INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) and its receptors HAE/HSL2 are required for cell expansion and cell separation during floral organ abscission in Arabidopsis thaliana (part 2)
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ABSTRACT: Plants have the ability to shed organs that are no longer in use. In Arabidopsis thaliana abscission of floral organs involves cell wall remodeling and cell expansion prior to cell wall dissolution. IDA encodes a secreted peptide that signals through the leucine-rich repeat receptor-like kinases (LRR-RLKs) HAESA (HAE) (At4g28490) and HASEA-LIKE2 (HSL2) (At5g65710).
Project description:Plants have the ability to shed organs that are no longer in use. In Arabidopsis thaliana abscission of floral organs involves cell wall remodeling and cell expansion prior to cell wall dissolution. IDA encodes a secreted peptide that signals through the leucine-rich repeat receptor-like kinases (LRR-RLKs) HAESA (HAE) (At4g28490) and HASEA-LIKE2 (HSL2) (At5g65710).
Project description:Plants have the ability to shed organs that are no longer in use. In Arabidopsis thaliana abscission of floral organs involves cell wall remodeling and cell expansion prior to cell wall dissolution. IDA encodes a secreted peptide that signals through the leucine-rich repeat receptor-like kinases (LRR-RLKs) HAESA (HAE) (At4g28490) and HASEA-LIKE2 (HSL2) (At5g65710). Arabidopsis thaliana (ecotype Colombia-0) plants were kept in growth chambers with a 16/8 h (light/dark) photoperiod at 22 M-BM-0C, and 100 mE m-2 s-1 light intensity. 4 biological replicates were prepared from each sample, each containing abscission zone regions of siliques position 4 to 8 (when counting from the flowe at anthesis at the top of the inflorescence) from plants with at least 20 siliques. Differences in transcriptional responses were measured by comparing genes expression in abscission zones of ida-2 plants (SALK_133209) against abscission zones from control plants.
Project description:Plants have the ability to shed organs that are no longer in use. In Arabidopsis thaliana abscission of floral organs involves cell wall remodeling and cell expansion prior to cell wall dissolution. IDA encodes a secreted peptide that signals through the leucine-rich repeat receptor-like kinases (LRR-RLKs) HAESA (HAE) (At4g28490) and HASEA-LIKE2 (HSL2) (At5g65710). Arabidopsis thaliana (ecotype Colombia-0) plants were kept in growth chambers with a 16/8 h (light/dark) photoperiod at 22 M-BM-0C, and 100 mE m-2 s-1 light intensity. 4 biological replicates were prepared from each sample, each containing abscission zone regions of siliques position 4 to 8 (when counting from the flowe at anthesis at the top of the inflorescence) from plants with at least 20 siliques. Differences in transcriptional responses were measured by comparing genes expression in abscission zones of hae hsl2 plants (SALK_021905 x SALK_030520) against abscission zones from control plants.
Project description:The receptor-like protein kinases encoded by HAESA (HAE) and HAESA-LIKE 2 (HSL2) are essential for floral organ abscission in Arabidopsis thaliana and the double hae hsl2 mutant fails to abscise. Expression of HAE and HSL2 is specific to Abscission Zone (AZ) cells and is higher in stage 15 flowers than in earlier developmental stages. By stage 16 floral organs have begun to abscise, suggesting that HAE HSL2 are most active in stage 15 flowers. Samples were enriched for AZ RNA by isolating RNA from flower receptacles, the region from the base of the flower to slightly above the base of attachment of the sepals, petals, and stamen. RNA-seq was then used to analyze and compare the transcriptomes of wild type and hae-3 hsl2-3 mutants. 2034 genes were differentially expressed with a False Discovery Rate adjusted p < 0.05, of which 349 genes 2 fold or greater change. Of these 349, 277 were lower in the mutant and 72 were higher. Differentially expressed genes with lower expression were enriched for hydrolytic enzymes, cell-wall modifying enzymes, and defense related genes. This suggests that HAE HSL2 signaling regulates gene expression of enzymes necessary for abscission.
