Abstract
Anaphalis hancockii Maxim is often used as an aromatic plant in China. To effectively protect and utilize the resources of A. hancockii, a completely rapid micropropagation system has been developed. Additionally, ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS) technology, coupled with multivariate statistical analysis and pathway enrichment analysis, was used to assess the regulatory influences of 6-benzylaminopurine (6-BA) and kinetin (KT) applied in A. hancockii. The results showed that when MS medium was supplemented with 4.0 mg/L 2,4-D and 2.0 mg/L KT, the effect of inducing callus from leaf, petiole, and root explants was the best. At this time, the leaf callus induction rate was 94.67%. Adding 3.0 mg/L KT to the medium alone had the best induction effect on adventitious shoots, and the induction rate in leaf tissue at this time was 91.96%. The optimal condition for rooting medium was MS supplement with 0.1 mg/L IBA. The use of 6-BA and KT made the endogenous metabolite content show a more up-regulated trend to a certain extent. In addition, this study found that 6-BA was advantageous to the synthesis and accumulation of phenolic acids, and KT was beneficial to the synthesis and proliferation of terpenoids. Cytokinins have a significant effect on the aromatic components of A. hancockii, and the effects of different cytokinins on aromatic components seem to be different. The results of this study indicate that plant tissue culture is an essential biotechnological means to change the pattern of aromatic components in aromatic plants.
Key message
The use of 6-BA and KT both made the endogenous metabolite content show a more up-regulated trend to a certain extent in the rapid micropropagation system of Anaphalis hancockii Maxim.
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Data availability
All data generated or analysed during this study are included in this published article (and its Supplementary Information files).
Abbreviations
- 6-BA:
-
6-Benzylaminopurine
- KT:
-
Kinetin
- TDZ:
-
N-Phenyl-N′-1,2,3-thiadiazol-5-ylurea
- 2,4-D:
-
2,4-Dichlorophenoxy acetic acid
- IAA:
-
Indole-3-acetic acid
- IBA:
-
4-(3-Indolyl) butyric acid
- NAA:
-
α-Napthaleneacetic acid
- CK:
-
Control check
- PGR:
-
Plant growth regulator
- MS:
-
Murashige and Skoog medium
References
An L, Ma JW, Wang H, Li FG, Qin DM, Wu JK, Zhu GY, Zhang JF, Yuan YL, Zhou L, Wu XJ (2018) NMR-based global metabolomics approach to decipher the metabolic effects of three plant growth regulators on strawberry maturation. Food Chem 269:559–566. https://doi.org/10.1016/j.foodchem.2018.07.061
Aremu AO, Gruz J, Subrtová M, Szücová L, Dolezal K, Bairu MW, Finnie JF, Van Staden J (2013) Antioxidant and phenolic acid profiles of tissue cultured and acclimatized Merwilla plumbea plantlets in relation to the applied cytokinins. J Plant Physiol 170:1303–1308. https://doi.org/10.1016/j.jplph.2013.04.008
Bao JY, Lu YC, Bai HS (2009) Quantitative and qualitative analysis on flavonoids in Anaphalis lactea Maxim. J Mol Sci 25:72–74. https://doi.org/10.3969/j.issn.1000-9035.2009.01.017
Bi HY, Shao Q, Sun Q, Yang ZR, Lin Y, Yun YX (2018) Lectotypifications of names in sympetalae from China—IV. Bull Bot Res 38:161–167. https://doi.org/10.7525/j.issn.1673-5102.2018.02.001
