菰腺忍冬组织培养快繁体系建立及总黄酮与绿原酸含量测定

HE Duo-xiu; ZHANG Zhi-yong; Chris Rey LITUANAS

doi:10.3969/j.issn.1009-7791.2024.04.004

亚热带植物科学 >

2024 , Vol. 53 >Issue 4: 315 - 324

DOI: https://doi.org/10.3969/j.issn.1009-7791.2024.04.004

研究论文

菰腺忍冬组织培养快繁体系建立及总黄酮与绿原酸含量测定

HE Duo-xiu ,
ZHANG Zhi-yong ,
Chris Rey LITUANAS

展开

(1. 湖南医药学院总医院，湖南怀化 418000；2. 湖南医药学院医学信息与工程学院，湖南怀化 418000；3. 中棉兰老大学生物系，菲律宾马拉马格 8710)

收稿日期: 2024-04-12

录用日期: 2024-05-14

网络出版日期: 2024-11-20

基金资助

湖南省教育厅重点项目(18A492)

收起

Establishment of an in vitro Tissue Culture and Rapid Propagation System using Lonicera hypoglauca and Content Assessment of Total Flavonoids and Chlorogenic Acid

何朵秀，张志勇，Chris Rey LITUANAS

Expand

(1. Hunan Medical College General Hospital, Hunan University of Medicine, Huaihua 418000, Hunan China; 2. School of Medical Information and Engineering, Hunan University of Medicine, Huaihua 418000, Hunan China; 3. Biology Department, Central Mindanao University, Musuan, Bukidnon 8710, Philippines)

Received date: 2024-04-12

Accepted date: 2024-05-14

Online published: 2024-11-20

Fold

摘要

菰腺忍冬是一种传统的中药材。本研究首次以菰腺忍冬嫩叶为外植体，建立了其体外快速繁殖和再生体系。结果表明，外植体消毒的最佳时间为8 min；外植体诱导愈伤组织的最佳培养基为MS + 2,4-D 4.0 mg·L–1 + 蔗糖30 g·L–1，平均诱导率为86.67%；诱导愈伤组织分化为芽的最佳培养基为MS + 6-BA 1.0 mg·L–1 + NAA 0.10 mg·L–1 + 蔗糖30 g·L–1，平均发芽率为83.33%。诱导芽增殖的最佳培养基为MS + 6-BA 1.5 mg·L–1 + NAA 0.05 mg·L–1 + 蔗糖30 g·L–1，增殖系数为5.42。生根的最佳培养基为1/2 MS + NAA 0.15 mg·L–1 + 活性炭0.3 g·L–1 + 蔗糖15 g·L–1，平均生根率为91.11%。再生植株幼苗在营养土壤培养基中的存活率高达100%。外植体、愈伤组织和再生植株中的总黄酮和绿原酸含量分别为1.83%、2.27%、1.33%和2.77%、1.83%、1.74%。本研究为菰腺忍冬再生植株的工业化快速繁殖和大规模生产提供了新的见解，具有良好的应用价值和未来前景。

关键词： 菰腺忍冬；快繁体系；组织培养；总黄酮；绿原酸

本文引用格式

HE Duo-xiu , ZHANG Zhi-yong , Chris Rey LITUANAS . 菰腺忍冬组织培养快繁体系建立及总黄酮与绿原酸含量测定[J]. 亚热带植物科学, 2024 , 53(4) : 315 -324 . DOI: 10.3969/j.issn.1009-7791.2024.04.004

