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Narciclasine, a potential allelochemical, affects subcellular trafficking of auxin transporter proteins and actin cytoskeleton dynamics in Arabidopsis roots

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Abstract

Main conclusion

The present study documented the action of a potential allelochemical, narciclasine, on auxin transport in Arabidopsis by mainly affecting subcellular trafficking of PIN and AUX1 proteins and through interfering actin cytoskeletal organization.

Narciclasine (NCS), an Amaryllidaceae alkaloid isolated from Narcissus tazetta bulbs, has potential allelopathic activity and affects auxin transport. However, little is known about the cellular mechanism of this inhibitory effect of NCS on auxin transport. The present study characterizes the effects of NCS at the cellular level using transgenic Arabidopsis plants harboring the promoters of PIN, in combination with PIN-GFP proteins or AUX1-YFP fusions. NCS treatment caused significant reduction in the abundance of PIN and AUX1 proteins at the plasma membrane (PM). Analysis of the subcellular distribution of PIN and AUX1 proteins in roots revealed that NCS induced the intracellular accumulation of auxin transporters, including PIN2, PIN3, PIN4, PIN7 and AUX1. However, other PM proteins, such as PIP2, BRI1, and low temperature inducible protein 6b (LTI6b), were insensitive to NCS treatment. NCS-induced PIN2 compartments were further defined using endocytic tracer FM 4-64 labeled early endosomes and suggested that this compound affects the endocytosis trafficking of PIN proteins. Furthermore, pharmacological analysis indicated that the brefeldin A (BFA)-insensitive pathway is employed in the cellular effects of NCS on PIN2 trafficking. Although NCS did not alter actin dynamics in vitro, it resulted in the depolymerization of the actin cytoskeleton in vivo. This disruption of actin filaments by NCS subsequently influences the actin-based vesicle motility. Hence, the elucidation of the specific role of NCS is useful for further understanding the mechanisms of allelopathy at the phytohormone levels.

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Abbreviations

ARF:

ADP-ribosylation factor family of GTP-binding proteins

AUX1/LAX:

AUXIN RESISTANT1/LIKE AUXIN RESISTANT

BFA:

Brefeldin A

eEF1A:

Elongation factor one alpha

GEF:

Guanine nucleotide exchange factor

LatB:

Latrunculin B

NCS:

Narciclasine

PIN:

PIN-FORMED auxin efflux carrier protein

PM:

Plasma membrane

TIBA:

2,3,5-Triiodobenzoic acid

TryA23:

Tyrphostin A23

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (31201145, 31170225), the Foundation of Science and Technology Program of Gansu Province (1208RJZA224), and Fundamental Research Funds for the Central Universities (lzujbky-2014-95, lzujbky-2013-bt05; lzujbky-2012-104).

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Authors and Affiliations

  1. Ministry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou, 730000, People’s Republic of China

    Yanfeng Hu, Xiaofan Na, Jiaolong Li, Lijing Yang, Xiaolei Liang, Jianfeng Wang, Liang Peng & Yurong Bi

  2. Key Laboratory of Mollisols Agroecology, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin, 150000, People’s Republic of China

    Yanfeng Hu

  3. School of Life Science, Northwest Normal University, Lanzhou, 730070, People’s Republic of China

    Jia You

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  1. Yanfeng Hu
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Correspondence to Yurong Bi.

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Hu, Y., Na, X., Li, J. et al. Narciclasine, a potential allelochemical, affects subcellular trafficking of auxin transporter proteins and actin cytoskeleton dynamics in Arabidopsis roots. Planta 242, 1349–1360 (2015). https://doi.org/10.1007/s00425-015-2373-6

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