Cao MD, Qin DC, Chen QC, Wang JJ (1998) Study on vanillin production by batch cell suspension culture of Vanilla planifolia. J Huazhong Univ Sci Technol 26:9–11. https://doi.org/10.1088/0256-307X/15/12/024
Castilho CVV, Leitao SG, Silva VD, Miranda CdO, Santos MCdS, Bizzo HR, da Silva NCB (2019) In vitro propagation of a carvacrol-producing type of Lippia origanoides Kunth: a promising oregano-like herb. Ind Crops Prod 130:491–498. https://doi.org/10.1016/j.indcrop.2019.01.006
Chen JR, Cao X (2019) Effects of different hormone treatments on the adventitious bud induction of ornamental pumpkin. China Cucurbits Veg 32:41–43. https://doi.org/10.16861/j.cnki.zggc.2019.0092
Chen FH, Ling R, Chen YL, Shi Z, Wang W (1966) De Genere Anaphalis DC. Familiae Compositarume Florae Sinicae. Acta Phytotaxon Sin 11:91–112
Cloarec O, Dumas ME, Trygg J, Craig A, Barton RH, Lindon JC, Nicholson JK, Holmes E (2005) Evaluation of the orthogonal projection on latent structure model limitations caused by chemical shift variability and improved visualization of biomarker changes in 1H NMR spectroscopic metabonomic studies. Anal Chem 77:517–526. https://doi.org/10.1021/ac048803i
Danova K, Čellárová E, Macková A, Daxnerová Z, Kapchina-Toteva V (2010) In vitro culture of Hypericum rumeliacum Boiss. and production of phenolics and flavonoids. In Vitro Cell Dev Biol Plant 46:422–429. https://doi.org/10.1007/s11627-010-9299-2
De Hsie BS, Bueno AIS, Bertolucci SKV, De Carvalho AA, Martins ER, Pinto JEBP (2019) Growth regulators induced shoot regeneration and volatile compound production in Lippia rotundifolia Cham., a threatened medicinal plant. Ind Crops Prod 137:401–409. https://doi.org/10.1016/j.indcrop.2019.05.050
Du JL, Ma WW, Li Y, Lu X, Geng ZP, Huang HJ, Yuan YY, Liu Y, Wang XD, Wang JL (2021) UPLC–MS-based non-targeted analysis of endogenous metabolite changes in the leaves of Scabiosa tschiliensis Grüning induced by 6-benzylaminopurine and kinetin. Front Plant Sci 12:700623. https://doi.org/10.3389/fpls.2021.700623
Fang HZ, Lu M, An HM (2019) Establishment of leaf callus culture system and contents of main active substance of Rosa roxburghii. Plant Physiol J 55:1147–1155. https://doi.org/10.13592/j.cnki.ppj.2019.0291
George EF, Hall MA, Klerk GJD (2008) Plant growth regulators II: cytokinins, their analogues and antagonists. In: George EF, Hall MA, Klerk GJD (eds) Plant propagation by tissue culture. Springer, Dordrecht, pp 205–226. https://doi.org/10.1007/978-1-4020-5005-3_6
Guo B, Gao M, Liu CZ (2007) In vitro propagation of an endangered medicinal plant Saussurea involucrata Kar. et Kir. Plant Cell Rep 26:261–265. https://doi.org/10.1007/s00299-006-0230-6
Hua Y, Wang HQ (2003) Analysis of the volatile oil components of Anaphalis sinica Hance produced in Gansu. Chin Tradit Herb Drugs 34:19–21. https://doi.org/10.3321/j.issn:0253-2670.2003.01.010
Jiao J, Gai QY, Wang X, Qin QP, Wang ZY, Lu J, Fu YJ (2018) Chitosan elicitation of Isatis tinctoria L. hairy root cultures for enhancing flavonoid productivity and gene expression and related antioxidant activity. Ind Crops Prod 124:28–35. https://doi.org/10.1016/j.indcrop.2018.07.056
Kirimer N, Mokhtarzadeh S, Demirci B, Goger F, Khawar KM, Demirci F (2017) Phytochemical profiling of volatile components of Lavandula angustifolia Miller propagated under in vitro conditions. Ind Crops Prod 96:120–125. https://doi.org/10.1016/j.indcrop.2016.11.061
