Abscission Zone Cells Research Articles

Arabidopsis uses a molecular grounding mechanism and a biophysical circuit breaker to limit floral abscission signaling

Abscission is the programmed separation of plant organs. It is widespread in the plant kingdom with important functions in development and environmental response. In Arabidopsis, abscission of floral organs (sepals, petals, and stamens) is controlled by two receptor-like protein kinases HAESA (HAE) and HAESA LIKE-2 (HSL2), which orchestrate the programmed dissolution of the abscission zone connecting floral organs to the developing fruit. In this work, we use single-cell RNA sequencing to characterize the core HAE/HSL2 abscission gene expression program. We identify the MAP KINASE PHOSPHATASE-1/MKP1 gene as a negative regulator of this pathway. MKP1 acts prior to activation of HAE/HSL2 signaling to establish a signaling threshold required for the initiation of abscission. Furthermore, we use single-cell data to identify genes expressed in two subpopulations of abscission zone cells: those proximal and those distal to the plane of separation. We identify INFLORESCENCE DEFICIENT IN ABSCISSION/IDA family genes, encoding activating ligands of HAE/HSL2, as enriched in distal abscission zone cells at the base of the abscising organs. We show how this expression pattern forms a biophysical circuit breaker whereby, when the organ is shed, the source of the IDA peptides is removed, leading to cessation of HAE/HSL2 signaling. Overall, this work provides insight into the multiple control mechanisms acting on the abscission-signaling pathway.

The single RRM domain-containing protein SARP1 is required for establishment of the separation zone in Arabidopsis.

Abscission is the shedding of plant organs in response to developmental and environmental cues. Abscission involves cell separation between two neighboring cell types, residuum cells (RECs) and secession cells (SECs) in the floral abscission zone (AZ) in Arabidopsis thaliana. However, the regulatory mechanisms behind the spatial determination that governs cell separation are largely unknown. The class I KNOTTED-like homeobox (KNOX) transcription factor BREVIPEDICELLUS (BP) negatively regulates AZ cell size and number in Arabidopsis. To identify new players participating in abscission, we performed a genetic screen by activation tagging a weak complementation line of bp-3. We identified the mutant ebp1 (enhancer of BP1) displaying delayed floral organ abscission. The ebp1 mutant showed a concaved surface in SECs and abnormally stacked cells on the top of RECs, in contrast to the precisely separated surface in the wild-type. Molecular and histological analyses revealed that the transcriptional programming during cell differentiation in the AZ is compromised in ebp1. The SECs of ebp1 have acquired REC-like properties, including cuticle formation and superoxide production. We show that SEPARATION AFFECTING RNA-BINDING PROTEIN1 (SARP1) is upregulated in ebp1 and plays a role in the establishment of the cell separation layer during floral organ abscission in Arabidopsis.

A multifaceted kinase axis regulates plant organ abscission through conserved signaling mechanisms

Plants have evolved mechanisms to abscise organs as they develop or when exposed to unfavorable conditions.1 Uncontrolled abscission of petals, fruits, or leaves can impair agricultural productivity.2,3,4,5 Despite its importance for abscission progression, our understanding of the IDA signaling pathway and its regulation remains incomplete. IDA is secreted to the apoplast, where it is perceived by the receptors HAESA (HAE) and HAESA-LIKE2 (HSL2) and somatic embryogenesis receptor kinase (SERK) co-receptors.6,7,8,9 These plasma membrane receptors activate an intracellular cascade of mitogen-activated protein kinases (MAPKs) by an unknown mechanism.10,11,12 Here, we characterize brassinosteroid signaling kinases (BSKs) as regulators of floral organ abscission in Arabidopsis. BSK1 localizes to the plasma membrane of abscission zone cells, where it interacts with HAESA receptors to regulate abscission. Furthermore, we demonstrate that YODA (YDA) has a leading role among other MAPKKKs in controlling abscission downstream of the HAESA/BSK complex. This kinase axis, comprising a leucine-rich repeat receptor kinase, a BSK, and an MAPKKK, is known to regulate stomatal patterning, early embryo development, and immunity.10,13,14,15,16 How specific cellular responses are obtained despite signaling through common effectors is not well understood. We show that the identified abscission-promoting allele of BSK1 also enhances receptor signaling in other BSK-mediated pathways, suggesting conservation of signaling mechanisms. Furthermore, we provide genetic evidence supporting independence of BSK1 function from its kinase activity in several developmental processes. Together, our findings suggest that BSK1 facilitates signaling between plasma membrane receptor kinases and MAPKKKs via conserved mechanisms across multiple facets of plant development.

