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The abscission zone (AZ) is a specialized tissue that usually develops at the base of an organ and is highly sensitive to phytohormones, e.g., abscisic acid (ABA), ethylene (ET), and gibberellins (GAs). A current model of organ abscission assumes that the formation of an auxin gradient around the AZ area determines the time of shedding; however, that thesis is supported by studies that are primarily concerned with auxin transporters. To better understand the events underlying the progression of abscission, we focused for the first time on indole-3-acetic acid (IAA) distribution following AZ activation. We performed a series of immunolocalization studies in proximal and distal regions of floral AZ cells in yellow lupine, which is an agriculturally important legume. The examined phytohormone was abundant in natural active AZ cells, as well as above and below parts of this structure. A similar gradient of IAA was observed during the early steps of abscission, which was induced artificially by flower removal. Surprisingly, IAA was not detected in inactive AZ cells. This paper is also a consequence of our comprehensive studies concerning the phytohormonal regulation of flower abscission in yellow lupine. We present new data on interactions between IAA and ET, previously pointed out as a strong modulator of flower separation. The detailed analysis shows that disruption of the natural auxin gradient around the AZ area through the application of synthetic IAA had a positive effect on ET biosynthesis genes. We proved that these changes are accompanied by an accumulation of the ET precursor. On the other hand, exposure to ET significantly affected IAA localization in the whole AZ area in a time-dependent manner. Our results provide insight into the existence of a spatio-temporal sequential pattern of the IAA gradient related to the abscission process; this pattern is maintained by interactions with ET. We present new valuable evidence for the existence of conservative mechanisms that regulate generative organ separation and can help to improve the yield of agronomically significant species in the future. [ABSTRACT FROM AUTHOR]

Drought causes the excessive abscission of flowers in yellow lupine, leading to yield loss and serious economic consequences in agriculture. The structure that determines the time of flower shedding is the abscission zone (AZ). Its functioning depends on the undisturbed auxin movement from the flower to the stem. However, little is known about the mechanism guiding cell–cell adhesion directly in an AZ under water deficit. Therefore, here, we seek a fuller understanding of drought-dependent reactions and check the hypothesis that water limitation in soil disturbs the natural auxin balance within the AZ and, in this way, modifies the cell wall structure, leading to flower separation. Our strategy combined microscopic, biochemical, and chromatography approaches. We show that drought affects indole-3-acetic acid (IAA) distribution and evokes cellular changes, indicating AZ activation and flower abortion. Drought action was manifested by the accumulation of proline in the AZ. Moreover, cell wall-related modifications in response to drought are associated with reorganization of methylated homogalacturonans (HG) in the AZ, and upregulation of pectin methylesterase (PME) and polygalacturonase (PG)—enzymes responsible for pectin remodeling. Another symptom of stress action is the accumulation of hemicelluloses. Our data provide new insights into cell wall remodeling events during drought-induced flower abscission, which is relevant to control plant production. [ABSTRACT FROM AUTHOR]

How auxin transport regulates organ abscission is a long-standing and intriguing question. Polar auxin transport across the abscission zone (AZ) plays a more important role in the regulation of abscission than a local concentration of this hormone. We recently reported the existence of a spatiotemporal sequential pattern of the indole-3-acetic acid (IAA) localization in the area of the yellow lupine AZ, which is a place of flower detachment. In this study, we performed analyses of AZ following treatment with an inhibitor of polar auxin transport (2,3,5-triiodobenzoic acid (TIBA)). Once we applied TIBA directly onto the AZ, we observed a strong response as demonstrated by enhanced flower abscission. To elucidate the molecular events caused by the inhibition of auxin movement, we divided the AZ into the distal and proximal part. TIBA triggered the formation of the IAA gradient between these two parts. The AZ-marker genes, which encode the downstream molecular components of the inflorescence deficient in abscission (IDA)-signaling system executing the abscission, were expressed in the distal part. The accumulation of IAA in the proximal area accelerated the biosynthesis of abscisic acid and ethylene (stimulators of flower separation), which was also reflected at the transcriptional level. Accumulated IAA up-regulated reactive oxygen species (ROS) detoxification mechanisms. Collectively, we provide new information regarding auxin-regulated processes operating in specific areas of the AZ. [ABSTRACT FROM AUTHOR]

