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Title: RALF peptide signaling controls the polytubey block in Arabidopsis
Signaling between pollen tube and female gametophyte ensures that only one pollen tube gets through but can re-establish access in case of failure.  more » « less
Award ID(s):
1645858
NSF-PAR ID:
10323429
Author(s) / Creator(s):
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Date Published:
Journal Name:
Science
Volume:
375
Issue:
6578
ISSN:
0036-8075
Page Range / eLocation ID:
290 to 296
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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  1. Abstract

    Differences in pollen performance, often revealed during pollen competition, have long been recognized as evolutionarily significant and agriculturally important. Though we have sophisticated models for the growth of individual pollen tubes, we have few robust models for larger-scale pollen competition, a process that has been linked with inbreeding avoidance, sexual selection, reproductive barrier reinforcement and speciation. Here we use existing data on pollen performance traits to develop an agent-based model of pollen competition. We calibrate our model parameters to empirical data found in the literature of seed siring proportions from mixed pollinations and pollen tube length distributions from single-accession pollinations. In this model, parameters that influence pollen tube movement and sensing of ovules were found to be primary factors in competition. Our model also demonstrates that interference competition emerges as a property of pollen competition, and suggests a potential mechanism for this phenomenon. This study integrates pollen performance measures with mathematical modelling conducted on a simplified and accessible system. This represents the first mechanistic agent-based model for pollen competition. Our model may be extended to predict seed siring proportions for other accessions of Arabidopsis thaliana given data on their pollen performance traits.

     
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    Cells employ multiple systems to maintain cellular integrity, including mechanosensitive ion channels and the cell wall integrity (CWI) pathway. Here, we use pollen as a model system to ask how these different mechanisms are interconnected at the cellular level. MscS-Like 8 (MSL8) is a mechanosensitive channel required to protect Arabidopsis thaliana pollen from osmotic challenges during in vitro rehydration, germination, and tube growth. New CRISPR/Cas9 and artificial miRNA-generated msl8 alleles produced unexpected pollen phenotypes, including the ability to germinate a tube after bursting, dramatic defects in cell wall structure, and disorganized callose deposition at the germination site. We document complex genetic interactions between MSL8 and two previously established components of the CWI pathway, MARIS and ANXUR1/2. Overexpression of MARISR240C-FP suppressed the bursting, germination, and callose deposition phenotypes of msl8 mutant pollen. Null msl8 alleles suppressed the internalized callose structures observed in MARISR240C-FP lines. Similarly, MSL8-YFP overexpression suppressed bursting in the anxur1/2 mutant background, while anxur1/2 alleles reduced the strong rings of callose around ungerminated pollen grains in MSL8-YFP overexpressors. These data show that mechanosensitive ion channels modulate callose deposition in pollen and provide evidence that cell wall and membrane surveillance systems coordinate in a complex manner to maintain cell integrity.

     
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    The collaborative non‐self‐recognition model for S‐RNase‐based self‐incompatibility predicts that multiple S‐locus F‐box proteins (SLFs) produced by pollen of a givenS‐haplotype collectively mediate ubiquitination and degradation of all non‐self S‐RNases, but not self S‐RNases, in the pollen tube, thereby resulting in cross‐compatible pollination but self‐incompatible pollination. We had previously used pollen extracts containingGFP‐fused S2SLF1 (SLF1 with anS2‐haplotype) ofPetunia inflatafor co‐immunoprecipitation (Co‐IP) and mass spectrometry (MS), and identified PiCUL1‐P (a pollen‐specific Cullin1), PiSSK1 (a pollen‐specific Skp1‐like protein) and PiRBX1 (a conventional Rbx1) as components of theSCFS2–SLF1complex. Using pollen extracts containing PiSSK1:FLAG:GFPfor Co‐IP/MS, we identified two additionalSLFs (SLF4 andSLF13) that were assembled intoSCFSLFcomplexes. As 17SLFgenes (SLF1toSLF17) have been identified inS2andS3pollen, here we examined whether all 17SLFs are assembled into similar complexes and, if so, whether these complexes are unique toSLFs. We modified the previous Co‐IP/MSprocedure, including the addition of style extracts from four differentS‐genotypes to pollen extracts containing PiSSK1:FLAG:GFP, to perform four separate experiments. The results taken together show that all 17SLFs and anSLF‐like protein,SLFLike1 (encoded by anS‐locus‐linked gene), co‐immunoprecipitated with PiSSK1:FLAG:GFP. Moreover, of the 179 other F‐box proteins predicted byS2andS3pollen transcriptomes, only a pair with 94.9% identity and another pair with 99.7% identity co‐immunoprecipitated with PiSSK1:FLAG:GFP. These results suggest thatSCFSLFcomplexes have evolved specifically to function in self‐incompatibility.

     
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