Process Of Leaf Development Research Articles

Advancements in Rice Leaf Development Research

Rice leaf morphology is a pivotal component of the ideal plant architecture, significantly impacting rice yield. The process of leaf development unfolds through three distinct stages: the initiation of leaf primordia, the establishment and maintenance of polarity, and leaf expansion. Genes regulating leaf morphology encompass transcription factors, hormones, and miRNAs. An in-depth synthesis and categorization of genes associated with leaf development, particularly those successfully cloned, hold paramount importance in unraveling the complexity of rice leaf development. Furthermore, it provides valuable insights into the potential for molecular-level manipulation of rice leaf types. This comprehensive review consolidates the stages of rice leaf development, the genes involved, molecular regulatory pathways, and the influence of plant hormones. Its objective is to establish a foundational understanding of the creation of ideal rice leaf forms and their practical application in molecular breeding.

Transcriptome Analyses Reveal the Key Regulators of Tomato Compound Leaf Development

Leaves are one of the organs involved in plant assimilation and transpiration. Different leaf development processes can result in different leaf shapes. Tomato plants have typical compound leaves. It is helpful to explore the regulatory factors affecting the leaf development and morphogenesis of tomatoes to cultivate varieties with high photosynthetic efficiency. We used the typical tomato leaf shape mutants Petroselinum (Pts), Trifoliate (tf2), and Entire (e), which showed a gradual decrease in leaflet number and compound leaf complexity. Transcriptome sequencing was performed to analyze the key differentially expressed genes (DEGs) among the 3 groups, which revealed 2393, 1366, and 1147 DEGs in Pts/VF36, tf2/CR, and e/AC, respectively. We found 86 overlapping DEGs among the 3 groups. In addition, we found that the mutation of Pts, tf2, and e affected not only leaf morphology but also the wax, fatty acid, and abscisic acid pathways during growth and development. An RT-qPCR analysis during leaf primordium development revealed three transcription factors (bHLH079, WRKY44, and WRKY76) and three hormone-regulated genes (IAA-amino acid hydrolase, Gibberellin2ox7, and Gibberellin20ox) that were differentially expressed in the transcriptome. Using virus-induced gene silencing (VIGS), we observed the leaf shape of VIGS plants and found that bHLH079, IAA-amino acid hydrolase, Gibberellin2ox7, Gibberellin20ox, WRKY44, and WRKY76 were the endogenous regulators influencing tomato compound leaf development. This study provides a promising direction for revealing the molecular regulation mechanism underlying compound leaf development in tomatoes.

The rubber tree RALF peptide hormone and its receptor protein kinase FER implicates in rubber production

RAPID ALKALINIZATION FACTORs (RALFs), which are secreted peptides serving as extracellular signals transduced to the inside of the cell, interact with the receptor-like kinase FERONIA (FER) and participates in various biological pathways. Here, we identified 23 RALF and 2 FER genes in Hevea brasiliensis (para rubber tree), and characterized their expression patterns in different tissues, across the process of leaf development, and in response to the rubber yield-stimulating treatments of tapping and ethylene. Four Hevea latex (the cytoplasm of rubber-producing laticifers)-abundant RALF isoforms, HbRALF19, HbRALF3, HbRALF22, and HbRALF16 were listed with descending expression levels. Of the four HbRALFs, expressions of HbRALF3 were markedly regulated in an opposite way by the treatments of tapping (depression) and ethylene (stimulation). All of the four latex-abundant RALFs specifically interacted with the extracellular domain of HbFER1. Transgenic Arabidopsis plants overexpressing these HbRALFs displayed phenotypes similar to those reported for AtRALFs, such as shorter roots, smaller plant architecture, and delayed flowering. The application of HbRALF3 and HbRALF19 recombinant proteins significantly reduced the pH of Hevea latex, an important factor regulating latex metabolism. An in vitro rubber biosynthesis assay in a mixture of latex cytosol (C-serum) revealed a positive role of HbFER1 in rubber biosynthesis. Taken together, these data provide evidence for the participation of the HbRALF-FER module in rubber production.

