In vitro Induction of Axillary Turion Production in Hydrilla Verticillata with the Potential for Short-Term Storage via Alginate Encapsulation
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Abstract
Hydrilla verticillata is an aquatic macrophyte that has attracted increasing attention due to its potential applications in ecological management and pharmaceutical research. To advance these applications, cultivating this plant using efficient large-scale operations under controlled conditions and germplasm storage technology, such as plant tissue culturing, are much more desirable than harvesting it from nature, which also has the potential to cause environmental damage. However, information regarding the tissue culture of this submerged vascular plant remains limited. Therefore, the present study aimed to establish an efficient in vitro stock culture system that could subsequently be utilized to investigate the effects of in vitro culture conditions, as well as the potential for synthetic seed production and short-term storage. In vitro cultivation of H. verticillata was successfully achieved using basal liquid Murashige and Skoog (MS) medium. The addition of either benzyladenine or paclobutrazol to the medium resulted in reduced shoot elongation and decreased lateral branching. Notably, when explants cultured in basal liquid MS basal medium were transferred to semi-solid MS medium, the development of axillary turions was observed in 55% of the jars, compared to 0% in those that remained submerged in the liquid medium, suggesting that water stress, reduced direct availability of nutrients and improved aeration conditions may stimulate axillary turion formation in vitro. Furthermore, these axillary turions were found to be a suitable explant source for synthetic seed production and enhanced plant survival during low-temperature storage compared with synthetic seeds derived from shoot tissues. The findings of this study provide a useful approach for H. verticillata germplasm conservation and facilitate the transportation of plant materials.
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References
Shrivastava M, Srivastava S. Application and research progress of Hydrilla verticillata in ecological restoration of water contaminated with metals and metalloids. Environ Chall. 2021;4:100177.
Al-Zurfi SKL, Al-Tabatabai HHM. Aquatic plant (Hydrilla verticillata) roles in bioaccumulation of heavy metals. Iraqi J Agric Sci. 2020;51(2):574-84.
Jain MS, Kalamdhad AS. A review on management of Hydrilla verticillata and its utilization as a potential nitrogen-rich biomass for compost or biogas production. Bioresour. Technol. Rep. 2018;1:69-78.
Behera B, Satapathy KB. Studies on in vitro evaluation of antibacterial and antioxidant activity of Hydrilla verticillata (L.f.) Royle. Plant Sci. Today 2023;10(4):66-71.
Prabha SP, Sadhana S, Karthik C, Caroline DG. In vitro evaluation of Hydrilla verticillata for anti-adipogenesis activity on 3T3 L1 cell lines. Phcog Mag. 2020;16:S498-505.
Roeswitawati D, Zamzami AF, Machmudi. The role of hydrilla (Hydrilla verticillata (L.f.) Royle) as soil protectant in improving soil physical, chemical, and biological properties. Ilmu Pertanian (Agric Sci) 2023;8(1):47-54.
Adamec L. Ecophysiological characteristics of turions of aquatic plants: A review. Aquat Bot. 2018;148:64-77.
Baniszewski J, Cuda JP, Gezan SA, Sharma S, Weeks ENI. Stem fragment regrowth of Hydrilla verticillata following desiccation. J Aquat Plant Manage. 2016;54:53-60.
Spencer DF, Ksander GG. Comparative growth and propagule production by Hydrilla verticillata grown from axillary turions or subterranean turions. Hydrobiologia 1991;222:153-8.
Netherland MD. Turion ecology of hydrilla. J Aquat Plant Manag. 1997;35:1-10.
Sudhersan C, AboEl-Nil M, Hussain J. Tissue culture technology for the conservation and propagation of certain native plants. J Arid Environ. 2003;54(1):133-47.
Rodboot N, Te-chato S, Yenchon S. Optimizing culture medium for commercial production of tropical day-blooming waterlily (Nymphaea colorata Peter). Thai J Agric Sci. 2024;57(4):224-35.
