Skip to main content

Chloroplast DNA sequencing and detailed microsatellite genotyping of all remnant populations suggests that only a single genet survives of the critically endangered plant Rehmannia japonica

Journal of Plant Research volume 130, pages 117–124 (2017)Cite this article

Abstract

Rehmannia japonica (Thunb.) Makino ex T. Yamaz. is an endangered perennial herb species in Japan. Although earlier the Japanese considered it a variety of R. glutinosa, recent Japanese taxonomists have consistently regarded it as an independent species. According to the historical literature, Rehmannia japonica seems to have been known in China and Japan in the past. However, Chinese taxonomists do not recognize R. japonica at present. In Japan, only two populations are known, and although these populations flower every year, seed reproduction has not been observed. In this study, we aimed to reveal the phylogenetic relationships and levels of genetic diversity of R. japonica. A haplotype network based on two chloroplast DNA regions (trnL-trnF and rps16) showed that the sequences of R. japonica were distinguishable by three or four sites of indels from the most closely related species, R. chingii, consistent with the separate species status of R. japonica. An analysis of genetic diversity using twelve microsatellite loci showed that all of the ramets of R. japonica collected from two geographically isolated populations had an identical multilocus genotype, including identical heterozygous genotypes at six loci. This result indicated asexual origin of all sampled ramets. This study also suggests that the absence of sexual reproduction of R. japonica is explained by self-incompatibility combined with only a single genet remaining in the R. japonica populations.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1

References

  • Albach DC, Li HQ, Zhao N, Jensen SR (2007) Molecular systematics and phytochemistry of Rehmannia (Scrophulariaceae). Biochem Syst Ecol 35:293–300

    Article  CAS  Google Scholar 

  • Bremer B, Bremer K, Heidari N, Erixon P, Olmstead RG, Anderberg AA, Källersjö M, Barkhordarian E (2002) Phylogenetics of asterids based on 3 coding and 3 non-coding chloroplast DNA markers and the utility of non-coding DNA at higher taxonomic levels. Mol Phylogenet Evol 24:274–301

    Article  PubMed  CAS  Google Scholar 

  • Callaghan TV, Carlsson BÅ, Jónsdóttir IS, Svensson BM, Jonasson S (1992) Clonal plants and environmental change: introduction to the proceedings and summary. Oikos 63:341–347

    Article  Google Scholar 

  • Chin CL (1998) Rehmannia. In: Wu ZY, Raven PH (eds) Scrophulariaceae through Gesneriaceae, Flora of China, vol 18. Science Press, Beijing and Missouri Botanical Garden Press, St. Louis, pp 53–55

    Google Scholar 

  • Clement M, Posada D, Crandall KA (2000) TCS: a computer program to estimate gene genealogies. Mol Ecol 9:1657–1659

    Article  PubMed  CAS  Google Scholar 

  • Goudet J (1995) FSTAT (version 1.2): a computer program to calculate F-statistics. J Hered 86:485–486

    Article  Google Scholar 

  • Hara H (1949) Enumeratio Spermatophytarum Japonicarum 1. Iwanami Shoten, Tokyo

    Google Scholar 

  • Hong DY, Yang HB, Jin CL, Holmgren NH (1998) Scrophulariaceae. In: Wu ZY, Raven PH (eds) Flora of China 18. Science Press, Beijing, pp 1–212

    Google Scholar 

  • Isagi Y, Kaneko S (2014) Ubiquitous genotyping for conservation of endangered plant species. In: Nakano S et al (eds) Integrative observations and assessments. Springer, Tokyo, pp 311–326

    Google Scholar 

  • Izuno A, Takamiya M, Kaneko S, Isagi Y (2011) Microsatellite loci in an endangered fern species, Athyrium viridescentipes (Woodsiaceae), and cross-species amplification. Am J Bot 98:e339–e341

    Article  PubMed  Google Scholar 

  • Kaneko S, Kondo T, Isagi Y (2011) Development of microsatellite markers for the northern Australian endemic fan palm Livistona rigida (Arecaceae), with cross-amplification in the five related species. Conserv Genet Resour 3:697–699

    Article  Google Scholar 

  • Lian C, Hogetsu T (2002) Development of microsatellite markers in black locust (Robinia pseudoacasia) using a dual-suppression-PCR technique. Mol Ecol Notes 2:211–213

    CAS  Google Scholar 

  • Lian C, Wadud MA, Geng Q, Shimatani K, Hougetsu T (2006) An improved technique for isolating codominant compound microsatellite markers. J Plant Res 119:415–417