The Formation of Shapes: Interplay of Genes during Leaf Development Processes

Leaf shape, as one of the clearest manifestations of plant morphology, shows considerable variation owing to genetics and the environment. Leaf initiation occurs in the peripheral zone of the SAM and goes through the three overlapping phases of leaf primordium initiation, leaf dorsiventral development, and leaf marginal meristem establishment. Transcription factors, such as KNOX, WOX, and CUC; hormone-regulating genes, such as GA2ox, GA20ox, and PIN1; and miRNAs such as miR164/165 are tightly involved in leaf shaping through the generation of intricate cooperative networks in different temporal phases and specific tissue zones. Here, we briefly discuss the critical interplay occurring between certain genes and the pivotal role these play in the leaf developmental network and phytohormone regulation, including AS1/AS2–KNOX–GA20ox–GA, miR164–NAM/CUC–PIN1–auxin, and CUC–BAS1/CYP734A–BR, and we attempt to summarize several basic insights into the mechanisms of leaf shape regulation.

The Eucalyptus grandis chloroplast proteome: Seasonal variations in leaf development.

Chloroplast metabolism is very sensitive to environmental fluctuations and is intimately related to plant leaf development. Characterization of the chloroplast proteome dynamics can contribute to a better understanding on plant adaptation to different climate scenarios and leaf development processes. Herein, we carried out a discovery-driven analysis of the Eucalyptus grandis chloroplast proteome during leaf maturation and throughout different seasons of the year. The chloroplast proteome from young leaves differed the most from all assessed samples. Most upregulated proteins identified in mature and young leaves were those related to catabolic-redox signaling and biogenesis processes, respectively. Seasonal dynamics revealed unique proteome features in the fall and spring periods. The most abundant chloroplast protein in humid (wet) seasons (spring and summer) was a small subunit of RuBisCO, while in the dry periods (fall and winter) the proteins that showed the most pronounced accumulation were associated with photo-oxidative damage, Calvin cycle, shikimate pathway, and detoxification. Our investigation of the chloroplast proteome dynamics during leaf development revealed significant alterations in relation to the maturation event. Our findings also suggest that transition seasons induced the most pronounced chloroplast proteome changes over the year. This study contributes to a more comprehensive understanding on the subcellular mechanisms that lead to plant leaf adaptation and ultimately gives more insights into Eucalyptus grandis phenology.

Dimorphic Leaf Development of the Aquatic Plant Callitriche palustris L. Through Differential Cell Division and Expansion.

Heterophylly, or phenotypic plasticity in leaf form, is a remarkable feature of amphibious plants. When the shoots of these plants grow underwater, they often develop surprisingly different leaves from those that emerge in air. Among aquatic plants, it is typical for two or more distinct leaf development processes to be observed in the same individual exposed to different environments. Here, we analyze the developmental processes of heterophylly in the amphibious plant Callitriche palustris L. (Plantaginaceae). First, we reliably cultured this species under laboratory conditions and established a laboratory strain. We also established a framework for molecular-based developmental analyses, such as whole-mount in situ hybridization. We observed several developmental features of aerial and submerged leaves, including changes in form, stomata and vein formation, and transition of the meristematic zone. Then we defined developmental stages for C. palustris leaves. We found that in early stages, aerial and submerged leaf primordia had similar forms, but became discriminable through cell divisions with differential direction, and later became highly distinct via extensive cell elongation in submerged leaf primordia.

The Analysis of Genes and Phytohormone Metabolic Pathways Associated with Leaf Shape Development in Liriodendron chinense via De Novo Transcriptome Sequencing.

The leaf, a photosynthetic organ that plays an indispensable role in plant development and growth, has a certain ability to adapt to the environment and exhibits tremendous diversity among angiosperms. Liriodendron chinense, an ancestral angiosperm species, is very popular in landscaping. The leaf of this species has two lobes and resembles a Qing Dynasty Chinese robe; thus, leaf shape is the most valuable ornamental trait of the tree. In this work, to determine the candidate genes associated with leaf development in L. chinense, scanning electron microscopy (SEM) was employed to distinguish the developmental stages of tender leaves. Four stages were clearly separated, and transcriptome sequencing was performed for two special leaf stages. Altogether, there were 48.23 G clean reads in the libraries of the two leaf developmental stages, and 48,107 assembled unigenes were annotated with five databases. Among four libraries, 3118 differentially expressed genes (DEGs) were enriched in expression profiles. We selected ten DEGs associated with leaf development and validated their expression patterns via quantitative real-time PCR (qRT-PCR) assays. Most validation results were closely correlated with the RNA-sequencing data. Taken together, we examined the dynamic process of leaf development and indicated that several transcription factors and phytohormone metabolism genes may participate in leaf shape development. The transcriptome data analysis presented in this work aims to provide basic insights into the mechanisms mediating leaf development, and the results serve as a reference for the genetic breeding of ornamental traits in L. chinense.

