基于响应面法优化白及原球茎液体培养基
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摘要
目的采用响应面优化(Box-Behnken design, BBD)法优选白及原球茎液体培养基,提高白及种子萌发形成原球茎的诱导率。方法采用3水平4因素设计BBD实验,在30 g/L的蔗糖溶液中添加大量元素(MS)、6-苄氨基腺嘌呤(6-BA)、萘乙酸(NAA)和玉米素(ZT)作为白及原球茎液体培养基,并通过响应面法优选4种添加物浓度配比。结果 BBD实验模型设计成功可用于响应面结果优化,经响应面优化后,在1 mg/L MS、2 mg/L 6-BA、0.5 mg/L NAA、0.07 mg/L ZT条件下,白及原球茎诱导率可达(93.13±0.34)%,与预测值95.38%差异无统计学意义(P>0.05)。白及原球茎直播种苗存活率良好,长出的叶片数、植株高度、茎长及茎宽均超过种子直播种苗的数值。结论在基本液体培养基中合理配比4种天然添加物可提高白及原球茎诱导率,进而提高种子萌发率。
Objective To increase the induction rate of Bletilla striata protocorms from seeds by optimizing Bletilla striata protocorm liquid medium based on Box-Behnken design(BBD) methodology. Methods The BBD experiment was designed with four factors and three levels. The Bletilla striata protocorm liquid medium was obtained by adding major elements(MS), 6-benzylamine adenine(6-BA), naphthylacetic acid(NAA), and zeatin(ZT) into 30 g/L sucrose solution. The concentrations of four additives(MS,6-BA, NAA, and ZT) were optimized by response surface methodology(RSM). Results The successfully established BDD model could be used for optimization by RSM. After optimization by RSM, the induction rate of Bletilla striata protocorms could reached(93.13 ±0.34)% under the conditions of 1 mg/L MS, 2 mg/L 6-BA, 0.5 mg/L NAA, and 0.07 mg/L ZT; there was no significant difference between the experimental value and the predicted one(93.13% 0.34% vs 95.38%, P>0.05). The seedlings from direct seeding of Bletilla striata protocorms had a good survival rate, and were superior to those from direct seeding of Bletilla striata seeds in terms of the number of leaves, the height of the plant, and the length and width of the stem. Conclusion An appropriate composition of four natural additives in the basic liquid medium results in an increase in the induction rate of Bletilla striata protocorms and thus promotes the seed germination rate.
Objective To increase the induction rate of Bletilla striata protocorms from seeds by optimizing Bletilla striata protocorm liquid medium based on Box-Behnken design(BBD) methodology. Methods The BBD experiment was designed with four factors and three levels. The Bletilla striata protocorm liquid medium was obtained by adding major elements(MS), 6-benzylamine adenine(6-BA), naphthylacetic acid(NAA), and zeatin(ZT) into 30 g/L sucrose solution. The concentrations of four additives(MS,6-BA, NAA, and ZT) were optimized by response surface methodology(RSM). Results The successfully established BDD model could be used for optimization by RSM. After optimization by RSM, the induction rate of Bletilla striata protocorms could reached(93.13 ±0.34)% under the conditions of 1 mg/L MS, 2 mg/L 6-BA, 0.5 mg/L NAA, and 0.07 mg/L ZT; there was no significant difference between the experimental value and the predicted one(93.13% 0.34% vs 95.38%, P>0.05). The seedlings from direct seeding of Bletilla striata protocorms had a good survival rate, and were superior to those from direct seeding of Bletilla striata seeds in terms of the number of leaves, the height of the plant, and the length and width of the stem. Conclusion An appropriate composition of four natural additives in the basic liquid medium results in an increase in the induction rate of Bletilla striata protocorms and thus promotes the seed germination rate.
引文
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[3]任风鸣,刘艳,李滢,等.白及属药用植物的资源分布及繁育[J].中草药,2016,47(24):4478-4487.
[4]周至明,黄程生,彭丽丽,等.白及人工种植初步研究[J].中药材,2006,29(1):7-8.
[5] KYLIN A. A new method for large-scale aseptic cultivationof higher plants[J].Physiologia Plantarum,2010,3(2):165-174.
[6] ZHANG A, WANG Y, HUANG H, et al. Efficient callus induc-tion and high-frequency regeneration system establishment ofstrawberry ‘Akihime'[J]. Guihaia, 2018, 38(4):482-491.
[7]徐德林,储士润,李树刚,等.四种天然添加物对白及组培苗诱导的影响[J].西南师范大学学报(自然科学版),2015,40(10):181-186.
[8]潘胤池,李林,肖世基,等.白及繁育技术研究进展[J].中成药,2018,40(5):1142-1149.
[9]聂宁,朱艳,田梅,等.白及种子萌发及原球茎发育过程的细胞组织学观察[J].中国中药杂志,2016,41(8):1446-1449.
[10]谢利,马晓娟,郭和蓉,等.杂交兰类原球茎增殖中芽分化的控制和快速繁殖[J].植物生理学报,2014,50(2):209-213.
[11]闫斌,潘超美,何洁,等.穿心莲组培苗的壮苗生根与移栽基质研究[J].时珍国医国药,2016,27(7):1730-1732.
[12]吕亚丽,郭顺星,孟志霞.天山雪莲组培苗生根培养体系的优化[J].中国药学杂志,2009,44(17):1291-1294.
[13]熊苏慧,唐洁,李诗卉,等.一种新型天然低共熔溶剂用于玉竹总黄酮的绿色提取[J].中草药,2018,49(10):2378-2386.
[14]宋晓丹,陈晓玲,尚丽,等.白及种子试管高频萌发的应用研究[J].中国现代中药,2014,16(9):751-754.
[15]崔瑞勤,陈科力,徐雷.基于组培快繁技术的白及种子萌发和幼苗形态观察[J].江苏农业科学,2015,43(2):238-240.