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.

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.

A Non-Shedding Fruit Elaeis oleifera Palm Reveals Perturbations to Hormone Signaling, ROS Homeostasis, and Hemicellulose Metabolism.

The developmentally programmed loss of a plant organ is called abscission. This process is characterized by the ultimate separation of adjacent cells in the abscission zone (AZ). The discovery of an American oil palm (Elaeis oleifera) variant that does not shed its has allowed for the study of the mechanisms of ripe fruit abscission in this species. A comparative transcriptome analysis was performed to compare the fruit AZs of the non-shedding E. oleifera variant to an individual of the same progeny that sheds its ripe fruit normally. The study provides evidence for widespread perturbation to gene expression in the AZ of the non-shedding variant, compared to the normal fruit-shedding control, and offers insight into abscission-related functions. Beyond the genes with known or suspected roles during organ abscission or indehiscence that were identified, a list of genes with hormone-related functions, including ethylene, jasmonic acid, abscisic acid, cytokinin and salicylic acid, in addition to reactive oxygen species (ROS) metabolism, transcriptional responses and signaling pathways, was compiled. The results also allowed a comparison between the ripe fruit abscission processes of the African and American oil palm species at the molecular level and revealed commonalities with environmental stress pathways.

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.

The PIP Peptide of INFLORESCENCE DEFICIENT IN ABSCISSION Enhances Populus Leaf and Elaeis guineensis Fruit Abscission.

The programmed loss of a plant organ is called abscission, which is an important cell separation process that occurs with different organs throughout the life of a plant. The use of floral organ abscission in Arabidopsis thaliana as a model has allowed greater understanding of the complexities of organ abscission, but whether the regulatory pathways are conserved throughout the plant kingdom and for all organ abscission types is unknown. One important pathway that has attracted much attention involves a peptide ligand-receptor signalling system that consists of the secreted peptide IDA (INFLORESCENCE DEFICIENT IN ABSCISSION) and at least two leucine-rich repeat (LRR) receptor-like kinases (RLK), HAESA (HAE) and HAESA-LIKE2 (HSL2). In the current study we examine the bioactive potential of IDA peptides in two different abscission processes, leaf abscission in Populus and ripe fruit abscission in oil palm, and find in both cases treatment with IDA peptides enhances cell separation and abscission of both organ types. Our results provide evidence to suggest that the IDA-HAE-HSL2 pathway is conserved and functions in these phylogenetically divergent dicot and monocot species during both leaf and fruit abscission, respectively.

SSR markers in transcripts of genes linked to post-transcriptional and transcriptional regulatory functions during vegetative and reproductive development of Elaeis guineensis.

BACKGROUND: The oil palm (Elaeis guineensis Jacq.) is a perennial monocotyledonous tropical crop species that is now the world's number one source of edible vegetable oil, and the richest dietary source of provitamin A. While new elite genotypes from traditional breeding programs provide steady yield increases, the long selection cycle (10-12 years) and the large areas required to cultivate oil palm make genetic improvement slow and labor intensive. Molecular breeding programs have the potential to make significant impacts on the rate of genetic improvement but the limited molecular resources, in particular the lack of molecular markers for agronomic traits of interest, restrict the application of molecular breeding schemes for oil palm. RESULTS: In the current study, 6,103 non-redundant ESTs derived from cDNA libraries of developing vegetative and reproductive tissues were annotated and searched for simple sequence repeats (SSRs). Primer pairs from sequences flanking 289 EST-SSRs were tested to detect polymorphisms in elite breeding parents and their crosses. 230 of these amplified PCR products, 88 of which were polymorphic within the breeding material tested. A detailed analysis and annotation of the EST-SSRs revealed the locations of the polymorphisms within the transcripts, and that the main functional category was related to transcription and post-transcriptional regulation. Indeed, SSR polymorphisms were found in sequences encoding AP2-like, bZIP, zinc finger, MADS-box, and NAC-like transcription factors in addition to other transcriptional regulatory proteins and several RNA interacting proteins. CONCLUSIONS: The identification of new EST-SSRs that detect polymorphisms in elite breeding material provides tools for molecular breeding strategies. The identification of SSRs within transcripts, in particular those that encode proteins involved in transcriptional and post-transcriptional regulation, will allow insight into the functional roles of these proteins by studying the phenotypic traits that cosegregate with these markers. Finally, the oil palm EST-SSRs derived from vegetative and reproductive development will be useful for studies on the evolution of the functional diversity within the palm family.

Transcriptome analysis during somatic embryogenesis of the tropical monocot Elaeis guineensis: evidence for conserved gene functions in early development.

With the aim of understanding the molecular mechanisms underlying somatic embryogenesis (SE) in oil palm, we examined transcriptome changes that occur when embryogenic suspension cells are initiated to develop somatic embryos. Two reciprocal suppression subtractive hybridization (SSH) libraries were constructed from oil palm embryogenic cell suspensions: one in which embryo development was blocked by the presence of the synthetic auxin analogue 2,4-dichlorophenoxyacetic acid (2,4-D: ) in the medium (proliferation library); and another in which cells were stimulated to form embryos by the removal of 2,4-D: from the medium (initiation library). A total of 1867 Expressed Sequence Tags (ESTs) consisting of 1567 potential unigenes were assembled from the two libraries. Functional annotation indicated that 928 of the ESTs correspond to proteins that have either no similarity to sequences in public databases or are of unknown function. Gene Ontology (GO) terms assigned to the two EST populations give clues to the underlying molecular functions, biological processes and cellular components involved in the initiation of embryo development. Macroarrays were used for transcript profiling the ESTs during SE. Hierarchical cluster analysis of differential transcript accumulation revealed 4 distinct profiles containing a total of 192 statistically significant developmentally regulated transcripts. Similarities and differences between the global results obtained with in vitro systems from dicots, monocots and gymnosperms will be discussed.

Pluripotent versus totipotent plant stem cells: dependence versus autonomy?

Little is known of the mechanisms that induce the dedifferentiation of a single somatic cell into a totipotent embryogenic cell that can either be regenerated or develop into an embryo and subsequently an entire plant. In this Opinion article, we examine the cellular, physiological and molecular similarities and differences between different plant stem cell types. We propose to extend the plant stem cell concept to include single embryogenic cells as a totipotent stem cell based on their capacity to regenerate or develop into an embryo under certain conditions. Our survey suggests that differences in chromatin structure might ensure that meristem-localized stem cells have supervised freedom and are pluripotent, and that embryogenic stem cells are unsupervised, autonomous and, hence, freely totipotent.