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Effects of a Coumarin Derivative, 4-Methylumbelliferone, on Seed Germination and Seedling Establishment in Arabidopsis

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Abstract

The root system is central for plant adaptation to soil heterogeneity and is organized primarily by root branching. To search for compounds that regulate root branching, a forward chemical genetics screen was employed, and 4-methylumbelliferone (4-MU), a coumarin derivative, was found to be a potent regulator of lateral root formation. Exogenous application of 4-MU to Arabidopsis thaliana seeds affected germination and led to reduced primary root growth, the formation of bulbous root hairs, and irregular detached root caps accompanied by reorganization of the actin cytoskeleton in root tips before seedling establishment. Abundant lateral roots formed after exposure to 125 μM 4-MU for 22 days. Molecular, biochemical, and phytochemical approaches were used to determine the effect of 4-MU on root growth and root branching. Arabidopsis seedlings grown in the presence of 4-MU accumulated this compound only in roots, where it was partially transformed by UDP-glycosyltransferases (UGTs) into 4-methylumbelliferyl-β-D-glucoside (4-MU-Glc). The presence of 4-MU-Glc in seedling roots was consistent with the upregulation of several genes that encode UGTs in the roots. This shows that UGTs play an integral role in the detoxification of 4-MU in plants. The increased expression of two auxin efflux facilitator genes (PIN2 and PIN3) in response to 4-MU and the lack of response of the auxin receptor TIR1 and the key auxin biosynthetic gene YUCCA1 suggest that auxin redistribution, rather than auxin biosynthesis, may directly or indirectly mediate 4-MU-induced root branching.

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Acknowledgements

X. Li was a recipient of a Visiting Fellowship to a Government Laboratory of Canada. The authors are grateful for technical assistance from Dr. Branimir Gjetvaj. We also thank Dr. Elison B. Blancaflor for providing the 35S::ABD2-GFP construct and Drs. Elizabeth Schultz and Tom Guilfoyle for providing Arabidopsis thaliana DR5::GUS transgenic lines. This work was partially supported by the Program for New Century Excellent Talents in University (XL, NCET-09-0423), Ministry of Education of the People’s Republic of China.

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Authors and Affiliations

  1. College of Plant Sciences, Jilin University, Changchun, 130062, China

    Xiang Li

  2. Agriculture and Agri-Food Canada, Saskatoon Research Centre, Saskatoon, Saskatchewan, S7N 0X2, Canada

    Xiang Li, Margaret Y. Gruber, Dwayne D. Hegedus, Derek J. Lydiate & Ming-Jun Gao

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  1. Xiang Li
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  2. Margaret Y. Gruber
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Correspondence to Ming-Jun Gao.

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Supplemental Fig. 1

Changes in auxin distribution and root branching in response to 4-MU as observed in roots of Arabidopsis plants transformed with DR5::GUS. (A) Root system of a 14-d-old DR5::GUS seedling treated with 125 μM 4-MU showing GUS expression (blue) in the meristematically active primordia. (B) Primary root of a 14-d-old untreated DR5::GUS seedling. Arrows indicate active DR5::GUS expression. (DOC 1223 kb)

Supplemental Fig. 2

600-MHz 1H-NMR spectrum of 4-methylumbelliferone (4-MU). (DOC 60 kb)

Supplemental Fig. 3

600-MHz 1H-NMR spectrum of product (4-MU-Glc) after in vivo uptake of 125 μM 4-MU by Arabidopsis roots from the growth medium. (DOC 62 kb)

Supplemental Fig. 4

1H-1H COSY (correlation spectroscopy) of product (4-MU-Glc) from Arabidopsis roots after treatment with 125 μM 4-MU. (DOC 3358 kb)

Supplemental Fig. 5

HMBC (heteronuclear multiple bond correlation) of product (4-MU-Glc) after uptake of 125 μM 4-MU by Arabidopsis roots from the medium. (DOC 3398 kb)

Supplemental Fig. 6

Functional categorization of microarray data showing genes regulated by 4-MU. (DOC 3834 kb)

Supplementary Table 1

List of compounds showing root phenotyps in this study. (DOC 42 kb)

Supplementary Table 2

Genes upregulated after exposure to 4-MU. (XLS 74 kb)

Supplementary Table 3

Genes downregulated after exposure to 4-MU. (XLS 71 kb)

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Li, X., Gruber, M.Y., Hegedus, D.D. et al. Effects of a Coumarin Derivative, 4-Methylumbelliferone, on Seed Germination and Seedling Establishment in Arabidopsis . J Chem Ecol 37, 880–890 (2011). https://doi.org/10.1007/s10886-011-9987-3

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