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A Novel Pathogenesis-Related Class 10 Protein Gly m 4l, Increases Resistance upon Phytophthora sojae Infection in Soybean (Glycine max [L.] Merr.).

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

Bottom Line: HQ913577.1).The recombinant Gly m 4l protein showed RNase activity and displayed directly antimicrobial activity that inhibited hyphal growth and reduced zoospore release in P. sojae.Further analyses showed that the RNase activity of the recombinant protein to degrading tRNA was significantly affected in the presence of zeatin.

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
Phytophthora root and stem rot of soybean, caused by Phytophthora sojae (P. sojae), is a destructive disease in many soybean planting regions worldwide. In a previous study, an expressed sequence tag (EST) homolog of the major allergen Pru ar 1 in apricot (Prunus armeniaca) was identified up-regulated in the highly resistant soybean 'Suinong 10' infected with P. sojae. Here, the full length of the EST was isolated using rapid amplification of cDNA ends (RACE). It showed the highest homology of 53.46% with Gly m 4 after comparison with the eight soybean allergen families reported and was named Gly m 4-like (Gly m 4l, GenBank accession no. HQ913577.1). The cDNA full length of Gly m 4l was 707 bp containing a 474 bp open reading frame encoding a polypeptide of 157 amino acids. Sequence analysis suggests that Gly m 4l contains a conserved 'P-loop' (phosphate-binding loop) motif at residues 47-55 aa and a Bet v 1 domain at residues 87-120 aa. The transcript abundance of Gly m 4l was significantly induced by P. sojae, salicylic acid (SA), NaCl, and also responded to methyl jasmonic acid (MeJA) and ethylene (ET). The recombinant Gly m 4l protein showed RNase activity and displayed directly antimicrobial activity that inhibited hyphal growth and reduced zoospore release in P. sojae. Further analyses showed that the RNase activity of the recombinant protein to degrading tRNA was significantly affected in the presence of zeatin. Over-expression of Gly m 4l in susceptible 'Dongnong 50' soybean showed enhanced resistance to P. sojae. These results indicated that Gly m 4l protein played an important role in the defense of soybean against P. sojae infection.

No MeSH data available.


Related in: MedlinePlus

Subcellular localization analysis of Gly m 4l-GFP protein in Arabidopsis protoplasts.(A) Flow chart of construction 35S: Gly m 4l-GFP for subcellular localization analysis. (B) Images visualized by a Confocal Laser Scanning Microscopy. The images of bright-field (a and b), the GFP fluorescence (green) only (c and d), the chlorophyll autofluorescence (red) only (e and f) and combined ones (g and h) are shown. All scale bars indicate 10 μm.
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pone.0140364.g004: Subcellular localization analysis of Gly m 4l-GFP protein in Arabidopsis protoplasts.(A) Flow chart of construction 35S: Gly m 4l-GFP for subcellular localization analysis. (B) Images visualized by a Confocal Laser Scanning Microscopy. The images of bright-field (a and b), the GFP fluorescence (green) only (c and d), the chlorophyll autofluorescence (red) only (e and f) and combined ones (g and h) are shown. All scale bars indicate 10 μm.

Mentions: To test the subcellular localization, the Gly m 4l protein was fused to the GFP coding sequences under the control of the CaMV 35S promoter (Fig 4A). Then, the newly constructed vector 35S: Gly m 4l-GFP was introduced into Arabidopsis protoplasts cells using PEG-mediated transient expression. As shown in Fig 4B, confocal microscopic observations showed that GFP was dispersed throughout the entire cells bombarded with the control plasmid 35S: GFP and the fusion Gly m 4l-GFP protein was localized exclusively to cell membrane, indicating that Gly m 4l was a cell membrane-localized protein.


A Novel Pathogenesis-Related Class 10 Protein Gly m 4l, Increases Resistance upon Phytophthora sojae Infection in Soybean (Glycine max [L.] Merr.).

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

Subcellular localization analysis of Gly m 4l-GFP protein in Arabidopsis protoplasts.(A) Flow chart of construction 35S: Gly m 4l-GFP for subcellular localization analysis. (B) Images visualized by a Confocal Laser Scanning Microscopy. The images of bright-field (a and b), the GFP fluorescence (green) only (c and d), the chlorophyll autofluorescence (red) only (e and f) and combined ones (g and h) are shown. All scale bars indicate 10 μm.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4608668&req=5

pone.0140364.g004: Subcellular localization analysis of Gly m 4l-GFP protein in Arabidopsis protoplasts.(A) Flow chart of construction 35S: Gly m 4l-GFP for subcellular localization analysis. (B) Images visualized by a Confocal Laser Scanning Microscopy. The images of bright-field (a and b), the GFP fluorescence (green) only (c and d), the chlorophyll autofluorescence (red) only (e and f) and combined ones (g and h) are shown. All scale bars indicate 10 μm.
Mentions: To test the subcellular localization, the Gly m 4l protein was fused to the GFP coding sequences under the control of the CaMV 35S promoter (Fig 4A). Then, the newly constructed vector 35S: Gly m 4l-GFP was introduced into Arabidopsis protoplasts cells using PEG-mediated transient expression. As shown in Fig 4B, confocal microscopic observations showed that GFP was dispersed throughout the entire cells bombarded with the control plasmid 35S: GFP and the fusion Gly m 4l-GFP protein was localized exclusively to cell membrane, indicating that Gly m 4l was a cell membrane-localized protein.

Bottom Line: HQ913577.1).The recombinant Gly m 4l protein showed RNase activity and displayed directly antimicrobial activity that inhibited hyphal growth and reduced zoospore release in P. sojae.Further analyses showed that the RNase activity of the recombinant protein to degrading tRNA was significantly affected in the presence of zeatin.

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
Phytophthora root and stem rot of soybean, caused by Phytophthora sojae (P. sojae), is a destructive disease in many soybean planting regions worldwide. In a previous study, an expressed sequence tag (EST) homolog of the major allergen Pru ar 1 in apricot (Prunus armeniaca) was identified up-regulated in the highly resistant soybean 'Suinong 10' infected with P. sojae. Here, the full length of the EST was isolated using rapid amplification of cDNA ends (RACE). It showed the highest homology of 53.46% with Gly m 4 after comparison with the eight soybean allergen families reported and was named Gly m 4-like (Gly m 4l, GenBank accession no. HQ913577.1). The cDNA full length of Gly m 4l was 707 bp containing a 474 bp open reading frame encoding a polypeptide of 157 amino acids. Sequence analysis suggests that Gly m 4l contains a conserved 'P-loop' (phosphate-binding loop) motif at residues 47-55 aa and a Bet v 1 domain at residues 87-120 aa. The transcript abundance of Gly m 4l was significantly induced by P. sojae, salicylic acid (SA), NaCl, and also responded to methyl jasmonic acid (MeJA) and ethylene (ET). The recombinant Gly m 4l protein showed RNase activity and displayed directly antimicrobial activity that inhibited hyphal growth and reduced zoospore release in P. sojae. Further analyses showed that the RNase activity of the recombinant protein to degrading tRNA was significantly affected in the presence of zeatin. Over-expression of Gly m 4l in susceptible 'Dongnong 50' soybean showed enhanced resistance to P. sojae. These results indicated that Gly m 4l protein played an important role in the defense of soybean against P. sojae infection.

No MeSH data available.


Related in: MedlinePlus