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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

Antimicrobial activity of the recombinant Gly m 4l protein.(A) The antimicrobial activity of the recombinant Gly m 4l protein against the hyphal growth of P. sojae. 1, 25 μg boiled Gly m 4l protein. 2, 15 μg Gly m 4l protein; 3, Elution buffer. 4, 25 μg Gly m 4l protein. (B) The antimicrobial activity of the recombinant Gly m 4l protein against zoospore release of P. sojae. The experiments were performed on three technical replicates and statistically analysed using Student’s t-test (*P<0.05, **P<0.01). Bars indicate standard error of the mean (SE).
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pone.0140364.g007: Antimicrobial activity of the recombinant Gly m 4l protein.(A) The antimicrobial activity of the recombinant Gly m 4l protein against the hyphal growth of P. sojae. 1, 25 μg boiled Gly m 4l protein. 2, 15 μg Gly m 4l protein; 3, Elution buffer. 4, 25 μg Gly m 4l protein. (B) The antimicrobial activity of the recombinant Gly m 4l protein against zoospore release of P. sojae. The experiments were performed on three technical replicates and statistically analysed using Student’s t-test (*P<0.05, **P<0.01). Bars indicate standard error of the mean (SE).

Mentions: In vitro, inhibition activity of the recombinant Gly m 4l protein against hyphal growth of P. sojae was assayed, and hyphal growth was monitored from 24 to 72 h. After incubation for 72 h, 2–3 mm and 4–6 mm zones with inhibited hyphal growth were detected with 15 μg and 25 μg of the recombinant protein, respectively (Fig 7A). No inhibition of the growth of P. sojae was observed using discs containing elution buffer or boiled protein (Fig 7A). Antimicrobial activity of the recombinant Gly m 4l protein against P. sojae zoospores was also assayed. The amount of zoospores was significantly decreased with the presence of 100 μg recombinant Gly m 4l protein, compared to that with the presence of 100 μg boiled Gly m 4l protein or buffer alone (Fig 7B). These results suggested that the recombinant Gly m 4l protein possessed antibiotic activity against P. sojae through the inhibition of hyphal growth and zoospore release.


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)

Antimicrobial activity of the recombinant Gly m 4l protein.(A) The antimicrobial activity of the recombinant Gly m 4l protein against the hyphal growth of P. sojae. 1, 25 μg boiled Gly m 4l protein. 2, 15 μg Gly m 4l protein; 3, Elution buffer. 4, 25 μg Gly m 4l protein. (B) The antimicrobial activity of the recombinant Gly m 4l protein against zoospore release of P. sojae. The experiments were performed on three technical replicates and statistically analysed using Student’s t-test (*P<0.05, **P<0.01). Bars indicate standard error of the mean (SE).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140364.g007: Antimicrobial activity of the recombinant Gly m 4l protein.(A) The antimicrobial activity of the recombinant Gly m 4l protein against the hyphal growth of P. sojae. 1, 25 μg boiled Gly m 4l protein. 2, 15 μg Gly m 4l protein; 3, Elution buffer. 4, 25 μg Gly m 4l protein. (B) The antimicrobial activity of the recombinant Gly m 4l protein against zoospore release of P. sojae. The experiments were performed on three technical replicates and statistically analysed using Student’s t-test (*P<0.05, **P<0.01). Bars indicate standard error of the mean (SE).
Mentions: In vitro, inhibition activity of the recombinant Gly m 4l protein against hyphal growth of P. sojae was assayed, and hyphal growth was monitored from 24 to 72 h. After incubation for 72 h, 2–3 mm and 4–6 mm zones with inhibited hyphal growth were detected with 15 μg and 25 μg of the recombinant protein, respectively (Fig 7A). No inhibition of the growth of P. sojae was observed using discs containing elution buffer or boiled protein (Fig 7A). Antimicrobial activity of the recombinant Gly m 4l protein against P. sojae zoospores was also assayed. The amount of zoospores was significantly decreased with the presence of 100 μg recombinant Gly m 4l protein, compared to that with the presence of 100 μg boiled Gly m 4l protein or buffer alone (Fig 7B). These results suggested that the recombinant Gly m 4l protein possessed antibiotic activity against P. sojae through the inhibition of hyphal growth and zoospore release.

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