Ethylene Treatment in Studying Leaf Senescence in Arabidopsis.

Plant leaf senescence is the final process of leaf development that involves the mobilization of nutrients from old leaves to newly growing tissues. Leaf senescence involves a coordinated action at the cellular and organism levels under the control of a highly regulated genetic program. However, leaf senescence is also influenced by multiple internal and environmental signals that are integrated into the age information. Among these internal factors, the simple gaseous phytohormone ethylene is well-known to be an important endogenous modulator of plant leaf senescence and fruit ripening. Ethylene and its biosynthetic precursor ACC (1-aminocyclopropane-l-carboxylic acid) as well as inhibitors of ethylene biosynthesis aminoethoxyvinylglycine (AVG) and action (AgNO3) or the inhibitor of ethylene perception 1-MCP (1-methylcyclopropene) have been widely used by the research community of plant leaf senescence. However, until now, no systemically experimental method about the usage of ethylene in studying the molecular mechanisms of leaf senescence in Arabidopsis is available. Here, we provide detailed methods for exogenous application of ethylene and its inhibitors in studying Arabidopsis leaf senescence, which has been successfully used in our laboratory.

Transcription factors NF-YA2 and NF-YA10 regulate leaf growth via auxin signaling in Arabidopsis

In plants, leaf is crucial for photosynthesis and respiration. Leaf area and quantity are important for leaf vegetables to increase biomass. The process of leaf development involves coordinated regulation among small RNAs, transcription factors and hormones. Here, we found leaf size were regulated by transcription factors NF-YA2 and NF-YA10 in Arabidopsis. NF-YA2 and NF-YA10 overexpression increased biomass accumulation through promoting leaf growth and cell expansion. NF-YA2 and NF-YA10 were expressed in SAM and leaf vasculature. Endogenous IAA content reduced by 20% and 24% in transgenic Arabidopsis plants overexpressing NF-YA2 and NF-YA10 compared to wild-type plants. Chromatin immunoprecipitation assays revealed that NF-YA2 and NF-YA10 bound directly to the cis-element CCAAT in the promoter of the YUC2, and decreased the expression of YUC2, a YUCCA family gene. The auxin transporter gene PIN1 and auxin response factor1 and 2 (ARF1 and ARF2) genes, transcriptional repressors, were downregulated. These findings showed leaf development was regulated by NF-YA2 and NF-YA10 through the auxin-signaling pathway and may provide a new insight into the genetic engineering of vegetables biomass and crop productivity.

A Regulatory Cascade Involving Class II ETHYLENE RESPONSE FACTOR Transcriptional Repressors Operates in the Progression of Leaf Senescence

Leaf senescence is the final process of leaf development that involves the mobilization of nutrients from old leaves to newly growing tissues. Despite the identification of several transcription factors involved in the regulation of this process, the mechanisms underlying the progression of leaf senescence are largely unknown. Herein, we describe the proteasome-mediated regulation of class II ETHYLENE RESPONSE FACTOR (ERF) transcriptional repressors and involvement of these factors in the progression of leaf senescence in Arabidopsis (Arabidopsis thaliana). Based on previous results showing that the tobacco (Nicotiana tabacum) ERF3 (NtERF3) specifically interacts with a ubiquitin-conjugating enzyme, we examined the stability of NtERF3 in vitro and confirmed its rapid degradation by plant protein extracts. Furthermore, NtERF3 accumulated in plants treated with a proteasome inhibitor. The Arabidopsis class II ERFs AtERF4 and AtERF8 were also regulated by the proteasome and increased with plant aging. Transgenic Arabidopsis plants with enhanced expression of NtERF3, AtERF4, or AtERF8 showed precocious leaf senescence. Our gene expression and chromatin immunoprecipitation analyses suggest that AtERF4 and AtERF8 targeted the EPITHIOSPECIFIER PROTEIN/EPITHIOSPECIFYING SENESCENCE REGULATOR gene and regulated the expression of many genes involved in the progression of leaf senescence. By contrast, an aterf4 aterf8 double mutant exhibited delayed leaf senescence. Our results provide insight into the important role of class II ERFs in the progression of leaf senescence.