Deciphering the auxin-ethylene crosstalk in petal abscission through auxin influx carrier IpAUX1 of Itoh peony ‘Bartzella’

Auxin gradient on either side of the abscission zone play a crucial role in regulating organ abscission during plant senescence. Yet, the impact of auxin on the opening and senescence of cut peony flowers, specifically Itoh peony ‘Bartzella’, remains elusive. We employed the frozen section method to investigate the cell morphology within the petal abscission zone. Additionally, we quantified auxin levels via enzyme immunoassay and assessed the expression of genes related to indole-3-acetic acid (IAA) levels. Through our analyses, we pinpointed the critical gene, IpAUX1. Further investigation of IpAUX1 involved virus-induced gene silencing (VIGS) and transient overexpression techniques, allowing us to assess its role both in vitro flowers, through vacuum application, and in vivo flowers, via injection. Significant alterations were observed in the structure and cell morphology of the abscission zone correlating with the process of petal abscission, alongside noticeable auxin gradient. Silencing IpAUX1 led to a noticeable delay in petal abscission for both in vitro flowers and vivo flowers, while its transient overexpression hastened this process. This silencing effect was accompanied by a reduction in IAA levels around the abscission zone and a subsequent delay in the erasure of the auxin gradient. It also resulted in the downregulation of several genes: the auxin response factor IpARF1, ethylene synthesis-related genes IpACS1 and IpACO1, and cell wall hydrolysis-related genes IpPME1 and IpPG1. Conversely, the expression of genes involved in auxin synthesis gene IpYUCCA10, auxin efflux carrier IpPIN1, and the ethylene receptor IpETR1 saw significant upregulation. This study concludes that the auxin influx carrier IpAUX1 enhances the abscission zone cells' responsiveness to ethylene by modulating the auxin gradient. This action promotes cell hydrolysis within the abscission zone, thereby encouraging petal abscission. Our findings underscore the pivotal role of IpAUX1 as a positive regulator during the process of petal abscission.

Influence of the Cellular Ultrastructure and Enzyme Activity of the Leaf Sheath on Spontaneous Defoliation in Sugarcane

Spontaneous defoliation improves the harvesting efficiency and yield of sugarcane. Here, we investigated the ultrastructural changes and pectinase and cellulase activity in the third, fifth, and seventh leaf sheaths in four sugarcane varieties with varying spontaneous defoliation performance during maturation. At the early and middle stages of spontaneous defoliation, the cells in the abscission zones of the third, fifth, and seventh leaf sheaths were degrading. At the late stage, no complete organelles and hollow or broken spots in the cell walls were observed in the abscission zone cells of defoliation-prone varieties, while complete organelles and intact cell walls were present in the abscission zone cells at the same leaf positions in defoliation-resistant varieties. From the early to late stages, defoliation-prone varieties had higher pectinase activity in the abscission zones of the fifth and seventh leaf sheaths. At the early stage of defoliation, defoliation-prone varieties had significantly higher cellulase activity in the abscission zones of the third, fifth, and seventh leaf sheaths. Correlation analysis showed that the spontaneous defoliation rate was significantly positively correlated with pectinase activity in the leaf sheaths. In conclusion, the spontaneous defoliation of sugarcane was closely related to changes in cell morphology and pectinase activites in leaf sheaths.

Abscisic acid- and ethylene-induced abscission of yellow lupine flowers is mediated by jasmonates