Abstract: The plant hormone auxin plays a critical role in regulating plant growth and development. Recent advances have been made that having improved our understanding of auxin response pathways, primarily by characterizing the genes encoding auxin response factors (ARFs) in Arabidopsis. In addition, the expression of some ARFs is regulated by microRNAs (miRNAs). In Arabidopsis thaliana, ARF6 and ARF8 are targeted by miR167, whereas ARF10, ARF16 and ARF17 are targeted by miR160. Nevertheless, little is known about any possible interactions between miRNAs and the auxin signaling pathway during plant development. In this study, we isolated the miR167 target gene InARF8 cDNA from the cotyledons of the short day plant (SDP) Ipomoea nil (named also Pharbitis nil). Additionally, the In-miR167 precursor was identified from the I. nil EST database and analyses of InARF8 mRNA, In-pre-miR167 and mature miR167 accumulation in the plant's vegetative and generative organs were performed. The identified cDNA of InARF8 contains a miR167 complementary sequence and shows significant similarity to ARF8 cDNAs of other plant species. The predicted amino acid sequence of InARF8 includes all of the characteristic domains for ARF family transcription factors (B3 DNA-binding domain, AUX/IAA-CTD and a glutamine-rich region). Quantitative RT-PCR reactions and in situ hybridization indicated that InARF8 was expressed primarily in the shoot apices, leaf primordia and hypocotyls of I. nil seedlings, as well as in flower pistils and petals. The InARF8 transcript level increased consistently during the entire period of pistil development, whereas in the stamens, the greatest transcriptional activity occurred only during the intensive elongation phase. Additionally, an expression analysis of both the precursor In-pre-miR167 molecules identified and mature miRNA was performed. We observed that, in most of the organs examined, the InARF8 expression pattern was opposite to that of MIR167, indicating that the gene's activity was regulated by mRNA cleavage. Our findings suggested that InARF8 and InMIR167 participated in the development of young tissues, especially the shoot apices and flower elements. The main function of MIR167 appears to be to regulate InARF8 organ localization. [Copyright &y& Elsevier]

Indole-3-acetic acid (IAA) conjugation is one of the mechanisms responsible for auxin homeostasis. IAA ester conjugates biosynthesis has been studied during development of maize seeds where IAA-inositol (IAInos) and its glycosidic forms make up about 50 % of its ester conjugates pool. 1- O -indole-3-acetyl-β- d -glucose (IAGlc) synthase and indole-3-acetyl transferase (IAInos synthase) are key enzymes in a two-step pathway of IAInos synthesis. In the first reaction, IAA is glucosylated to a high energy acetal, 1- O -indole-3-acetyl-β- d -glucose by IAGlc synthase, whereas in the second step, IAInos synthase transfers IAA moiety to myo -inositol forming a stable auxin ester, indole-3-acetyl- myo -inositol (IAInos). It should be mentioned that IAGlc synthase catalyzes a reversible reaction with unfavourable equilibrium that delivers IAGlc for favourable transacylation to IAInos. This is the first study where IAGlc synthase and IAInos synthase are simultaneously analyzed by enzymatic activity assay and quantitative RT-PCR in maize seeds at four stages of development (13, 26, 39 and 52 Days After Flowering). Activity of IAGlc/IAInos synthases as well as their expression profiles during seed development were different. While both enzymatic activities and ZmIAIn expression were the highest in seeds at 26 DAF, the highest expression of ZmIAGlc was observed at 13 DAF. Protein gel blot analysis showed that IAInos synthase exists as a mixture of several isoforms at a similar protein level at particular stages of seed development. Neither of other ester conjugates of IAA (IAA-mannose) nor IAA-amino acids were detected at the stages studied. Catalytic activity of l -tryptophan aminotransferase involved in IAA biosynthesis as well as UDPG pyrophosphorylase, synthesizing UDPG as a substrate for IAGlc synthase, were also analyzed. l -tryptophan aminotransferase activity was the highest at 26 DAF. Changes in enzyme activity of UDPG pyrophosphorylase are difficult to interpret. Expression levels of ZmIPS and ZmIPP encoding two enzymes of myo -inositol biosynthesis pathway: inositol-x-phosphate synthase (IPS) and inositol-x-phosphate phosphatase (IPP), respectively, were analyzed. 26 DAF seeds displayed the highest expression level of ZmIPS , whereas transcription of ZmIPP was the highest at 13 DAF. [ABSTRACT FROM AUTHOR]