We investigated a suitable method for in vitro germination of spores, propagation of prothalli,
and the formation of sporophytes in the fern Dryopteris nipponensis Koidz. Spore germination
rate was relatively high regardless of culture medium. Prothallus development was faster in
Knop medium than in Murashige and Skoog (MS) media. Prothalli used in all experiments
were obtained from germinated spores, and were cultivated in different concentrations of media
components. The active formation of sexual organs such as antheridium made 1MS medium
suitable for prothallus propagation, although there was a lower propagation ratio compared to
Knop medium. Growth and morphogenesis of prothalli were most effective on 1MS medium
containing 2% sucrose, and 60 mM of total nitrogen source with 20:40 mM ratio of NH4+:NO3-
To select a suitable soil composition for sporophyte formation, ground prothalli were cultivated
on single and mixed soils using bed soil, peat moss, perlite, and decomposed granite for 14
weeks. Bed soil promoted sporophyte formation and growth regardless of single or mixed use.
In particular, a mixture of bed soil and decomposed granite in a 2:1 ratio (v:v) led to accelerated
sporophyte formation (0.83/cm2).
Keywords
antheridium
bed soil
decomposed granite
peat moss
perlite
본 연구는 참지네고사리의 기내 포자발아, 전엽체 증식 및 포자체 형성에 적합한 배지 및 배양토를 구명하고 균일묘를 생산
하고자 수행되었다. 포자는 배지의 종류와 농도에 관계없이 모든 처리구에서 발아율이 우수하였다. 그 중 Knop배지에서는 MS
계열 배지에 비해 전엽체로의 발달이 빠르게 진행되었다. 포자를 발아시켜 획득한 전엽체를 재료로 배지의 구성물질을 달리하
여 배양하였다. 전엽체의 증식율은 Knop배지에 비해 낮았으나, 장정기 등 생식기관의 형성이 활발한 1MS배지가 전엽체 증식
용 배지로 적합한 것으로 생각되었다. 또한 MS배지의 sucrose 농도는 2%, 총 질소급원의 농도는 60mM, NH4+:NO3-의 비율
을 20:40mM로 조절하는 것이 전엽체의 생육 및 형태형성에 효과적이었다. 포자체 형성에 적합한 배양토를 선발하기 위하여
믹서기로 분쇄한 전엽체를 인공토양(원예상토, 피트모스, 펄라이트, 마사토)을 단용 또는 혼용한 토양에 분주하여 14주간 재배
하였다. 그 결과, 원예상토를 사용한 처리구에서 포자체의 형성 및 생육이 우수하였으며, 특히 원예상토와 마사토를 2:1(v:v)로
혼용하였을 때 포자체의 형성(0.83개/cm2)이 가장 왕성하였다.
Chi LW, Lee CH (2015) Alpha-glucosidase inhibition activity of methanol extracts obtained from three Dryopteris species and fractions obtained from D. nipponensis . J Korean Soc People Plants Environ 18:137-141. doi:10.11628/ksppe.2015.18.2.137
2
Cho JS, Kwon HJ, Kim SY, Lee CH (2015) Effect of medium and environmental condition of spore germination of Dryopteris
crassirhizoma Nakai in vitro. Korean J Hortic Sci Technol 33:195-195
3
Dewir YH, El-Mahrouk ME, Murthy HN, Paek KY (2015) Micropropagation of Cattleya : Improved in vitro rooting and acclimatization.
