Interaction between Meristem Tissue Layers Controls Phyllotaxis

Phyllotaxis and vein formation are among the most conspicuous patterning processes in plants. The expression and polarization of the auxin efflux carrier PIN1 is the earliest marker for both processes, with mathematical models indicating that PIN1 can respond to auxin gradients and/or auxin flux. Here, we use cell-layer-specific PIN1 knockouts and partial complementation of auxin transport mutants to examine the interaction between phyllotactic patterning, which occurs primarily in the L1 surface layer of the meristem, and midvein specification in the inner tissues. We show that PIN1 expression in the L1 is sufficient for correct organ positioning, as long as the L1-specific influx carriers are present. Thus, differentiation of inner tissues can proceed without PIN1 or any of the known polar transporters. On theoretical grounds, we suggest that canalization of auxin flux between an auxin source and an auxin sink may involve facilitated diffusion rather than polar transport.

Regulation of phyllotaxis by polar auxin transport

The regular arrangement of leaves around a plant's stem, called phyllotaxis, has for centuries attracted the attention of philosophers, mathematicians and natural scientists; however, to date, studies of phyllotaxis have been largely theoretical. Leaves and flowers are formed from the shoot apical meristem, triggered by the plant hormone auxin. Auxin is transported through plant tissues by specific cellular influx and efflux carrier proteins. Here we show that proteins involved in auxin transport regulate phyllotaxis. Our data indicate that auxin is transported upwards into the meristem through the epidermis and the outermost meristem cell layer. Existing leaf primordia act as sinks, redistributing auxin and creating its heterogeneous distribution in the meristem. Auxin accumulation occurs only at certain minimal distances from existing primordia, defining the position of future primordia. This model for phyllotaxis accounts for its reiterative nature, as well as its regularity and stability.

Phyllotaxis involves auxin drainage through leaf primordia

The spatial arrangement of leaves and flowers around the stem, known as phyllotaxis, is controlled by an auxin-dependent reiterative mechanism that leads to regular spacing of the organs and thereby to remarkably precise phyllotactic patterns. The mechanism is based on the active cellular transport of the phytohormone auxin by cellular influx and efflux carriers, such as AUX1 and PIN1. Their important role in phyllotaxis is evident from mutant phenotypes, but their exact roles in space and time are difficult to address due to the strong pleiotropic phenotypes of most mutants in phyllotaxis. Models of phyllotaxis invoke the accumulation of auxin at leaf initials and removal of auxin through their developing vascular strand, the midvein. We have developed a precise microsurgical tool to ablate the midvein at high spatial and temporal resolution in order to test its function in leaf formation and phyllotaxis. Using amplified femtosecond laser pulses, we ablated the internal tissues in young leaf primordia of tomato (Solanum lycopersicum) without damaging the overlying L1 and L2 layers. Our results show that ablation of the future midvein leads to a transient accumulation of auxin in the primordia and to an increase in their width. Phyllotaxis was transiently affected after midvein ablations, but readjusted after two plastochrons. These results indicate that the developing midvein is involved in the basipetal transport of auxin through young primordia, which contributes to phyllotactic spacing and stability.

Regulation of phyllotaxis

Plant architecture is characterized by a high degree of regularity. Leaves, flowers and floral organs are arranged in regular patterns, a phenomenon referred to as phyllotaxis. Regular phyllotaxis is found in virtually all higher plants, from mosses, over ferns, to gymnosperms and angiosperms. Due to its remarkable precision, its beauty and its accessibility, phyllotaxis has for centuries been the object of admiration and scientific examination. There have been numerous hypotheses to explain the nature of the mechanistic principle behind phyllotaxis, however, not all of them have been amenable to experimental examination. This is due mainly to the delicacy and small size of the shoot apical meristem, where plant organs are formed and the phyllotactic patterns are laid down. Recently, the combination of genetics, molecular tools and micromanipulation has resulted in the identification of auxin as a central player in organ formation and positioning. This paper discusses some aspects of phyllotactic patterns found in nature and summarizes our current understanding of the regulatory mechanism behind phyllotaxis.

Phyllotaxis — a new chapter in an old tale about beauty and magic numbers

Phyllotaxis, the regular arrangement of leaves and flowers around the stem, is one of the most fascinating patterning phenomena in biology. Numerous theoretical models, that are based on biochemical, biophysical and other principles, have been proposed to explain the development of the patterns. Recently, auxin has been identified as the inducer of organ formation. An emerging model for phyllotaxis states that polar auxin transport in the plant apex generates local peaks in auxin concentration that determine the site of organ formation and thereby the different phyllotactic patterns found in nature. The PIN proteins play a primary role in auxin transport. These proteins are localized in a polar fashion, reflecting the directionality of polar auxin transport. Recent evidence shows that most aspects of phyllotaxis can be explained by the expression pattern and the dynamic subcellular localization of PIN1.

Auxin and phyllotaxis

Our understanding of phyllotaxis is still largely based on surgical and pharmacological experiments carried out before 1970. Recent experiments implicate the plant hormone auxin in the regulation of phyllotaxis. A recent paper shows how the polar auxin transport mutant, pin1-1, which fails to make flowers, affects the expression of well known meristem genes. This work opens the door for the genetic analysis of phyllotaxis.

