The Ecological Society of Japan (Japanese Journal of Conservation Ecology) 20 : (2015) 調査報告 Habitats, floral morphs and escape of Ny

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1 (Japanese Journal of Conservation Ecology) 20 : (2015) 調査報告 Habitats, floral morphs and escape of Nymphoides peltata and re-examination of its genetic diversity in Japan hinji Fujii 1, Ryuji Uesugi 2 and Masumi Yamamuro 3 1 University of Human Environments, 2 NARO Agricultural Research Center, 3 Graduate chool of Frontier ciences, The University of Tokyo Abstract: Habitats, flowering characters and information of escape from cultivation were studied by investigating the populations in the field, observing plants under cultivation, examining herbarium specimens, hearing investigation and literature survey. Genetic diversity of Nymphoides peltata was re-examined by R polymorphism. Though the analysis of genetic diversity was not successful because some loci were difficult to identify, genetic polymorphism was found within the population of Lake Inawashiro and Lake Biwa, respectively. Non-flowering populations were found to grow scattered in Japan and have genetic polymorphism. The most populations of N. peltata in Lake Biwa were recorded from the attached small ponds and connected drains or rivers. Escaping populations were revealed to scatter various places. Keywords: Lake Biwa, non-flowering population, R polymorphism Nymphoides peltata (.G.Gmel.) Kuntze 1993 NT 4 I alvinia natans (L.) All University of Human Environments, 6-2 Kami-sanbonmatsu, Okazaki City , Japan shinji@uhe.ac.jp

2 DNA DNA Uesugi et al DNA DNeasy Plant Mini Kit QIAGEN, Hilden, Germany Kikuta and Hayashi 2002 R6GddCTP Fw 5 GTTRv 5 C PCR BIOTAQ Bioline, London, UK 5 µl 10 NH 4 Buffer 0.5 µl dntp 2.5 mm 0.4 µl MgCl 2 50 mm 0.3 µl50 µm 0.04 µl µl BIOTAQ 5U/µL µl DNA 2 µl PCR Genecan ize tandard 500 Liz, Applied Biosystems, Foster City, CA DNA ABI 3730xl, Applied Biosystems, Foster City, CA DNA NA H F 4 PI Probability of identity GenAlEx6.5 Peakall and mouse

3 1 N NA H F Np Np ** Np ** Np306* ** Np ** Np641* Np694* ** Np ** Np ** Np382-2* NAHF * 4 ** P < populations Langella 1999 DAJin and Chakraborty tructure2.3 Pritchard et al MCMC burn-in period tructure Harvester Evanno et al K K 遺伝子型 1 N PCR PCR F NP274 NP382 2 NP152 NP207 3 NP NP NP306 NP382-2 NP641 NP694 PCR PI Probability Identity Nuphar fluminalis PI

4 2K=2 5 PI [] ca. 50** PI > * * <30** * ca AFHJ BC K L M N O P * ca * < * ca * * ca [] ca.1000?** * ca.10** [] > * [] []* []* * * ca.100** [] *** [] []* * * * * ** []* * 10** < ca * < > < * M * * I 1* I 3* H * A * T * [] A P * ** *** m

5 : : : : 34& : 32 1: 33& : 遺伝的な類縁関係 4 ΔK K = 同一産地での多型 4 PI 2 NP NP NP306 NP NP694 & NP NP L

6 1 6 NP730 DNA bp a No.18 b No.22 c No.26 d No.19 e No.24 f No bp 144 bp a-c 146 bp d-f 2 NP730 76

7 m m 花型 mizu03/dai11.html 逸出 H.Marui 3318 TN Fujii KYO 77

8 H H.Muramatsu s.n. TI H L 1890 T.Makino s.n. MAK1937 M.Honda 2 TI1971 C.Kimura and T.Kyogoku s.n. TI H 1898 U.Faurie 2489 KYO1959 H.ase FKE2008 L 1 L T.Makino s.n. MAK Matsuda s.n. KYO 1954 T.Haginiwa s.n. TN L 1939 T.Makino s.n. MAK H 1934.Miki s.n. OA K.Hamasaka s.n. MAK 1902 K.Umemura s.n. MAK 5 2 L C.Takemura s.n. MAK 1981 Y.Kadono 1325 KYO Y.Kadono 591 KYO 1989.Hamajima s.n. OA H H 1923.Miki s.n. OA L H L H 1 HL

