Simulation of organ patterning on the floral meristem using a polar auxin transport model

An intriguing phenomenon in plant development is the timing and positioning of lateral organ initiation, which is a fundamental aspect of plant architecture. Although important progress has been made in elucidating the role of auxin transport in the vegetative shoot to explain the phyllotaxis of leaf formation in a spiral fashion, a model study of the role of auxin transport in whorled organ patterning in the expanding floral meristem is not available yet. We present an initial simulation approach to study the mechanisms that are expected to play an important role. Starting point is a confocal imaging study of Arabidopsis floral meristems at consecutive time points during flower development. These images reveal auxin accumulation patterns at the positions of the organs, which strongly suggests that the role of auxin in the floral meristem is similar to the role it plays in the shoot apical meristem. This is the basis for a simulation study of auxin transport through a growing floral meristem, which may answer the question whether auxin transport can in itself be responsible for the typical whorled floral pattern. We combined a cellular growth model for the meristem with a polar auxin transport model. The model predicts that sepals are initiated by auxin maxima arising early during meristem outgrowth. These form a pre-pattern relative to which a series of smaller auxin maxima are positioned, which partially overlap with the anlagen of petals, stamens, and carpels. We adjusted the model parameters corresponding to properties of floral mutants and found that the model predictions agree with the observed mutant patterns. The predicted timing of the primordia outgrowth and the timing and positioning of the sepal primordia show remarkable similarities with a developing flower in nature.     

The role of floral meristems in patterning

The flower is one of the most complex and varied structures found in plants. Over the past decade, we have begun to understand how floral patterning is established in a handful of model species. Recent studies have identified the presence of several potential pathways for organ patterning. Many genes that are involved in these pathways have been cloned, providing opportunities for further fruitful investigations into the genetic components of flower development.     

The role of auxin level and sensitivity in floral induction

Flower initiation takes place during a rise of peroxidase activity following a peak of minimum activity which marked the completion of the flowering inductive phase. Since basic isoperoxidases underwent an inverse variation of activity in the course of successive inductive and initiative phases, it was hypothesized that the induction of flowering led to a temporary peak of maximum auxin level in the leaves. Our analyses and available literature data support the view. They also show the different capacity of non-induced and induced material to respond to external auxin application. Since some aspects of the physiological state characterizing induced plants can be simultaneously obtained in all plant parts as a result of rapid interorgan communication, the classical florigen theory is seriously challenged. On leave from University of Liège-Start Tilman, Institute of Botany B22, B-4000 Liège, Belgium.     

A role for auxin response factor 19 in auxin and ethylene signaling in Arabidopsis

Although auxin response factors (ARFs) are the first well-characterized proteins that bind to the auxin response elements, elucidation of the roles of each ARF gene in auxin responses and plant development has been challenging. Here we show that ARF19 and ARF7 not only participate in auxin signaling, but also play a critical role in ethylene responses in Arabidopsis (Arabidopsis thaliana) roots, indicating that the ARFs serve as a cross talk point between the two hormones. Both arf19 and arf7 mutants isolated from our forward genetic screens are auxin resistant and the arf19arf7 double mutant had stronger auxin resistance than the single mutants and displayed phenotypes not seen in the single mutants. Furthermore, we show that a genomic fragment of ARF19 not only complements arf19, but also rescues arf7. We conclude that ARF19 complements ARF7 at the protein level and that the ARF7 target sequences are also recognized by ARF19. Therefore, it is the differences in expression level/pattern and not the differences in protein sequences between the two ARFs that determines the relative contribution of the two ARFs in auxin signaling and plant development. In addition to being auxin resistant, arf19 has also ethylene-insensitive roots and ARF19 expression is induced by ethylene treatment. This work provides a sensitive genetic screen for uncovering auxin-resistant mutants including the described arf mutants. This study also provides a likely mechanism for coordination and integration of hormonal signals to regulate plant growth and development.     

SEUSS and AINTEGUMENTA mediate patterning and ovule initiation during gynoecium medial domain development

The Arabidopsis (Arabidopsis thaliana) gynoecium, the female floral reproductive structure, requires the action of genes that specify positional identities during its development to generate an organ competent for seed development and dispersal. Early in gynoecial development, patterning events divide the primordium into distinct domains that will give rise to specific tissues and organs. The medial domain of the gynoecium gives rise to the ovules, and several other structures critical for reproductive competence. Here we report a synergistic genetic interaction between seuss and aintegumenta mutants resulting in a complete loss of ovule initiation and a reduction of the structures derived from the medial domain. We show that patterning events are disrupted early in the development of the seuss aintegumenta gynoecia and we identify PHABULOSA (PHB), REVOLUTA, and CRABS CLAW (CRC) as potential downstream targets of SEUSS (SEU) and AINTEGUMENTA (ANT) regulation. Our genetic data suggest that SEU additionally functions in pathways that are partially redundant and parallel to PHB, CRC, and ANT. Thus, SEU and ANT are part of a complex and robust molecular system that coordinates patterning cues and cellular proliferation along the three positional axes of the developing gynoecium.     

