Abstract
Phytophthora root rot, caused by Phytophthora sojae is a destructive disease of soybean ( Glycine max ) worldwide. We previously confirmed that the bHLH transcription factor GmPIB1 ( P. sojae -inducible bHLH transcription factor) reduces accumulation of reactive oxygen species (ROS) in cells by inhibiting expression of the peroxidase-related gene GmSPOD thus improving the resistance of hairy roots to P. sojae . To identify proteins interacting with GmPIB1 and assess their participation in the defense response to P. sojae , we obtained transgenic soybean hairy roots overexpressing GmPIB1 by Agrobacterium rhizogenes mediated transformation and examined GmPIB1 protein–protein interactions using immunoprecipitation combined with mass spectrometry. We identified 392 proteins likely interacting with GmPIB1 and selected 20 candidate genes, and only 26S proteasome regulatory subunit GmPSMD (Genbank accession no. XP_014631720 ) interacted with GmPIB1 in luciferase complementation and pull-down experiments and yeast two-hybrid assays. Overexpression of GmPSMD ( GmPSMD- OE) in soybean hairy roots remarkably improved resistance to P. sojae and RNA interference of GmPSMD ( GmPSMD -RNAi) increased susceptibility. In addition, accumulation of total ROS and hydrogen peroxide (H 2 O 2 ) in GmPSMD- OE transgenic soybean hairy roots were remarkably lower than those of the control after P. sojae infection. Moreover, in GmPSMD -RNAi transgenic soybean hairy roots, H 2 O 2 and the accumulation of total ROS exceeded those of the control. There was no obvious difference in superoxide anion (O 2 – ) content between control and transgenic hairy roots. Antioxidant enzymes include peroxidase (POD), glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT) are responsible for ROS scavenging in soybean. The activities of these antioxidant enzymes were remarkably higher in GmPSMD -OE transgenic soybean hairy roots than those in control, but were reduced in GmPSMD -RNAi transgenic soybean hairy roots. Moreover, the activity of 26S proteasome in GmPSMD -OE and GmPIB1 -OE transgenic soybean hairy roots was significantly higher than that in control and was significantly lower in PSMD -RNAi soybean hairy roots after P. sojae infection. These data suggest that GmPSMD might reduce the production of ROS by improving the activity of antioxidant enzymes such as POD, SOD, GPX, CAT, and GmPSMD plays a significant role in the response of soybean to P. sojae . Our study reveals a valuable mechanism for regulation of the pathogen response by the 26S proteasome in soybean.
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Claims 13
Claim · model-written summary
Immunoprecipitation combined with mass spectrometry identified 392 proteins likely interacting with GmPIB1.
- Taxon
- Soybean
Glycine maxt0 - Gene
- GmPIB1e0
- Outcome
- 392 proteins likely interacting with GmPIB1e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We identified 392 proteins likely interacting with GmPIB1”
t0“Soybean”
Claim · model-written summary
Of 20 candidate genes, only 26S proteasome regulatory subunit GmPSMD interacted with GmPIB1 in luciferase complementation, pull-down and yeast two-hybrid assays.
- Taxon
- SoybeanGlycine maxt0
- Gene
- GmPSMDe0
- Outcome
- interacted with GmPIB1e0
- Setting
- luciferase complementation and pull-down experiments and yeast two-hybrid assayse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“only 26S proteasome regulatory subunit GmPSMD (Genbank accession no. XP_014631720 ) interacted with GmPIB1 in luciferase complementation and pull-down experiments and yeast two-hybrid assays”
t0“Soybean”
Claim · model-written summary
Overexpression of GmPSMD in soybean hairy roots remarkably improved resistance to P. sojae.
- Taxon
- soybeanGlycine maxe0
- Treatment
- Overexpression of GmPSMDe0
- Interactor
- P. sojaePhytophthora sojaee0pathogen
- Gene
- GmPSMDe0
- Outcome
- resistance to P. sojaee0
- Direction
- improvede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Overexpression of GmPSMD ( GmPSMD- OE) in soybean hairy roots remarkably improved resistance to P. sojae”
Claim · model-written summary
RNA interference of GmPSMD increased susceptibility of soybean hairy roots.
- Taxon
- SoybeanGlycine maxt0
- Treatment
- RNA interference of GmPSMDe0
- Gene
- GmPSMDe0
- Outcome
- susceptibilitye0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“RNA interference of GmPSMD ( GmPSMD -RNAi) increased susceptibility”
t0“Soybean”
Claim · model-written summary
After P. sojae infection, accumulation of total ROS and hydrogen peroxide in GmPSMD-OE soybean hairy roots was remarkably lower than in the control.