Effect of metabolic enzyme on organic acids in developing ‘Dangshansuli’ pear leaf

Changes in the content of citric and malic acids and the activities of enzymes involved in the metabolism of these two organic acids, including citrate synthase (CS), cytoplast aconitase (Cyt-ACO), NAD-isocitrate dehydrogenase (NAD-IDH), phosphoenolpyruvate carboxylase (PEPC), NAD-malate dehydrogenase (NAD-MDH) and NADP-malic enzyme (NADP-ME), were examined during the process of leaf development in ‘Dangshansuli’ pear. The citric acid content exhibited an overall slightly decreasing trend and the malic acid content exhibited an overall appreciably increasing trend in developing leaf. The activities of CS, Cyt-ACO, NAD-IDH, PEPC and NAD-MDH all showed an overall decreasing trend, whereas the activities of NADP-ME indicated an overall increasing trend. The correlation analysis of the change in the organic acid content and related metabolic enzyme activities during the development of the leaf revealed that the activities of CS, Cyt-ACO and NAD-IDH showed a negative correlation with the content of citrc acid, the activities of PEPC exhibited a positive correlation with the content of malic acid, the activities of NAD-MDH and NADP-ME indicated a negative correlation with the content of malic acid. Key words: Pear, leaf, development, organic acid, metabolic enzyme, correlation.

The volumetric component of individual leaf expansion: Taking into account sub-epidermal tissues in the description of leaf expansion over time

Most leaf development studies at the cell and organ levels have been limited to the leaf surface, with data referring to the leaf surface area and to the number and surface area of epidermal cells. However, leaf sub-epidermal tissues, the palisade and spongy mesophyll, contain the main actors in photosynthesis. The number and thickness of palisade cell layers and the volume occupied by spongy mesophyll (cells and intercellular spaces) affect the accumulation of photosynthates and, as such, whole plant growth. Studies into the leaf phenotype of growth-affected Arabidopsis thaliana mutants have revealed a higher variability in leaf thickness than in leaf surface area. In general, there is no correlation between these two variables, which means that to describe a leaf phenotype, leaf volume has to be taken into account. A method has been developed for high-resolution imaging of leaves in three dimensions usingmultiphoton laser scanning microscopy, and for the analysis of images, providing data on volumes and volumetric proportions of cells and tissues and cell density. The method has been used in the study of A. thaliana leaf expansion from emergence to the onset of senescence for leaves located at different nodal positions in the rosette, completing our knowledge of individual leaf development processes with their volumetric component. The method will further be applied in the study of leaf plasticity in response to the environment for both A. thaliana and apple tree, a model and an agronomic species, respectively. (Texte integral)

애기장대의 세포 크기와 세포 수를 조절하는 LANCEOLATA1 유전자

쌍자엽 식물에 있어서 잎의 발달에 관한 과정은 매우 복잡하게 얽혀있지만, 이를 조절하는 메커니즘에 대해서는 아직 잘 알려져 있지 않다. 모델식물인 애기장대의 잎의 발달의 기초과정을 해명하기 위하여 잎의 형태이상 돌연변이체를 이용한 접근방법을 시도하였다. 본 연구에서 잎이 작고 가는 lanceolata1 돌연변이체를 선별하였고, 이원인유전자의 기능을 밝히기 위하여 유전학적 및 해부학적인 연구를 실시하였다. lan1-7 돌연변이체는 잎 뿐만아니라 줄기도 가늘어지는 표현형을 나타내었고, 이는 세포수준에서 세포크기와 세포수의 감소에 의한 조절되고 있음이 밝혀졌다. 이 결과는 LAN1 유전자가 잎의 발달과정에서 세포분열과 세포신장을 제어하고 있다는 사실을 시사하고 있다. lan1-7 돌연변이체와 35S-AG 형질전환체와의 F1 세대의 표현형 분석을 통하여, AG 유전자가 잎이 가늘고 위로 말리는 모양을 제어하고 있다는 사실이 판명되었다. 이 결과로 MADS-box유전자가 LAN1 유전자에 의해서 제어되고 잎의 발달과정에 관여하고 있다는 사실이 판명되었다. The Precesses for leaf development in dicotyledonous plants are surprisingly complex, while the mechanism of controlling and coordinating them is poorly understood. To characterize the fundamental features of the leaf development of Arabidopsis, we first attempted to isolate mutants that alter leaf morphology. Here, leaf morphological mutant of Arabidopsis, lanceolatal (lan1) which has small and narrow leaves have isolated and characterized. To clarify the function of LAN1 in organ development, we characterized lan1-7 mutant using an anatomical and genetic approach. The lan1-7 mutant had reduced size of foliage leaves and reduced dimensions of stems. A reduction both in cell size and in cell number was evident at the cellular level in the lan1 mutant, revealing that LAN1 gene appears to affect cell division at an earlier stage and cell elongation throughout the development of leaf primordia. from the analysis of heterogeneous plant with lan1 mutation and 35S-AG transgenic plant, AG gene is revealed to regulate leaf morphology under the control of 35S promoter. Thus, MADS-box gene was revealed to have some relationship to that of LAN1 gene at certain stage in leaf development processes.

