Limits...
Isolation and Characterization of a Novel Pathogenesis-Related Protein Gene (GmPRP) with Induced Expression in Soybean (Glycine max) during Infection with Phytophthora sojae.

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

Bottom Line: GmPRP was localized in the cell plasma membrane and cytoplasm.Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro.Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively.

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
Pathogenesis-related proteins (PR proteins) play crucial roles in the plant defense system. A novel PRP gene was isolated from highly resistant soybean infected with Phytophthora sojae (P. sojae) and was named GmPRP (GenBank accession number: KM506762). The amino acid sequences of GmPRP showed identities of 74%, 73%, 72% and 69% with PRP proteins from Vitis vinifera, Populus trichocarpa, Citrus sinensis and Theobroma cacao, respectively. Quantitative real-time reverse transcription PCR (qRT-PCR) data showed that the expression of GmPRP was highest in roots, followed by the stems and leaves. GmPRP expression was upregulated in soybean leaves infected with P. sojae. Similarly, GmPRP expression also responded to defense/stress signaling molecules, including salicylic acid (SA), ethylene (ET), abscisic acid (ABA) and jasmonic acid (JA). GmPRP was localized in the cell plasma membrane and cytoplasm. Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro. Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively. These results indicated that the GmPRP protein played an important role in the defense of soybean against P. sojae infection.

No MeSH data available.


Related in: MedlinePlus

Antimicrobial activity ribonuclease activity and assays for the recombinant GmPRP protein.(A) Inhibition of P. sojae race 1 growth by purified recombinant GmPRP. 1, 15 μg renatured recombinant GmPRP protein; 2, 25 μg renatured recombinant GmPRP protein; 3, 35μg renatured recombinant GmPRP protein; CK, 35 μg boiled renatured recombinant GmPRP protein. (B) Ribonuclease activities of recombinant GmPRP proteins on soybean RNA. Gel electrophoresis using 1.0% agarose gel was performed to separate hydrolyzed RNAs. Each reaction mixture containing total RNA from soybean was incubated for 2 h at 37°C. Lane 1, 5 μg RNA+ Elution buffer; Lane 2, 5 μg RNA+ 6 μg of boiled dialytically renatured GmPRP protein. Lane 3, 5 μg RNA+ 2 μg dialytically renatured GmPRP protein; Lane 4, 5 μg RNA+ 4 μg dialytically renatured GmPRP protein; Lane 5, 5 μg RNA+ 6 μg dialytically renatured GmPRP protein.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4482714&req=5

pone.0129932.g005: Antimicrobial activity ribonuclease activity and assays for the recombinant GmPRP protein.(A) Inhibition of P. sojae race 1 growth by purified recombinant GmPRP. 1, 15 μg renatured recombinant GmPRP protein; 2, 25 μg renatured recombinant GmPRP protein; 3, 35μg renatured recombinant GmPRP protein; CK, 35 μg boiled renatured recombinant GmPRP protein. (B) Ribonuclease activities of recombinant GmPRP proteins on soybean RNA. Gel electrophoresis using 1.0% agarose gel was performed to separate hydrolyzed RNAs. Each reaction mixture containing total RNA from soybean was incubated for 2 h at 37°C. Lane 1, 5 μg RNA+ Elution buffer; Lane 2, 5 μg RNA+ 6 μg of boiled dialytically renatured GmPRP protein. Lane 3, 5 μg RNA+ 2 μg dialytically renatured GmPRP protein; Lane 4, 5 μg RNA+ 4 μg dialytically renatured GmPRP protein; Lane 5, 5 μg RNA+ 6 μg dialytically renatured GmPRP protein.

Mentions: To examine the antimicrobial activity effect of the recombinant GmPRP protein on the growth of P. sojae 1, filter-paper discs containing recombinant GmPR10 protein (15, 25, 35 μg) were placed near the front of the growing hyphae of P. sojae 1. After incubation, a 2- to 3-mm zone with inhibited hyphal growth was detected when 25 μg of recombinant protein was applied to the filter-paper discs, and the bacteriostatic effect was enhanced by the application of 35 μg of the protein (Fig 5A). A control filter-paper disc containing boiled recombinant protein did not show an inhibitory effect.