Abstract

Lonicera hypoglauca is a traditional Chinese medicinal plant. In this study, the tender young leaves of L. hypoglauca were used for the first time as the explants to establish a rapid in vitro propagation and regeneration system. The results revealed that the optimal time for disinfection of the explants was 8 min and the optimal medium for callus induction was MS + 2,4-D 4.0 mg·L–1 + sucrose 30 g·L–1, with an average callus induction rate of 86.67%. The optimal medium to induce differentiation of callus to bud was MS + 6-BA 1.0 mg·L–1 + NAA 0.10 mg·L–1 + sucrose 30 g·L–1, with an average germination rate of 83.33%. The optimal medium to induce multiplication was MS + 6-BA 1.5 mg·L–1 + NAA 0.05 mg·L–1 + sucrose 30 g·L–1, with a multiplication coefficient of 5.42. The optimal medium for root induction was 1/2 MS + NAA 0.15 mg·L–1 + activated carbon 0.3 g·L–1 + sucrose 15 g·L–1, with an average rooting rate of 91.11%. The survival rate of tissue-cultured seedlings in nutrient soil cultivation medium was as high as 100%. The total flavonoid content and chlorogenic acid content in the explant, callus tissue and regenerated plant were 1.83%, 2.27%, 1.33% and 2.77%, 1.83%, 1.74% respectively. This study provides novel insights into the rapid propagation and mass production of L. hypoglauca seedlings at an industrial scale and that it exhibits important application value and future prospects.

Key words： Lonicera hypoglauca; rapid propagation system; tissue culture; total flavonoids; chlorogenic acid

