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Genetic interactions between SUPPRESSOR OF GENE SILENCING3 and ABSCISIC ACID HYPERSENSITIVE1/CAP-BINDING PROTEIN80 affecting fertilization, embryogenesis, germination, flowering time, and powdery mildew resistance

ABSTRACT: Arabidopsis ABA HYPERSENSITIVE1 (ABH1) / CAP-BINDING-PROTEIN80 functions in cap-binding, splicing, ABA and stress response signaling and in the biogenesis of microRNAs (miRNAs). SUPPRESSOR OF GENE SILENCING3 (SGS3) plays a crucial role in post transcriptional gene silencing (PTGS). We show that abh1 sgs3 double mutant manifests pleiotropic phenotypes including altered ABA sensitivity in seed germination, fused cotyledons, delayed growth in root initiation and leaf production, accelerated juvenile-to-adult phase transition and flowering time, short and bent siliques with ectopic carpelloid tissues, reduced seed set due to altered carpel structure and pollen tube growth, and increased resistance to powdery mildew infection. A systems approach using meta-analysis of whole genome tiling microarray datasets for mutants of PTGS, RNA processing, and responses to environmental stress was taken to design a 135k feature custom microarray to probe the molecular bases of these phenotypes. Results confirmed documented effects of ABA or abh1 on precursor MIRNA expression (pre-MIR159, pre-MIR168) and identified novel miRNAs and their targets regulated by ABA (pre-MIR408, LACCASE12 and -13; pre-MIR824) or elevated in abh1 sgs3 double mutant (pre-MIR156A,C; pre-MIR164B, pre-MIR167A, pre-MIR408, At5g15845/CONSTANS-LIKE1-cis-NAT-RNA). miR156 targets SQUAMOSA-PROMOTER-BINDING-PROTEIN-LIKE genes that spawn siRNAs and control phase change; miR164 targets CUP-SHAPED-COTYLEDON1 and -2 genes; miR166 methylates target PHABULOSA genomic DNA in trans; miR167 targets AUXIN RESPONSE FACTOR 6 (ARF6) and ARF8 which regulate both female and male reproduction; miR408 targets a plantacyanin which is expressed predominantly in the transmitting tract of the pistil and affects pollen tube guidance on the stigma. We also identified sgs3 by ABA interactions on gene expression which reveal potential nodes in regulatory networks linking ABA, mRNA processing and PTGS with development.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE42796 | GEO | 2014/11/19

SECONDARY ACCESSION(S): PRJNA183215

REPOSITORIES: GEO

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Project description:As in animals, cell-cell communication plays pivotal role in male-female recognition during plant sexual reproduction. Prelaid peptides secreted from the female reproductive tissues guide pollen tubes towards ovules for fertilization. However, the elaborate mechanisms for this dialogue have remained elusive, particularly from the male perspective. We perform genome-wide quantitative liquid chromatography coupled tandem mass spectrometry of a pistil-stimulated pollen tube secretome and identify 801 pollen tube-secreted proteins. Interestingly, in silico analysis reveals that the pollen tube-secretome is dominated by unconventional-type secreted proteins representing 57% of the total secretome. In support, we show that unconventional-type protein, translationally controlled tumor protein, is secreted to the apoplast. Remarkably, we discover that this protein could be secreted by infiltrating through the initial phases of the conventional secretory pathway and could reach the apoplast via exosomes as demonstrated by co-localization with Oleisin1 exosome marker. We demonstrate that Arabidopsis thaliana translationally controlled tumor protein-knockdown plants have pollen tubes that poorly navigate to the target ovule, and theÂ knocked down allele is poorly transmitted through the male. We show that regulators of the endoplasmic reticulum-trans-golgi network protein secretory pathway control secretion of pollen tube-secreted cysteine-rich proteins, including pollen tube attractants, and are essential for pollen tube growth and guidance, as well as ovule-targeting competence. This work, the first pollen tube secretome study, identifies novel genome-wide pollen tube-secreted proteins with potential function in pollen tube-ovule guidance for sexual reproduction. Functional analysis highlights a potential mechanism for pollen tube unconventional protein secretion and reveals likely regulators of pollen tube protein secretion. The association of pollen tube-secreted proteins with marker proteins shown to be secreted via exosomes in other species suggest secretion via exosomes as a possible mechanism for cell-cell communication between the pollen tube and female reproductive cells. For processed dataset with quantitative information, see Hafidh S, Potesil D, Fila J et al. Genome Bilogy 2016.

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Genetic interactions between SUPPRESSOR OF GENE SILENCING3 and ABSCISIC ACID HYPERSENSITIVE1/CAP-BINDING PROTEIN80 affecting fertilization, embryogenesis, germination, flowering time, and powdery mildew resistance

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