Abstract
Macleaya microcarpa are rich in benzylisoquinoline alkaloids (BIAs). Hairy roots can effectively produce natural secondary metabolites. A reliable and efficient hairy root induction system for M. macrocarpa was established. The highest rate of transformation (96.7%) was obtained in 37 days leaves. The levels of BIAs in hairy roots and wild roots of M. cordata and M. microcarpa were compared. It was found that the content of sanguinarine (SAN) and chelerythrine (CHE) in hairy roots of M. microcarpa was higher than that in wild roots, but the content of protopine (PRO) and allocryptopine (ALL) was lower than wild roots. This finding has important implications to produce SAN and CHE by using the hairy roots of M. microcarpa as an additional source of SAN and CHE to conserve M. cordata plant resources.
Key message
Macleaya microcarpa hairy roots has been established. Benzylisoquinoline alkaloids (BIAs) was identified. Among BIAs, sanguinarine (SAN), chelerythrine (CHE), protopine (PRO) and allocryptopine (ALL) are the M. microcarpa main active ingredients. The content of SAN((4.28 mg/g))、CHE(2.26mg/g) level in the hairy roots is higher than wild roots M. macrocarpa. The content of PRO(0.63mg/g)、ALL(1.50mg/g) in the hairy roots is lower than wild roots M. mIcrocarpa.
Data availability
The data supporting the findings of this study are available from the corresponding author, Jianguo Zeng, upon request.
References
Altamura MM (2004) Agrobacterium rhizogenes rolB and rolD genes: regulation and involvement in plant development. Plant Cell Tissue Org Cult 77:89–101. https://doi.org/10.1023/B:TICU.0000016609.22655.33
Biswas T, Mathur AK, Mathur A (2017) A literature update elucidating production of Panax ginsenosides with a special focus on strategies enriching the anti-neoplastic minor ginsenosides in ginseng preparations. Appl Microbiol Biotechnol 101:4009–4032. https://doi.org/10.1007/s00253-017-8279-4
Chandra S (2012) Natural plant genetic engineer Agrobacterium rhizogenes: role of T-DNA in plant secondary metabolism. Biotechnol Lett 34:407–415. https://doi.org/10.1007/s10529-011-0785-3
Chandra S, Chandra R (2011) Engineering secondary metabolite production in hairy roots. Phytochem Rev 10(3):371–395. https://doi.org/10.1007/s11101-011-9210-8
Dehio C, Grossmann K, Schell J, Schmülling T (1993) Phenotype and hormonal status of transgenic tobacco plants overexpressing the rolA gene of Agrobacterium rhizogenes T-DNA. Plant Mol Biol 23:1199–1210. https://doi.org/10.1007/BF00042353
Dhakulkar S, Ganapathi TR, Bhargava S, Bapat VA (2005) Induction of hairy roots in Gmelina arborea Roxb. and production of verbascoside in hairy roots. Plant Sci 169:812–818. https://doi.org/10.1016/j.plantsci.2005.05.014
Gantait S, Mukherjee E (2021) Hairy root culture technology: applications, constraints and prospect. Appl Microbiol Biotechnol 105(1):35–53. https://doi.org/10.1007/s00253-020-11017-9
Guan G, Ding S, Yin Y, Duraipandiyan V, Al-Dhabi NA, Liu G (2019) Macleaya cordata extract alleviated oxidative stress and altered innate immune response in mice challenged with enterotoxigenic Escherichia coli. Sci China Life Sci 62:1019–1027. https://doi.org/10.1007/s11427-018-9494-6
Huang P, Xia L, Liu W, Jiang R, Liu X, Tang Q, Xu M, Yu L, Tang Z, Zeng JG (2018) Hairy root induction and benzylisoquinoline alkaloid production in Macleaya Cordata. Sci Rep 8:11986. https://doi.org/10.1038/s41598-018-30560-0
Jiao J, Gai QY, Wang X, Liu J, Lu Y, Wang ZY, Xu XJ, Fu YJ (2020) Effective production of phenolic compounds with health benefits in pigeon pea [Cajanus cajan (L.) Millsp.] hairy root cultures. J Agric Food Chem 68:8350–8361. https://doi.org/10.1021/acs.jafc.0c02600
Khalili S, Moieni A, Abdoli M (2014) Influence of different strains of Agrobacterium rhizogenes, culture medium, age and type of explant on hairy root induction in Echinacea angustifolia. Iran J Genet Plant Breed 3:49–56
Kosina P, Gregorova J, Gruz J, Vacek J, Kolar M, Vogel M, Roos W, Naumann K, Simanek V, Ulrichova J (2010) Phytochemical and antimicrobial characterization of Macleaya cordata herb. Fitoterapia 81:1006–1012. https://doi.org/10.1016/j.fitote.2010.06.020
Li Y, Xu F, Tong X, Chen RN, Shen CY, Liang TT, Chu QP, Zhou B (2020) Effects of Macleaya cordata extract on small intestinal morphology and gastrointestinal microbiota diversity of weaned pigs. Livest Sci 237:104040. https://doi.org/10.1016/j.livsci.2020.104040
