Abstract
Reprogramming of differentiated root cortical cells into proliferative stem cells is a prerequisite for legume nodule organogenesis, yet the molecular trigger that confers stem-cell identity upon these cortical cells remains elusive. Here we demonstrate that, in soybean (Glycine max), the canonical root stem-cell regulator WUSCHEL-RELATED HOMEOBOX gene WOX5 is activated by rhizobia specifically in cortical cells that will give rise to nodule primordia. CRISPR-Cas9-mediated knockout of the three WOX5 homologs, wox5abc mutants, reduced nodule number and attenuated nitrogenase activity, attributable to a decrease in primordium density rather than impaired rhizobial infection. Promoter dissection identified a 442 bp legume-specific promoter fragment within the WOX5a promoter that is both necessary and sufficient for primordium-specific expression. Chromatin immunoprecipitation and dual-luciferase assays revealed that this promoter fragment is directly bound by the symbiosis-responsive transcription factor NF-YAc to activate expression of WOX5a. Loss of NF-YAc phenocopied wox5abc, and NF-YAc overexpression failed to rescue nodulation in wox5abc mutants. Collectively, our findings reveal that NF-YAc-mediated activation of WOX5 initiates a de novo stem-cell niche in root cortical cells, providing the critical developmental trigger for nodule primordium initiation in soybean.
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Claims 10
Claim · model-written summary
WOX5 is activated by rhizobia specifically in cortical cells that give rise to nodule primordia in soybean.
- Taxon
- soybean
Glycine maxt0 - Organ / stage
- cortical cellse1
- Interactor
- rhizobiae1symbiont
- Gene
- WOX5e1
- Outcome
- WOX5e1
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e1“the canonical root stem-cell regulator WUSCHEL-RELATED HOMEOBOX gene WOX5 is activated by rhizobia specifically in cortical cells that will give rise to nodule primordia”
Claim · model-written summary
Knockout of the three WOX5 homologs reduced nodule number in soybean.
- Taxon
- soybeanGlycine maxt0
- Material
- wox5abc mutantse2
- Treatment
- CRISPR-Cas9-mediated knockout of the three WOX5 homologse2
- Gene
- WOX5e2
- Outcome
- nodule numbere2
- Direction
- reducede2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e2“CRISPR-Cas9-mediated knockout of the three WOX5 homologs, wox5abc mutants, reduced nodule number and attenuated nitrogenase activity”
Claim · model-written summary
Knockout of the three WOX5 homologs attenuated nitrogenase activity in soybean.
- Taxon
- soybeanGlycine maxt0
- Material
- wox5abc mutantse2
- Treatment
- CRISPR-Cas9-mediated knockout of the three WOX5 homologse2
- Outcome
- nitrogenase activitye2
- Direction
- attenuatede2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e2“CRISPR-Cas9-mediated knockout of the three WOX5 homologs, wox5abc mutants, reduced nodule number and attenuated nitrogenase activity”
Claim · model-written summary
The reduced nodule number in wox5abc mutants is attributable to a decrease in primordium density.
- Taxon
- soybeanGlycine maxt0
- Organ / stage
- primordiume3
- Outcome
- primordium densitye3
- Direction
- decreasee3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e3“attributable to a decrease in primordium density rather than impaired rhizobial infection”
Claim · model-written summary
Loss of WOX5 function does not impair rhizobial infection in soybean.
- Taxon
- soybeanGlycine maxt0
- Outcome
- rhizobial infectione3
- Direction
- rather than impairede3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e3“attributable to a decrease in primordium density rather than impaired rhizobial infection”
Claim · model-written summary
A 442 bp legume-specific fragment of the WOX5a promoter is necessary and sufficient for primordium-specific expression.
- Taxon
- soybeanGlycine maxt0
- Organ / stage
- primordiume4
- Gene
- WOX5ae4
- Outcome
- primordium-specific expressione4
- Qualifier
- necessary and sufficiente4
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e4“a 442 bp legume-specific promoter fragment within the WOX5a promoter that is both necessary and sufficient for primordium-specific expression”
Claim · model-written summary
NF-YAc directly binds the WOX5a promoter fragment and activates expression of WOX5a.
- Taxon
- soybeanGlycine maxt0
- Gene
- NF-YAce5
- Outcome
- expression of WOX5ae5
- Direction
- activatee5
- Qualifier
- directly bounde5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e5“Chromatin immunoprecipitation and dual-luciferase assays revealed that this promoter fragment is directly bound by the symbiosis-responsive transcription factor NF-YAc to activate expression of WOX5a”
Claim · model-written summary
Loss of NF-YAc phenocopies the wox5abc mutant in soybean.
- Taxon
- soybeanGlycine maxt0
- Treatment
- Loss of NF-YAce6
- Outcome
- phenocopiede6
- Direction
- phenocopiede6
- Comparator
- wox5abce6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e6“Loss of NF-YAc phenocopied wox5abc”
Claim · model-written summary
NF-YAc overexpression fails to rescue nodulation in wox5abc mutants.
- Taxon
- soybeanGlycine maxt0
- Treatment
- NF-YAc overexpressione7
- Outcome
- nodulatione7
- Direction
- failed to rescuee7
- Comparator
- wox5abc mutantse7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e7“NF-YAc overexpression failed to rescue nodulation in wox5abc mutants”
Claim · model-written summary
NF-YAc-mediated activation of WOX5 initiates a de novo stem-cell niche in root cortical cells.
- Taxon
- soybeanGlycine maxt0
- Organ / stage
- root cortical cellse2
- Gene
- WOX5e2
- Outcome
- de novo stem-cell nichee2
- Direction
- initiatese2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in soybean”
e2“NF-YAc-mediated activation of WOX5 initiates a de novo stem-cell niche in root cortical cells”