Seasonal variations in water relations in current-year leaves of evergreen trees with delayed greening

We investigated seasonal patterns of water relations in current-year leaves of three evergreen broad-leaved trees (Ilex pedunculosa Miq., Ligustrum japonicum Thunb., and Eurya japonica Thunb.) with delayed greening in a warm-temperate forest in Japan. We used the pressure-volume method to: (1) assess the extent to which seasonal variation in leaf water relations is attributable to leaf development processes in delayed greening leaves versus seasonal variation in environmental variables; and (2) investigate variation in leaf water relations during the transition from the sapling to the adult tree stage. Leaf mass per unit leaf area was generally lowest just after completion of leaf expansion in May (late spring), and increased gradually throughout the year. Osmotic potential at full turgor (Psi(o) (ft)) and leaf water potential at the turgor loss point (Psi(w) (tlp)) were highest in May, and lowest in midwinter in all species. In response to decreasing air temperature, Psi(o) (ft) dropped at the rate of 0.037 MPa degrees C(-1). Dry-mass-based water content of leaves and the symplastic water fraction of total leaf water content gradually decreased throughout the year in all species. These results indicate that reductions in the symplastic water fraction during leaf development contributed to the passive concentration of solutes in cells and the resulting drop in winter Psi(o) (ft). The ratio of solutes to water volume increased in winter in current-year leaves of L. japonicum and E. japonica, indicating that osmotic adjustment (active accumulation of solutes) also contributed to the drop in winter in Psi(o) (ft). Bulk modulus of elasticity in cell walls fluctuated seasonally, but no general trend was found across species. Over the growing season, Psi(o) (ft) and Psi(w) (tlp) were lower in adult trees than in saplings especially in the case of I. pedunculosa, suggesting that adult-tree leaves are more drought and cold tolerant than sapling leaves. The ontogenetic increase in the stress resistance of I. pedunculosa may be related to its characteristic life form because I. pedunculosa grows taller than the other species studied.

Fine mapping of an incomplete recessive gene for leaf rolling in rice (Oryza sativa L.)

Genetic analysis and fine mapping of genes controlling leaf rolling were conducted using two backcrossed generations (BC4F2, BC4F3) derived from a cross between QMX, a non-rolled leaf cultivar as a recurrent parent, and JZB, a rolled leaf NIL of ZB as a donor parent. Results indicated that leaf rolling was mainly controlled by an incompletely recessive major gene, namelyrl (t), and at the same time, affected by quantitative trait loci (QTLs) and/or the environment. A genetic linkage map was constructed using MAPMAKER/EXP3.0 with eight polymorphic markers on chromosome 2, which were screened by BAS method from 500 SSR markers and 15 newly developed insertion/deletion (InDel) markers. The position ofrl (t) was estimated with composite interval mapping (CIM) method using WinQTLcart2.5. Generl (t) was mapped between markers InDel 112 and RM3763, and 1.0 cM away from InDel 112 using 241 plants in BC4F2 population. To fine maprl (t), one BC4F3 line with 855 plants was generated from one semi-rolled leaf plant in BC4F2. Four new polymorphic InDel markers were developed, including InDel 112.6 and InDel 113 located between markers InDel112 and RM3763. Based on the information of recombination offered by 191 rolled leaf plants and 185 non-rolled leaf plants from the BC4F3 line, we mappedrl (t) to a 137-kb region between markers InDel 112.6 and InDel 113. Homologous gene analysis suggested thatrl (t) was probably related to the process of leaf development regulated by microRNA.

