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Physiology and Molecular Biology of Plants

, Volume 25, Issue 2, pp 511–522 | Cite as

Regulation of α-expansins genes in Arabidopsis thaliana seeds during post-osmopriming germination

  • Alessandra Ferreira RibasEmail author
  • Nathalia Volpi e Silva
  • Tiago Benedito dos Santos
  • Fabiana Lima Abrantes
  • Ceci Castilho Custódio
  • Nelson Barbosa Machado-Neto
  • Luiz Gonzaga Esteves Vieira
Research Article
  • 75 Downloads

Abstract

Seed osmopriming is a pre-sowing treatment that involves limitation of the seed water imbibition, so that pre-germinative metabolic activities proceed without radicular protrusion. This technique is used for improving germination rate, uniformity of seedling growth and hastening the time to start germination. In Arabidopsis thaliana, seed germination has been associated with the induction of enzymes involved in cell wall modifications, such as expansins. The α-expansins (EXPAs) are involved in cell wall relaxation and extension during seed germination. We used online tools to identify AtEXPA genes with preferential expression during seed germination and RT-qPCR to study the expression of five EXPA genes at different germination stages of non-primed and osmoprimed seeds. In silico promoter analysis of these genes showed that motifs similar to cis-acting elements related to abiotic stress, light and phytohormone responses are the most overrepresented in promoters of these AtEXPA genes, showing that their expression is likely be regulated by intrinsic developmental and environmental signals during Arabidopsis seed germination. The osmopriming conditioning had a decreased time and mean to 50% germination without affecting the percentage of final seed germination. The dried PEG-treated seeds showed noticeable high mRNA levels earlier at the beginning of water imbibition (18 h), showing that transcripts of all five EXPA isoforms were significantly produced during the osmopriming process. The strong up-regulation of these AtEXPA genes, mainly AtEXPA2, were associated with the earlier germination of the osmoprimed seeds, which qualifies them to monitor osmopriming procedures and the advancement of germination.

Keywords

Arabidopsis thaliana α-Expansins Osmopriming Seed germination 

Notes

Acknowledgements

We are grateful by the suggestions provided by Dr. Peter E. Toorop. All authors read and approved the final manuscript.

Author contributions

LGEV and AFR conceived and designed the experiments. FLA, CCC and NBMN performed the germination and priming experiments. AFR and NVS carried out the bioinformatics analysis. AFR and TBS carried out the RT-qPCR assays and analyzed the data. LGEV, AFR and NVC wrote the manuscript.

Compliance with ethical standards

Conflict of interest

NVS has received a grant from São Paulo Research Foundation (FAPESP) and Coordination for the Improvement of Higher Education Personnel (CAPES) for the fellowship during her doctor degree and National Council for Scientific and Technological Development—CNPq for the fellowship during her master degree. TBS has received a grant from Coordination for the Improvement of Higher Education Personnel (CAPES) for postdoctoral fellowship. LGEV has received a grant from National Council for Scientific and Technological Development—CNPq for the research fellowship. AFR, FLA, CCC, NBM declare that they have no conflict of interest.

Supplementary material

12298_2018_620_MOESM1_ESM.tiff (1.4 mb)
Fig. S1 Heatmap produced by GENEVESTIGATOR (https://www.genevestigator.com) showing the expression pattern of 23 α-expansins during developmental stages of A. thaliana. The stages of development from seed germination to senescence are grouped arbitrarily based on Boyes et al. (2001). Data were retrieved from GENEVESTIGATOR (Zimmermann et al. 2004) (TIFF 1478 kb)
12298_2018_620_MOESM2_ESM.tiff (1.4 mb)
Fig. S2 Expression pattern of eight α-expansins during developmental stages of A. thaliana produced by GENEVESTIGATOR (https://www.genevestigator.com). The stages of development from seed germination to senescence are grouped arbitrarily based on Boyes et al. (2001). Data were retrieved from GENEVESTIGATOR (Zimmermann et al. 2004) (TIFF 1415 kb)
12298_2018_620_MOESM3_ESM.tiff (586 kb)
Fig. S3 Transcriptional profiles of five AtEXPAs during seed germination. The data were generated in the database BAR eFP Browser - http://bar.utoronto.ca/efp_arabidopsis/cgi-bin/efpWeb.cgi (Winter et al. 2007) - based on experiments for (A) non-dormant, non-stratified after-ripened wild-type seeds (Nakabayashi et al. 2005), (B) cold-stratified wild-type seeds (Yamauchi et al. 2004) (TIFF 586 kb)
12298_2018_620_MOESM4_ESM.tiff (1.5 mb)
Fig. S4 Distribution of the identified α-expansins genes on five chromosomes of A. thaliana. The chromosomal position of each gene was mapped according to the A. thaliana genome. The location of each gene was indicated by a line (TIFF 1576 kb)
12298_2018_620_MOESM5_ESM.tiff (1.8 mb)
Fig. S5 Putative cis-regulatory elements mapped onto the promoters of five EXPAs of A. thaliana up-regulated during seed germination. A Gibberellic acid motifs; B Abscisic acid/Abiotic stress motifs; C Light related motifs and D Seed related motifs. The size of each promoter region was one kb in relation to the translation start codon (ATG). A color key of cis-elements is given at the left of the image. Horizontal black lines in each block represent the promoter sequences and colored squares show the position of the mapped cis-element occurrences found in sense (above line) or anti-sense orientation (below line) (TIFF 1871 kb)

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Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  • Alessandra Ferreira Ribas
    • 1
    Email author
  • Nathalia Volpi e Silva
    • 2
  • Tiago Benedito dos Santos
    • 1
  • Fabiana Lima Abrantes
    • 3
  • Ceci Castilho Custódio
    • 3
  • Nelson Barbosa Machado-Neto
    • 3
  • Luiz Gonzaga Esteves Vieira
    • 1
  1. 1.Agronomy Graduate Program, Molecular Genetic LaboratoryUniversidade do Oeste Paulista (UNOESTE)Presidente PrudenteBrazil
  2. 2.Department of Plant Biology, Institute of BiologyUniversity of Campinas (UNICAMP)CampinasBrazil
  3. 3.Agronomy Graduate Program, Seed Reserach LaboratoryUniversidade do Oeste Paulista (UNOESTE)Presidente PrudenteBrazil

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