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Isolation and Characterization of a Novel Pathogenesis-Related Protein Gene (GmPRP) with Induced Expression in Soybean (Glycine max) during Infection with Phytophthora sojae.

Jiang L, Wu J, Fan S, Li W, Dong L, Cheng Q, Xu P, Zhang S - PLoS ONE (2015)

Bottom Line: GmPRP was localized in the cell plasma membrane and cytoplasm.Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro.Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively.

View Article: PubMed Central - PubMed

Affiliation: Soybean Research Institute, Key Laboratory of Soybean Biology of Chinese Education Ministry, Northeast Agricultural University, Harbin, 150030, Heilongjiang, People's Republic of China.

ABSTRACT
Pathogenesis-related proteins (PR proteins) play crucial roles in the plant defense system. A novel PRP gene was isolated from highly resistant soybean infected with Phytophthora sojae (P. sojae) and was named GmPRP (GenBank accession number: KM506762). The amino acid sequences of GmPRP showed identities of 74%, 73%, 72% and 69% with PRP proteins from Vitis vinifera, Populus trichocarpa, Citrus sinensis and Theobroma cacao, respectively. Quantitative real-time reverse transcription PCR (qRT-PCR) data showed that the expression of GmPRP was highest in roots, followed by the stems and leaves. GmPRP expression was upregulated in soybean leaves infected with P. sojae. Similarly, GmPRP expression also responded to defense/stress signaling molecules, including salicylic acid (SA), ethylene (ET), abscisic acid (ABA) and jasmonic acid (JA). GmPRP was localized in the cell plasma membrane and cytoplasm. Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro. Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively. These results indicated that the GmPRP protein played an important role in the defense of soybean against P. sojae infection.

No MeSH data available.


Related in: MedlinePlus

Nucleotide and amino acid sequences of GmPRP cDNA.(A) Nucleotide and amino acid sequences of GmPRP cDNA. Putative phosphorylation sites are marked in bold italics. (B) Analysis of the 3D structure and conserved motifs of GmPRP. The 3D structure of GmPRP. The N-terminal, C-terminal, α-helix, and β-strand are marked in bold italics. (C) The conserved motif of the GmPRP protein. The predicted GmPRP protein contained a conserved motif at residues 131–211 aa that belonged to the NTF2-like superfamily. (D) Alignments of the amino acid sequences of the GmPRP with other plant PRP proteins. Comparison of the predicted amino acid sequence of GmPRP with other plant PRPs from Vitis vinifera (XP002262980), Populus trichocarpa (XP002306682), Citrus sinensis (XP006472936) and Theobroma cacao (XP007019416). Conserved residues are shaded in black.
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pone.0129932.g001: Nucleotide and amino acid sequences of GmPRP cDNA.(A) Nucleotide and amino acid sequences of GmPRP cDNA. Putative phosphorylation sites are marked in bold italics. (B) Analysis of the 3D structure and conserved motifs of GmPRP. The 3D structure of GmPRP. The N-terminal, C-terminal, α-helix, and β-strand are marked in bold italics. (C) The conserved motif of the GmPRP protein. The predicted GmPRP protein contained a conserved motif at residues 131–211 aa that belonged to the NTF2-like superfamily. (D) Alignments of the amino acid sequences of the GmPRP with other plant PRP proteins. Comparison of the predicted amino acid sequence of GmPRP with other plant PRPs from Vitis vinifera (XP002262980), Populus trichocarpa (XP002306682), Citrus sinensis (XP006472936) and Theobroma cacao (XP007019416). Conserved residues are shaded in black.

Mentions: The full-length cDNA sequence was obtained from ‘Suinong 10’ soybean using RACE. The clone, designated GmPRP (GenBank accession no. KM506762), was chosen for further functional analysis. It comprised 952 bp with a single open reading frame (ORF) of 717 nucleotides, encoding a polypeptide of 238 amino acids with a calculated molecular mass of 27.225 kDa and a theoretical PI of 7.07. The nucleotide sequence also showed a 5’ untranslated region (5’ UTR) of 13 nucleotides and a 3’ UTR of 222 nucleotides along with a poly-A signal (AAAAAAAAAAAAAAAA) at 936–952 bp. A database search (http://www.cbs.dtu.dk/services/signalp/) indicated that GmPRP contained no signal peptide. Searches of the NCBI and Phytozome databases (http://www.n.nihcbi.nlm.gov/; http://www.phytozome.net/soybean) revealed that the GmPRP gene had a 694-bp intron and was located on chromosome 6 with two copies. The software NetPhos (http://www.cbs.dtu.dk/services/NetPhos) predicted that GmPRP had eight serines (Ser 5, 19, 52, 79, 96, 131, 132, 223, in bold italics), five tyrosines (Tyr 11, 33, 85, 125, 167, in bold italics), and five lysines (Lys 45, 59, 128, 138, 191, in bold italics) as potential phosphorylation sites (Fig 1A). The 3D structure of the GmPRP protein consisted of a 10-amino-acid C-terminal α-helix (α3) surrounded by a six-stranded antiparallel β-sheet (from β1 to β6) and three N-terminal α-helices (α1, α2 and α3), which are two short α-helices and a long α-helix between the β2 and β3 sheets. The connection sequences between the α-helix and β-strand were short loop structures (Fig 1B). The predicted GmPRP protein contained a conserved motif at residues 131–211 aa that belonged to the NTF2-like superfamily (Fig 1C).


