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Journal of Plant Research

, Volume 130, Issue 2, pp 407–416 | Cite as

Detection of chlorophylls in spores of seven ferns

  • Mei-Hwei Tseng
  • Kuei-Huei Lin
  • Yi-Jia Huang
  • Ya-Lan Chang
  • Sheng-Cih Huang
  • Li-Yaung Kuo
  • Yao-Moan Huang
Regular Paper

Abstract

Fern spores were traditionally classified into chlorophyllous (green) and nonchlorophyllous (nongreen) types based on the color visible to the naked eye. Recently, a third type, “cryptochlorophyllous spores”, is recognized, and these spores are nongreen under white light but contain chlorophylls. Epifluorescence microscopy was previously used to detect chlorophylls in cryptochlorophyllous spores. In addition to epifluorescence microscopy, current study performed some other approaches, including spore-squash epifluorescence, absorption spectra, laser-induced fluorescence emission spectra, thin layer chromatography (TLC), and ultra-high performance liquid chromatography with ultraviolet and mass spectrometric detection (UHPLC-UV-MS) in order to detect chlorophylls of spores of seven ferns (Sphaeropteris lepifera, Ceratopteris thalictroides, Leptochilus wrightii, Leptochilus pothifolius, Lepidomicrosorum buergerianum, Osmunda banksiifolia, and Platycerium grande). Destructive methods, such as TLC and UHPLC-UV-MS, successfully detected chlorophylls inside the spores when their signals of red fluorescence under epifluorescence microscope were masked by spore wall. Although UHPLC-UV-MS analysis was the most sensitive and reliable for determining the chlorophylls of spores, spore-squash epifluorescence is not only reliable but also cost- and time-effective one among our study methods. In addition, we first confirmed that Lepidomicrosorium buergerianum, Leptochilus pothifolius, Leptochilus wrightii, and Platycerium grande, produce cryptochlorophyllous spores.

Keywords

Chlorophyll Cryptochlorophyllous spore Spore-squash epifluorescence UHPLC-UV-MS 

Notes

Acknowledgements

We thank Dr. Wen-Liang Chiou, Tzu-Tong Kao, and Daniel P. Chamberlin for fruitful discussions and English editing assistance, respectively. We also thank Hajime Ikeda and two anonymous reviews for providing constructive comments on the manuscript. This study was supported by the Ministry of Science and Technology, Taiwan (MOST 102-2313-B-845-001-MY3).

Supplementary material

10265_2016_901_MOESM1_ESM.pdf (72 kb)
Online resource 1 Spectrum of laser-induced fluorescence of Sphaeropteris lepifera spores (PDF 71 KB)
10265_2016_901_MOESM2_ESM.pdf (2.2 mb)
Online resource 2 LC UV ESI MS fragment ions of spore acetone extracts (PDF 2205 KB)

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Copyright information

© The Botanical Society of Japan and Springer Japan 2016

Authors and Affiliations

  • Mei-Hwei Tseng
    • 1
  • Kuei-Huei Lin
    • 1
  • Yi-Jia Huang
    • 1
  • Ya-Lan Chang
    • 2
  • Sheng-Cih Huang
    • 3
  • Li-Yaung Kuo
    • 4
  • Yao-Moan Huang
    • 4
  1. 1.Department of Applied Physics and ChemistryUniversity of TaipeiTaipeiTaiwan
  2. 2.Dr. Cecilia Koo Botanic Conservation CenterPingtungTaiwan
  3. 3.Department of Applied ChemistryNational Chiao Tung UniversityHsinchuTaiwan
  4. 4.Division of SilvicultureTaiwan Forestry Research InstituteTaipeiTaiwan

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