Isolation and Characterization of a Novel Pathogenesis-Related Protein Gene (GmPRP) with Induced Expression in Soybean (Glycine max) during Infection with Phytophthora sojae.

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

Antimicrobial activity ribonuclease activity and assays for the recombinant GmPRP protein.(A) Inhibition of P. sojae race 1 growth by purified recombinant GmPRP. 1, 15 μg renatured recombinant GmPRP protein; 2, 25 μg renatured recombinant GmPRP protein; 3, 35μg renatured recombinant GmPRP protein; CK, 35 μg boiled renatured recombinant GmPRP protein. (B) Ribonuclease activities of recombinant GmPRP proteins on soybean RNA. Gel electrophoresis using 1.0% agarose gel was performed to separate hydrolyzed RNAs. Each reaction mixture containing total RNA from soybean was incubated for 2 h at 37°C. Lane 1, 5 μg RNA+ Elution buffer; Lane 2, 5 μg RNA+ 6 μg of boiled dialytically renatured GmPRP protein. Lane 3, 5 μg RNA+ 2 μg dialytically renatured GmPRP protein; Lane 4, 5 μg RNA+ 4 μg dialytically renatured GmPRP protein; Lane 5, 5 μg RNA+ 6 μg dialytically renatured GmPRP protein.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4482714&req=5

pone.0129932.g005: Antimicrobial activity ribonuclease activity and assays for the recombinant GmPRP protein.(A) Inhibition of P. sojae race 1 growth by purified recombinant GmPRP. 1, 15 μg renatured recombinant GmPRP protein; 2, 25 μg renatured recombinant GmPRP protein; 3, 35μg renatured recombinant GmPRP protein; CK, 35 μg boiled renatured recombinant GmPRP protein. (B) Ribonuclease activities of recombinant GmPRP proteins on soybean RNA. Gel electrophoresis using 1.0% agarose gel was performed to separate hydrolyzed RNAs. Each reaction mixture containing total RNA from soybean was incubated for 2 h at 37°C. Lane 1, 5 μg RNA+ Elution buffer; Lane 2, 5 μg RNA+ 6 μg of boiled dialytically renatured GmPRP protein. Lane 3, 5 μg RNA+ 2 μg dialytically renatured GmPRP protein; Lane 4, 5 μg RNA+ 4 μg dialytically renatured GmPRP protein; Lane 5, 5 μg RNA+ 6 μg dialytically renatured GmPRP protein.
Mentions: To examine the antimicrobial activity effect of the recombinant GmPRP protein on the growth of P. sojae 1, filter-paper discs containing recombinant GmPR10 protein (15, 25, 35 μg) were placed near the front of the growing hyphae of P. sojae 1. After incubation, a 2- to 3-mm zone with inhibited hyphal growth was detected when 25 μg of recombinant protein was applied to the filter-paper discs, and the bacteriostatic effect was enhanced by the application of 35 μg of the protein (Fig 5A). A control filter-paper disc containing boiled recombinant protein did not show an inhibitory effect.

Bottom Line: GmPRP was localized in the cell plasma membrane and cytoplasm.Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro.Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively.

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
Pathogenesis-related proteins (PR proteins) play crucial roles in the plant defense system. A novel PRP gene was isolated from highly resistant soybean infected with Phytophthora sojae (P. sojae) and was named GmPRP (GenBank accession number: KM506762). The amino acid sequences of GmPRP showed identities of 74%, 73%, 72% and 69% with PRP proteins from Vitis vinifera, Populus trichocarpa, Citrus sinensis and Theobroma cacao, respectively. Quantitative real-time reverse transcription PCR (qRT-PCR) data showed that the expression of GmPRP was highest in roots, followed by the stems and leaves. GmPRP expression was upregulated in soybean leaves infected with P. sojae. Similarly, GmPRP expression also responded to defense/stress signaling molecules, including salicylic acid (SA), ethylene (ET), abscisic acid (ABA) and jasmonic acid (JA). GmPRP was localized in the cell plasma membrane and cytoplasm. Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro. Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively. These results indicated that the GmPRP protein played an important role in the defense of soybean against P. sojae infection.

No MeSH data available.


Related in: MedlinePlus