Bodhipadma K, Noichinda S, Kluiprasert P, Luekklin P, Raksri T, Leung DWM. Promotive effect of ginger rhizome extract supplement on in vitro rooting of Persian violet shoot tip culture. Curr Appl Sci Technol. 2025;25(6):e0264756.
Wijerathna-Yapa A, Hiti-Bandaralage J. Tissue culture—A sustainable approach to explore plant stresses. Life (Basel) 2023;13(3):780.
Mazri MA, Koufan M, Rham I, Radi H, Belkoura I. Use of tissue culture methods to improve stress tolerance in plants. In: Aftab T, editor. New frontiers in plant environment interactions (Innovative technologies and developments). Cham, Switzerland: Springer; 2023. p. 425-60.
Bodhipadma K, Noichinda S, Maneeruang T, Nathalang K, Punnakanta L, Leung DWM. Effect of paclobutrazol on three different aquatic macrophytes under in vitro monoculture or polyculture conditions. Afr J Biotechnol. 2013;12(39):5809-13.
Muktawapai, K., & Wongchaochant, S. Micropropagation and in vitro short-term storage of Globba sherwoodiana W.J. Kress & V. Gowda. Agr Nat Resour. 2020;54(4):405-14.
Chauhan R, Singh V, Quraishi A. In vitro conservation through slow-growth storage. In: Faisal M, Alatar AA, editors. Synthetic seeds (Germplasm regeneration, preservation and prospects). Cham, Switzerland: Springer; 2019. p. 397-416.
Cruz-Cruz CA, González-Arnao MT, Bautista-Aguilar JR, Ramírez-Mosqueda MA. In vitro short-term storage of Stanhopea tigrina Bateman ex Lind. S Afr J Bot. 2022;151(Part A):334-8.
Sundararaj SG, Agrawal A, Tyagi RK. Encapsulation for in vitro short term storage and exchange of ginger (Zingiber officinale Rosc.) germplasm. Sci. Hort. 2010;125,761-6.
Klaocheed S, Rittirat S, Thammasiri K, Prasertsongskun S. Alginate-encapsulation, short-term storage and plantlet regeneration from encapsulated protocorm-like bodies (PLBs) of Cymbidium finlaysonianum Lindl.: An endangered orchid of Thailand. Walailak J Sci & Tech. 2018;15(10):725-37.
Phanomchai S, Bodhipadma K, Noichinda S, Leung DWM. Short-term storability of alginate-encapsulated Persian violet microshoots for germplasm exchange. Plants (Basel) 2022;11(2):185.
Ghosh T, Samai I, Debnath S, Singh AP, Atta K. Synthetic seeds: Innovation in plant propagation and conservation for sustainable agriculture. In: Roy B, Shukla G, Dunna V, Sharma P, Shukla PS, editors. Advances in seed quality evaluation and improvement. Singapore: Springer; 2025. p. 431-446.
Kiran VE, Aswathi NV, Thomas TD. Micropropagation and conservation of Rotala malabarica Pradeep Joseph and Shivarajan, a critically endangered aquatic herb, endemic to Western Ghats. Plant Cell Tiss Organ Cult. 2025;160:26.
Micheli M, Benelli C. Synthetic seeds of two aquatic plants. In: Faisal M, Alatar AA, editors. Synthetic seeds (Germplasm regeneration, preservation and prospects). Cham, Switzerland: Springer; 2019. p. 233-9.
Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 1962;15:473-
Sukthavornthum W, Bodhipadma K, Noichinda S, Phanomchai S, Deelueak U, Kachonpadungkitti Y, Leung DWM. UV-C irradiation induced alterations in shoot proliferation and in vitro flowering in plantlets developed from encapsulated and non-encapsulated microshoots of Persian violet. Sci Hortic. 2018;233:9-13.
Laksana C, Sophiphun O, Kaewhom P, Chanprame S. Light spectrum effects on micropropagation and gene expression of Bucephalandra sp. in a temporary immersion system for sustainable production. Front Plant Sci. 2025;16:1660632.
Lin F, Kang Y, Li Y, Guo Y, Wang W, Yang G, Yin J, Tang F, Eissa MA. In vitro propagation technology for the endangered aquatic species Nymphoides coronata. Plants (Basel) 2024;13(11):1508.