The appropriate timing of organ abscission determines plant growth, development, reproductive success, and yield in relation to crop species. Among these, yellow lupine is an example of a crop species that loses many fully developed flowers, which limits the formation of pods with high-protein seeds and affects its economic value. Lupine flower abscission, similarly to the separation of other organs, depends on a complex regulatory network functioning in the cells of the abscission zone (AZ). In the present study, genetic, biochemical, and cellular methods were used to highlight the complexity of the interactions among strong hormonal stimulators of abscission, including abscisic acid (ABA), ethylene, and jasmonates (JAs) precisely in the AZ cells, with all results supporting that the JA-related pathway has an important role in the phytohormonal cross-talk leading to flower abscission in yellow lupine. Based on obtained results, we conclude that ABA and ET have positive influence on JAs biosynthesis and signaling pathway in time-dependent manner. Both phytohormones changes lipoxygenase (LOX) gene expression, affects LOX protein abundance, and JA accumulation in AZ cells. We have also shown that the signaling pathway of JA is highly sensitive to ABA and ET, given the accumulation of COI1 receptor and MYC2 transcription factor in response to these phytohormones. The results presented provide novel information about the JAs-dependent separation of organs and provide insight and details about the phytohormone-related mechanisms of lupine flower abscission.

Grain shattering by cell death and fracture in Eragrostis tef.

Abscission, known as shattering in crop species, is a highly regulated process by which plants shed parts. Although shattering has been studied extensively in cereals and a number of regulatory genes have been identified, much diversity in the process remains to be discovered. Teff (Eragrostis tef) is a crop native to Ethiopia that is potentially highly valuable worldwide for its nutritious grain and drought tolerance. Previous work has suggested that grain shattering in Eragrostis might have little in common with other cereals. In this study, we characterize the anatomy, cellular structure, and gene regulatory control of the abscission zone (AZ) in E. tef. We show that the AZ of E. tef is a narrow stalk below the caryopsis, which is common in Eragrostis species. X-ray microscopy, scanning electron microscopy, transmission electron microscopy, and immunolocalization of cell wall components showed that the AZ cells are thin walled and break open along with programmed cell death (PCD) at seed maturity, rather than separating between cells as in other studied species. Knockout of YABBY2/SHATTERING1, documented to control abscission in several cereals, had no effect on abscission or AZ structure in E. tef. RNA sequencing analysis showed that genes related to PCD and cell wall modification are enriched in the AZ at the early seed maturity stage. These data show that E. tef drops its seeds using a unique mechanism. Our results provide the groundwork for understanding grain shattering in Eragrostis and further improvement of shattering in E. tef.

Transcriptome Analysis of the Development of Pedicel Abscission Zone in Tomato

Plant organ abscission is a common phenomenon that occurs at a specific position called the abscission zone (AZ). The differentiation and development of the pedicel AZ play important roles in flower and fruit abscission, which are of great significance for abscission in tomatoes before harvest. Previous studies have reported some genes involved in AZ differentiation; however, the genes regulating pedicel AZ cell development in tomatoes after AZ differentiation remain poorly understood. In this study, transcriptome analyses of tomato pedicel AZ samples were performed at 0, 5, 15, and 30 days post-anthesis (DPA). Pedicel AZ growth was mainly observed before 15 DPA. A principal component analysis and a correlation analysis were carried out in order to compare the repeatability and reliability for different samples. We observed 38 up-regulated and 31 down-regulated genes that were significantly altered during 0 to 5 DPA, 5 to 15 DPA, and 0 to 15 DPA, which may play key roles in AZ cell enlargement. GO and KEGG enrichment analyses of the selected DEGs under all three different comparisons were conducted. Auxin-signaling-related genes were analyzed, as well as AUX/IAA, GH3, and small auxin up-regulated RNA (SAUR) gene expression patterns. The presented results provide information on pedicel AZ development, which might help in regulating flower or fruit pedicel abscission in tomato production facilities.

Floral organ abscission in Arabidopsis requires the combined activities of three TALE homeodomain transcription factors.

Floral organ abscission is a separation process in which sepals, petals, and stamens detach from the plant at abscission zones. Here, we investigated the collective role of three amino-acid-loop-extension (TALE) homeobox genes ARABIDOPSIS THALIANA HOMEOBOX GENE1 (ATH1), KNAT6 (for KNOTTED LIKE from Arabidopsis thaliana) and KNAT2, which form a module that patterns boundaries under the regulation of BLADE-ON-PETIOLE 1 and 2 (BOP1/2) co-activators. These TALE homeodomain transcription factors were shown to maintain boundaries in the flower, functioning as a unit to coordinate the growth, patterning, and activity of abscission zones. Together with BOP1 and BOP2, ATH1 and its partners KNAT6 and KNAT2 collectively contribute to the differentiation of lignified and separation layers of the abscission zone. The genetic interactions of BOP1/2 and ATH1 with INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) were also explored. We showed that BOP1/2 co-activators and ATH1 converge with the IDA signalling pathway to promote KNAT6 and KNAT2 expression in the abscission zone and cell separation. ATH1 acts as a central regulator in floral organ abscission as it controls the expression of other TALE genes in abscission zone cells.