参考文献

[1] Wu Z Y, Raven P H, Hong D Y. Flora of China [M]. Beijing: Science Press &St. Louis: Missouri Botanical Garden Press, 2007: 239.
[2] China Pharmacopeia Commission. Pharmacopoeia of the People’s Republic of China [M]. Beijing: The Medicine Science and Technology Press of China, 2020: 32.
[3] Xiong L W, Jin Y, Wang Y Y, Zhao H W, Li J, Pu G B, Zhang L, Yang H B, Zhang Y Q, Zhang L F. Progress on chemical composition, pharmacological activity and in vivo metabolism of Lonicera japonica [J]. Chinese Traditional Patent Medicine, 2022, 44: 864–871.
[4] Wu Q, Huang B. Report on the germplasm resources of Flos Lonicerae in Guangxi [J]. Lishizhen Medicine and Materia Medica Research, 2008, 19: 394–395.
[5] Wu Q, Chen L, Fu J, Wan L, Pan L, Wei S. Cultivation techniques of Lonicera hypoglauca Miq. [J]. Chinese Journal of Tropical Agriculture, 2019, 39: 35–39.
[6] Sun X, Chen S, Cheng R, Li Y, Xin N. Drought stress’s impact on physiological and biochemical indexes of seedlings of Lonicera hypoglauca Miq [J]. Journal of Guangxi Agriculture, 2013, 28: 9–13.
[7] Nong Y, Yang M, Su X, Cen X. Tissue culture and rapid propagation of Lonicera hypoglauca [J]. Journal of Southern Agriculture, 2013, 44: 717–721.
[8] Shi F, Tan M, Mo Q, Pu Z. Induction of adventitious buds from petiole of sterile tissue culture seedlings of Lonicera hypoglauca Miq. [P], CN: 105409778A. 2016.
[9] Zhou J, Yang M, Cen X, Wei P. Influence of explants and its sterilization time on induction of Lonicera hypoglauca buds [J]. Journal of Southern Agriculture, 2011, 42: 124–127.
[10] Li K, Huang C, Zhou S, Yang M. Germination and induction of axillary buds in micropropagated Lonicerae hypoglauca, Guangxi [J]. Agriculture Science, 2010, 41: 1029–1032.
[11] Liao J, Tan M, Mo Q, Pu Z, Shi F. Adventitious root induction of petiole callus from sterile tissue culture seedlings of Lonicera hypoglauca Miq. [P]. CN: 104170733A. 2014.
[12] Wang L, Xin N, Qiu Y, Yang B. Antioxidative effects of total flavonoids from Guangxi Lonicera hypoglauca Miq. and Shandong Lonicera japonica in vivo [J]. Journal of Guangxi Medical University, 2010, 27: 681–683.
[13] Ge S, Zhu Y, Guo X, Shi S. Content determination of total polyphenol in antioxidant effective fraction of leaves of Lonicera hypoglauca Miq [J]. Journal of Zhejiang TCM University, 2014, 38: 316–320,326.
[14] Zhu Y, Li J, Ren J. The study of total flavonoids in leaves of Lonicera hypoglauca Miq [J]. Journal of Medical Research, 2009, 38: 43–45.
[15] Zhu Y, Xu Z. Studies on correlation between bacteriostasis and total flavonoids, chlorogenic acid content in leaves of Lonicera hypoglauca Miq [J]. Chinese Journal of Traditional Medicine Science Technology, 2014, 21: 39–41.
[16] Zhu Y, Li S. Experimental study of anti-inflammatory and antipyretic effect on leaves of Lonicera hypoglauca Miq. [J]. Journal of Gansu College of Traditional Chinese Medicine, 2014, 31: 12–17.
[17] Shi S. Screening anti-tumor effective fraction from the leaf of Lonicera hypoglauca Miq. and its mechanism [D]. Hangzhou, China: Master dissertation of Zhejiang Chinese Medical University, 2015.
[18] Komatsu T, Kondo N. Winter habitat of Xylophilus ampelinus, the cause of bacterial blight of grapevine, in Japan [J]. Journal of General Plant Pathology, 2015, 81: 237–242.
[19] Lei T G, He Y R, Zou X P, Wang X F, Fu S M, Peng A H, Xu L Z, Yao L X, Chen S C, Zhou C Y. A rapid multiplication system for ‘Candidatus Liberibacter asiaticus’ through regeneration of axillary buds in vitro [J]. Journal of Integrative Agriculture, 2022, 21: 1683–1693.
[20] Ahmed E U, Hayashi T, Zhu Y, Hosokawa M, Yazawa S. Lower incidence of variants in Caladium bicolor Ait. plants propagated by culture of explants from younger tissue [J]. Scientia Horticulturae, 2002, 96: 187–194.
[21] Mazumdar P, Basu A, Paul A, Mahanta C, Sahoo L. Age and orientation of the cotyledonary leaf explants determine the efficiency of de novo plant regeneration and agrobacterium tumefaciens-mediated transformation in Jatropha curcas L. [J]. South African Journal of Botany, 2010, 76: 337–344.
[22] Lv B, Yan Z, Tian H, Zhang X, Ding Z. Local auxin biosynthesis mediates plant growth and development [J]. Trends in Plant Science, 2019, 24: 6–9.
[23] Sarabi V, Arjmand-Ghajur E. Exogenous plant growth regulators/plant growth promoting bacteria roles in mitigating water-deficit stress on chicory (Cichorium pumilum Jacq.) at a physiological level [J]. Agricultural Water Manage, 2021, https://doi.org/10.1016/j.agwat.2020.106439.
[24] Li X, Yang M C, Quan Q, Wu Q H. Tissue culture and rapid propagation of Lonicera confuse [J]. Guihaia, 2008, 28: 823–826.
[25] Wang H, Jiang X. Tissue culture of Lonicera macranthoides Hand. Mazz [J]. Journal of Anhui Agricultural Sciences, 2013, 41: 11952–11953,11964.
[26] Ye L T, Peng S X, Xiang H T, Chen F, Shi S C, Xiang Y, Fu W J, Ning W J. Tissue culture and rapid propagation technologies of Lonicera macranthoides Hand. Mazz. [J]. Hunan Agricultural Sciences, 2013(5): 27–29.
[27] Sugimoto K. Plant cell reprogramming as an adaptive strategy [J]. Journal of Plant Research, 2015, 128: 345–347.
[28] Pasternak T, Dudits D. Epigenetic clues to better understanding of the asexual embryogenesis in planta and in vitro [J]. Front Plant Science, 2019, https://doi.org/10.3389/fpls.2019.00778.
[29] Chen L. Studies on the establishment of rapid propagation system in vitro of Lonicera confuse [D]. Guangzhou, China: Master dissertation of Guangzhou University of Chinese Medicine, 2009.
[30] Yu K W, Murhty H N, Hahn E J, Paek K Y. Ginsenoside production by hairy root cultures of Panax ginseng: influence of temperature and light quality [J]. Biochemical Engineering Journal, 2005, 23: 53–56.

Options

文章导航

〈

〉

闽ICP备11008787号-5　

模态框（Modal）标题

摘要

本文引用格式

Abstract

参考文献