Hortic Environ Biotechnol 56:89-93
4
Dewir YH, Murthy HN, Ammar MH, Alghamdi SS, Al-Suhaibani NA, Alsadon AA, Paek KY (2016) In vitro rooting of leguminous plants:
Difficulties, alternatives, and strategies for improvement. Hortic Environ Biotechnol 57:311-322
5
Fernandez H, Bertrand AM, Sanchez-Tames R (1997) Germination in cultured gametophytes of Osmunda regalis . Plant Cell
Rep16:358-362. doi:10.1007/BF01088297
6
Fernandez H, Revilla MA (2003) In vitro culture of ornamental ferns. Plant Cell Tissue Organ Cult 73:1-13 doi:10.1023/A:1022650701341
7
Gabriels R, Verdonck O, Mekers O (1986) Substrate requirements for pot plants in recirculating water culture. Acta Hortic 93-100
doi:10.17660/ActaHortic. 1986.178.11
8
George EF, Puttock DJM, George HJ (1987) Plant culture media. Vol 1. Formulations and uses. Exegetics Limited, Edington Westbury,
UK, pp 306-333
9
Gruda N, Schnitzler WH (2004) Suitability of wood fiber substrate for production of vegetable transplants: I. Physical properties of
wood fiber substrates. Sci Hortic 100:309-322. doi:10.1016/j.scienta.2003.10.001
10
Jang BK, Han JH, Kim SY, Lee CH (2015) Effect of medium composition on in vitro spore germination of four Pteridophyte species
native to Korea. Korean J Plants Res p 44 (Abstr)
11
Jeong JA, Lee CH (2006) Several factors affecting on in vitro culture of prothallus and ex vitro sporophyte formation from prothallus of
Dryopteris varia (L.) O. Kuntze. Korean J Plant Res 19:252-258
12
Kang UC, Jung SS (2012) Composition of Pteridophyta’s life-form spectra in Korea. Korean J Plant Res 25:433-446. doi:10.7732/
kjpr.2012.25.4.433
13
Kim HS, Kim KH (2011) Physical properties of the horticultural substrate according to mixing ratio of peatmoss, perlite and vermiculite.
Korean J Soil Sci Fert 44:321-330. doi:10.7745/KJSSF.2011.44.3.321
14
Kim NR, Kwon HJ, Cho JS, Lee CH (2012) Antioxidant activities of fractions obtained from Dryopteris crassirhizoma , D. nipponensis
and Polystichum lepidocaulon . Korean J Plant Res 25:176-183
15
Knop W (1865) Quantitative untersuchungen uber die ernahrungsprozesse der pflanzen. Landwirtsch Vers Stn 7:93-107.
doi:10.7732/kjpr.2012.25.2.176
16
Korea Biodiversity Information System (KBIS) (2016) Korea National Arboretum. Pocheon, Korea. Available via http://www.nature.go.kr/
index.jsp
17
Korea Fern Society (2005) Fern and fern allies of Korea. Geobook, Seoul, Korea
18
Korea National Arboretum (2008) Illustrated Pteridophytes of Korea. Korea National Arboretum, Pocheon, Korea
19
Lee HH, Kim KH, Kang JY (2006) Comparison of the European standard methods and the rural development administration methods
for determining chemical properties of horticultural substrates. Korean J Hortic Sci Technol 24:425-430
20
Melan MA, Whittier DP (1990) Effects of inorganic nitrogen sources on spore germnination and gametophyte growth in Botrychium
dissectum . Plant Cell Environ 13:477-482. doi:10.1111/j.1365-3040.1990.tb01325.x
21
Miller JH (1968) Fern gametophytes as experimental material. Bot Rev 34:361-440. doi:10.1007/BF02859133
22
Moran RC (2004) A Natural History of Ferns. Timber Press, Portland, OR, USA
23
Oh HI, Lee CH (2002) Effect of media and media components on prothallus propagation of Osmunda cinnamomea var. forkiensis
Cople. and Asplenium vidalii. J Plant Biotechnol p 130 (Abstr)
24
Ramirez-Trejo MDR, Perez-Garcia B, Mendoza-Ruiz A, Valdez-Avila R, Espinosa-Matias S (2013) Observations of the spore,
gametophyte and young sporophyte of Pteridium caudatum (L.) Maxon using scanning electron microscopy. Micron 45:37-44.
doi:10.1016/j.micron.2012.10.011
25
Ravi BX, Robert J, Gabriel M (2014) In vitro spore germination and gametophytic growth development of a critically endangered fern
Pteris tripartita Sw. African J Biotechnol 13:2350-2358. doi:10.5897/AJB2013.13419
26
Shim MS, Kim YJ, Lee DS, Kwon YH, Kim SS, Kang UC (2011) Growth responses of various ferns on shading and fertilizer application.
J Bio-Environ Control 20:109-115
27
Shin SL (2007) Several factors affecting in vitro masspropagation of eight fern species. PhD Diss, Chungbuk Univ, Cheongju, Korea
28
Shin SL, Lee CH (2011) Effect of medium components and culture methods on shoots regeneration from Athyrium niponicum . Korean
J Plant Res 24:113-120. doi:10.7732/kjpr.2011.24.2.113