花器官形成的基因调控

主要论述了花发育过程中花器官同源异形基因及其相关基因的调控机理。基因调控是一个复杂的 系统,花同源异形基因既受到上游基因的调控,同时又决定了下游基因的表达。对花发育基因调控的研究,不 仅可以从微观水平了解植物花发育的分子机制,同时对花卉等作物的遗传育种也具有重要的指导意义。     

新疆野杏开花物候与花器官对海拔的响应

新疆野杏(Prunus armeniaca Lam.)是天山野果林的优势种,具有重要的生态与资源价值。野果林的生境条件与其分布及生长密切相关。为了明确新疆野杏在不同海拔下的开花物候与花器官变化规律,于2021年3月—4月,选择新疆新源县吐尔根杏花沟野杏林为研究区,在野杏集中分布的1000—1500m的山地,由低到高划分Ⅰ—Ⅴ级海拔梯度设置样地,监测环境条件,对野杏群体开花物候期与花器官发育特征进行调查。结果表明：(1)新疆野杏群体开花物候期历时32d左右,各海拔梯度最长相差2d,第Ⅰ级与第Ⅱ级海拔的开花物候期差异不明显,其他海拔梯度间均存在显著差异,开花最晚的第Ⅴ级比最早的第Ⅰ级晚9d,但群体开花期长4d,海拔梯度与开花物候期呈显著正相关,而温度与开花物候期呈显著负相关;(2)新疆野杏的花萼长度和宽度、子房高度和宽度均是第Ⅱ级海拔的最大;花冠直径、花瓣纵径和横径均是第Ⅰ级的最大,花药长度和宽度均是第Ⅳ级的最大;花柱长度是第Ⅴ级的最大。海拔与花的外部器官、雌蕊呈显著负相关,与雄蕊呈显著正相关;光照强度与花的外部器官、雌蕊呈显著负相关;(3)新疆野杏开花期气候因子,第Ⅳ级、第Ⅴ级与第Ⅰ级存...     

