High light intensity stress as the limiting factor in micropropagation of sugar maple (Acer saccharum Marsh.)

Abstract

Attempts of clonal propagation of sugar maple through conventional and micropropagation techniques have been largely unsuccessful and the basis for this longstanding problem has remained unknown. Typical symptoms of stress in in vitro cultures of sugar maple and its intrinsic tolerance to highly shaded growing conditions indicated that optimization of light intensity might overcome its recalcitrance. To test this hypothesis we evaluated the growth, morphology, chlorophyll content and various chlorophyll fluorescence parameters of in vitro plantlets cultured under three intensities (4, 16 and 40 µmol m−2 s−1) using two spectra of light (red-blue and full spectrum white). Higher light intensities increased fresh and dry mass, but had no significant effect on the number of nodes or plant height. Anthocyanin accumulation in tissues, reduction in chlorophyll content, and significantly increased physiological stress quantified by chlorophyll fluorescence parameters, indicated increasing levels of light induced damage to the tissues with increasing light intensities. The critical factor limiting sugar maple’s response to in vitro propagation was light intensity and the levels commonly used for in vitro propagation are not suitable for sugar maple. The current study demonstrates the unique low light requirement, between 4 and 16 µmol m−2 s−1, of this species to overcome the recalcitrance to in vitro propagation. Spectral differences in quality of light and its effect on in vitro growth and stress in cultures are also discussed. In general light induced stress symptoms begin to appear at lower intensities with RB spectrum as compared to the full spectrum lights. While this study is limited to maples, requirement of specific light intensity is likely the cause of recalcitrance in other woody species with similar ecological roles.

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Acknowledgements

The authors acknowledge motivation from Todd Leuty, Specialist—Agroforestry and Speciality Crops, Ontario Ministry of Agriculture and Food (OMAFRA), Ontario; Ray Fortune, Fortune Farms, Almonte, ON and the Ontario Maple Syrup Producers Association. The authors also acknowledge the help of Bob Nichols, Scott Belton, Shuping Li and Sarita Khanal for their assistance in the lab and field work.

Author contributions

AS conducted the experiments, collected the in vitro growth data, collected chlorophyll fluorescence imaging data, and analyzed the data. AS and AMPJ, conceptualized and designed the experiments. MS provided guidance and inputs at various stages of in vitro culturing and experiments. AS and AMPJ prepared the manuscript. AMPJ conceived the project. PKS managed, organized, acquired funding, supervised and guided the whole study. All authors read and approved the final manuscript.

Funding

The authors acknowledge the Gosling Research Institute for Plant Preservation (GRIPP), University of Guelph, ON, Canada and Natural Sciences and Engineering Research Council of Canada (NSERC) for financial support.

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Correspondence to Praveen K. Saxena.

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Singh, A.S., Jones, A.M.P., Shukla, M.R. et al. High light intensity stress as the limiting factor in micropropagation of sugar maple (Acer saccharum Marsh.). Plant Cell Tiss Organ Cult 129, 209–221 (2017). https://doi.org/10.1007/s11240-017-1170-2

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Keywords

  • Light intensity
  • In vitro stress
  • Sugar maple
  • Acer saccharum
  • Micropropagation
  • Chlorophyll fluorescence imaging