14-3-3 proteins SGF14C and SGF14L play critical roles during soybean nodulation

Osman Radwan, Xia Wu, Manjula Govindarajulu, Marc Libault, David J. Neece, Man Ho Oh, R. Howard Berg, Gary Stacey, Christopher G. Taylor, Steven C Huber, Steven J Clough

Research output: Contribution to journalArticle

Abstract

The soybean (Glycine max) genome contains 18 members of the 14-3-3 protein family, but little is known about their association with specific phenotypes. Here, we report that the Glyma0529080 Soybean G-box Factor 14-3-3c (SGF14c) and Glyma08g12220 (SGF14l) genes, encoding 14-3-3 proteins, appear to play essential roles in soybean nodulation. Quantitative reverse transcription-polymerase chain reaction and western-immunoblot analyses showed that SGF14c mRNA and protein levels were specifically increased in abundance in nodulated soybean roots 10, 12, 16, and 20 d after inoculation with Bradyrhizobium japonicum. To investigate the role of SGF14c during soybean nodulation, RNA interference was employed to silence SGF14c expression in soybean roots using Agrobacterium rhizogenes-mediated root transformation. Due to the paleopolyploid nature of soybean, designing a specific RNA interference sequence that exclusively targeted SGF14c was not possible. Therefore, two highly similar paralogs (SGF14c and SGF14l) that have been shown to function as dimers were silenced. Transcriptomic and proteomic analyses showed that mRNA and protein levels were significantly reduced in the SGF14c/ SGF14l-silenced roots, and these roots exhibited reduced numbers of mature nodules. In addition, SGF14c/SGF14l-silenced roots contained large numbers of arrested nodule primordia following B. japonicum inoculation. Transmission electron microscopy further revealed that the host cytoplasm and membranes, except the symbiosome membrane, were severely degraded in the failed nodules. Altogether, transcriptomic, proteomic, and cytological data suggest a critical role of one or both of these 14-3-3 proteins in early development stages of soybean nodules.

Original languageEnglish (US)
Pages (from-to)2125-2136
Number of pages12
JournalPlant physiology
Volume160
Issue number4
DOIs
StatePublished - Dec 1 2012

Fingerprint

14-3-3 Proteins
nodulation
Soybeans
soybeans
proteins
Bradyrhizobium japonicum
RNA interference
transcriptomics
RNA Interference
proteomics
Proteomics
Bradyrhizobium
Rhizobium rhizogenes
Messenger RNA
Agrobacterium
Membranes

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

Cite this

Radwan, O., Wu, X., Govindarajulu, M., Libault, M., Neece, D. J., Oh, M. H., ... Clough, S. J. (2012). 14-3-3 proteins SGF14C and SGF14L play critical roles during soybean nodulation. Plant physiology, 160(4), 2125-2136. https://doi.org/10.1104/pp.112.207027

14-3-3 proteins SGF14C and SGF14L play critical roles during soybean nodulation. / Radwan, Osman; Wu, Xia; Govindarajulu, Manjula; Libault, Marc; Neece, David J.; Oh, Man Ho; Berg, R. Howard; Stacey, Gary; Taylor, Christopher G.; Huber, Steven C; Clough, Steven J.

In: Plant physiology, Vol. 160, No. 4, 01.12.2012, p. 2125-2136.

Research output: Contribution to journalArticle

Radwan, O, Wu, X, Govindarajulu, M, Libault, M, Neece, DJ, Oh, MH, Berg, RH, Stacey, G, Taylor, CG, Huber, SC & Clough, SJ 2012, '14-3-3 proteins SGF14C and SGF14L play critical roles during soybean nodulation', Plant physiology, vol. 160, no. 4, pp. 2125-2136. https://doi.org/10.1104/pp.112.207027
Radwan O, Wu X, Govindarajulu M, Libault M, Neece DJ, Oh MH et al. 14-3-3 proteins SGF14C and SGF14L play critical roles during soybean nodulation. Plant physiology. 2012 Dec 1;160(4):2125-2136. https://doi.org/10.1104/pp.112.207027
Radwan, Osman ; Wu, Xia ; Govindarajulu, Manjula ; Libault, Marc ; Neece, David J. ; Oh, Man Ho ; Berg, R. Howard ; Stacey, Gary ; Taylor, Christopher G. ; Huber, Steven C ; Clough, Steven J. / 14-3-3 proteins SGF14C and SGF14L play critical roles during soybean nodulation. In: Plant physiology. 2012 ; Vol. 160, No. 4. pp. 2125-2136.
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abstract = "The soybean (Glycine max) genome contains 18 members of the 14-3-3 protein family, but little is known about their association with specific phenotypes. Here, we report that the Glyma0529080 Soybean G-box Factor 14-3-3c (SGF14c) and Glyma08g12220 (SGF14l) genes, encoding 14-3-3 proteins, appear to play essential roles in soybean nodulation. Quantitative reverse transcription-polymerase chain reaction and western-immunoblot analyses showed that SGF14c mRNA and protein levels were specifically increased in abundance in nodulated soybean roots 10, 12, 16, and 20 d after inoculation with Bradyrhizobium japonicum. To investigate the role of SGF14c during soybean nodulation, RNA interference was employed to silence SGF14c expression in soybean roots using Agrobacterium rhizogenes-mediated root transformation. Due to the paleopolyploid nature of soybean, designing a specific RNA interference sequence that exclusively targeted SGF14c was not possible. Therefore, two highly similar paralogs (SGF14c and SGF14l) that have been shown to function as dimers were silenced. Transcriptomic and proteomic analyses showed that mRNA and protein levels were significantly reduced in the SGF14c/ SGF14l-silenced roots, and these roots exhibited reduced numbers of mature nodules. In addition, SGF14c/SGF14l-silenced roots contained large numbers of arrested nodule primordia following B. japonicum inoculation. Transmission electron microscopy further revealed that the host cytoplasm and membranes, except the symbiosome membrane, were severely degraded in the failed nodules. Altogether, transcriptomic, proteomic, and cytological data suggest a critical role of one or both of these 14-3-3 proteins in early development stages of soybean nodules.",
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