EPIP as an abscission promoting agent in the phytohormonal pathway

Understanding the mechanisms underlying the activation of the abscission zone (AZ) responsible for organ separation from plant body in crop species will help improve their yielding and economic importance. Special attention has been given recently to the role of the INFLORESCENCE DEFICIENT IN ABSCISSION protein, particularly its functional fragment, EPIP peptide. Its stimulatory effect on abscission in different crops has been demonstrated. Recently we described the role of EPIP in the redox, lipid, and pectin-related events taking place in AZ of Lupinus luteus flowers, which undergo massive abscission in natural conditions. To further examine EPIP contribution in AZ functioning, here, we analyze its impact on the ultrastructural changes, synthesis of two hormonal abscission stimulators – abscisic acid (ABA) and ethylene (ET), and the appearance of phosphoproteins. As our results show, the response of flower AZ to exogenous EPIP involves the induction of distinct modifications related to the one hand with upregulation of cell activity but on the other hand degradation processes and possible autophagy. Furthermore, the EPIP stimulated biosynthesis pathways of ABA and ET precisely in AZ cells. In addition, progressive phosphorylation of proteins has been observed under EPIP influence. The highly accumulated ones were identified as those, related to primary metabolism and reactive oxygen species homeostasis, and their role in abscission has been discussed. To summarizing, the presented detailed description of EPIP action in AZ cells in combination with our previous data offers new insights into its regulatory function and provides opportunities to counteract excessive flower abscission in lupine.

The acceleration of yellow lupine flower abscission by jasmonates is accompanied by lipid-related events in abscission zone cells

Yellow lupine is an economically important crop. This species has been used as a great model for abscission processes for several years due to extreme flower abortion, which takes place in the abscission zone (AZ). AZ activation involves modifications of cell walls, membranes, and cellular structure. In this paper, we applied physiological, molecular, biochemical, and instrumental methods to explore lipid-associated changes and the possible involvement of lipid-derived phytohormones – jasmonates (JAs) - in flower AZ activation. Our comprehensive analyses revealed that natural abscission is accompanied by the upregulation of peroxidase, which reflects a disruption of redox balance and/or lipids peroxidation in AZ cell membranes. Redox imbalance was confirmed by appearance of malondialdehyde. Lipid-related processes involved the specific localization and increased level and activity of lipase and LOX, enzymes associated with cell membrane rupture, and JA biosynthesis. Lipid-hydrolyzing phospholipase D, implicated previously in abscission, is also found in naturally active AZs. Observed changes are accompanied by the accumulation of jasmonates, both free jasmonic acid and its methyl ester. The JA derivative exhibited higher biological activity than the nonconjugated form. Overall, our study shed new light on the lipid and phytohormonal regulation of AZ functioning supporting a role of JAs during abscission-associated events.

Cold stress triggers premature fruit abscission through ABA-dependent signal transduction in early developing apple.

Fruit abscission is a complex physiological process that is regulated by internal and environmental factors. During early development, apple fruit are exposed to extreme temperature fluctuations that are associated with premature fruit drop; however, their effect on fruit abscission is largely unknown. We hypothesized that fruit abscission is triggered by cold stress and investigated the molecular basis of premature fruit drop using RNA-Seq and metabolomics data from apple fruit undergoing abscission following cold stress in the field. Genes responsive to abscisic acid signaling and cell wall degradation were upregulated during abscission, consistent with the increased abscisic acid concentrations detected by liquid chromatography-mass spectrometry. We performed ex vivo cold shock experiments with excised tree subunits consisting of a branch, pedicel, and fruit. Abscission induction occurred in the cold-stressed subunits with concurrent upregulation of abscisic acid biosynthesis (MdNCED1) and metabolism (MdCYP707A) genes, and ethylene biosynthesis (MdACS1) and receptor (MdETR2) genes in the pedicel. Another key finding was the activation of cytoplasmic streaming in abscission-zone cells detected by electron microscopy. Our results provide a novel insight into the molecular basis of fruit abscission physiology in response to cold stress in apple.

