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Light-dependent gravitropism and negative phototropism of inflorescence stems in a dominant Aux/IAA mutant of Arabidopsis thaliana, axr2

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Abstract

Gravitropism and phototropism of the primary inflorescence stems were examined in a dominant Aux/IAA mutant of Arabidopsis, axr2/iaa7, which did not display either tropism in hypocotyls. axr2-1 stems completely lacked gravitropism in the dark but slowly regained it in light condition. Though wild-type stems showed positive phototropism, axr2 stems displayed negative phototropism with essentially the same light fluence-response curve as the wild type (WT). Application of 1-naphthaleneacetic acid-containing lanolin to the stem tips enhanced the positive phototropism of WT, and reduced the negative phototropism of axr2. Decapitation of stems caused a small negative phototropism in WT, but did not affect the negative phototropism of axr2. p-glycoprotein 1 (pgp1) pgp19 double mutants showed no phototropism, while decapitated double mutants exhibited negative phototropism. Expression of auxin-responsive IAA14/SLR, IAA19/MSG2 and SAUR50 genes was reduced in axr2 and pgp1 pgp19 stems relative to that of WT. These suggest that the phototropic response of stem is proportional to the auxin supply from the shoot apex, and that negative phototropism may be a basal response to unilateral blue-light irradiation when the levels of auxin or auxin signaling are reduced to the minimal level in the primary stems. In contrast, all of these treatments reduced or did not affect gravitropism in wild-type or axr2 stems. Tropic responses of the transgenic lines that expressed axr2-1 protein by the endodermis-specific promoter suggest that AXR2-dependent auxin response in the endodermis plays a more crucial role in gravitropism than in phototropism in stems but no significant roles in either tropism in hypocotyls.

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Acknowledgments

We thank Prof. K. Shimazaki (Kyushu University) for seeds of phot1 phot2 double mutants, Prof. T. Sakai (Niigata University) for pgp1-101 and pgp19-101, and Arabidopsis Biological Resource Center (Ohio State University) for seeds of the other T-DNA insertion and mutant lines. We also thank Dr. M. K. Watahiki (Hokkaido University) for ACTIN2 primers. This work was supported in part by Grants-in-Aid from Ministry of Education, Culture, Sports, Science and Technology, Japan to K. T. Y. (19060008). S. S. was supported by Research Fellowship for Young Scientists of Japan Society for the Promotion of Science.

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  1. Atsuko Sato

    Present address: National Institute of Advanced Industrial Science and Technology Hokkaido, 2-17-2-1 Tsukisamu-Higashi, Toyohira-Ku, Sapporo, 062-8517, Japan

  2. Jun Matsuzaki

    Present address: National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, 305-8602, Japan

Authors and Affiliations

  1. Division of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, 060-0810, Japan

    Atsuko Sato, Jun Matsuzaki & Kotaro T. Yamamoto

  2. Biosystems Science Course, Graduate School of Life Science, Hokkaido University, Sapporo, 060-0810, Japan

    Shu Sasaki & Kotaro T. Yamamoto

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10265_2014_643_MOESM1_ESM.tif

Supplementary material 1 (TIFF 2925 kb). Fig. S1 Longitudinal sections of the primary stem of the wild type and axr2-1 stained for amyloplasts. The wild type (left) and axr2-1 (right) stem segments about 1 cm long were fixed with 3 % paraformaldehyde in phosphate-buffered saline overnight, and embedded in 3 % agarose. About 60-μm-thick longitudinal sections were prepared with a vibrating blade microtome (VT1200S, Leica), and stained with 5 % I2–KI solution for a few min. en, endodermis

10265_2014_643_MOESM2_ESM.tif

Supplementary material 2 (TIFF 1243 kb). Fig. S2 Effects of decapitation or application of NAA or NPA on the maximum bending rate of gravitropism in wild-type inflorescence stems in the dark or under light conditions. Plants were placed in a horizontal position ~5 h after decapitation (upper panel) or after application of lanolin paste containing 0.5 mM NAA or NPA to apical 1-cm-long portions of stem (lower panel), and the time course of the gravitropic response determined for 18 h thereafter in the dark or under light conditions. Each of the data represents mean and SD of three to 14 measurements

10265_2014_643_MOESM3_ESM.tif

Supplementary material 3 (TIFF 532 kb). Fig. S3 Time-course of the gravitropic response of eal1 and sgr2 (SALK_098981) inflorescence stems under different light conditions. eal1 (upper panel) or sgr2 (lower panel) were placed in the dark (closed symbols) and white-light conditions (open symbols) after changing the position of plants by ~90°. Two independent measurements are shown for each genotype (circles and triangles)

10265_2014_643_MOESM4_ESM.tif

Supplementary material 4 (TIFF 1639 kb). Fig. S4 Effects of application of NAA or decapitation on gravitropism of inflorescence stems of axr2-1 in white-light conditions. a Gravitropic response was determined with 0.5 mM NAA (triangles) or mock treatment (circles). b Gravitropic response was determined after decapitation and removal of all the lateral organs (triangles). Circles show response of intact inflorescences. Each point represents mean and SD of three to 14 measurements. For more details, see the legend to Fig. 3

10265_2014_643_MOESM5_ESM.tif

Supplementary material 5 (TIFF 2400 kb). Fig. S5 Effects of application of NPA on phototropism of inflorescence stems of the wild type and axr2-1. Lanolin paste containing 0.5 mM NPA was applied to wild-type (circles) or axr2-1 (triangles) stems. Data of mock treatment are shown with grey symbols, which are the same as those in Fig. 3a. Each point represents mean and SD of seven measurements. For more details, see a legend to Fig. 3

10265_2014_643_MOESM6_ESM.tif

Supplementary material 6 (TIFF 1268 kb). Fig. S6 Phototropic responses of inflorescence stems of pin1-1 induced by unilateral irradiation with blue light (57 μmol m−2 s−1). Each point represents mean and SD of four measurements

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Sato, A., Sasaki, S., Matsuzaki, J. et al. Light-dependent gravitropism and negative phototropism of inflorescence stems in a dominant Aux/IAA mutant of Arabidopsis thaliana, axr2 . J Plant Res 127, 627–639 (2014). https://doi.org/10.1007/s10265-014-0643-1

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