Kong DM (2010) Medicinal Compositae resources and their diversity in Pangquangou Nature Reserve, Shanxi. J Shanxi Univ (Nat Sci Ed) 3:302–305. https://doi.org/10.13451/j.cnki.shanxi.univ(nat.sci.).2010.02.028
Lan W, Xu PP, Xu SC (2013) Conservation of Dracocephalum moldavica L. in vitro. Chin J Trop Crops 34:675–680. https://doi.org/10.3969/j.issn.1000-2561.2013.04.016
Li Q, Sun D, Li HB, Zhang WQ, Piao ZY (2014) The study of callus proliferation and embryogenic callus induction of Schisandra chinensis (Turcz.) Baillon. Seed 33:5–9. https://doi.org/10.3969/j.issn.1001-4705.2014.06.002
Liang ZX, Huang Q, Zhang XY, Li CB (2021) Stem tip tissue culture and aroma components analysis in fruits of “Benihoppequot” strawberry. J Dalian Natl Univ 23:393–398. https://doi.org/10.13744/j.cnki.cn21-1431/g4.2021.05.003
Lin J, Wu LH, Liang J, Wang JH (2010) Effect of different plant growth regulators on callus induction in Catalpa bungei. Afr J Agric Res 5:2699–2704. https://doi.org/10.5897/AJAR.9000202
Liu CF, Li L, Shi HP, Pan RZ (1999) Effects of auxin and cytokinin substances on organogenesis of Pueraria lobata in vitro. J South China Normal Univ 2:100–104. https://doi.org/10.3969/j.issn.1000-5463.1999.02.022
Liu HJ, Zong RX, Liu JP, You MH, Gou R (2018) Impact on slope position about population characteristics and morphological plasticity of the dioecious herb Anaphalis lactea. Pratacult Sci 35:2105–2114. https://doi.org/10.11829/j.issn.1001-0629.2017-0608
Lu FG, Jin LZ, Xie GP, Li QL (1998) Preliminary study on the geographical distribution and utilization value of grassland plants in Hainan Prefecture. Qinghai Pratacult 7:28–31
Lu JQ, He DL, Xu JQ, Ruan XY, Wang Q (2011) Analysis of the chemical constituents of essential oil from Anaphalis aureopunctata by GC–MS. J Chin Med Mater 34:392–395. https://doi.org/10.13863/j.issn1001-4454.2011.03.023
Makowczyńska J, Sliwinska E, Kalemba D, Piątczak E, Wysokińska H (2016) In vitro propagation, DNA content and essential oil composition of Teucrium scorodonia L. Plant Cell Tissue Organ Cult 127:1–13. https://doi.org/10.1007/s11240-016-1024-3
Monfort LEF, Bertolucci SKV, Lima AF, De Carvalho AA, Mohammed A, Blank AF, Pinto JEBP (2018) Effects of plant growth regulators, different culture media and strength MS on production of volatile fraction composition in shoot cultures of Ocimum basilicum. Ind Crops Prod 116:231–239. https://doi.org/10.1016/j.indcrop.2018.02.075
Mu HM, Ci ZJ, Mamat A, Liang YP, Liu XH, Du XY, Yu Q, Li QY, Li YJ (2021) Analysis of metabolite differences in skin between Clapp’s Favorite and its mutant Red Clapp’s Favorite through non-targeted metabolomics. Chin J Chromatogr 39:1203–1212. https://doi.org/10.3724/SP.J.1123.2021.05002
Newman DJ, Cragg GM, Snader KM (2003) Natural products as sources of new drugs over the period 1981–2002. J Nat Prod 66:1022–1037. https://doi.org/10.1021/np030096l
Nie ZL, Funk V, Sun H, Deng T, Meng Y, Wen J (2013) Molecular phylogeny of Anaphalis (Asteraceae, Gnaphalieae) with biogeographic implications in the Northern Hemisphere. J Plant Res 126:17–32. https://doi.org/10.1007/s10265-012-0506-6