Project description:The receptor-like protein kinases encoded by HAESA (HAE) and HAESA-LIKE 2 (HSL2) are essential for floral organ abscission in Arabidopsis thaliana and the double hae hsl2 mutant fails to abscise. Expression of HAE and HSL2 is specific to Abscission Zone (AZ) cells and is higher in stage 15 flowers than in earlier developmental stages. By stage 16 floral organs have begun to abscise, suggesting that HAE HSL2 are most active in stage 15 flowers. Samples were enriched for AZ RNA by isolating RNA from flower receptacles, the region from the base of the flower to slightly above the base of attachment of the sepals, petals, and stamen. RNA-seq was then used to analyze and compare the transcriptomes of wild type and hae-3 hsl2-3 mutants. 2034 genes were differentially expressed with a False Discovery Rate adjusted p < 0.05, of which 349 genes 2 fold or greater change. Of these 349, 277 were lower in the mutant and 72 were higher. Differentially expressed genes with lower expression were enriched for hydrolytic enzymes, cell-wall modifying enzymes, and defense related genes. This suggests that HAE HSL2 signaling regulates gene expression of enzymes necessary for abscission. 6 samples were sequenced, 3 biological replicates of Col-0 wild type and 3 biological replicates of the hae-3 hsl2-3 double mutant. Samples were barcoded and all 6 samples multiplexed and sequenced on 3 lanes, each lane on a separate flow cell, of an Illumina HiSeq 2000.
Project description:Throughout their life cycle, plants produce new organs, such as leaves, flowers, and lateral roots. Organs that have served their purpose may be shed after breakdown of primary cell walls between adjacent cell files at the site of detachment. In Arabidopsis, floral organs abscise after pollination, and this cell separation event is controlled by the peptide INFLORESCENCE DEFICIENT IN ABSCISSION (IDA), which signals through the leucine-rich repeat receptor-like kinases HAESA (HAE) and HAESA-LIKE2 (HSL2). Emergence of new lateral root primordia, initiated deep inside the root under the influence of auxin, is similarly dependent on cell wall dissolution between cells in the overlaying endodermal, cortical, and epidermal tissues. Here we show that this process requires IDA, HAE, and HSL2. Mutation in these genes constrains the passage of the growing lateral root primordia through the overlaying layers, resulting in altered shapes of the lateral root primordia and of the overlaying cells. The HAE and HSL2 receptors are redundant in function during floral organ abscission, but during lateral root emergence they are differentially involved in regulating cell wall remodeling genes. In the root, IDA is strongly auxin-inducible and dependent on key regulators of lateral root emergence--the auxin influx carrier LIKE AUX1-3 and AUXIN RESPONSE FACTOR7. The expression levels of the receptor genes are only transiently induced by auxin, suggesting they are limiting factors for cell separation. We conclude that elements of the same cell separation signaling module have been adapted to function in different developmental programs.
Project description:The peptide INFLORESCENCE DEFICIENT IN ABSCISSION (IDA), which signals through the leucine-rich repeat receptor-like kinases HAESA (HAE) and HAESA-LIKE2 (HSL2), controls different cell separation events in Arabidopsis thaliana. We hypothesize the involvement of this signaling module in abscission processes in other plant species even though they may shed other organs than A. thaliana. As the first step toward testing this hypothesis from an evolutionarily perspective we have identified genes encoding putative orthologs of IDA and its receptors by BLAST searches of publically available protein, nucleotide and genome databases for angiosperms. Genes encoding IDA or IDA-LIKE (IDL) peptides and HSL proteins were found in all investigated species, which were selected as to represent each angiosperm order with available genomic sequences. The 12 amino acids representing the bioactive peptide in A. thaliana have virtually been unchanged throughout the evolution of the angiosperms; however, the number of IDL and HSL genes varies between different orders and species. The phylogenetic analyses suggest that IDA, HSL2, and the related HSL1 gene, were present in the species that gave rise to the angiosperms. HAE has arisen from HSL1 after a genome duplication that took place after the monocot-eudicots split. HSL1 has also independently been duplicated in the monocots, while HSL2 has been lost in gingers (Zingiberales) and grasses (Poales). IDA has been duplicated in eudicots to give rise to functionally divergent IDL peptides. We postulate that the high number of IDL homologs present in the core eudicots is a result of multiple whole genome duplications (WGD). We substantiate the involvement of IDA and HAE/HSL2 homologs in abscission by providing gene expression data of different organ separation events from various species.
