Limits...
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

Analysis of the purified recombinant Gly m 4l protein.(A) The recombinant Gly m 4l protein induced with 0.5 mM IPTG at 37°C for 1–5 h in E.coli BL21 (DE3). (B) SDS-PAGE analysis of the purified recombinant Gly m 4l protein using His-Bind Kits. (C) The purified recombinant Gly m 4l protein analyzed by immunoblotting using anti-His antibody.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4608668&req=5

pone.0140364.g005: Analysis of the purified recombinant Gly m 4l protein.(A) The recombinant Gly m 4l protein induced with 0.5 mM IPTG at 37°C for 1–5 h in E.coli BL21 (DE3). (B) SDS-PAGE analysis of the purified recombinant Gly m 4l protein using His-Bind Kits. (C) The purified recombinant Gly m 4l protein analyzed by immunoblotting using anti-His antibody.

Mentions: The expression of the recombinant Gly m 4l protein was remarkably enhanced after induction with 0.5 mM IPTG at 37°C for 1–5 h, reaching the maximum expression at 5 h, and the recombinant Gly m 4l protein was not detected in the control groups (Fig 5A). The recombinant Gly m 4l protein was purified using His-Bind Kits (EMD Millipore, USA), and the molecular weight of the purified protein was about 17 kDa in SDS-PAGE, consistent with the calculated molecular mass (17.14 kDa) (Fig 5B). Meanwhile, the Gly m 4l protein was also immunoprecipitated using anti-His antibody, and a single band appeared on the X-ray film in the regular western blotting (Fig 5C).


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)

Analysis of the purified recombinant Gly m 4l protein.(A) The recombinant Gly m 4l protein induced with 0.5 mM IPTG at 37°C for 1–5 h in E.coli BL21 (DE3). (B) SDS-PAGE analysis of the purified recombinant Gly m 4l protein using His-Bind Kits. (C) The purified recombinant Gly m 4l protein analyzed by immunoblotting using anti-His antibody.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140364.g005: Analysis of the purified recombinant Gly m 4l protein.(A) The recombinant Gly m 4l protein induced with 0.5 mM IPTG at 37°C for 1–5 h in E.coli BL21 (DE3). (B) SDS-PAGE analysis of the purified recombinant Gly m 4l protein using His-Bind Kits. (C) The purified recombinant Gly m 4l protein analyzed by immunoblotting using anti-His antibody.
Mentions: The expression of the recombinant Gly m 4l protein was remarkably enhanced after induction with 0.5 mM IPTG at 37°C for 1–5 h, reaching the maximum expression at 5 h, and the recombinant Gly m 4l protein was not detected in the control groups (Fig 5A). The recombinant Gly m 4l protein was purified using His-Bind Kits (EMD Millipore, USA), and the molecular weight of the purified protein was about 17 kDa in SDS-PAGE, consistent with the calculated molecular mass (17.14 kDa) (Fig 5B). Meanwhile, the Gly m 4l protein was also immunoprecipitated using anti-His antibody, and a single band appeared on the X-ray film in the regular western blotting (Fig 5C).

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