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Identification and characterization of a matrix metalloproteinase (Pta1-MMP) expressed during Loblolly pine (Pinus taeda) seed development, germination completion, and early seedling establishment

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Abstract

Extracellular matrix (ECM) modifications occur during plant growth, development, and in response to environmental stimuli. Key modulators of ECM modification in vertebrates, the extracellular matrix metalloproteinases (MMPs), have also been described in a few plants. Here, we report the identification of Loblolly pine (Pinus taeda) Pta1-MMP and its characterization during seed development and germination. Pta1-MMP protein has the structural characteristics of other plant MMPs, the recombinant protein exhibits Zn2+-dependent protease activity, and is inhibited by EDTA and the active site-binding hydroxamate inhibitor GM6001. The Pta1-MMP gene is expressed in both embryo and megagametophyte, with transcript levels increasing in both during the period from proembryo to early cotyledonary stage, then declining during late embryogenesis and maturation drying. Protein extracts exhibited similar developmental-stage MMP-like activity. Seed germination was stimulated by GA3 and inhibited by ABA, and the timing of germination completion was mirrored by the presence of MMP-like protease activity in both water- and GA3-imbibed embryos. Pta1-MMP gene transcript levels increased in association with radicle protrusion for both GA3- and water-treated embryos, in agreement with MMP-like activity. In contrast, by 11 days after imbibition, Pta1-MMP gene transcripts in ABA-treated embryos were at levels similar to the other treatments, although MMP-like activity was not observed. The application of GM6001 during Loblolly pine seed germination inhibited radicle protrusion. Our results suggest that MMP activity may be involved in ECM modification, facilitating the cell division and expansion required during seed development, germination completion, and subsequent seedling establishment.

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Abbreviations

ECM:

Extracellular matrix

MMP:

Matrix metalloproteinase

GA3 :

Gibberellic acid 3

ABA:

Abscisic acid

AGP:

Arabinogalactan protein

HRGP:

Hydroxyproline-rich glycoprotein

TIMP:

Tissue inhibitor of matrix metalloproteinase

DE:

Dry embryo

DM:

Dry megagametophyte

DAI:

Days after imbibition

DMSO:

Dimethyl sulfoxide

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Acknowledgments

This work represents a portion of S. Ratnaparkhe’s doctoral dissertation. Many thanks to Dr. Amy Brunner and Dr. Ian Clark for their comments during the course of this study, Kelly Merricks for her help in gene cloning, and Dr. Jerzy Nowak for his comments on the article. This work was funded through the “High Value Horticulture and Forestry – Virginia” Special Grants (Project No. 2003-38891-02112) and HATCH funds (Project No. VA-135816) from the United States Department of Agriculture—CSREES, and operating funds from the Commonwealth of Virginia to the Institute for Advanced Learning and Research.

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Authors and Affiliations

  1. Department of Forestry, Virginia Polytechnic Institute and State University, Blacksburg, VA, 24061-0002, USA

    Supriya M. Ratnaparkhe, E. M. Ulrika Egertsdotter & Barry S. Flinn

  2. Institute for Sustainable and Renewable Resources, Institute for Advanced Learning and Research, 150 Slayton Avenue, Danville, VA, 24540, USA

    Supriya M. Ratnaparkhe & Barry S. Flinn

  3. School of Biology, Georgia Institute of Technology, 500 Tenth Street, NW, Atlanta, GA, 30332-0620, USA

    E. M. Ulrika Egertsdotter

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  1. Supriya M. Ratnaparkhe
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  2. E. M. Ulrika Egertsdotter
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Correspondence to Barry S. Flinn.

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E. M. Ulrika Egertsdotter and Barry S. Flinn contributed equally to project development, support, and supervision.

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Ratnaparkhe, S.M., Egertsdotter, E.M.U. & Flinn, B.S. Identification and characterization of a matrix metalloproteinase (Pta1-MMP) expressed during Loblolly pine (Pinus taeda) seed development, germination completion, and early seedling establishment. Planta 230, 339–354 (2009). https://doi.org/10.1007/s00425-009-0949-8

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