Pollen tubes (PTs) grow by the targeted secretion of new cell wall material to their expanding tip region. Sec1/Munc18 (SM) proteins promote membrane fusion through regulation of the SNARE complex. We have previously shown that disruption of protein glycosylation in the
VPS45 belongs to the Sec1/Munc18 family of proteins, which interact with and regulate Qa-SNARE function during membrane fusion. We have shown previously that
As
We thus show that VPS45 is essential for proper tip growth and propose that the observed vacuolar defects lead to loss of the turgor pressure needed for tip growth.
- Award ID(s):
- 2040582
- NSF-PAR ID:
- 10485505
- Publisher / Repository:
- Frontiers
- Date Published:
- Journal Name:
- Frontiers in Plant Science
- Volume:
- 14
- ISSN:
- 1664-462X
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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SUMMARY Arabidopsis thaliana hpat1 hpat3 double mutant leads to PT growth defects that can be suppressed by reducing secretion. Here, we identified five point mutant alleles of the SM proteinSEC1A ashpat1/3 suppressors. The suppressors increased seed set, reduced PT growth defects and reduced the rate of glycoprotein secretion. In the absence of thehpat mutations,sec1a reduced pollen germination and PT elongation producing shorter and wider PTs. Consistent with a defect in membrane fusion,sec1a PTs accumulated secretory vesicles. Thoughsec1a had significantly reduced male transmission, homozygoussec1a plants maintained full seed set, demonstrating thatSEC1A was ultimately dispensable for pollen fertility. However, when combined with a mutation in anotherSEC1 ‐likeSM gene,keule , pollen fertility was totally abolished. Mutation insec1b , the final member of the Arabidopsis SEC1 clade, did not enhance thesec1a phenotype. Thus, SEC1A is the major SM protein promoting pollen germination and tube elongation, but in its absence KEULE can partially supply this activity. When we examined the expression of the SM protein family in other species for which pollen expression data were available, we found that at least one Sec1‐like protein was highly expressed in pollen samples, suggesting a conserved role in pollen fertility in other species. -
Summary HYDROXYPROLINE
O ‐ARABINOSYLTRANSFERASEs (HPATs) initiate a post‐translational protein modification (Hyp‐Ara) found abundantly on cell wall structural proteins. InArabidopsis thaliana ,HPAT1 andHPAT3 are redundantly required for full pollen fertility. In addition to the lack of Hyp‐Ara inhpat1/3 pollen tubes (PTs), we also found broadly disrupted cell wall polymer distributions, particularly the conversion of the tip cell wall to a more shaft‐like state. Mutant PTs were slow growing and prone to rupture and morphological irregularities. In a forward mutagenesis screen for suppressors of thehpat1/3 low seed‐set phenotype, we identified a missense mutation inexo70a2 , a predicted member of the vesicle‐tethering exocyst complex. The suppressed pollen had increased fertility, fewer morphological defects and partially rescued cell wall organization. A transcriptional null allele ofexo70a2 also suppressed thehpat1/3 fertility phenotype, as did mutants of core exocyst complex membersec15a , indicating that reduced exocyst function bypassed the PT requirement for Hyp‐Ara. In a wild‐type background,exo70a2 reduced male transmission efficiency, lowered pollen germination frequency and slowed PT elongation. EXO70A2 also localized to the PT tip plasma membrane, consistent with a role in exocyst‐mediated secretion. To monitor the trafficking of Hyp‐Ara modified proteins, we generated an HPAT‐targeted fluorescent secretion reporter. Reporter secretion was partially dependent onEXO70A2 and was significantly increased inhpat1/3 PTs compared with the wild type, but was reduced in the suppressedexo70a2 hpat1/3 tubes. -
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