Fabio Tebaldi Silveira Nogueira

Tomato ( Solanum lycopersicum ) is classified as a day-neutral plant, whereas its wild relatives exhibit delayed flowering under long day (LD) conditions due to higher activity of the SELF-PRUNING 5G ( SP5G ). In Arabidopsis thaliana , CONSTANS (CO) activates FLOWERING LOCUS T ( FT ), a homolog of SP5G , but whether and how CO integrates with SP5G in tomato flowering was unclear. Here, we demonstrate that SlCOL1 (the tomato CO homolog) delays flowering by directly activating SP5G in a PHYTOCHROME B1 ( PHYB1 )-dependent manner. Importantly, genetic and molecular analyses combining a photoperiod-responsive tomato line carrying the wild SP5G allele from S. pennellii, together with SlCOL1 and flowering-pathway mutants, revealed synergistic crosstalk among the photoperiodic SlCOL1–SP5G module, age-dependent pathway (mediated mainly by the microRNA156– SlSBP module), gibberellin (GA) pathway, and SINGLE FLOWER TRUSS ( SFT ) pathway. Mechanistically, we show that SP5G forms a complex with miR156-targeted SlSBP13 to directly regulate SFT expression, and that GA may interfere with SP5G activity. Together, these findings revealed a coordinated network that integrates multiple flowering signals to modulate both shared and pathway-specific targets. Our findings provide a significant advance in understanding the molecular regulation of tomato flowering and offer promising avenues for breeding strategies optimized for diverse environmental conditions and latitudes.

Phytohormone signaling is fine-tuned by regulatory feedback loops. The phytohormone abscisic acid (ABA) plays key roles in plant development and abiotic stress tolerance. PYRABACTIN RESISTENCE 1/PYR1-LIKE/REGULATORY COMPONENT OF ABA RECEPTOR (PYR/PYL/RCAR) receptors sense ABA, and in turn, ABA represses their expression. Conversely, ABA induces expression of Type 2C PROTEIN PHOSPHATASES (PP2C) genes, which negatively regulate the ABA signaling pathway. This regulatory feedback scheme is likely important for modulating ABA signaling. Here, we provide insight into the mechanisms underlying the ABA-induced repression of PYR/PYL/RCAR expression in Arabidopsis (Arabidopsis thaliana). ABA time course analyses revealed strong and sustained repression of PYR/PYL/RCARs, suggesting that receptor gene regulation is an important step in resetting the ABA signaling pathway. Cordycepin-induced transcription inhibition showed that PYL1/4/5/6 mRNA destabilization is involved in the ABA-induced repression of these genes. Furthermore, genetic evidence indicated that decapping may play a role in PYL4/5/6 mRNA decay. We also provide evidence that the Arabidopsis-specific microRNA5628 (miR5628), which is transiently induced by the ABA core signaling pathway, guides PYL6 transcript cleavage in response to ABA. After cleavage, the resulting 5′- and 3′-cleaved fragments of PYL6 mRNA may be degraded by the XRN4 exoribonuclease. miR5628 is an evolutionary novelty that may enhance PYL6 mRNA degradation, along with decapping and XRN4 activity. Thus, regulating the stability of PYR/PYL/RCAR transcripts maintains ABA signaling homeostasis.

Summary: Passion flower extrafloral nectaries (EFNs) protrude from leaves and facilitate mutualistic interactions with insects; however, how age cues control EFN growth remains poorly understood. Here, we examined leaf and EFN morphology and development of two Passiflora species with distinct leaf shapes, and compared the phenotype of these to transgenics with manipulated activity of the age‐dependent miR156, which targets several SQUAMOSA PROMOTER‐BINDING PROTEIN‐LIKE (SPL) transcription factors. Low levels of miR156 correlated with leaf maturation and EFN formation in Passiflora edulis and P. cincinnata. Accordingly, manipulating miR156 activity affected leaf heteroblasty and EFN development. miR156‐overexpressing leaves exhibited less abundant and tiny EFNs in both Passiflora species. EFN abundance remained mostly unchanged when miR156 activity was reduced, but it led to larger EFNs in P. cincinnata. Transcriptome analysis of young leaf primordia revealed that miR156‐targeted SPLs may be required to properly express leaf and EFN‐associated genes. Importantly, altered miR156 activity impacted sugar profiles of the nectar and modified ecological relationships between EFNs and ants. Our work provides evidence that the miR156/SPL module indirectly regulates EFN development in an age‐dependent manner and that the EFN development program is closely associated with the heteroblastic developmental program of the EFN‐bearing leaves.

SUMMARY: Fruit morphogenesis is determined by the coordination of cell division and expansion, which are fundamental processes required for the development of all plant organs. Here, we show that the regulation of TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCP) LANCEOLATE (TCP2/LA) by miR319 is crucial for tomato fruit morphology. The loss of miR319 regulation in the semi‐dominant La mutant led to a premature SlTCP2/LA expression during gynoecium patterning, which results in modified cell division during carpel development. As a consequence, La mutants exhibited elongated ovary and fruit shape, and a reduced number of ovules and seeds. Elongated fruit shape in La may be partially due to the SlTCP2/LA‐mediated repression of OVATE activity in young floral buds. Further analysis showed that the de‐repression of SlTCP2/LA decreases auxin responses in young floral buds by directly repressing SlYUCCA4 expression, but SlTCP2/LA also acts in parallel with ENTIRE (E) to orchestrate fruit morphology and seed production. Our study defines a novel miRNA‐based molecular link between the domestication‐associated OVATE gene and auxin responses. Given the striking variation in fruit morphology among members of the Solanaceae family, fine‐tuning regulation of gene expression by miRNA coupled with modulation of auxin dynamics may be a common driver in the evolution of fruit shape diversity.

