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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

Nucleotide and amino acid sequence of Gly m 4l.The P-loop motif and Bet v 1 motif are shown in shadow. The α-helices and β-sheets are marked underlined.
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pone.0140364.g001: Nucleotide and amino acid sequence of Gly m 4l.The P-loop motif and Bet v 1 motif are shown in shadow. The α-helices and β-sheets are marked underlined.

Mentions: The full-length cDNA sequence was obtained from ‘Suinong 10’ soybean using RACE. Alignment and phylogenetic tree analysis of the full-length amino acids sequence with the eight soybean allergen families reported showed identities of 53.46% homology with Gly m 4 protein, representing the highest homology (S1 Fig), and it was designated Gly m 4-like (Gly m 4l, GenBank accession no. HQ913577.1). As shown in Fig 1, the 707 bp Gly m 4l gene contains a single open reading frame (ORF) of 474 nucleotides encoding a polypeptide of 157 amino acids, with a calculated molecular mass of 17.14 kDa and a theoretical PI of 4.98. The nucleotide sequence showed a 5’ untranslated region (5’ UTR) of 88 nucleotides and a 3’ UTR of 145 nucleotides, along with a poly A signal (AATAAA) at 656–661 bp. The genomic Gly m 4l DNA sequence was also amplified, showing that Gly m 4l had an intron of 341 bps at 184 bp. The results of the database search revealed that Gly m 4l contained no signal peptide. The predicted structure of Gly m 4l protein included a conserved ‘P-loop’ (phosphate-binding loop) motif (GXGGXGXXK at residues 47–55 aa) and a Bet v 1 domain comprising 34 amino acids (at residues 87–120 aa), present in many PR 10 proteins. A neighbor-joining (NJ) phylogram was used to construct a phylogenetic tree based on the deduced sequence of Gly m 4l that contained other members of the PR10 family (Fig 2A), indicating that these proteins might share a common ancestor and display similar functions. The homology analysis of Gly m 4l with five nearby PR10 protein sequences showed that it shared 56.9% identity with PdPR10 (Prunus domestica) (ABW99634.1), representing the highest homology (Fig 2B). The prediction of the three dimensional (3D) structure of Gly m 4l based on the data from Phyre (http://www.sbg.bio.jc.ac.uk/phyre/), showed that this protein was very similar to that of the other PR10 proteins (Fig 2C). Gly m 4l contained a 24-amino-acid long α-helix (α2) at the C-terminus and a short α-helix (α1) at the N- terminus. The α2 was surrounded by seven anti-parallel β-sheets (β1 to β7) and the α1 was located between the β1 and β2 strands. The short loop structures were named L1 to L8 located between the α-helix and β- sheet (Fig 2A). The results suggested that Gly m 4l belonged to PR 10 protein family.


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)

Nucleotide and amino acid sequence of Gly m 4l.The P-loop motif and Bet v 1 motif are shown in shadow. The α-helices and β-sheets are marked underlined.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140364.g001: Nucleotide and amino acid sequence of Gly m 4l.The P-loop motif and Bet v 1 motif are shown in shadow. The α-helices and β-sheets are marked underlined.
Mentions: The full-length cDNA sequence was obtained from ‘Suinong 10’ soybean using RACE. Alignment and phylogenetic tree analysis of the full-length amino acids sequence with the eight soybean allergen families reported showed identities of 53.46% homology with Gly m 4 protein, representing the highest homology (S1 Fig), and it was designated Gly m 4-like (Gly m 4l, GenBank accession no. HQ913577.1). As shown in Fig 1, the 707 bp Gly m 4l gene contains a single open reading frame (ORF) of 474 nucleotides encoding a polypeptide of 157 amino acids, with a calculated molecular mass of 17.14 kDa and a theoretical PI of 4.98. The nucleotide sequence showed a 5’ untranslated region (5’ UTR) of 88 nucleotides and a 3’ UTR of 145 nucleotides, along with a poly A signal (AATAAA) at 656–661 bp. The genomic Gly m 4l DNA sequence was also amplified, showing that Gly m 4l had an intron of 341 bps at 184 bp. The results of the database search revealed that Gly m 4l contained no signal peptide. The predicted structure of Gly m 4l protein included a conserved ‘P-loop’ (phosphate-binding loop) motif (GXGGXGXXK at residues 47–55 aa) and a Bet v 1 domain comprising 34 amino acids (at residues 87–120 aa), present in many PR 10 proteins. A neighbor-joining (NJ) phylogram was used to construct a phylogenetic tree based on the deduced sequence of Gly m 4l that contained other members of the PR10 family (Fig 2A), indicating that these proteins might share a common ancestor and display similar functions. The homology analysis of Gly m 4l with five nearby PR10 protein sequences showed that it shared 56.9% identity with PdPR10 (Prunus domestica) (ABW99634.1), representing the highest homology (Fig 2B). The prediction of the three dimensional (3D) structure of Gly m 4l based on the data from Phyre (http://www.sbg.bio.jc.ac.uk/phyre/), showed that this protein was very similar to that of the other PR10 proteins (Fig 2C). Gly m 4l contained a 24-amino-acid long α-helix (α2) at the C-terminus and a short α-helix (α1) at the N- terminus. The α2 was surrounded by seven anti-parallel β-sheets (β1 to β7) and the α1 was located between the β1 and β2 strands. The short loop structures were named L1 to L8 located between the α-helix and β- sheet (Fig 2A). The results suggested that Gly m 4l belonged to PR 10 protein family.

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