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Micropropagation and HPLC-DAD, UPLC MS/MS analysis of oenothein B and phenolic acids in shoot cultures and in regenerated plants of fireweed ( Chamerion angustifolium (L.) Holub)
Plant Cell, Tissue and Organ Culture ( IF 2.3 ) Pub Date : 2020-10-19 , DOI: 10.1007/s11240-020-01949-5
Mariola Dreger , Agnieszka Gryszczyńska , Milena Szalata , Karolina Wielgus

In this study, a micropropagation protocol using nodal explants from in vitro grown plants of Chamerion angustifolium (L.) Holub was developed and analysis of oenothein B and selected phenolic acids in shoot cultures was performed for the first time. For shoot induction and multiplication Murashige and Skoog’s (MS) basal medium supplemented with 2-isopentenyladenine (2iP), zeatin (Z) and 6-benzyloaminopurine (BAP) was used. 2iP was the most responsive in terms of promoting shoots per explant with the maximum (6.57 ± 1.14) recorded at a concentration of 2.0 mg L−1 after 6 weeks of culture. After two subcultures the multiplication rate was increased up to 19 shoots per explant on medium with 2iP (1.0 mg L−1). To prevent tissue browning, ascorbic acid and casein hydrolysate were added to the induction medium, resulting in a reduction of browning by 30%. The rooted plantlets were successfully transferred to soil and acclimatized with 97% frequency. Quantitative and qualitative assessments of oenothein B and phenolic acid contents in in vitro regenerated shoots as well as in ex vitro plants were performed using high-performance liquid chromatography with a diode-array detector (HPLC-DAD) and ultra-performance liquid chromatography–tandem mass spectrometry (UPLC MS/MS) methods. Oenothein B (1.62‒4.55 g 100 g−1 DW), ellagic acid, gallic and caffeic acids were identified in in vitro regenerated plants. The results of this study confirm that the oenothein B-producing plantlets can be obtained using the micropropagation method with axillary shoots being a valuable source of oenothein B and phenolic acids.



中文翻译:

杂草(Chamerion angustifolium(L.)Holub)的芽培养和再生植物中的卵磷脂,酚醛酸的微繁殖和HPLC-DAD,UPLC MS / MS分析

在这项研究中,开发了使用来自Chamerion angustifolium(L.)Holub的离体生长植物的节点外植体的微繁殖方案,并且首次进行了茎培苗中皂苷B和所选酚酸的分析。对于芽的诱导和繁殖,使用补充有2-异戊烯腺嘌呤(2iP),玉米蛋白(Z)和6-苄基氨基嘌呤(BAP)的Murashige和Skoog's(MS)基础培养基。在培养6周后,就每个外植体的促生芽而言,2iP的响应最强,在2.0 mg L -1的浓度下记录的最大值为(6.57±1.14)。经过两次传代培养,在2iP(1.0 mg L -1)。为防止组织褐变,将抗坏血酸和酪蛋白水解物添加到诱导培养基中,从而使褐变减少30%。生根的小苗成功地转移到土壤中,并以97%的频率适应环境。高效液相色谱-二极管阵列检测器(HPLC-DAD)和超高效液相色谱-串联质谱法对离体再生芽和离体植物中的皂苷B和酚酸含量进行了定量和定性评估质谱(UPLC MS / MS)方法。皂甙B(1.62‒4.55 g 100 g -1在体外再生植物中鉴定出鞣花酸,没食子酸,没食子酸和咖啡酸。这项研究的结果证实,使用微繁繁殖法可以得到产卵泡蛋白B的小植株,而腋生芽是卵泡蛋白B和酚酸的重要来源。

更新日期:2020-10-19
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