Summary: Plant pigments are diverse natural molecules involved in numerous biological functions such as development, growth, and metabolism. As plants age, not only new organs will be formed, but also, they will acquire the necessary pigments in response to the environment and endogenous programming in order to achieve reproductive success. Among the endogenous cues, the small RNAs (sRNAs), an endogenous group of ubiquitous regulatory molecules, may regulate the pigments‐associated biosynthetic pathways at posttranscriptional level. Although plant pigments and sRNAs have been comprehensively studied in several processes throughout the entire plant cycle in model and nonmodel species, connections among these central players must be revised. Studying these complex networks allow us not only to know the progress that has been made in this area, but also generate research questions to be explored in order to unravel novel mechanisms for improving plant yield; therefore, in this review we have summarized the emerging roles of sRNAs‐regulated nodes in mediating plant pigmentation‐associated biosynthetic pathways, focused on chlorophylls, flavonoids, carotenoids, and betalains. In addition, we discuss perspectives related to the manipulation of those genes associated with plant pigments for obtaining genetically improved plants.

Auxin coordinates cell growth by promoting or inhibiting cell expansion during etiolated seedling development, but whether and how microRNA modules participate in this process remains unclear. Here, we show the miRNA156/ SQUAMOSA PROMOTER-BINDING-PROTEIN-LIKE9 (miR156/ SPL9 ) module is critical for skotomorphogenesis. Perturbation of the miR156/ SPL9 module affected skotomorphogenesis, as the loss of miR156 function or SPL9 de-repression led to shorter hypocotyl, higher hook angle, and delayed hook opening. Opposing phenotypes were observed in dark-grown spl9 and miR156-overexpressing seedlings. Importantly, loss of miR156-dependent SPL9 regulation triggered apical hook formation even under reduced levels of endogenous auxin. miR156-targeted SPL9 arrested cell expansion by repressing small auxin-up RNA19 ( SAUR19 ) gene in a FRUITFULL ( FUL )-dependent and independent manner. The conserved miR156/ SPL9/15 module also affects skotomorphogenesis in tomato, impacting its successful soil emergence. Our findings unravel how the miR156/ SPL9 module plays a pivotal role in the auxin network coordinating apical hook development to enable appropriate seedling emergence.

Strigolactones are a class of phytohormones with various functions in plant development, stress responses, and in the interaction with (micro)organisms in the rhizosphere. While their effects on vegetative development are well studied, little is known about their role in reproduction. We investigated the effects of genetic and chemical modification of strigolactone levels on the timing and intensity of flowering in tomato ( Solanum lycopersicum L.) and the molecular mechanisms underlying such effects. Results showed that strigolactone levels in the shoot, whether endogenous or exogenous, correlate inversely with the time of anthesis and directly with the number of flowers and the transcript levels of the florigen-encoding gene SINGLE FLOWER TRUSS ( SFT ) in the leaves. Transcript quantifications coupled with metabolite analyses demonstrated that strigolactones promote flowering in tomato by inducing the activation of the microRNA319- LANCEOLATE module in leaves. This, in turn, decreases gibberellin content and increases the transcription of SFT . Several other floral markers and morpho-anatomical features of developmental progression are induced in the apical meristems upon treatment with strigolactones, affecting floral transition and, more markedly, flower development. Thus, strigolactones promote meristem maturation and flower development via the induction of SFT both before and after floral transition, and their effects are blocked in plants expressing a miR319-resistant version of LANCEOLATE . Our study positions strigolactones in the context of the flowering regulation network in a model crop species.

How does a plant detect the changing seasons and make important developmental decisions accordingly? How do they incorporate daylength information into their routine physiological processes? Photoperiodism, or the capacity to measure the daylength, is a crucial aspect of plant development that helps plants determine the best time of the year to make vital decisions, such as flowering. The protein CONSTANS (CO) constitutes the central regulator of this sensing mechanism, not only activating florigen production in the leaves but also participating in many physiological aspects in which seasonality is important. Recent discoveries place CO in the center of a gene network that can determine the length of the day and confer seasonal input to aspects of plant development and physiology as important as senescence, seed size, or circadian rhythms. In this review, we discuss the importance of CO protein structure, function, and evolutionary mechanisms that embryophytes have developed to incorporate annual information into their physiology.

SUMMARY: - Passion flower extrafloral nectaries (EFNs) protrude from adult leaves and facilitate mutualistic interactions with insects, but how age cues control EFN establishment remains poorly understood. - Here, we combined genetic and molecular studies to investigate how leaf development and EFN patterning are regulated through the age-dependent miR156- SQUAMOSA PROMOTER BINDING PROTEIN LIKE ( SPL ) module in two EFN-containing Passiflora species with distinct leaf shapes. - Low levels of miR156 correlate with leaf maturation and EFN formation in Passiflora edulis and P. cincinnata . Consistently, overexpression of miR156 (miR156-OE), which leads to low levels of SPLs , affected leaf ontogeny and EFN development in both species. Laminar EFNs were underdeveloped and less abundant in both P. edulis and P. cincinnata miR156-OE leaves. Importantly, the ecological relationships established by EFNs and their sugar profiles were negatively regulated by high levels of miR156. Moreover, transcriptome analysis of young leaf primordia revealed that miR156-targeted SPLs may be required for proper expression of leaf and EFN development- associated genes in P. edulis and P. cincinnata . - Our work provides the first evidence that the highly conserved miR156/ SPL module regulates EFN development in an age-dependent manner and that the program responsible for EFN development is closely associated with the heteroblastic developmental program of the EFN-bearing leaves.