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Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula
Small, post-translationally modified and secreted peptides regulate diverse plant developmental processes. Due to low natural abundance, it is difficult to isolate and identify these peptides. Using an improved peptide isolation protocol and Orbitrap mass spectrometry, nine 15-amino-acid CEP peptide...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4526912/ https://www.ncbi.nlm.nih.gov/pubmed/25711701 http://dx.doi.org/10.1093/jxb/erv008 |
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author | Mohd-Radzman, Nadiatul A. Binos, Steve Truong, Thy T. Imin, Nijat Mariani, Michael Djordjevic, Michael A. |
author_facet | Mohd-Radzman, Nadiatul A. Binos, Steve Truong, Thy T. Imin, Nijat Mariani, Michael Djordjevic, Michael A. |
author_sort | Mohd-Radzman, Nadiatul A. |
collection | PubMed |
description | Small, post-translationally modified and secreted peptides regulate diverse plant developmental processes. Due to low natural abundance, it is difficult to isolate and identify these peptides. Using an improved peptide isolation protocol and Orbitrap mass spectrometry, nine 15-amino-acid CEP peptides were identified that corresponded to the two domains encoded by Medicago truncatula CEP1 (MtCEP1). Novel arabinosylated and hydroxylated peptides were identified in root cultures overexpressing MtCEP1. The five most abundant CEP peptides were hydroxylated and these species were detected also in low amounts in vector control samples. Synthetic peptides with different hydroxylation patterns differentially affected root development. Notably, the domain 1 peptide hydroxylated at Pro4 and Pro11 (D1:HyP4,11) imparted the strongest inhibition of lateral root emergence when grown with 5mM KNO(3) and stimulated the highest increase in nodule number when grown with 0mM KNO(3). Inhibition of lateral root emergence by D1:HyP4,11 was not alleviated by removing peptide exposure. In contrast, the domain 2 peptide hydroxylated at Pro11 (D2:HyP11) increased stage III–IV lateral root primordium numbers by 6-fold (P < 0.001) which failed to emerge. Auxin addition at levels which stimulated lateral root formation in wild-type plants had little or no ameliorating effect on CEP peptide-mediated inhibition of lateral root formation or emergence. Both peptides increased and altered the root staining pattern of the auxin-responsive reporter GH3:GUS suggesting CEPs alter auxin sensitivity or distribution. The results showed that CEP primary sequence and post-translational modifications influence peptide activities and the improved isolation procedure effectively and reproducibly identifies and characterises CEPs. |
format | Online Article Text |
id | pubmed-4526912 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-45269122015-08-10 Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula Mohd-Radzman, Nadiatul A. Binos, Steve Truong, Thy T. Imin, Nijat Mariani, Michael Djordjevic, Michael A. J Exp Bot Research Paper Small, post-translationally modified and secreted peptides regulate diverse plant developmental processes. Due to low natural abundance, it is difficult to isolate and identify these peptides. Using an improved peptide isolation protocol and Orbitrap mass spectrometry, nine 15-amino-acid CEP peptides were identified that corresponded to the two domains encoded by Medicago truncatula CEP1 (MtCEP1). Novel arabinosylated and hydroxylated peptides were identified in root cultures overexpressing MtCEP1. The five most abundant CEP peptides were hydroxylated and these species were detected also in low amounts in vector control samples. Synthetic peptides with different hydroxylation patterns differentially affected root development. Notably, the domain 1 peptide hydroxylated at Pro4 and Pro11 (D1:HyP4,11) imparted the strongest inhibition of lateral root emergence when grown with 5mM KNO(3) and stimulated the highest increase in nodule number when grown with 0mM KNO(3). Inhibition of lateral root emergence by D1:HyP4,11 was not alleviated by removing peptide exposure. In contrast, the domain 2 peptide hydroxylated at Pro11 (D2:HyP11) increased stage III–IV lateral root primordium numbers by 6-fold (P < 0.001) which failed to emerge. Auxin addition at levels which stimulated lateral root formation in wild-type plants had little or no ameliorating effect on CEP peptide-mediated inhibition of lateral root formation or emergence. Both peptides increased and altered the root staining pattern of the auxin-responsive reporter GH3:GUS suggesting CEPs alter auxin sensitivity or distribution. The results showed that CEP primary sequence and post-translational modifications influence peptide activities and the improved isolation procedure effectively and reproducibly identifies and characterises CEPs. Oxford University Press 2015-08 2015-02-22 /pmc/articles/PMC4526912/ /pubmed/25711701 http://dx.doi.org/10.1093/jxb/erv008 Text en © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Paper Mohd-Radzman, Nadiatul A. Binos, Steve Truong, Thy T. Imin, Nijat Mariani, Michael Djordjevic, Michael A. Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula |
title | Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula
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title_full | Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula
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title_fullStr | Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula
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title_full_unstemmed | Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula
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title_short | Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula
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title_sort | novel mtcep1 peptides produced in vivo differentially regulate root development in medicago truncatula |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4526912/ https://www.ncbi.nlm.nih.gov/pubmed/25711701 http://dx.doi.org/10.1093/jxb/erv008 |
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