Drynaria bonii spore culture: optimization of culture conditions and analysis of gametophyte and sporophyte development
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Plant Biotechnology Reports https://doi.org/10.1007/s11816-020-00632-7
ORIGINAL ARTICLE
Drynaria bonii spore culture: optimization of culture conditions and analysis of gametophyte and sporophyte development Quyen Van Nguyen1,2 · Hang Thi Khuat1 · Yen‑Ngoc Thi Nguyen1 · Dung Thi Vu1 · Thu‑Ha Bui1 · Kyung‑Hwan Boo3,4 Received: 21 August 2019 / Accepted: 10 June 2020 © Korean Society for Plant Biotechnology 2020
Abstract Drynaria bonii H. Christ is a medicinal plant that has traditionally been used for the treatment of chronic diseases and tuberculosis. However, D. bonii has not yet been cultivated or propagated. In this study, we optimized the conditions for the spore culture of D. bonii to produce sporophytes. First, we evaluated the effects of various factors, such as temperature, light intensity, potting material, and pH, on spore germination and early gametophyte development. Under optimal culture conditions, 95–99% of the spores germinated within 2 weeks of culture. Next, based on these results, we established a culture system for gametophyte development and sporophyte production. Mature gametophytes first appeared 5 months post-germination (mpg), and at 12 mpg, the rate of production of heart-shaped gametophytes reached 56.6%. Sporophytes at the early first-leaf stage first appeared at 6 mpg, and at 12 mpg, the rate of formation of sporophytes reached 15.4%. Interestingly, 6.3% of all gametophytes produced multiple sporophytes. Additionally, 42–62% of the gametophytes also produced multiple secondary gametophytes, indicating a high potential of D. bonii gametophytes to generate new gametophytes and, subsequently, sporophytes. When transferred to new trays or pots, sporophytes grew well and showed 100% survival. Overall, we conclude that this spore culture system can be successfully used for the propagation of D. bonii sporophytes. Keywords Culture condition · Drynaria bonii · Multiple sporophytes · Secondary gametophyte · Spore culture · Spore germination
Introduction The fern Drynaria bonii H. Christ has traditionally been used in Asian medicine to treat bone diseases and disorders, such as osteoporosis and fracture, kidney diseases, and tuberculosis (Elkington et al. 2009; Loi 2004). Crude * Quyen Van Nguyen [email protected] * Kyung‑Hwan Boo [email protected] 1
Faculty of Biology, Hanoi National University of Education (HNUE), 136 Xuanthuy, Caugiay, Hanoi, Vietnam
2
Center for Environmental Research and Education (CERE), Hanoi National University of Education, 136 Xuanthuy, Caugiay, Hanoi, Vietnam
3
Department of Biotechnology, College of Applied Life Science (SARI), Jeju National University, Jeju 63243, Republic of Korea
4
Subtropical/Tropical Organism Gene Bank, Jeju National University, Jeju 63243, Republic of Korea
extracts of D. bonii plants and secondary metabolites isolated from these plants promote the proliferation of human osteoblast-like cells (Trinh et al. 2016) and exhibit antibacterial and antioxidant activities (Trinh et al. 2015). Plants of D. bonii are naturally
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