Ou Yang GC, Xue YL (1988) Physiological role and regulation of phenylpropanoid metabolism in plant. Plant Physiol Commun 3:9–16. https://doi.org/10.13592/j.cnki.ppj.1988.03.003
Ruta C, Morone-Fortunato I (2010) In vitro propagation of Cistus clusii Dunal, an endangered plant in Italy. In Vitro Cell Dev Biol Plant 46:172–179. https://doi.org/10.1007/s11627-010-9284-9
Salem MA, De Souza LP, Serag A, Fernie A, Fernie AR, Farag MA, Ezzat SM, Alseekh S (2020) Metabolomics in the context of plant natural products research: from sample preparation to metabolite analysis. Metabolites 10:37. https://doi.org/10.3390/metabo10010037
Song HJ, Huang JY (2007) Application of aromatic medicinal plants in bonsai. J Henan Sci Technol 27:18–19
Sun LY, Yu ZJ, Li HY, Li JB, Liu HZ, Lin XF, Ma R (2009) Advances in secondary metabolites of medicinal plant. J Jilin Agric Sci 34:4–10. https://doi.org/10.3969/j.issn.1003-8701.2009.04.002
Sun DQ, Lu XH, Liang GL, Guo QG, Mo YW, Xie JH (2011) Production of triploid plants of papaya by endosperm culture. Plant Cell Tissue Organ Cult 104:23–29. https://doi.org/10.1007/s11240-010-9795-4
Tan YT, Shang JK, Luo XL, Zhang H, Ding Y, Zhao P, Yan XH, Tang JR (2020) Effects of auxin on the content of secondary metabolites in vitro culture of Vaccinium dunalianum. J West China For Sci 49:133–139. https://doi.org/10.16473/j.cnki.xblykx1972.2020.01.021
Trauger M, Hile A, Sreenivas K, Shouse EM, Bhatt J, Lai T, Mohandass R, Tripathi L, Ogden AJ, Curtis WR (2022) CO2 supplementation eliminates sugar-rich media requirement for plant propagation using a simple inexpensive temporary immersion photobioreactor. Plant Cell Tissue Organ Cult 150:57–71. https://doi.org/10.1007/s11240-021-02210-3
Tripathi PK, Awasthi S, Kanojiya S, Tripathi V, Mishra DK (2013) Callus culture and in vitro biosynthesis of cardiac glycosides from Calotropis gigantea (L.) Ait. In Vitro Cell Dev Biol Plant 49:455–460. https://doi.org/10.1007/s11627-012-9481-9
Wang L, Shi LL, Zhang YX, Liu YJ (2007a) Biosynthesis and regulation of the secondary metabolites in plants. J Wuhan Bot Res 25:500–508. https://doi.org/10.1016/S1872-2075(07)60055-7
Wang XL, Jiang LL, Li N, Zhao YL, Li TF (2007b) Gas chromatographic–mass spectra analysis of Ionone. J Anshan Univ Sci Technol 30:573–575. https://doi.org/10.3969/j.issn.1674-1048.2007.06.004
Wang JL, Liu K, Xu DT, Wang Q, Bi KL, Song YF, Li JF, Zhang L (2013a) Rapid micropropagation system in vitro and antioxidant activity of Scabiosa tschiliensis Grunning. Plant Growth Regul 69:305–310. https://doi.org/10.1007/s10725-012-9765-4
Wang LJ, Fang X, Yang CQ, Li JR, Chen XY (2013b) Biosynthesis and regulation of secondary terpenoid metabolism in plants. Sci Sin (Vitae) 43:1030–1046. https://doi.org/10.1360/052013-307
Wang Y, Zhang BY, Dang J, Tao YD, Mei LJ, Wang QL (2013c) Determination of total flavones and antioxidant activity of Anaphalis lactea Maxim. and Anaphalis margaritacea Benth. Chin J Spectrosc Lab 30:2104–2108. https://doi.org/10.3969/j.issn.1004-8138.2013.05.012
Wang T, Xu QH, Lu JY, Shen W, Sun YH, Li YW, Li YC, Yang XL, Zhang RC, Wei HC (2018) Pollen morphology of summer-flowering species of alpine meadows in northeastern Qinghai-Tibet Plateau. Acta Palaeontol Sin 57:524–542. https://doi.org/10.19800/j.cnki.aps.2018.04.011
