Plant and Soil

, Volume 444, Issue 1–2, pp 299–314 | Cite as

Transcriptome analysis of Medicago lupulina seedlings leaves treated by high calcium provides insights into calcium oxalate formation

  • Xi-Min Zhang
  • Lun-Xian Liu
  • Zhi-Meng Su
  • Zhi-Jun Shen
  • Gui-Feng Gao
  • Yin YiEmail author
  • Hai-Lei ZhengEmail author
Regular Article


Background and aim

Calcium oxalate (CaOx) is a common biomineral found in the plant kingdom. Crystals of CaOx occur in different plant tissues, such as leaves and stems. However, little is known about the biosynthesis of CaOx in oxalate-accumulating plants. Moreover, the literature on genes related to CaOx formation under high-calcium environment is scarce. In the present study, the physiological parameters and the transcript profiles of Medicago lupulina leaves treated with 0.1 and 25 mM Ca2+ were analyzed to study the genes involved in the biosynthesis of CaOx.


We demonstrated that exposure to high external calcium concentration induced H2O2 production, ascorbic acid degradation, and CaOx accumulation in M. lupulina leaves. Moreover, we identified 1715 differentially expressed genes (DEGs) (1322 up-regulated and 393 down-regulated genes) in leaves treated with 25 mM Ca2+ compared with the leaves treated with 0.1 mM Ca2+. We further demonstrated the involvement of DEGs in oxalic acid production, calcium transport, and calcium buffering. These results revealed that a high calcium promoted oxalic acid biosynthesis by inducing the expression of NADPH oxidase and ascorbate oxidase genes. In addition, several genes encoding cyclic nucleotide-gated channel, Ca2+-ATPase, H+/Ca2+ exchangers, and calcium-binding proteins were found to be differentially expressed and involved in calcium transport and calcium buffering.


Our transcriptome analyses provide a comprehensive insight into the biosynthesis of CaOx in oxalate-accumulating plants.


Calcium oxalate High calcium Medicago Oxalate-accumulating plants Transcriptome 



This study was financially supported by the Joint Fund of the Natural Science Foundation of China and the Karst Science Research Center of Guizhou Province (Grant No. U1812401), the Programme for Changjiang Scholars and Innovative Research Teams in Universities (PCSIRT–1227), the Initial Fund for Key Laboratory of Guizhou Province (2011-4005), the Major Science and Technology Project of the Education Department of Guizhou Province during the “12th Five-year Plan” (2012-005), the Joint Fund for the Department of Science and Technology of Guizhou Province and Guizhou Normal University ([2016]7209, [2016]7210), the National Key Research and Development Program of China (2017YFC0506102), and the Natural Science Foundation of China (NSFC) (31570586, 31870581).

Author contributions

X. M. Z. designed the experiments. X. M. Z., L. X. L. and Z. M. S performed the experiments. Z. J. S. and G. F. G. analyzed transcriptome data. X. M. Z. wrote the paper. Y. Y. and H. L. Z. revised this paper. All authors have read and approved the manuscript.

Compliance with ethical standards

Competing financial interests

We declare no competing financial interests.

Supplementary material

11104_2019_4283_MOESM1_ESM.docx (672 kb)
ESM 1 (DOCX 671 kb)


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© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Key Laboratory for Subtropical Wetland Ecosystem Research of MOE, College of the Environment and EcologyXiamen UniversityXiamenPeople’s Republic of China
  2. 2.Key Laboratory of State Forestry Administration on Biodiversity Conservation in Karst Area of SouthwestGuizhou Normal UniversityGuiyangPeople’s Republic of China

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