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

Response of Gly m 4l transgenic soybean plants to P. sojae.(A) Quantitative real-time PCR of the T2 transgenic soybean plants. (B) Disease symptoms on the leaves of the transgenic lines and non-transgenic lines treated with a P. sojae race 1 inoculum at 48 h and 96 h. (C) The lesion area of the transgenic lines and non-transgenic lines were detected after 96 h of incubation with P. sojae. (D) Quantitative real-time PCR of the T3 transgenic soybean plants. (E) Quantitative real-time PCR analysis of P. sojae relative biomass based on the transcript level of the P. sojae TEF1 gene. The experiment was performed on three biological replicates with their respective 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
getmorefigures.php?uid=PMC4608668&req=5

pone.0140364.g008: Response of Gly m 4l transgenic soybean plants to P. sojae.(A) Quantitative real-time PCR of the T2 transgenic soybean plants. (B) Disease symptoms on the leaves of the transgenic lines and non-transgenic lines treated with a P. sojae race 1 inoculum at 48 h and 96 h. (C) The lesion area of the transgenic lines and non-transgenic lines were detected after 96 h of incubation with P. sojae. (D) Quantitative real-time PCR of the T3 transgenic soybean plants. (E) Quantitative real-time PCR analysis of P. sojae relative biomass based on the transcript level of the P. sojae TEF1 gene. The experiment was performed on three biological replicates with their respective 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: To investigate whether over-expression of Gly m 4l enhances resistance in transgenic plants, the T1 transgenic soybean plants, confirmed through PCR and Southern hybridization (S2 Fig), were developed to T2 transgenic soybean plants and selected by qPCR (Fig 8A) to assay the pathogen response (S3 Fig). After 96 h incubation with P. sojae, the leaves of the non-transgenic soybean plants exhibited clear and large lesions compared to those of the transgenic plants (Fig 8B), and the lesion area of the transgenic soybean lines is significantly (P<0.01) smaller than that of non-transgenic soybean plants after 96 h incubation with P. sojae (Fig 8C). Moreover, the T3 transgenic soybean plants were confirmed by qPCR (Fig 8D), and the relative biomass of P. sojae in infected cotyledons after 48 h incubation with zoospores suspension of P. sojae was also analyzed. The results indicated that the biomass of P. sojae based on the transcript level of the P. sojae TEF1 gene was significantly (P<0.01) lower in Gly m 4l-overexpressing transgenic plants than that in non-transgenic ones (Fig 8E). These results indicated that the over-expression of Gly m 4l in soybean plants improved resistance to P. sojae.


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)

Response of Gly m 4l transgenic soybean plants to P. sojae.(A) Quantitative real-time PCR of the T2 transgenic soybean plants. (B) Disease symptoms on the leaves of the transgenic lines and non-transgenic lines treated with a P. sojae race 1 inoculum at 48 h and 96 h. (C) The lesion area of the transgenic lines and non-transgenic lines were detected after 96 h of incubation with P. sojae. (D) Quantitative real-time PCR of the T3 transgenic soybean plants. (E) Quantitative real-time PCR analysis of P. sojae relative biomass based on the transcript level of the P. sojae TEF1 gene. The experiment was performed on three biological replicates with their respective 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.g008: Response of Gly m 4l transgenic soybean plants to P. sojae.(A) Quantitative real-time PCR of the T2 transgenic soybean plants. (B) Disease symptoms on the leaves of the transgenic lines and non-transgenic lines treated with a P. sojae race 1 inoculum at 48 h and 96 h. (C) The lesion area of the transgenic lines and non-transgenic lines were detected after 96 h of incubation with P. sojae. (D) Quantitative real-time PCR of the T3 transgenic soybean plants. (E) Quantitative real-time PCR analysis of P. sojae relative biomass based on the transcript level of the P. sojae TEF1 gene. The experiment was performed on three biological replicates with their respective 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: To investigate whether over-expression of Gly m 4l enhances resistance in transgenic plants, the T1 transgenic soybean plants, confirmed through PCR and Southern hybridization (S2 Fig), were developed to T2 transgenic soybean plants and selected by qPCR (Fig 8A) to assay the pathogen response (S3 Fig). After 96 h incubation with P. sojae, the leaves of the non-transgenic soybean plants exhibited clear and large lesions compared to those of the transgenic plants (Fig 8B), and the lesion area of the transgenic soybean lines is significantly (P<0.01) smaller than that of non-transgenic soybean plants after 96 h incubation with P. sojae (Fig 8C). Moreover, the T3 transgenic soybean plants were confirmed by qPCR (Fig 8D), and the relative biomass of P. sojae in infected cotyledons after 48 h incubation with zoospores suspension of P. sojae was also analyzed. The results indicated that the biomass of P. sojae based on the transcript level of the P. sojae TEF1 gene was significantly (P<0.01) lower in Gly m 4l-overexpressing transgenic plants than that in non-transgenic ones (Fig 8E). These results indicated that the over-expression of Gly m 4l in soybean plants improved resistance to P. sojae.

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