EPIP-Evoked Modifications of Redox, Lipid, and Pectin Homeostasis in the Abscission Zone of Lupine Flowers.

Yellow lupine is a great model for abscission-related research given that excessive flower abortion reduces its yield. It has been previously shown that the EPIP peptide, a fragment of LlIDL (INFLORESCENCE DEFICIENT IN ABSCISSION) amino-acid sequence, is a sufficient molecule to induce flower abortion, however, the question remains: What are the exact changes evoked by this peptide locally in abscission zone (AZ) cells? Therefore, we used EPIP peptide to monitor specific modifications accompanied by early steps of flower abscission directly in the AZ. EPIP stimulates the downstream elements of the pathway—HAESA and MITOGEN-ACTIVATED PROTEIN KINASE6 and induces cellular symptoms indicating AZ activation. The EPIP treatment disrupts redox homeostasis, involving the accumulation of H2O2 and upregulation of the enzymatic antioxidant system including superoxide dismutase, catalase, and ascorbate peroxidase. A weakening of the cell wall structure in response to EPIP is reflected by pectin demethylation, while a changing pattern of fatty acids and acyl lipids composition suggests a modification of lipid metabolism. Notably, the formation of a signaling molecule—phosphatidic acid is induced locally in EPIP-treated AZ. Collectively, all these changes indicate the switching of several metabolic and signaling pathways directly in the AZ in response to EPIP, which inevitably leads to flower abscission.

SlERF52 regulates SlTIP1;1 expression to accelerate tomato pedicel abscission.

Abscission of plant organs is induced by developmental signals and diverse environmental stimuli and involves multiple regulatory networks, including biotic or abiotic stress-impaired auxin flux in the abscission zone (AZ). Depletion of auxin activates AZ ethylene (ETH) production and triggers acceleration of abscission, a process that requires hydrogen peroxide (H2O2). However, the interaction between these networks and the underlying mechanisms that control abscission are poorly understood. Here, we found that expression of tonoplast intrinsic proteins, which belong to the aquaporin (AQP) family in the AZ was important for tomato (Solanum lycopersicum) pedicel abscission. Liquid chromatography-tandem mass spectrometry and in situ hybridization revealed that SlTIP1;1 was most abundant and specifically present in the tomato pedicel AZ. SlTIP1;1 localized in the plasma membrane and tonoplast. Knockout of SlTIP1;1 resulted in delayed abscission, whereas overexpression of SlTIP1;1 accelerated abscission. Further analysis indicated that SlTIP1;1 mediated abscission via gating of cytoplasmic H2O2 concentrations and osmotic water permeability (Pf). Elevated cytoplasmic levels of H2O2 caused a suppressed auxin signal in the early abscission stage and enhanced ETH production during abscission. Furthermore, we found that increasing Pf was required to enhance the turgor pressure to supply the break force for AZ cell separation. Moreover, we observed that SlERF52 bound directly to the SlTIP1;1 promoter to regulate its expression, demonstrating a positive loop in which cytoplasmic H2O2 activates ETH production, which activates SlERF52. This, in turn, induces SlTIP1;1, which leads to elevated cytoplasmic H2O2 and water influx.

Expression Kinetics of Regulatory Genes Involved in the Vesicle Trafficking Processes Operating in Tomato Flower Abscission Zone Cells during Pedicel Abscission.

The abscission process occurs in a specific abscission zone (AZ) as a consequence of the middle lamella dissolution, cell wall degradation, and formation of a defense layer. The proteins and metabolites related to these processes are secreted by vesicle trafficking through the plasma membrane to the cell wall and middle lamella of the separating cells in the AZ. We investigated this process, since the regulation of vesicle trafficking in abscission systems is poorly understood. The data obtained describe, for the first time, the kinetics of the upregulated expression of genes encoding the components involved in vesicle trafficking, occurring specifically in the tomato (Solanum lycopersicum) flower AZ (FAZ) during pedicel abscission induced by flower removal. The genes encoding vesicle trafficking components included soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), SNARE regulators, and small GTPases. Our results clearly show how the processes of protein secretion by vesicle trafficking are regulated, programmed, and orchestrated at the level of gene expression in the FAZ. The data provide evidence for target proteins, which can be further used for affinity purification of plant vesicles in their natural state. Such analyses and dissection of the complex vesicle trafficking networks are essential for further elucidating the mechanism of organ abscission.

