Abstract
Potassium (K(+)) influx into pollen tubes via K(+) transporters is essential for pollen tube growth; however, the mechanism by which K(+) transporters are regulated in pollen tubes remains unknown. Here, we report that Arabidopsis thaliana Ca(2+)-dependent protein kinase11 (CPK11) and CPK24 are involved in Ca(2+)-dependent regulation of the inward K(+) (K(+)in) channels in pollen tubes. Using patch-clamp analysis, we demonstrated that K(+)in currents of pollen tube protoplasts were inhibited by elevated [Ca(2+)]cyt. However, disruption of CPK11 or CPK24 completely impaired the Ca(2+)-dependent inhibition of K(+)in currents and enhanced pollen tube growth. Moreover, the cpk11 cpk24 double mutant exhibited similar phenotypes as the corresponding single mutants, suggesting that these two CDPKs function in the same signaling pathway. Bimolecular fluorescence complementation and coimmunoprecipitation experiments showed that CPK11 could interact with CPK24 in vivo. Furthermore, CPK11 phosphorylated the N terminus of CPK24 in vitro, suggesting that these two CDPKs work together as part of a kinase cascade. Electrophysiological assays demonstrated that the Shaker pollen K(+)in channel is the main contributor to pollen tube K(+)in currents and acts as the downstream target of the CPK11-CPK24 pathway. We conclude that CPK11 and CPK24 together mediate the Ca(2+)-dependent inhibition of K(+)in channels and participate in the regulation of pollen tube growth in Arabidopsis.
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Claims 8
Claim · model-written summary
Elevated cytosolic calcium inhibited inward potassium currents.
- Taxon
- Arabidopsis thaliana
Arabidopsis thalianat0 - Organ / stage
- pollen tube protoplastse0
- Treatment
- elevated [Ca(2+)]cyte0
- Outcome
- K(+)in currentse0
- Direction
- inhibitede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“K(+)in currents of pollen tube protoplasts were inhibited by elevated [Ca(2+)]cyt.”
t0“Arabidopsis thaliana”
Claim · model-written summary
Disrupting CPK11 impaired calcium dependent current inhibition.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Treatment
- disruption of CPK11e0
- Gene
- CPK11e0
- Outcome
- Ca(2+)-dependent inhibition of K(+)in currentse0
- Direction
- impairede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“disruption of CPK11 or CPK24 completely impaired the Ca(2+)-dependent inhibition of K(+)in currents”
t0“Arabidopsis thaliana”
Claim · model-written summary
Disrupting CPK24 impaired calcium dependent current inhibition.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Treatment
- CPK24e0
- Gene
- CPK24e0
- Outcome
- Ca(2+)-dependent inhibition of K(+)in currentse0
- Direction
- impairede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“disruption of CPK11 or CPK24 completely impaired the Ca(2+)-dependent inhibition of K(+)in currents”
t0“Arabidopsis thaliana”
Claim · model-written summary
Kinase disruption enhanced pollen tube growth.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Treatment
- disruption of CPK11 or CPK24e0
- Outcome
- pollen tube growthe0
- Direction
- enhancede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“disruption of CPK11 or CPK24 completely impaired the Ca(2+)-dependent inhibition of K(+)in currents and enhanced pollen tube growth.”
t0“Arabidopsis thaliana”
Claim · model-written summary
The double mutant resembled single mutants.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Material
- cpk11 cpk24 double mutante0
- Outcome
- similar phenotypese0
- Comparator
- corresponding single mutantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the cpk11 cpk24 double mutant exhibited similar phenotypes as the corresponding single mutants”
t0“Arabidopsis thaliana”
Claim · model-written summary
CPK11 interacted with CPK24 in vivo.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Gene
- CPK11e0
- Outcome
- interact with CPK24e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“CPK11 could interact with CPK24 in vivo”
t0“Arabidopsis thaliana”
Claim · model-written summary
CPK11 phosphorylated CPK24 in vitro.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Gene
- CPK11e0
- Outcome
- phosphorylated the N terminus of CPK24e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“CPK11 phosphorylated the N terminus of CPK24 in vitro”
t0“Arabidopsis thaliana”
Claim · model-written summary
Shaker contributed most inward potassium current and was downstream of CPK11–CPK24.
- Taxon
- Arabidopsis thalianaArabidopsis thalianat0
- Gene
- Shaker pollen K(+)in channele0
- Outcome
- main contributor to pollen tube K(+)in currentse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the Shaker pollen K(+)in channel is the main contributor to pollen tube K(+)in currents and acts as the downstream target of the CPK11-CPK24 pathway.”
t0“Arabidopsis thaliana”
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