Studies of aberrant phyllotaxy1 Mutants of Maize Indicate Complex Interactions between Auxin and Cytokinin Signaling in the Shoot Apical Meristem

One of the most fascinating aspects of plant morphology is the regular geometric arrangement of leaves and flowers, called phyllotaxy. The shoot apical meristem (SAM) determines these patterns, which vary depending on species and developmental stage. Auxin acts as an instructive signal in leaf initiation, and its transport has been implicated in phyllotaxy regulation in Arabidopsis (Arabidopsis thaliana). Altered phyllotactic patterns are observed in a maize (Zea mays) mutant, aberrant phyllotaxy1 (abph1, also known as abphyl1), and ABPH1 encodes a cytokinin-inducible type A response regulator, suggesting that cytokinin signals are also involved in the mechanism by which phyllotactic patterns are established. Therefore, we investigated the interaction between auxin and cytokinin signaling in phyllotaxy. Treatment of maize shoots with a polar auxin transport inhibitor, 1-naphthylphthalamic acid, strongly reduced ABPH1 expression, suggesting that auxin or its polar transport is required for ABPH1 expression. Immunolocalization of the PINFORMED1 (PIN1) polar auxin transporter revealed that PIN1 expression marks leaf primordia in maize, similarly to Arabidopsis. Interestingly, maize PIN1 expression at the incipient leaf primordium was greatly reduced in abph1 mutants. Consistently, auxin levels were reduced in abph1, and the maize PIN1 homolog was induced not only by auxin but also by cytokinin treatments. Our results indicate distinct roles for ABPH1 as a negative regulator of SAM size and a positive regulator of PIN1 expression. These studies highlight a complex interaction between auxin and cytokinin signaling in the specification of phyllotactic patterns and suggest an alternative model for the generation of altered phyllotactic patterns in abph1 mutants. We propose that reduced auxin levels and PIN1 expression in abph1 mutant SAMs delay leaf initiation, contributing to the enlarged SAM and altered phyllotaxy of these mutants.

Semi-hypogeal germination in Pachyrhizus ahipa (Wedd.) parodi (Fabaceae: Phaseoleae): seedling and sapling morphology

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morfologia de plântulas e plantas jovens de 30 espécies arbóreas de Leguminosae

Tendo em vista a carência de informações a respeito das formas jovens das plantas e a importância desses dados como subsídios para trabalhos taxonômicos, filogenéticos e ecológicos, foram estudadas morfologicamente as plântulas e plantas jovens de 30 espécies arbóreas de Leguminosae, ocorrentes no Estado de São Paulo, Brasil, visando à apresentação de características úteis para a identificação das espécies selecionadas. O desenvolvimento das plântulas e plantas jovens foi acompanhado diariamente, em germinador e em casa de vegetação. São apresentados dados relativos à morfologia da plântula, número de catáfilos, época de diferenciação do primeiro eófilo e sua filotaxia, época da abscisão cotiledonar e da formação do primeiro metáfilo, bem como a ocorrência de nodulação radicular. A análise das plântulas e plantas jovens demonstrou a grande variação que existe nas Leguminosae. Nessa família, plântulas epígeo-foliáceas e epígeo-carnosas ocorrem em 80% das espécies estudadas. em Caesalpinioideae, todas as plântulas se mostraram epígeas, 20% delas com cotilédones carnosos. em Mimosoideae, 66,7% das espécies produziram plântulas epígeo-foliáceas, 22,2% epígeo-carnosas e 11,1% semi-hipógeas. As espécies de Faboideae apresentaram dois tipos de plântulas: epígeo-carnosas em 54,5% e hipógeas em 45,5%.

Aspectos morfológicos de frutos, sementes, germinação e plântulas de Hymenolobium petraeum

Hymenolobium petraeum Ducke é uma espécie arbórea pertencente à família Leguminosae conhecida popularmente por angelim-pedra. Apresenta alto valor comercial, com madeira muito utilizada na construção civil e marcenaria. Este trabalho teve por objetivo descrever morfologicamente o fruto, a semente e as plântulas, assim como, determinar as temperaturas cardeais para a germinação de sementes de angelim-pedra. Foram determinados o comprimento, a largura e a massa fresca de frutos e sementes. Para os testes de germinação foram utilizadas três repetições de 50 sementes, colocadas em placas de Petri e mantidas em germinadores nas temperaturas de 15, 20, 25, 30, 35 e 40°C e fotoperíodo de 12 horas. Os frutos são legumes-samaróides, indeiscentes, oblongos e unicarpelares. As sementes são de coloração castanho-escura, oblongas, estenospérmicas, exalbuminosas e com plúmula inconspícua. A raiz primária é branca e pubescente na região próxima ao colo; a parte aérea das plântulas possui pilosidade branca, protófilos compostos imparipinados e com inserção oposta, epicótilo verde, ereto, cilíndrico e piloso e os metáfilos imparipinados e com inserção alterna-espiralada. A germinação é semi-hipógea criptocotiledonar. Para de sementes de angelim-pedra as temperaturas máximas de germinação estão acima de 35°C e a mínima abaixo de 15°C, enquanto a faixa de temperatura ótima para germinação está entre 25 e 35°C.