- Taxon
- soybeanGlycine maxe0
- Treatment
- GmPSMD- OEe0
- Interactor
- P. sojaePhytophthora sojaee0pathogen
- Outcome
- accumulation of total ROS and hydrogen peroxide (H 2 O 2 )e0
- Direction
- lowere0
- Comparator
- the controle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“accumulation of total ROS and hydrogen peroxide (H 2 O 2 ) in GmPSMD- OE transgenic soybean hairy roots were remarkably lower than those of the control after P. sojae infection”
Claim · model-written summary
In GmPSMD-RNAi soybean hairy roots, H2O2 and total ROS accumulation exceeded those of the control.
- Taxon
- soybeanGlycine maxe0
- Treatment
- GmPSMD -RNAie0
- Outcome
- H 2 O 2 and the accumulation of total ROSe0
- Direction
- exceedede0
- Comparator
- the controle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“in GmPSMD -RNAi transgenic soybean hairy roots, H 2 O 2 and the accumulation of total ROS exceeded those of the control”
Claim · model-written summary
There was no obvious difference in superoxide anion content between control and transgenic soybean hairy roots.
- Taxon
- SoybeanGlycine maxt0
- Treatment
- transgenice0
- Outcome
- superoxide anion (O 2 – ) contente0
- Direction
- no obvious differencee0
- Comparator
- controle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“There was no obvious difference in superoxide anion (O 2 – ) content between control and transgenic hairy roots”
t0“Soybean”
Claim · model-written summary
Activities of antioxidant enzymes (POD, GPX, SOD, CAT) were remarkably higher in GmPSMD-OE soybean hairy roots than in the control.
- Taxon
- soybeanGlycine maxe0
- Treatment
- GmPSMD -OEe0
- Outcome
- activities of these antioxidant enzymese0
- Direction
- highere0
- Comparator
- controle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The activities of these antioxidant enzymes were remarkably higher in GmPSMD -OE transgenic soybean hairy roots than those in control, but were reduced in GmPSMD -RNAi transgenic soybean hairy roots”
Claim · model-written summary
Activities of antioxidant enzymes were reduced in GmPSMD-RNAi soybean hairy roots.
- Taxon
- soybeanGlycine maxe0
- Treatment
- GmPSMD -RNAie0
- Outcome
- activities of these antioxidant enzymese0
- Direction
- reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The activities of these antioxidant enzymes were remarkably higher in GmPSMD -OE transgenic soybean hairy roots than those in control, but were reduced in GmPSMD -RNAi transgenic soybean hairy roots”
Claim · model-written summary
After P. sojae infection, 26S proteasome activity was significantly higher in GmPSMD-OE and GmPIB1-OE soybean hairy roots than in the control.
- Taxon
- soybeanGlycine maxe0
- Treatment
- GmPSMD -OE and GmPIB1 -OEe0
- Interactor
- P. sojaePhytophthora sojaee0pathogen
- Outcome
- activity of 26S proteasomee0
- Direction
- highere0
- Comparator
- controle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the activity of 26S proteasome in GmPSMD -OE and GmPIB1 -OE transgenic soybean hairy roots was significantly higher than that in control and was significantly lower in PSMD -RNAi soybean hairy roots after P. sojae infection”
Claim · model-written summary
After P. sojae infection, 26S proteasome activity was significantly lower in PSMD-RNAi soybean hairy roots.
- Taxon
- soybeanGlycine maxe0
- Treatment
- PSMD -RNAie0
- Interactor
- P. sojaePhytophthora sojaee0pathogen
- Outcome
- activity of 26S proteasomee0
- Direction
- lowere0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the activity of 26S proteasome in GmPSMD -OE and GmPIB1 -OE transgenic soybean hairy roots was significantly higher than that in control and was significantly lower in PSMD -RNAi soybean hairy roots after P. sojae infection”
Claim · model-written summary
GmPSMD might reduce ROS production by improving the activity of antioxidant enzymes such as POD, SOD, GPX and CAT.
- Taxon
- SoybeanGlycine maxt0
- Gene
- GmPSMDe0
- Outcome
- production of ROSe0
- Qualifier
- improving the activity of antioxidant enzymes such as POD, SOD, GPX, CATe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GmPSMD might reduce the production of ROS by improving the activity of antioxidant enzymes such as POD, SOD, GPX, CAT”
t0“Soybean”
Claim · model-written summary
GmPSMD plays a significant role in the response of soybean to P. sojae.
- Taxon
- soybeanGlycine maxe0
- Interactor
- P. sojaePhytophthora sojaee0pathogen
- Gene
- GmPSMDe0
- Outcome
- response of soybean to P. sojaee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GmPSMD plays a significant role in the response of soybean to P. sojae”
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