Spatio-temporal immunolocalization of extensin protein and hemicellulose polysaccharides during olive fruit abscission.

Immunocytochemical and molecular analyses reveal that the disassembly of the cell wall may be mediated by changes in the level and subcellular location of extensin protein and hemicelluloses during olive-fruit abscission. Although cell-wall modification is believed to underlie the changes in organ abscission, information concerning the changes in cell-wall proteins and hemicellulose polysaccharides is still limited. The aim of this work was to analyze the spatio-temporal patterns of the distribution of different extensin proteins and hemicelluloses in the abscission zone (AZ) during natural ripe-fruit abscission in olive (Olea europaea L.). In this study, we employed immunogold labeling in the ripe-fruit AZ during olive AZ cell separation, using an expanded set of monoclonal antibodies that recognize different types of hemicelluloses (LM11, LM15, and LM21), callose (anti-(1,3)-β-D-glucan) and extensin (JIM19) epitopes, and transmission electron microscopy imaging. Our data demonstrate that AZ cell separation was accompanied by a loss of the JIM19 extensin epitopes and a reduction in the detection of the LM15 xyloglucan epitopes in AZ cell walls, whereas AZ cells were found to be enriched with respect to the xylan and callose levels of the cell wall during olive ripe-fruit abscission. By contrast, AZ cell-wall polysaccharide remodeling did not involve mannans. Moreover, in ripe-fruit AZ, quantitative RT-PCR analysis revealed that OeEXT1, OeEXT2, OeXTH9, and OeXTH13 genes were downregulated during abscission, whereas the expression of OeXTH1, OeXTH5, and OeXTH14 genes increased during abscission. Taken together, the results indicate that AZ cell-wall dynamics during olive ripe-fruit abscission involves extensin protein and hemicellulose modifications, as well as related expressed genes. This is the first study available demonstrating temporal degradation of extensin protein and hemicelluloses in the AZ at the subcellular level.

Cell Wall Composition and Ultrastructural Immunolocalization of Pectin and Arabinogalactan Protein during Olea europaea L. Fruit Abscission.

Cell wall modification is integral to many plant developmental processes where cells need to separate, such as abscission. However, changes in cell wall composition during natural fruit abscission are poorly understood. In olive (Olea europaea L.), some cultivars such as 'Picual' undergo massive natural fruit abscission after fruit ripening. This study investigates the differences in cell wall polysaccharide composition and the localization of pectins and arabinogalactan protein (AGP) in the abscission zone (AZ) during cell separation to understand fruit abscission control in 'Picual' olive. To this end, immunogold labeling employing a suite of monoclonal antibodies to cell wall components (JIM13, LM5, LM6, LM19 and LM20) was investigated in olive fruit AZ. Cell wall polysaccharide extraction revealed that the AZ cell separation is related to the de-esterification and degradation of pectic polysaccharides. Moreover, ultrastructural localization showed that both esterified and unesterified homogalacturonans (HGs) localize mainly in the AZ cell walls, including the middle lamella and tricellular junction zones. Our results indicate that unesterified HGs are likely to contribute to cell separation in the olive fruit AZ. Similarly, immunogold labeling demonstrated a decrease in both galactose-rich and arabinose-rich pectins in AZ cell walls during ripe fruit abscission. In addition, AGPs were localized in the cell wall, plasma membrane and cytoplasm of AZ cells with lower levels of AGPs during ripe fruit abscission. This detailed temporal profile of the cell wall polysaccharide composition, and the pectins and AGP immunolocalization in the olive fruit AZ, offers new insights into cell wall remodeling during ripe fruit abscission.

Hydrogen sulfide inhibits ethylene-induced petiole abscission in tomato (Solanum lycopersicum L.)

