1.1 folds greater than the control, respectively. The results also showed that the growth of roots was certain positively correlated with the change of IBA concentration. Therefore, 3 mg·L~(-1) IBA was the most suitable for the production of podophyllotoxin in the in vitro roots of D. versipellis, and the stimulating effect of Z. syzygii fungal elicitor was between 1.5 mg·L~(-1) and 1 mg·L~(-1) IBA, which was a potential natural elicitor to induce the accumulation of podophyllotoxin in future production.
Using the White as basic medium, the effects of the exogenous IBA and endophytic fungal elicitor on the growth of in vitro roots cultures of Dysosma versipellis and production of podophyllotoxin were investigated in this study. The results showed that the IBA and the endophytic fungus Zasmidium syzygii elicitor could increase the content of podophyllotoxin of in vitro roots of D. versipellis after 3 weeks. The White medium added with 3 mg·L~(-1) IBA induced the highest increase of podophyllotoxin(1 830.86 μg·g~(-1)), which was 2.07 folds greater than the control, and followed by 1.5 mg·L~(-1) IBA, fungal elicitor, 1 mg·L~(-1) IBA, 0.5 mg·L~(-1) IBA and 4.5 mg·L~(-1) IBA, which was 1.82, 1.71, 1.63, 1.43 and 1.1 folds greater than the control, respectively. The results also showed that the growth of roots was certain positively correlated with the change of IBA concentration. Therefore, 3 mg·L~(-1) IBA was the most suitable for the production of podophyllotoxin in the in vitro roots of D. versipellis, and the stimulating effect of Z. syzygii fungal elicitor was between 1.5 mg·L~(-1) and 1 mg·L~(-1) IBA, which was a potential natural elicitor to induce the accumulation of podophyllotoxin in future production.
引文
[1] 黄家云,黄慧莲.液体振荡培养八角莲愈伤组织及其鬼臼毒素的含量测定[J].现代中药研究与实践,2008(2):20.
[2] 陈日道,段瑞刚,邹建华,等.八角莲愈伤组织中黄酮苷类化学成分研究[J].中国中药杂志,2016,41(1):87.
[3] 殷梦龙,陈仲良.云南八角莲和广西八角莲的化学成分[J].中国中药杂志,1989,14(7):36.
[4] Xu X Q,Gao X H,Jin L H,et al.Antiproliferation and cell apoptosis inducing bioactivities of constituents from Dysosma versipellis in PC3 and Bcap-37 cell lines[J].Cell Division,2011,6:14.
[5] 段瑞刚,李军伟,邹建华,等.八角莲愈伤组织中木脂素类化学成分研究[J].中国药学杂志,2014,49(15):1306.
[6] Anbazhagan V R,Ahn C H,Harada E,et al.Podophyllotoxin production via cell and adventitious root cultures of Podophyllum peltatum[J].In Vitro Cell Dev Biol-Plant,2008,44:494.
[7] 姜银姬,朴炫春,李贺,等.几种因素对生物反应器培养东北刺人参不定根中皂苷积累的影响[J].林业工程学报,2016,1(3):68.
[8] 周亦,赵桐,李文月,等.白木香离体根液体培养条件优化以及色酮类物质诱导[J].中药材,2018,41(3):522.
[9] 严硕,高文远,王娟,等.不同生物反应器对丹参不定根培养过程中有效成分的影响[J].中国中药杂志,2009,34(16):2027.
[10] 李琰,崔蕾,杨钰琪,等.雷公藤不定根培养体系的建立及中试放大研究[J].中国中药杂志,2015,40(1):53.
[11] 韩献忠,张治国,刘骤,等.条叶龙胆离体根培养条件的初步研究[J].植物学通报,1990,7(3):49.
[12] 张焕欣,董春娟,尚庆茂.外源IBA对辣椒下胚轴不定根发生的调控作用[J].园艺学报,2017,44(10):1937.
