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RAP2.6L and jasmonic acid–responsive genes are expressed upon Arabidopsis hypocotyl grafting but are not needed for cell proliferation related to healing

Abstract

Key message

Jasmonic acid and RAP2.6L are induced upon wounding but are not involved in cell proliferation during healing in Arabidopsis hypocotyls.

Abstract

Plants produce jasmonic acid in response to wounding, but its role in healing, if any, has not been determined. Previously, the jasmonic acid–induced transcription factor, RAP2.6L, related to APETALA 2.6-like, was identified as a spatially expressed factor involved in tissue reunion in partially incised flowering stems of Arabidopsis. In the present study, we investigated the function of JA and RAP2.6L on wound healing using an Arabidopsis hypocotyl-grafting system, in which separated tissues are reattached by vascular tissue cell proliferation. The jasmonic acid–responsive genes AOS and JAZ10 were transiently expressed immediately after grafting. We confirmed that the endogenous content of jasmonic acid-Ile, which is the bioactive form of jasmonic acid, increased in hypocotyls 1 h after grafting. Morphological analysis of the grafted tissue revealed that vascular tissue cell proliferation occurred in a similar manner in wild-type Arabidopsis, the jasmonic acid–deficient mutant aos, the jasmonic acid–insensitive mutant coi1, and in Arabidopsis that had been exogenously treated with jasmonic acid. RAP2.6L expression was also induced during graft healing. Because RAP2.6L expression occurred during graft healing in aos and coi1, its expression must be regulated via a jasmonic acid–independent pathway. The rap2.6L mutant and dominant repressor transformants for RAP2.6L showed normal cell proliferation during graft healing. Taken together, our results suggest that JA and RAP2.6L, induced by grafting, are not necessary for cell proliferation process in healing.

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Abbreviations

AOS:

Allene oxide synthase

COI1:

Coronatine insensitive 1

CYCB:

Cyclin B

DMSO:

Dimethyl sulfoxide

GUS:

β-glucuronidase

DAG:

Days after grafting

HAG:

Hours after grafting

IAA:

Indole-3-acetic acid

JA:

Jasmonic acid

JAZ:

Jasmonate ZIM-domain

MeJA:

Methyl jasmonate

OPDA:

12-oxo-phytodienoic acid

qRT-PCR:

Quantitative reverse transcription PCR

RAP2.6:

Related to APETALA2.6

RAP2.6L:

Related to APETALA2.6-like

SRDX:

Superman repression domain X

TIBA:

Triiodobenzoic acid

WT:

Wild type

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Acknowledgements

This work was supported in part by the Japan Society for the Promotion of Science (Grant-in-Aid for Young Scientists B No. 16K18572 to M.A.), Promotion and Mutual aid corporation for Private School of Japan (to M.A.) and the Ministry of Education, Culture, Sports, Science, and Technology Program for the Strategic Research Foundation at Private Universities (Grant No. S1311014 to T.Y., H.Y., and M.A.). We thank Dr. Ines Kubigsteltig (Ruhr University Bochum, Bochum, Germany; Kubigsteltig et al. 1999) for providing the pAOS::GUS vector, and Drs. Nobutaka Mitsuda (National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan) and Masaru Ohme-Takagi (Saitama University, Saitama, Japan) for the kind gift of the RAP2.6L-SRDX seeds.

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Contributions

KM contributed to all experiments and manuscript preparation. KM and MA designed the overall study. RY performed the microscopic experiments. EY and TY performed the phytohormone assays. MA, SS, and HY helped develop the manuscript text.

Corresponding author

Correspondence to Masashi Asahina.

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The authors have no conflicts of interest to declare.

Research involving human and animal rights

This study did not involve human and animal subjects.

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Matsuoka, K., Yanagi, R., Yumoto, E. et al. RAP2.6L and jasmonic acid–responsive genes are expressed upon Arabidopsis hypocotyl grafting but are not needed for cell proliferation related to healing. Plant Mol Biol 96, 531–542 (2018). https://doi.org/10.1007/s11103-018-0702-4

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  • DOI: https://doi.org/10.1007/s11103-018-0702-4

Keywords

  • Arabidopsis
  • Grafting
  • Wound healing
  • Jasmonic acid
  • RAP2.6L