9 5 1 H 1997 K.Takita 7225 TN 1934 H.Iwamoto 1596 OAT.asaki TN bluelist.ies.hro.or.jp L m 1 HL MAK OA T.higa 3747 OA 生育環境 Miki s.n. June 24, 1923 OA m 1 km Pearsall 1920 Hutchinson 1975 Keddy 2000 Lacoul and Freedman

10 Fujii OA Miki s.n. ept. 14, 1927 OA.Miki s.n. Nov. 13, 1932 OA * 1968.Kitamura s.n. Aug KYO *.Miki s.n. Nov. 10, 1932 OA C.Takemura s.n. July 15, 1901 MAK C.Hashimoto 9445 May 29, 1942 OA C.Hashimoto s.n. Aug. 29, 1925 KYO Fujii OA 1994 G.Koidzumi s.n, June 7, 1921 KYO.Miki s.n. Jun. 24, 1923 OA * 2 m Phragmites australis (Cav.) Trin. ex teud

11 Y.Tsurumachi s.n. Aug. 25, 1934 OA.Matsuda s.n. no date KYO Y.Narita s.n. Aug. 26, 1924 TI T.Makino s.n MAK M.uzuki June 19, upper from akaijima* upper from Kawamukae Fukushima, Tsukisu & Ipponmatsu Mishima, Nasanden Ikisu * 2001 Table 1 Fig Takagawa et al Takagawa et al Takagawa et al Uesugi et al FKE TNKYO OAMAK

12 B B Evanno G, Regnaut, Goudet J (2005) Detecting the number of clusters of individuals using the software TRUCTURE: A simulation study. Molecular Ecology 14: (1994) (2). Nature tudy 40: (1995) : (2002). (). pp ,. (2004). Nature tudy 50: (2010). 33:7-22. Hutchinson GE (1975) A Treatise on Limnology. Volume 3. Limnological Botany. John Wiley, New York. Jin L, Chakraborty R (1993) Estimation of genetic distance and coefficient of gene diversity from single-probe multilocus DNA fingerprinting data. Molecular Biology and Evolution 11: (1994).,. (2000) 8 I ( ).,. Keddy PA (2000) Wetland Ecology: Principles and Conservation. Cambridge University Press, Cambridge. (2008).,. Kukita Y, Hayashi K (2002) Multicolor post-pcr labeling of DNA fragments with fluorescent dideoxynucleotides. BioTechniques 33: (2012). 24: Lacoul P, Freedman B (2006) Environmental influences on aquatic plants in freshwater ecosystems. Environmental Reviews 14: Langella O (1999) Populations, copyright (C) 1999, Olivier Langella, CNR UPR9034 (Population genetic software). (1993). 17: (1995) (). 57: (2004). 27: (2008).,. (2010). (). pp ,. (2001). 4: Peakall R, mouse PE (2012) GenAlEx 6.5: Genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics 28: Pearsall WH (1920) The aquatic vegetation of the English lakes. Journal of Ecology 8: Pritchard JK, tephens M, Donnelli P (2000) Inference of population structure from multilocus genotype data. Genetics 155: (1995). ( ) 30:1-52. (2009) 41.,. (2008). Nature tudy 54: (2013) Nuphar submersan. fluminalis. 18: (1984). 18:2-6. Takagawa, Nishihiro J, Washitani I (2005) afe sites for establishment of Nymphoides peltata seedlings for recovering the population from the soil seed bank. Ecological Research 20: Takagawa, Washitani I, Uesugi R, Tsumura Y (2006) Influence of inbreeding depression on a lake population of Nymphoides peltata after restoration from the soil seed bank. Conservation Genetics 7: (2013) (). 94:1-2. Uesugi R, Nishihiro J, Tsumura Y, Washitani I (2007) Restoration of genetic diversity from soil seed banks in a 82

13 threatened aquatic plant, Nymphoides peltata. Conservation Genetics 8: (2009). 14: Uesugi R, Tani N, Goka K, Nishihiro J, Tsumura Y, Washitani I (2005) Isolation and characterization of highly polymorphic microsatellites in the aquatic plant, Nymphoides peltata (Menyanthaceae). Molecular Ecology Notes 5: (2005) 114.,. 83

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