Lin L, Li XY, Liu SS, Qing ZX, Liu XB, Zeng JG, Liu ZY (2020) Systematic identification of compounds in Macleaya Microcarpa by high-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry combined with mass spectral fragmentation behavior of Macleaya alkaloids. Rapid Commun Mass Spectrom (RCM) 34:e8715. https://doi.org/10.1002/rcm.8715
Liu M, Lin YL, Chen XR, Liao CC, Poo WK (2013) vitro assessment of Macleaya cordata crude extract bioactivity and anticancer properties in normal and cancerous human lung cells. Exp Toxicol Pathol 65:775–787. https://doi.org/10.1016/j.etp.2012.11.004
Liu X, Liu Y, Huang P, Ma Y, Qing Z, Tang Q, Cao H, Cheng P, Zheng Y, Yuan Z, Zhou Y, Liu J, Tang Z, Zhuo Y, Zhang Y, Yu L, Huang J, Yang P, Peng Q, Zhang J, Jiang W, Zhang Z, Lin K, Ro DK, Chen X, Xiong X, Shang Y, Huang S, Zeng J (2017) The genome of medicinal plant Macleaya cordata provides new insights into benzylisoquinoline alkaloids metabolism. Mol Plant 10:975–989. https://doi.org/10.1016/j.molp.2017.05.007
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Park SU, Facchini PJ (2000) Agrobacterium rhizogenes-mediated transformation of opium poppy, Papaver somniferum L., and California poppy, Eschscholzia californica Cham., root cultures. J Exp Bot 51:1005–1016. https://doi.org/10.1093/jexbot/51.347.1005
Park H, Bergeron E, Senta H, Guillemette K, Beauvais S, Blouin R, Sirois J, Faucheux N (2010) Sanguinarine induces apoptosis of human osteosarcoma cells through the extrinsic and intrinsic pathways. Biochem Biophys Res Commun 399:446–451. https://doi.org/10.1016/j.bbrc.2010.07.114
Pavlova OA, Matveyeva TV, Lutova LA (2014) rol-Genes of Agrobacterium rhizogenes. Russ J Genet Appl Res 4:137–145. https://doi.org/10.1134/S2079059714020063
Sai C, Wang J, Li B, Ding L, Wang H, Wang Q, Hua H, Zhang F, Ren Q (2020) Isolation and identification of alkaloids from Macleaya microcarpa by UHPLC-Q-TOF-MS and their cytotoxic activity in vitro, antiangiogenic activity in vivo. BMC Chem 14:5. https://doi.org/10.1186/s13065-020-0660-1
Sato F, Hashimoto T, Hachiya A, Tamura K, Choi KB, Morishige T, Fujimoto H, Yamada Y (2001) Metabolic engineering of plant alkaloid biosynthesis. Proc Natl Acad Sci U S A 98(1):367–372. https://doi.org/10.1073/pnas.011526398
Shkryl YN, Veremeichik GN, Bulgakov VP, Tchernoded GK, Mischenko NP, Fedoreyev SA, Zhuravlev YN (2008) Individual and combined effects of the rolA, B, and C genes on anthraquinone production in Rubia cordifolia transformed calli. Biotechnol Bioeng 100:118–125. https://doi.org/10.1002/bit.21727
Sudha CG, Sherina TV, Anu Anand VP, Reji JV, Padmesh P, Soniya EV (2013) Agrobacterium rhizogenes mediated transformation of the medicinal plant Decalepis arayalpathra and production of 2-hydroxy-4-methoxy benzaldehyde. Plant Cell Tissue Org Cult (PCTOC) 112:217–226. https://doi.org/10.1007/s11240-012-0226-6
Tani C, Takao N (1962) Studies on the alkaloids of fumariaceous plants. V. Alkaloids of Bocconia cordata (Macleya cordata). Yakugaku Zasshi 82:755–759. https://doi.org/10.1248/yakushi1947.82.5_755
Yao SC, Bai LH, Lan ZZ, Tang MQ, Zhai YJ, Huang H, Wei RC (2016) Hairy root induction and polysaccharide production of medicinal plant Callerya speciosa Champ. Plant Cell Tissue Org Cult (PCTOC) 126:177–186. https://doi.org/10.1007/s11240-016-0988-3
Funding
This study was financially supported by National Key R&D Program of China (2017YFD0501500), Hunan Provincial Natural Science Foundation of China (2020JJ4353) and Hunan Provincial Key Research and Development Project (2020NK2031).
Author information
Authors and Affiliations
Contributions
Q-YZ and PH conceived and designed the research. Q-YZ, M-SS, Z-XX, HL, Y-YW, PH and J-GZ performed the experiments. Q-YZ and PH analyzed the data. Q-YZ and J-GZ wrote the manuscript. All authors have read and agreed to the final version of the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest or competing interests.
Additional information
Communicated by Konstantin V. Kiselev.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Zheng, Q., Xu, Z., Sun, M. et al. Hairy root induction and benzylisoquinoline alkaloid production in Macleaya microcarpa. Plant Cell Tiss Organ Cult 147, 189–196 (2021). https://doi.org/10.1007/s11240-021-02109-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-021-02109-z