Temperature compensation of auxin dependent developmental patterning

The establishment of localized auxin gradients plays a central role in developmental patterning in plants. Auxin levels and responses have been shown to increase with temperature although developmental patterning is not affected. This suggests the existence of a homeostatic mechanism that ensures that patterning occurs normally over a range of temperatures. We recently described the cloning and characterization of BOBBER1 (BOB1), an Arabidopsis gene which encodes a small heat shock protein. BOB1 is required for the establishment of auxin gradients and for normal developmental patterning. BOB1 is also required for organismal thermotolerance and localizes to heat shock granules at elevated temperatures. Since BOB1 functions in both temperature responses and developmental patterning we propose that BOB1 may encode a component of a developmental temperature compensation mechanism.Key words: BOBBER1, BOB1, temperature, development, homeostasis, heat shock granule, sHSPPolar auxin transport and the resulting establishment of local auxin gradients is a central theme in plant developmental patterning. Auxin acts as a morphogen which establishes patterning and cell identity in the female gametophyte and is subsequently required for the establishment of polarity and patterning during embryogenesis.^1,2 During vegetative plant development the correct establishment of auxin gradients is required for the establishment of phyllotaxy and for sculpting the shape of leaf margins among many other patterning functions.³ These patterning events occur in a stereotypical manner regardless of changes in temperature.Given that these important developmental functions proceed normally over a range of temperatures it is perhaps surprising that auxin levels and responses are significantly influenced by changing temperatures. Gray et al. showed that there is a ∼1.75x increase in free IAA levels when Arabidopsis plants are grown at 29°C instead of at 20°C.⁴ Increased IAA levels at elevated temperatures are accompanied by auxin dependent hypocotyl elongation as well as changes in the patterns of auxin responsive gene expression. However, changes in developmental patterning were not reported. This suggests the existence of a robust homeostatic mechanism which normalizes auxin mediated developmental patterning events in response to changing temperatures.We recently characterized BOBBER1 (BOB1) which encodes a non-canonical small heat shock protein (sHSP) with functions both in developmental patterning and in high temperature responses.^5,6 BOB1 is an essential gene and bob1-1 null mutants arrest as globular embryos. In bob1-1 mutants a DR5:GFP reporter was used to show that there is a failure to establish auxin gradients during embryogenesis with resulting patterning abnormalities. A similar lack of auxin gradients is observed in embryos treated with 2,4-D, which, like bob1-1 embryos, arrest at the globular stage.⁷ In the apical half of bob1-1 embryos the central meristematic domain is expanded to encompass the lateral domains where cotyledons would normally form. Similarly, in the basal half of bob1-1 embryos auxin maxima are not established and the expression of SCARECROW (SCR) at the root pole is never observed. This observation is consistent with a requirement for a localized auxin maxima in the formation of the root meristem.⁸During post-embryonic development BOB1 is required for normal growth as well as for leaf and floral patterning. When bob1 activity is genetically reduced pin-formed inflorescence meristems are observed. Pinformed meristems reflect a lack of organ primordium initiation. This phenotype is specifically observed in mutants including pinoid (pid), pin-formed1 (pin1) and yucca1,4,6 triple mutants in which the ability to establish auxin maxima has been disrupted.^9–11 While the presence of phenotypes that phenocopy auxin deficiencies is not evidence that BOB1 directly modulates auxin mediated patterning, these observation are certainly suggestive. Given the extensive overlap between bob1 phenotypes and auxin phenotypes a plausible hypothesis is that BOB1 may be required for normal auxin transport or signal transduction.bob1-3 is a partial loss of function allele which has, among other phenotypes, serrated leaf margins on early leaves.⁶ Normal (Col-O) plants do not produce serrations on the margins of their first two leaves, however bob1-3 mutant plants have one or two leaf serrations on leaves one and two. The formation of leaf margin serrations has been shown to require the activity of the auxin efflux carrier PIN1.¹² In order to investigate whether the leaf margin phenotype observed in bob1-3 mutants requires normal PIN1 activity we crossed bob1-3 plants to pin1-1 mutant plants to generate bob1-3; pin1-1 double mutants. The margins of the first leaves of double mutant plants are smooth. This demonstrates that the serrated leaf phenotype of bob1-3 mutants depends on, and modulates, normal PIN1 activity (Fig. 1).Open in a separate windowFigure 1Leaf morphology of col-O, bob1-3, pin1-1 and bob1-3; pin1-1 plants. the margins of leaf 2 of col-O plants but are completely smooth, while serrations are present on the margins of bob1-3 leaves (arrow heads). no serrations are present on pin1-1 leaves, and pin1-1 is epistatic to bob1-3 in bob1-3; pin1-1 double mutants. Scale bar is 1 cm.In addition to characterizing BOB1’s developmental functions we also demonstrated that BOB1 encodes a small heat shock protein (sHSP) with protein chaperone activity. BOB1 protein can prevent the thermal denaturation of a model substrate in vitro and co-localizes to heat shock granules with canonical sHSPs. bob1-3 mutants also have thermotolerance defects.⁶ sHSP chaperones function by binding to partially unfolded substrates and preventing their irreversible aggregation.¹³ Assuming that bob1 developmental defects are due to a change in chaperone activity this would suggest a model where BOB1 modulates the function of developmental pathways at normal temperatures by ensuring the correct folding of target proteins. One possible target of BOB1 activity could be signal transduction molecules, perhaps in the auxin signaling pathway, which require stabilization in the absence of ligands or interaction partners, similar to Hsp90 targets.¹⁴Developmental homeostasis of auxin mediated events over a range of temperatures requires a mechanism which responds to temperature and also modulates auxin responses. We propose that BOB1 might be a component of this homeostatic mechanism as it has both of these characteristics. sHSPs like BOB1 are known to undergo temperature dependent conformational changes that result in altered substrate interactions.¹³ In addition to a role in temperature responses we have shown that BOB1 is required for the establishment of auxin gradients and appears to modulate auxin mediated developmental events throughout the plant life cycle. We are currently investigating this hypothesis in order to obtain a better understanding of how plants develop normally over a wide range of temperatures.     

