Abstract
Echinodorus ‘Indian Red’ is an underwater plant, used worldwide for aquarium ornamentation. An efficient method for in vitro propagation and plantlet acclimatization of this popular aquarium plant was standardized. Surface-disinfected shoot-tips were cultured in submerged conditions in a solid–liquid bilayer medium, consisting of an upper, liquid layer (sterile distilled water) and a lower, solid layer Murashige and Skoog (MS) basal medium supplemented with 3.0% (w/v) sucrose, 0.8% (w/v) agar-agar, and plant growth regulators (PGRs) in different combinations and concentrations. The combination of 2.5 mg L−1 6-benzylaminopurine and 1.0 mg L−1 α-naphthaleneacetic acid improved the multiplication rate to a maximum of 26.8 ± 0.51 shoots per explant after 60 d of culture. The number of multiplied shoots increased with each regeneration cycle, thus from only 26.8 ± 0.51 shoots per explant (first regeneration cycle), this number increased to 33.5 ± 0.58 (second regeneration cycle), and to 38.3 ± 0.62 for the third regeneration cycle with the same medium composition. The highest number of roots (8.3 ± 0.28) per shoot was induced in the presence of 1.0 mg L−1 indole-3-butyric acid, but further growth of these roots was stunted. The best rooting was achieved on PGR-free ½-strength MS medium, where 6.1 ± 0.21 roots per shoot were induced with 5.8 ± 0.35 cm length after 30 d of culture. The regenerated plantlets were successfully acclimatized to submerged underwater conditions, with 100% survival rate. The present protocol is suitable for the commercial propagation of Echinodorus ‘Indian Red’ for aquarium-industries.
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References
Barth H (1998) Echinodorus plant named ‘Indian Red’, united state patent, patent number: plant 10726. Date of Patent: 15th December 1998
Bolyard M (2018) In vitro regeneration of Artemisia abrotanum L. by means of somatic organogenesis. In Vitro Cell Dev Biol-Plant 54:127–130. https://doi.org/10.1007/s11627-017-9878-6
Brunel S (2009) Pathway analysis: aquatic plants imported in 10 EPPO countries. Eur Mediterr Plant Prot Org Bull 39:201–213. https://doi.org/10.1111/j.1365-2338.2009.02291.x
Dissanayake C, Hettiarachchi M, Iqbal MCM (2007) Sustainable use of Cryptocoryne wendtii and Echinodorus cordifolius in the aquaculture industry of Sri Lanka by micropropagation. Sri Lanka J Aquat Sci 12:89–101 https://sljas.sljol.info/articles/abstract/10.4038/sljas.v12i0.2216/
Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42
Haque SM, Ghosh B (2013a) Micropropagation, in vitro flowering and cytological studies of Bacopa chamaedryoides, an ethno-medicinal plant. Env Exp Biol 11:59–68 http://eeb.lu.lv/EEB/201303/EEB_11_Haque.pdf
Haque SM, Ghosh B (2013b) High frequency microcloning of Aloe vera and their true-to-type conformity by molecular cytogenetic assessment of two years old field growing regenerated plants. Bot Stud 54:46 http://www.as-botanicalstudies.com/content/54/1/46
Haque SM, Ghosh B (2016) High-frequency somatic embryogenesis and artificial seeds for mass production of true-to-type plants in Ledebouria revoluta: an important cardioprotective plant. Plant Cell Tissue Organ Cult 127:71–83. https://doi.org/10.1007/s11240-016-1030-5
Haque SM, Paul S, Ghosh B (2016) High-frequency in vitro flowering, hand-pollination and fruit setting in ten different cultivars of Capsicum spp. (C. annuum, C. chinense, and C. frutescens): an initial step towards in vitro hybrid production. Plant Cell Tissue Organ Cult 127:161–173. https://doi.org/10.1007/s11240-016-1039-9
Jabir T, George S, Raj A, Sree Lakshmi S, Joseph A (2016) Micropropagation and in vitro flowering of an ornamental aquarium plant Lindernia antipoda L. (Alston). Int J Aquacult 6:8 http://biopublisher.ca/index.php/ija/article/html/2456/
Jenks MA, Kane ME, McConnell DB (2000) Shoot organogenesis from petiole explants in the aquatic plant Nymphoides indica. Plant Cell Tissue Organ Cult 63:1–8. https://doi.org/10.1023/A:1006471027372