Abscission is a dynamic physiological process that is ubiquitous in plants and can also be an essential agronomic trait in crops, thus attracting attention from plant growers and breeders. In general, the process of plant organ abscission can be divided into four steps, among which the step to obtain the competence to respond to abscission signals (step 2) is the most complex; however, the molecular mechanism underlying this process remains unclear. In this study, we found that hydrogen sulfide (H2S) inhibited the abscission of the tomato petiole in a dose-dependent manner, and the abscission of the petiole was accelerated when an H2S scavenger was applied. Further enzymatic activity and gene expression analyses showed that H2S suppressed the activity of enzymes capable of modifying the cell wall by inhibiting the usual upregulation of the transcription of the corresponding genes during the abscission process but not by affecting the activities of these enzymes by direct posttranslational modification. H2S treatment upregulated the expression levels of SlIAA3 and SlIAA4 but downregulated the transcription of ILR-L3 and ILR-L4 in the earlier stages of the abscission process, indicating that H2S probably functioned in the second step of the abscission process by preventing the abscission zone cells from obtaining the competence to respond to abscission signals by modulating the content of the bioactive-free auxin in these cells. Moreover, similar H2S inhibitory effects were also demonstrated in the process of floral organ abscission and anther dehiscence in other plant species, suggesting a ubiquitous role for H2S in cell separation processes.

Molecular and Hormonal Aspects of Drought-Triggered Flower Shedding in Yellow Lupine.

The drought is a crucial environmental factor that determines yielding of many crop species, e.g., Fabaceae, which are a source of valuable proteins for food and feed. Herein, we focused on the events accompanying drought-induced activation of flower abscission zone (AZ)—the structure responsible for flower detachment and, consequently, determining seed production in Lupinus luteus. Therefore, detection of molecular markers regulating this process is an excellent tool in the development of improved drought-resistant cultivars to minimize yield loss. We applied physiological, molecular, biochemical, immunocytochemical, and chromatography methods for a comprehensive examination of changes evoked by drought in the AZ cells. This factor led to significant cellular changes and activated AZ, which consequently increased the flower abortion rate. Simultaneously, drought caused an accumulation of mRNA of genes inflorescence deficient in abscission-like (LlIDL), receptor-like protein kinase HSL (LlHSL), and mitogen-activated protein kinase6 (LlMPK6), encoding succeeding elements of AZ activation pathway. The content of hydrogen peroxide (H2O2), catalase activity, and localization significantly changed which confirmed the appearance of stressful conditions and indicated modifications in the redox balance. Loss of water enhanced transcriptional activity of the abscisic acid (ABA) and ethylene (ET) biosynthesis pathways, which was manifested by elevated expression of zeaxanthin epoxidase (LlZEP), aminocyclopropane-1-carboxylic acid synthase (LlACS), and aminocyclopropane-1-carboxylic acid oxidase (LlACO) genes. Accordingly, both ABA and ET precursors were highly abundant in AZ cells. Our study provides information about several new potential markers of early response on water loss, which can help to elucidate the mechanisms that control plant response to drought, and gives a useful basis for breeders and agronomists to enhance tolerance of crops against the stress.

Control of Organ Abscission and Other Cell Separation Processes by Evolutionary Conserved Peptide Signaling.

Plants both generate and shed organs throughout their lifetime. Cell separation is in function during opening of anthers to release pollen; floral organs are detached after pollination when they have served their purpose; unfertilized flowers are shed; fruits and seeds are abscised from the mother plant to secure the propagation of new generations. Organ abscission takes place in specialized abscission zone (AZ) cells where the middle lamella between adjacent cell files is broken down. The plant hormone ethylene has a well-documented promoting effect on abscission, but mutation in ethylene receptor genes in Arabidopsis thaliana only delays the abscission process. Microarray and RNA sequencing have identified a large number of genes differentially expressed in the AZs, especially genes encoding enzymes involved in cell wall remodelling and disassembly. Mutations in such genes rarely give a phenotype, most likely due to functional redundancy. In contrast, mutation in the INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) blocks floral organ abscission in Arabidopsis. IDA encodes a small peptide that signals through the leucine-rich repeat receptor-like kinases HAESA (HAE) and HAE-LIKE2 (HSL2) to control floral organ abscission and facilitate lateral root emergence. Untimely abscission is a severe problem in many crops, and in a more applied perspective, it is of interest to investigate whether IDA-HAE/HSL2 is involved in other cell separation processes and other species. Genes encoding IDA and HSL2 orthologues have been identified in all orders of flowering plants. Angiosperms have had enormous success, with species adapted to all kinds of environments, adaptations which include variation with respect to which organs they shed. Here we review, from an evolutionary perspective, the properties of the IDA-HAE/HSL2 signaling module and the evidence for its hypothesized involvement in various cell separation processes in angiosperms.