    Article  PubMed  CAS  Google Scholar 

  • Mabberley DJ (2008) Mabberley’s plant-book: a portable dictionary of plants, their classification and uses. Cambridge University Press, Cambridge

    Google Scholar 

  • Makino T (1932) A contribution to the knowledge of the flora of Nippon. J Jpn Bot 8:24

    Google Scholar 

  • Matsuda S (1918) Dziwau-zoku-oyobi-hanadziau-ni-tsuite (On Rehmannia and R. glutinosa Libosch. var. makinoi Matsuda, var. nov.). Bot Mag Tokyo 32:140–142 (in Japanese)

    Article  Google Scholar 

  • Milligan B (1992) Plant DNA isolation. In: Hoelzel AR (ed) Molecular genetic analysis of populations: a practical approach. IRL Press, Oxford, pp 59–88

    Google Scholar 

  • Ministry of Environment of Japan (2015) Red Data Book 2014-Threatened Wildlife of Japan-Vol. 8, Vascular Plants. GYOSEI, Tokyo (in Japanese)

    Google Scholar 

  • Mori K, Kaneko S, Isagi Y, Murakami N, Kato H (2008) Isolation and characterization of 10 microsatellite loci in Callicarpa subpubescens (Verbenaceae), an endemic species of the Bonin Islands. Mol Ecol Resour 8:1423–1425

    Article  PubMed  CAS  Google Scholar 

  • Olmstead RG, Wolfe AD, Young ND, Elisons W, Reeves PA (2001) Disintegration of the Scrophulariaceae. Am J Bot 88:348–361

    Article  PubMed  CAS  Google Scholar 

  • Oxelman B, Kornhall P, Olmstead RG, Bremer B (2005) Further disintegration of Scrophulariaceae. Taxon 54:411–425

    Article  Google Scholar 

  • Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics Methods and Protocols: Methods in Molecular Biology. Humana Press, Totowa, pp 365–386

    Google Scholar 

  • Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol Biol 17:1105–1109

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Xia Z, Wang YZ, Smith JF (2009) Familial placement and relations of Rehmannia and Triaenophora (Scrophulariaceae sl) inferred from five gene regions. Am J Bot 96:519–530

    Article  PubMed  CAS  Google Scholar 

  • Yamazaki T (1961) Meeting with Rehmannia japonica Makino. J Jpn Bot 36:351–352 (in Japanese)

    Google Scholar 

  • Yamazaki T (1993) Scrophulariaceae 17. Rehmannia. In: Iwatsuki K, Yamazaki T, Goufford DE, Ohba H (eds) Flora of Japan IIIa. Kodansha, Tokyo, pp 343–344

    Google Scholar 

  • Yonekura K (2012) An enumeration of the vascular plants of Japan. Hokuryukan, Tokyo

    Google Scholar 

  • Zhou Y, Gu F, Zhou C, Chu S (2010) Genetic diversity of Rehmannia glutinosa cultivars based on sequence-related amplified polymorphism markers. Sci Hortic 125:789–794

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors wish to acknowledge T. Kurosawa for manuscript preparation. We also thank S. Ohba, J. Fukui, and S. Sakaguchi for sampling. This work was supported by the Global Environment Research Fund (D-0903) of the Ministry of the Environment, Japan.

Author information

Affiliations

  1. Faculty of Symbiotic Systems Science, Fukushima University, 1 Kanayagawa, Fukushima, Fukushima, 960-1296, Japan

    Shingo Kaneko

  2. Filed Science Center, Graduate School of Agricultural Science, Tohoku University, Osaki, Miyagi, 989-6711, Japan

    Yu Matsuki

  3. Department of Biology, College of Life Sciences, Zijingang Campus of Zhejiang University, Yuhangtang Road No. 388, Hangzhou, 310058, Zhejiang, People’s Republic of China

    Ying-Xiong Qiu

  4. Laboratory of Forest Biology, Division of Forest and Biomaterials Science, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku Kyoto, Kyoto, 606-8502, Japan

    Yuji Isagi

Authors
  1. Shingo Kaneko
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yu Matsuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ying-Xiong Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yuji Isagi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shingo Kaneko.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Kaneko, S., Matsuki, Y., Qiu, YX. et al. Chloroplast DNA sequencing and detailed microsatellite genotyping of all remnant populations suggests that only a single genet survives of the critically endangered plant Rehmannia japonica . J Plant Res 130, 117–124 (2017). https://doi.org/10.1007/s10265-016-0873-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10265-016-0873-5

Keywords

  • Clonal plant
  • Conservation
  • Historical transplantation
  • Microsatellite
  • Rehmannia chingii
  • Rehmannia japonica