Kam MYY, Chai LC, Chin CF (2016) The biology and in vitro propagation of the ornamental aquatic plant, Aponogeton ulvaceus. SpringerPlus 5:1657. https://doi.org/10.1186/s40064-016-3041-4
Kane ME, Gilman EF, Jenks MA, Sheehan TJ (1990) Micropropagation of the aquatic plant Cryptocoryne lucens. HortSci 25:687–689 http://hortsci.ashspublications.org/content/25/6/687.full.pdf+html
Lehtonen S, Falck D (2011) Watery varieties: aquarium plant diversity from aesthetic, commercial, and systematic prospective. In: Aquino JC (ed) Ornamental plants: types, cultivation and nutrition. Nova Science Publishers, New York, pp 1–46
Lunardi RF, Wohlenberg M, Medeiros N, Agostini F, Funchal C, Dani C (2014) In vitro antioxidant capacity of tea of Echinodorus grandiforus, “leather hat,” in Wistar rat liver. An Acad Bras Ciênc 86:1451–1461. https://doi.org/10.1590/0001-3765201420130507
Marques AM, Provance DW Jr, Kapla MAC, Figueiredo MR (2017) Echinodorus grandiflorus: Ethnobotanical, phytochemical and pharmacological overview of a medicinal plant used in Brazil. Food Chem Toxicol 109(Pt 2):1032–1047 https://www.ncbi.nlm.nih.gov/pubmed/28322968
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:495–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Öztürk M, Khawar KM, Atar HH, Sancak C, Özcan S (2004) In vitro micropropagation of the aquarium plant Ludwigia repens. Asia Pacific J Mol Biol Biotechnol 12:21–25
Ridings WH, Zettler FW (1973) Aphanomyces blight of Amazon sword plants. Phytopathology 63:289–295. https://doi.org/10.1094/Phyto-63-289
Rieder A, Figueiredo GC, Bonilla MG (2011) Plant known as “Leather hat” (Echinodorus spp.) (Alismataceae) and its medicinal use in southwestern Mato Grosso, Brazil. Planta Med 77:PF86. https://doi.org/10.1055/s-0031-1282474
Sahai A, Shahzad A (2013) High frequency in vitro regeneration system for conservation of Coleus forskohlii: a threatened medicinal herb. Acta Physiol Plant 35:473–481. https://doi.org/10.1007/s11738-012-1090-z
Sheeja GE, Joseph A, Korath A (2015) In vitro propagation of an ornamental aquatic plant, Anubias barterii Var. Nana petite. Int J Curr Sci 18:E1–E12 http://www.currentsciencejournal.info/issuespdf/Anubias-Sheeja%203.pdf
Shekhawat MS, Manokari M, Revathi J (2017) In vitro propagation, micromorphological studies and ex vitro rooting of Alternanthera philoxeroides (Mart.) Griseb.: an important aquatic plant. Aquacult Int 25:423–435. https://doi.org/10.1007/s10499-016-0039-4
Shibayama Y, Kadono Y (2007) Reproductive success and genetic structure of populations of the heterostylous aquatic plant Nymphoides indica (L.) Kuntze (Menyanthaceae). Aquat Bot 86:1–8. https://doi.org/10.1016/j.aquabot.2006.06.002
Winkelmann T, Geier T, Preil W (2006) Commercial in vitro plant production in Germany. Plant Cell Tissue Organ Cult 86:319–327. https://doi.org/10.1007/s11240-006-9125-z1
Acknowledgements
SMH acknowledges the Ministry of Minority Affairs and the University Grant Commission (UGC) of India for providing Maulana Azad National Fellowship (MANF). The authors acknowledge Swami Kamalasthananda, Principal, Ramakrishna Mission Vivekananda Centenary College, Rahara, Kolkata, India for the facilities provided for this study. Further, the authors are thankful to aquarium hobbyist Mr. Diptarup Das for gifting us initial plant materials and Mr. Anupam Das for providing consent to use his aquariums during acclimatization of the regenerated plants. The DST-FIST program is also acknowledged hereby for infrastructural facilities.
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Editor: Masaru Nakano
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Haque, S.M., Ghosh, B. A submerged culture system for rapid micropropagation of the commercially important aquarium plant, ‘Amazon sword’ (Echinodorus ‘Indian Red’). In Vitro Cell.Dev.Biol.-Plant 55, 81–87 (2019). https://doi.org/10.1007/s11627-018-9938-6
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DOI: https://doi.org/10.1007/s11627-018-9938-6