Isolation and Characterization of a Novel Pathogenesis-Related Protein Gene (GmPRP) with Induced Expression in Soybean (Glycine max) during Infection with Phytophthora sojae.

Jiang L, Wu J, Fan S, Li W, Dong L, Cheng Q, Xu P, Zhang S - PLoS ONE (2015)

Nucleotide and amino acid sequences of GmPRP cDNA.(A) Nucleotide and amino acid sequences of GmPRP cDNA. Putative phosphorylation sites are marked in bold italics. (B) Analysis of the 3D structure and conserved motifs of GmPRP. The 3D structure of GmPRP. The N-terminal, C-terminal, α-helix, and β-strand are marked in bold italics. (C) The conserved motif of the GmPRP protein. The predicted GmPRP protein contained a conserved motif at residues 131–211 aa that belonged to the NTF2-like superfamily. (D) Alignments of the amino acid sequences of the GmPRP with other plant PRP proteins. Comparison of the predicted amino acid sequence of GmPRP with other plant PRPs from Vitis vinifera (XP002262980), Populus trichocarpa (XP002306682), Citrus sinensis (XP006472936) and Theobroma cacao (XP007019416). Conserved residues are shaded in black.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4482714&req=5

pone.0129932.g001: Nucleotide and amino acid sequences of GmPRP cDNA.(A) Nucleotide and amino acid sequences of GmPRP cDNA. Putative phosphorylation sites are marked in bold italics. (B) Analysis of the 3D structure and conserved motifs of GmPRP. The 3D structure of GmPRP. The N-terminal, C-terminal, α-helix, and β-strand are marked in bold italics. (C) The conserved motif of the GmPRP protein. The predicted GmPRP protein contained a conserved motif at residues 131–211 aa that belonged to the NTF2-like superfamily. (D) Alignments of the amino acid sequences of the GmPRP with other plant PRP proteins. Comparison of the predicted amino acid sequence of GmPRP with other plant PRPs from Vitis vinifera (XP002262980), Populus trichocarpa (XP002306682), Citrus sinensis (XP006472936) and Theobroma cacao (XP007019416). Conserved residues are shaded in black.
Mentions: The full-length cDNA sequence was obtained from ‘Suinong 10’ soybean using RACE. The clone, designated GmPRP (GenBank accession no. KM506762), was chosen for further functional analysis. It comprised 952 bp with a single open reading frame (ORF) of 717 nucleotides, encoding a polypeptide of 238 amino acids with a calculated molecular mass of 27.225 kDa and a theoretical PI of 7.07. The nucleotide sequence also showed a 5’ untranslated region (5’ UTR) of 13 nucleotides and a 3’ UTR of 222 nucleotides along with a poly-A signal (AAAAAAAAAAAAAAAA) at 936–952 bp. A database search (http://www.cbs.dtu.dk/services/signalp/) indicated that GmPRP contained no signal peptide. Searches of the NCBI and Phytozome databases (http://www.n.nihcbi.nlm.gov/; http://www.phytozome.net/soybean) revealed that the GmPRP gene had a 694-bp intron and was located on chromosome 6 with two copies. The software NetPhos (http://www.cbs.dtu.dk/services/NetPhos) predicted that GmPRP had eight serines (Ser 5, 19, 52, 79, 96, 131, 132, 223, in bold italics), five tyrosines (Tyr 11, 33, 85, 125, 167, in bold italics), and five lysines (Lys 45, 59, 128, 138, 191, in bold italics) as potential phosphorylation sites (Fig 1A). The 3D structure of the GmPRP protein consisted of a 10-amino-acid C-terminal α-helix (α3) surrounded by a six-stranded antiparallel β-sheet (from β1 to β6) and three N-terminal α-helices (α1, α2 and α3), which are two short α-helices and a long α-helix between the β2 and β3 sheets. The connection sequences between the α-helix and β-strand were short loop structures (Fig 1B). The predicted GmPRP protein contained a conserved motif at residues 131–211 aa that belonged to the NTF2-like superfamily (Fig 1C).

Bottom Line: GmPRP was localized in the cell plasma membrane and cytoplasm.Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro.Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively.

View Article: PubMed Central - PubMed

Affiliation: Soybean Research Institute, Key Laboratory of Soybean Biology of Chinese Education Ministry, Northeast Agricultural University, Harbin, 150030, Heilongjiang, People's Republic of China.

ABSTRACT
Pathogenesis-related proteins (PR proteins) play crucial roles in the plant defense system. A novel PRP gene was isolated from highly resistant soybean infected with Phytophthora sojae (P. sojae) and was named GmPRP (GenBank accession number: KM506762). The amino acid sequences of GmPRP showed identities of 74%, 73%, 72% and 69% with PRP proteins from Vitis vinifera, Populus trichocarpa, Citrus sinensis and Theobroma cacao, respectively. Quantitative real-time reverse transcription PCR (qRT-PCR) data showed that the expression of GmPRP was highest in roots, followed by the stems and leaves. GmPRP expression was upregulated in soybean leaves infected with P. sojae. Similarly, GmPRP expression also responded to defense/stress signaling molecules, including salicylic acid (SA), ethylene (ET), abscisic acid (ABA) and jasmonic acid (JA). GmPRP was localized in the cell plasma membrane and cytoplasm. Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro. Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively. These results indicated that the GmPRP protein played an important role in the defense of soybean against P. sojae infection.

No MeSH data available.


Related in: MedlinePlus