SEUSS and SEUSS‐LIKE 2 coordinate auxin distribution and KNOXI activity during embryogenesis

In Arabidopsis, SEUSS (SEU) and SEUSS‐LIKE 2 (SLK2) are components of the LEUNIG (LUG) repressor complex that coordinates various aspects of post‐embryonic development. The complex also plays a critical role during embryogenesis, as seu slk2 double mutants have small, narrow cotyledons and lack a shoot apical meristem (SAM). Here we show that seu slk2 double mutant embryos exhibit delayed cotyledon outgrowth and that this is associated with altered PIN‐FORMED1 (PIN1) expression and localisation during the early stages of embryogenesis. These observations suggest that SEU and SLK2 promote the transition to bilateral symmetry by modulating auxin distribution in the embryonic shoot. This study also shows that loss of SAM formation in seu slk2 mutants is associated with reduced expression of the class I KNOX (KNOXI) genes SHOOTMERISTEMLESS (STM), BREVIPEDICELLUS and KNAT2. Furthermore, elevating STM expression in seu slk2 mutant embryos was sufficient to restore SAM formation but not post‐embryonic activity, while both SAM formation and activity were rescued when SLK2 expression was restored in either the cotyledons or boundary regions. These results demonstrate that SEU and SLK2 function redundantly to promote embryonic shoot development and likely act through a non‐cell autonomous pathway to promote KNOXI activity.     

Maintenance of embryonic auxin distribution for apical-basal patterning by PIN-FORMED-dependent auxin transport in Arabidopsis

Molecular mechanisms of pattern formation in the plant embryo are not well understood. Recent molecular and cellular studies, in conjunction with earlier microsurgical, physiological, and genetic work, are now starting to define the outlines of a model where gradients of the signaling molecule auxin play a central role in embryo patterning. It is relatively clear how these gradients are established and interpreted, but how they are maintained is still unresolved. Here, we have studied the contributions of auxin biosynthesis, conjugation, and transport pathways to the maintenance of embryonic auxin gradients. Auxin homeostasis in the embryo was manipulated by region-specific conditional expression of indoleacetic acid-tryptophan monooxygenase or indoleacetic acid-lysine synthetase, bacterial enzymes for auxin biosynthesis or conjugation. Neither manipulation of auxin biosynthesis nor of auxin conjugation interfered with auxin gradients and patterning in the embryo. This result suggests a compensatory mechanism for buffering auxin gradients in the embryo. Chemical and genetic inhibition revealed that auxin transport activity, in particular that of the PIN-FORMED1 (PIN1) and PIN4 proteins, is a major factor in the maintenance of these gradients.     

The role of auxin in inductive phenomena

The current status of our knowledge of auxin effects on floral induction is summarized. The most general effect is inhibition, although the concentration of synthetic auxins added to. plants tends to be too high for us to be certain that the inhibitory effects are truly physiological. Studies of endogenous levels of auxin have focused almost entirely on IAA-like bioassay activity. Chemical identifications of endogenous IAA are needed and feasible. In addition, a search for-other auxins involved in vegetative to floral transitions, their chemical identification, and measurement of their changing levels in the plant are urgently needed.     

A novel role of BELL1-like homeobox genes, PENNYWISE and POUND-FOOLISH, in floral patterning

Flowers are determinate shoots comprised of perianth and reproductive organs displayed in a whorled phyllotactic pattern. Floral organ identity genes display region-specific expression patterns in the developing flower. In Arabidopsis, floral organ identity genes are activated by LEAFY (LFY), which functions with region-specific co-regulators, UNUSUAL FLORAL ORGANS (UFO) and WUSCHEL (WUS), to up-regulate homeotic genes in specific whorls of the flower. PENNYWISE (PNY) and POUND-FOOLISH (PNF) are redundant functioning BELL1-like homeodomain proteins that are expressed in shoot and floral meristems. During flower development, PNY functions with a co-repressor complex to down-regulate the homeotic gene, AGAMOUS (AG), in the outer whorls of the flower. However, the function of PNY as well as PNF in regulating floral organ identity in the central whorls of the flower is not known. In this report, we show that combining mutations in PNY and PNF enhance the floral patterning phenotypes of weak and strong alleles of lfy, indicating that these BELL1-like homeodomain proteins play a role in the specification of petals, stamens and carpels during flower development. Expression studies show that PNY and PNF positively regulate the homeotic genes, APETALA3 and AG, in the inner whorls of the flower. Moreover, PNY and PNF function in parallel with LFY, UFO and WUS to regulate homeotic gene expression. Since PNY and PNF interact with the KNOTTED1-like homeodomain proteins, SHOOTMERISTEMLESS (STM) and KNOTTED-LIKE from ARABIDOPSIS THALIANA2 (KNAT2) that regulate floral development, we propose that PNY/PNF-STM and PNY/PNF-KNAT2 complexes function in the inner whorls to regulate flower patterning events.