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PsPMEP, a pollen-specific pectin methylesterase of pea (Pisum sativum L.)

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Abstract

Pectin methylesterases (PMEs) are a family of enzymes involved in plant reproductive processes such as pollen development and pollen tube growth. We have isolated and characterized PsPMEP, a pea (Pisum sativum L.) pollen-specific gene that encodes a protein with homology to PMEs. Sequence analysis showed that PsPMEP belongs to group 2 PMEs, which are characterized by the presence of a processable amino-terminal PME inhibitor domain followed by the catalytic PME domain. Moreover, PsPMEP contains several motifs highly conserved among PMEs with the essential amino acid residues involved in enzyme substrate binding and catalysis. Northern blot and in situ hybridization analyses showed that PsPMEP is expressed in pollen grains from 4 days before anthesis till anther dehiscence and in pollinated carpels. In the PsPMEP promoter region, we have identified several conserved cis-regulatory elements that have been associated with gene pollen-specific expression. Expression analysis of PsPMEP promoter fused to the uidA reporter gene in Arabidopsis thaliana plants showed a similar expression pattern when compared with pea, indicating that this promoter is also functional in a non-leguminous plant. GUS expression was detected in mature pollen grains, during pollen germination, during pollen tube elongation along the transmitting tract, and when the pollen tube reaches the embryo sac in the ovule.

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Acknowledgments

This work was funded by grants BIO2006-09374 and BIO2009-08134 from the Spanish Ministry of Science and Innovation (MICINN). The collaboration and assistance of Julia Marín-Navarro in the catalytic activity assays of PsPMEP in yeast and Rafael Martínez-Pardo in the greenhouse is gratefully acknowledged. We would like to thank the HAPRECI consortium (COST Action FA0903) to bring us the opportunity to collaborate with other European research groups working in the field of Plant Reproduction and to select our manuscript to be published in this special issue.

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Authors and Affiliations

  1. Instituto de Biología Molecular y Celular de Plantas (CSIC-UPV), Ciudad Politécnica de la Innovación, Edf. 8E. C/, Ingeniero Fausto Elio s/n, 46011, Valencia, Spain

    María Dolores Gómez, Begoña Renau-Morata, Edelín Roque, José Pío Beltrán & Luis A. Cañas

  2. Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Avda. Agustín Escardino, 7, 46980, Paterna, Valencia, Spain

    Julio Polaina

Authors
  1. María Dolores Gómez
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  2. Begoña Renau-Morata
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  3. Edelín Roque
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  4. Julio Polaina
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  5. José Pío Beltrán
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  6. Luis A. Cañas
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Corresponding author

Correspondence to Luis A. Cañas.

Additional information

Communicated by T. Dresselhaus.

A contribution to the Special Issue “HAPRECI—Plant Reproduction Research in Europe.”

María Dolores Gómez and Begoña Renau-Morata contributed equally to this work.

PsPMEP GenBank accession no.: KC964536.

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497_2013_220_MOESM1_ESM.tif

Figure S1. Neighbor-Joining Tree of selected pollen-specific PMEs. The numbers next to the nodes refer to bootstrap values from 1000 pseudo-replicates. Accession numbers for the PME proteins used in this N-J tree are: O80722 (AT2G47040), Q9FJ21 (AT5G49180), AEE35001(AT1G69940) and ABI97858.1 (AT5G55590.1) Arabidopsis thaliana; P41510 Brassica napus; Q42608 Brassica rapa; Q43043 Petunia integrifolia subsp. inflata; AAX13972 Nicotiana tabacum; Q9MBB6 Salix gilgiana; Q24596 Zea mays; Q9SC89, Q9SC90 Medicago truncatula; Q42920 Medicago sativa and KC964536 (PsPMEP) Pisum sativum. We inferred the phylogeny based on the alignment of mature enzyme sequences of 14 representative PMEs from pollen, which are clustered into four clades (A to D). Branch lengths are proportional to sequence divergence

Table S1. Primers used to sequence the PsPMEP genomic clone

Table S2. Putative pollen-specific motifs identified in the PsPMEP promoter region (-1 to -2133)

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Gómez, M.D., Renau-Morata, B., Roque, E. et al. PsPMEP, a pollen-specific pectin methylesterase of pea (Pisum sativum L.). Plant Reprod 26, 245–254 (2013). https://doi.org/10.1007/s00497-013-0220-0

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