Rittirat S, Klaocheed S, Thammasiri K. Thidiazuron induces high-frequency plant regeneration from shoot tip explants of an ornamental aquatic plant, Anubias barteri var. nana. Acta Hortic. 2022;1339:273-80.
Song Y, He X-J, Chen M, Zhang L-L, Li J, Deng Y. Effects of pH on the submerged macrophyte Hydrilla verticillata. Russ J Plant Physiol. 2018;65(4):611-9.
Priyadharshini S, Kannan N, Manokari M, Shekhawat MS. In vitro regeneration using twin scales for restoration of critically endangered aquatic plant Crinum malabaricum Lekhak & Yadav: a promising source of galanthamine. Plant Cell Tiss Organ Cult. 2020;141:593-604.
Kanchanapoom K, Chunui P, Kanchanapoom K. Micropropagation of Anubias barteri var. nana from shoot tip culture and the analysis of ploidy stability. Not Bot Horti Agrobo. 2012;40(2):148-151.
Abdalla N, Taha N, Bayoumi Y, El-Ramady H, Shalaby TA. Paclobutrazol applications in agriculture, plant tissue cultures and its potential as stress ameliorant: A mini review. Env. Biodivers. Soil Secur. 2021;5:245-257.
Tauk C, Chuaduangpui P, Khawniam T. Effects of explants on plant regeneration and concentration of paclobutrazol on morphological responses of dwarf water hyssop (Bacopa monnieri). Asian Health Sci Tech Rep. 2021;29(3):56-66.
Spencer D, Anderson L, Ksander G, Klaine S, Bailey F. Vegetative propagule production and allocation of carbon and nitrogen by monoecious Hydrilla verticillata (L.f.) Royle grown at two photoperiods. Aquat Bot. 1994;48(2):121-32.
Płachno BJ, Adamec L, Feldo M, Stolarczyk P, Kapusta M. Can aquatic plant turions serve as a source of arabinogalactans?
immunohistochemical detection of AGPs in turion cells. Molecules 2025;30(24):4689.
Bhattacharjee S, Saha AK. Plant waterstress response mechanisms. In: Gaur RK, Sharma P, editors. Approaches to plant stress and their management. New Delhi: Springer; 2014. p. 149-72.
Marcin´ska I, Czyczyło-Mysza I, Skrzypek E, Filek M, Grzesiak S, Grzesiak MT, Janowiak F, Hura T, Dziurka M, Dziurka K, Nowakowska A, Quarrie SA. Impact of osmotic stress on physiological and biochemical characteristics in droughtsusceptible and drought-resistant wheat genotypes. Acta Physiol Plant 2013;35:451-61.
Gopal J, Iwama K, Jitsuyama Y. Effect of water stress mediated through agar on in vitro growth of potato. In Vitro Cell Dev Biol-Plant 2008;44:221-8.
Turhan H, Baser I. In vitro and in vivo water stress in sunflower (Helianthus annuus L.). Helia 2004;27(40):227-36.
Derelli E, Allahverdikhan Vaziri P, Mirzapour M, Yildiz, M. The effect of water deficiency originated stress in tissue on shoot regeneration of flax (Linum usitatissimum L.) hypocotyl explants. Res J Agric Sci. 2012; 5(1): 153-6.
Jang BK, Lee CH. Synthetic seed production using eastern bracken (Pteridium aquilinum var. latiusculum) gametophyte: Effect of gametophyte density and short-term storage conditions. S Afr J Bot. 2021;139:267-72.
Regni L, Micheli M, Pino AMD, Facchin SL, Rabica E, Camilloni L, Cesarini A, Proietti P. Blackberry synthetic seeds storage: effects of temperature, time, and sowing substrate. Plant Cell Tiss Organ Cult. 2024;158:17.
Chotikadachanarong K. Artificial seed production from nodal explant of Stevia rebaudiana Bertoni. BUU SCI J. 2018;23(1):247-54.