Project description:BackgroundAbscission is a mechanism by which plants shed entire organs in response to both developmental and environmental signals. Arabidopsis thaliana, in which only the floral organs abscise, has been used extensively to study the genetic, molecular and cellular processes controlling abscission. Abscission in Arabidopsis requires two genes that encode functionally redundant receptor-like protein kinases, HAESA (HAE) and HAESA-LIKE 2 (HSL2). Double hae hsl2 mutant plants fail to abscise their floral organs at any stage of floral development and maturation.ResultsUsing RNA-Seq, we compare the transcriptomes of wild-type and hae hsl2 stage 15 flowers, using the floral receptacle which is enriched for abscission zone cells. 2034 genes were differentially expressed with a False Discovery Rate adjusted p < 0.05, of which 349 had two fold or greater change in expression. Differentially expressed genes were enriched for hydrolytic, cell wall modifying, and defense related genes. Testing several of the differentially expressed genes in INFLORESCENCE DEFICIENT IN ABSCISSION (ida) mutants shows that many of the same genes are co-regulated by IDA and HAE HSL2 and support the role of IDA in the HAE and HSL2 signaling pathway. Comparison to microarray data from stamen abscission zones show distinct patterns of expression of genes that are dependent on HAE HSL2 and reveal HAE HSL2- independent pathways.ConclusionHAE HSL2-dependent and HAE HSL2-independent changes in genes expression are required for abscission. HAE and HSL2 affect the expression of cell wall modifying and defense related genes necessary for abscission. The HAE HSL2-independent genes also appear to have roles in abscission and additionally are involved in processes such as hormonal signaling, senescence and callose deposition.
Project description:The unwanted detachment of organs such as flowers, leaves, and fruits from the main body of a plant (abscission) has significant effects on agricultural practice. Both timely and precise regulation of organ abscission from a plant is crucial as it influences the agricultural yield. The tomato (Solanum lycopersicum) has become a model system for research on organ abscission. Here, we characterized four tomato natural abscission variants named jointless (j), functionally impaired jointless (fij), functionally impaired jointless like (fij like), and normal joint (NJ), based on their cellular features within the flower abscission zones (AZ). Using eight INFLORESCENCE DEFICIENT IN ABSCISSION (SlIDA) genes and eight HAESA genes (SlHAE) identified in the genome sequence of tomato, we analyzed the pattern of gene expression during flower abscission. The AZ-specific expression for three tomato abscission polygalacturonases (SlTAPGs) in the development of flower AZ, and the progression of abscission validated our natural abscission system. Compared to that of j, fij, and fij like variants, the AZ-specific expression for SlIDA, SlIDL2, SlIDL3, SlIDL4, and SlIDL5 in the NJ largely corelated and increased with the process of abscission. Of eight SlHAE genes examined, the expression for SlHSL6 and SlHSL7 were found to be AZ-specific and increased as abscission progressed in the NJ variant. Unlike the result of gene expression obtained from natural abscission system, an in silico analysis of transcriptional binding sites uncovered that SlIDA genes (SlIDA, SlIDL6, and SlIDL7) are predominantly under the control of environmental stress, while most of the SlHSL genes are affiliated with the broader context in developmental processes and stress responses. Our result presents the potential bimodal transcriptional regulation of the tomato IDA-HAE module associated with flower abscission in tomatoes.
Project description:BackgroundAbscission is an active, organized, and highly coordinated cell separation process enabling the detachment of aerial organs through the modification of cell-to-cell adhesion and breakdown of cell walls at specific sites on the plant body known as abscission zones. In Arabidopsis thaliana, abscission of floral organs and cauline leaves is regulated by the interaction of the hormonal peptide INFLORESCENCE DEFICIENT IN ABSCISSION (IDA), a pair of redundant receptor-like protein kinases, HAESA (HAE) and HAESA-LIKE2 (HSL2), and SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) co-receptors. However, the functionality of this abscission signaling module has not yet been demonstrated in other plant species.ResultsThe expression of the pair of NbenIDA1 homeologs and the receptor NbenHAE.1 was supressed at the base of the corolla tube by the inoculation of two virus-induced gene silencing (VIGS) constructs in Nicotiana benthamiana. These gene suppression events arrested corolla abscission but did not produce any obvious effect on plant growth. VIGS plants retained a higher number of corollas attached to the flowers than control plants, an observation related to a greater corolla breakstrength. The arrest of corolla abscission was associated with the preservation of the parenchyma tissue at the base of the corolla tube that, in contrast, was virtually collapsed in normal corollas. In contrast, the inoculation of a viral vector construct that increased the expression of NbenIDA1A at the base of the corolla tube negatively affected the growth of the inoculated plants accelerating the timing of both corolla senescence and abscission. However, the heterologous ectopic overexpression of citrus CitIDA3 and Arabidopsis AtIDA in N. benthamiana did not alter the standard plant phenotype suggesting that the proteolytic processing machinery was unable to yield active peptides.ConclusionHere, we demonstrate that the pair of NbenIDA1 homeologs encoding small peptides of the IDA-like family and the receptor NbenHAE.1 control cellular breakdown at the base of the corolla tube awhere an adventitious AZ should be formed and, therefore, corolla abscission in N. benthamiana flowers. Altogether, our results provide the first evidence supporting the notion that the IDA-HAE/HSL2 signaling module is conserved in angiosperms.