Xiong WX, Huang QJ (2022) Tissue culture and establishment of regeneration of Flickingeria albopurpurea. Plant Physiol J 58:1537–1542. https://doi.org/10.13592/j.cnki.ppj.100085
Xu R, Yu YY, Shi J, Mao GL, Yao XL, Zheng R (2016) Callus induction from different explants of wolfberry ‘Ningqi No. 1.’ North Hortic 24:91–95. https://doi.org/10.11937/bfyy.201624023
Yan LN, Ren LQ, Jiao Y (2011) Investigation on wild medicinal plant resources in Zhangshiyan area, Hebei Province. Lishizhen Med Mater Med Res 22:2752–2754. https://doi.org/10.3969/j.issn.1008-0805.2011.11.081
Yang Q, Wang J, Da W, Yang CJ, Quan X, Tao K, Hou TP (2011) Antimicrobial activity of the extracts of fourteen plants from the Qinghai-Tibet Plateau against phytopathogen. J Sichuan Univ (Nat Sci Ed) 48:720–724. https://doi.org/10.3969/j.issn.0490-6756.2011.03.044
Yang J, Mu WW, Cao YX, Liu GY (2020) Synthesis and biological evaluation of β-ionone oriented proapoptosis agents by enhancing the ROS generation. Bioorg Chem 104:104273. https://doi.org/10.1016/j.bioorg.2020.104273
Yu D, Kong FL, Qi J (2019) Determination of phenylacetaldehyde in model reaction system by high performance liquid chromatography. Food Ind 40:298–301
Zhang QG, Bi LW, Zhao ZD, Bo CY, Li DM, Gu Y, Wang J (2008a) Research progress in preparation and application of p-cymene. Mod Chem Ind 28(S2):401–406. https://doi.org/10.3321/j.issn:0253-4320.2008.z2.110
Zhang YM, Zhu SX, Li JM, Li QB, Gu ZY (2008b) A comparative study on characters of leaf morphology and anatomical structure of genus Anaphalis. J Henan Agric Univ 42:646–649. https://doi.org/10.16445/j.cnki.1000-2340.2008.06.001
Zhang HQ, Yang Y, She JY, Liu YQ, Jiang WQ, Zhang B, Deng YH, Xiong XY (2019) Composition and antitumor activities of essential oils from two Anaphalis DC. species. Nat Prod Res Dev 31(2087–2092):2076. https://doi.org/10.16333/j.1001-6880.2019.12.011
Zhang BB, Ye YY, Zhou JS, Tang YP, Yin YL, Luo SC (2022) Tissue culture and rapid propagation technology of Trichosanthes kirilowii Maxim. Seed 41:127–131. https://doi.org/10.16590/j.cnki.1001-4705.2022.06.127
Zheng XQ, Tan PP, Feng G, Cao F, Zhou F, Peng FR, Li YR (2020) Optimization of callus induction in pecan. Mol Plant Breed 18:3704–3712. https://doi.org/10.13271/j.mpb.018.003704
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This work was financially supported by The National Key Research and Development Program of China (2018YFD0201300).
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WJL designed the experiments and manuscript. WXD developed the UPLC–MS methods. GZP performed the metabolomics analysis, interpreted the results, and prepared the manuscript. HN cultivated the experimental materials. DJL, LX, and LY developed the UPLC–MS methods and performed the experiments. YYY, LJL and WMJ contributions to the acquisition of data. All authors have read and approved the manuscript for publication.
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Geng, Z., Hao, N., Du, J. et al. Micropropagation system of Anaphalis hancockii Maxim in vitro and exploration of endogenous metabolome differences coursed by plant growth regulators. Plant Cell Tiss Organ Cult 152, 81–98 (2023). https://doi.org/10.1007/s11240-022-02390-6
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DOI: https://doi.org/10.1007/s11240-022-02390-6