Establishment of an Efficient Tissue Culture and Regeneration System for Lycium chinense var. macrophyllum and Its Callus Culture Characteristics 

Wei Guanghui¹  Zhu Jiayue¹  Wei Shijing¹  Shamu Buayi¹  Yao Ye ¹  Zhang Conghao¹  Wang Hemeng^{1,2（Correspondence）}

1 Key Laboratory of Conservation and Utilization of Biological Resources in the Tarim Basin, Alar, Xinjiang / College of Life Science and Technology, Tarim University, Alar, Xinjiang China，843300；

2 College of Agriculture, Northeast Agricultural University, Harbin China，150030；

Abstract：Lycium chinense var. macrophyllum, a unique Chinese medicinal and edible plant with high nutritional value and broad application prospects, is limited by inefficient traditional breeding methods. Taking different tissur as explants, this study explored 6-BA and NAA ratio effects on tissue culture and optimized disinfection. The results showed that the optimal disinfection procedure was rinsing with running water for 2 h soaking in 75% ethanol for 60 s, after rinsing with sterile water, sterilizing with 1% sodium hypochlorite for 20 min. The optimal media for callus induction and bud differentiation were MS+6-BA 0.3 mg/L+NAA 1.0 mg/L and MS+6-BA 0.3 mg/L +NAA 0.5 mg/L, respectively. After acclimatization, the survival rate of transplanted regenerated plants reached 95%. This study established an efficient tissue culture and regeneration system, providing technical support for its germplasm preservation and genetic improvement.

Keywords: Lycium chinense var. macrophyllum.; callus; tissue culture; plant regeneration

References

[1] Jee W, Cho HS, Kim SW, Bae H, Chung WS, Cho JH, Kim H, Song MY, Jang HJ. Lycium chinense Mill Induces Anti-Obesity and Anti-Diabetic Effects In Vitro and In Vivo. Int J Mol Sci. 2024 Aug 6;25(16):8572.

[2] Ryu MJ, Kim M, Ji M, Lee C, Yang I, Hong SB, Chin J, Seo EK, Paik MJ, Lim KM, Nam SJ. Discrimination of Lycium chinense and L. barbarum Based on Metabolite Analysis and Hepatoprotective Activity. Molecules. 2020 Dec 10;25(24):5835.

[3] Potterat O. Goji (Lycium barbarum and L. chinense): Phytochemistry, pharmacology and safety in the perspective of traditional uses and recent popularity. Planta Med. 2010 Jan;76(1):7-19.

[4] Sathuphan P, Vajrodaya S, Sanevas N, Wongkantrakorn N. In Vitro Propagation of Clausena lenis Drake. Plants (Basel). 2025 Apr 4;14(7):1123. doi: 10.3390/plants14071123. PMID: 40219192; PMCID: PMC11991351.

International Symposiumon Botanical Gardens and Landscapes1298; ISHS:Leuven,Belgium,2019;pp. 249–256.

[5] Noor,W.;Lone,R.;Kamili,A.N.;Husaini,A.M.Callus induction and regeneration in high-altitude Himalayan ric egenotype SR4 via seed explant. Biotechnol.Rep. 2022, 36, e00762.

[6] Beveridge,C.A.; Rameau,C.;Wijerathna-Yapa,A. Lessons froma century of apical dominance research. J. Exp. Bot. 2023,74, 3903–3922.

[7] Abdel-Rahman,S.;Abdul-Hafeez,E.;Saleh,A. Improving rooting and growth of Conocarpus erectus stem cutting susingindole-3-butyric acid(IBA) and some biostimulants. Sci. J.FlowersOrnam. Plants2020, 7, 109–129.

[8] Nakasha JJ, Sinniah UR, Kemat N, Mallappa KS. Induction, Subculture Cycle, and Regeneration of Callus in Safed Musli (Chlorophytum borivilianum) using Different Types of Phytohormones. Pharmacogn Mag. 2016 Jul;12(Suppl 4):S460-S464. doi: 10.4103/0973-1296.

[9] Li H, Wang H, Guan L, Li Z, Wang H, Luo J. Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar. Life (Basel). 2023 Sep 11;13(9):1896.

[10] Wang,Z.;Yeo,D.;Kwon,H.;Zhang,Y.;Yoon,D.;Jung,S.;Hossain,Md.I.;Jeong,M.-I.;Choi,C.Disinfection efficiency of chlorine dioxide and peracetic acid against MNV-1 and HAV in simulated soil-rich wash water. J.Food Res.Int. 2024,175,113772.

[11] Nazih,A.;Baghour,M.;Maatougui,A.;Aboukhalid,K.;Chiboub,B.;Bazile,D.Effect of Gibberellic Acid and Mechanical Scarification on the Germination and Seedling Stages of Chenopodium quinoa Willd. under Salt Stress. Plants 2024,13(10),1330.

[12]AL-Huqail,A.A.;Alghanem,S.M.S.;Alghamdi,S.A.;Alhaithloul,H.A.S.;Al-Robai,S.A.;Alalawy,A.I.;Alzuaibr,F.M.;Alasmari,A.;Ali,B.;Saleem,M.H.;Fahad,S.;Ali,S.;Abeed,A.H.A.Coactive Application of Bacillus Mycoides PM35 and Calcium Oxide Nanoparticles Stimulate Gene Expression Responses in Maize (Zea Mays L.) under Chromium Stress. J.Soil Sci.Plant Nutr. 2024,24(3),4224–4242.