[13] 孙晶,高珂,王玲,等.不同培养基、外源激素和真菌诱导子对北柴胡毛状根生长及柴胡皂苷含量的影响[J].生物技术通讯,2015,26(4):569.
[14] 陈巍,郭肖红,高文远,等.丹参不定根离体培养的研究[J].中国中药杂志,2006,31(17):1409.
[15] Li J,Li H,Liu D,et al.Analysis of ginsenoside content,functional genes involved in ginsenosides biosynthesis,and activities of antioxidant enzymes in Panax quinquefolium L.adventitious roots by fungal elicitors[J].Res Chem Intermed,2016,43(4):2415.
[16] 何梦玲,何芳,孟京兰,等.3种诱导子对白木香根悬浮培养细胞中2-(2-苯乙基)色酮化合物形成的影响[J].中成药,2013,35(7):1367.
[17] Zhai X,Jia M,Chen L,et al.The regulatory mechanism of fungal elicitor-induced secondary metabolite biosynthesis in medical plants[J].Crit Rev Microbiol,2017,43(2):24.
[2] 陈日道,段瑞刚,邹建华,等.八角莲愈伤组织中黄酮苷类化学成分研究[J].中国中药杂志,2016,41(1):87.
[3] 殷梦龙,陈仲良.云南八角莲和广西八角莲的化学成分[J].中国中药杂志,1989,14(7):36.
[4] Xu X Q,Gao X H,Jin L H,et al.Antiproliferation and cell apoptosis inducing bioactivities of constituents from Dysosma versipellis in PC3 and Bcap-37 cell lines[J].Cell Division,2011,6:14.
[5] 段瑞刚,李军伟,邹建华,等.八角莲愈伤组织中木脂素类化学成分研究[J].中国药学杂志,2014,49(15):1306.
[6] Anbazhagan V R,Ahn C H,Harada E,et al.Podophyllotoxin production via cell and adventitious root cultures of Podophyllum peltatum[J].In Vitro Cell Dev Biol-Plant,2008,44:494.
[7] 姜银姬,朴炫春,李贺,等.几种因素对生物反应器培养东北刺人参不定根中皂苷积累的影响[J].林业工程学报,2016,1(3):68.
[8] 周亦,赵桐,李文月,等.白木香离体根液体培养条件优化以及色酮类物质诱导[J].中药材,2018,41(3):522.
[9] 严硕,高文远,王娟,等.不同生物反应器对丹参不定根培养过程中有效成分的影响[J].中国中药杂志,2009,34(16):2027.
[10] 李琰,崔蕾,杨钰琪,等.雷公藤不定根培养体系的建立及中试放大研究[J].中国中药杂志,2015,40(1):53.
[11] 韩献忠,张治国,刘骤,等.条叶龙胆离体根培养条件的初步研究[J].植物学通报,1990,7(3):49.
[12] 张焕欣,董春娟,尚庆茂.外源IBA对辣椒下胚轴不定根发生的调控作用[J].园艺学报,2017,44(10):1937.
[13] 孙晶,高珂,王玲,等.不同培养基、外源激素和真菌诱导子对北柴胡毛状根生长及柴胡皂苷含量的影响[J].生物技术通讯,2015,26(4):569.
[14] 陈巍,郭肖红,高文远,等.丹参不定根离体培养的研究[J].中国中药杂志,2006,31(17):1409.
[15] Li J,Li H,Liu D,et al.Analysis of ginsenoside content,functional genes involved in ginsenosides biosynthesis,and activities of antioxidant enzymes in Panax quinquefolium L.adventitious roots by fungal elicitors[J].Res Chem Intermed,2016,43(4):2415.
[16] 何梦玲,何芳,孟京兰,等.3种诱导子对白木香根悬浮培养细胞中2-(2-苯乙基)色酮化合物形成的影响[J].中成药,2013,35(7):1367.
[17] Zhai X,Jia M,Chen L,et al.The regulatory mechanism of fungal elicitor-induced secondary metabolite biosynthesis in medical plants[J].Crit Rev Microbiol,2017,43(2):24.