Enhanced photoprotection at the early stages of leaf expansion in field-grown soybean plants

Gas exchange, chlorophyll a fluorescence kinetics, chloroplast pigments and antioxidant enzymes were investigated to explore the development of photoprotective mechanisms in soybean leaves from emergence to full expansion under field conditions. During leaf development, photosynthesis ( P n) gradually increased. Although the maximum quantum yield of PSII photochemistry ( F v/ F m) was quite high at the initial stages of leaf development, it was appreciably lower than that in fully expanded leaves. During daily courses, reversible decrease in F v/ F m was noted at various stages, implying that no photodamage occurred. When exposed to irradiance, a substantial elevation in the actual PSII efficiency ( Φ PSII) together with a remarkable decrease in non-photochemical quenching (NPQ) was observed with the process of leaf development. In this study, we observed that newly initiating leaves possessed relative larger xanthophyll pool size (V + A + Z)/Chl, but it began to decrease with leaf expanding until fully expanded. Further experiments revealed that the decline of relative xanthophyll cycle pool size during leaf expanding was owing to the fact that the chlorophyll contents per area increased faster than that of the xanthophyll cycle pigments. In addition, activities of the antioxidative enzymes, such as SOD, APX, CAT, POD and GR, were also enhanced at the early stages of leaf expansion. We suggested that photoprotective mechanisms, including xanthophyll cycle and antioxidant enzymic system, have developed firstly as soon as leaf emerged. It was the timely development of the mechanisms in young leaves that dissipate excess excitation energy protecting leaves against photodamage at the early stages of leaf development. The relative leaf water content increased with the process of leaf development and exhibited significant negative linear correlation with NPQ, xanthophyll cycle pigments and antioxidant enzyme activities in soybean leaves. We proposed that the changes in leaf turgor might be involved in triggering the activation of photoprotective mechanisms together with high irradiance at the initiating stages of leaf expanding.

CDNA cloning and expression of the phytoene synthase gene in sunflower

A PCR approach was used to isolate a phytoene synthase (Psy) cDNA from sunflower (Helianthus annuus L.). A reconstructed full-length sequence (1598 bp) of the Psy cDNA was obtained; it contained a 1242 bp CDS, 172-nucleotides of 5'-untranslated region (UTR), and 170-nucleotides of 3'-UTR. The predicted protein (46.8kDa) displayed a sequence of 414 amino acid residues with a putative transit sequence for plastid targeting in the N-terminal region. Phylogenetic analysis demonstrated that the sunflower Psy (HaPsy) clustered with the marigold (Tagetes erecta) Psy gene, with which it showed an overall amino acid identity of 93.7%. Moreover, the HaPsy sequence relates closely with other Psy sequences of higher plants. HaPsy was highly expressed in cotyledons, and young and mature leaves. In contrast, HaPsy transcript levels were comparatively lower in the stem and almost absent in the roots. The HaPsy transcript levels were influenced by leaf expansion, which suggested that the expression of the HaPsy gene is regulated during the process of leaf development. The role of HaPsy in controlling carotenoid biosynthesis is demonstrated by the concurrent increase of HaPsy transcript levels with the light-dependent enhanced carotenoid production in green tissues of sunflower.

Leaf development.

The shoot system is the basic unit of development of seed plants and is composed of a leaf, a stem, and a lateral bud that differentiates into a lateral shoot. The most specialized organ in angiosperms, the flower, can be considered to be part of the same shoot system since floral organs, such as the sepal, petal, stamen, and carpel, are all modified leaves. Scales, bracts, and certain kinds of needle are also derived from leaves. Thus, an understanding of leaf development is critical to an understanding of shoot development. Moreover, leaves play important roles in photosynthesis, respiration and photoperception. Thus, a full understanding of leaves is directly related to a full understanding of seed plants.The details of leaf development remain unclear. The difficulties encountered in studies of leaf development, in particular in dicotyledonous plants such as Arabidopsis thaliana (L.) Henyn., are derived from the complex process of leaf development, during which the division and elongation of cells occur at the same time and in the same region of the leaf primordium (Maksymowych, 1963; Poethig and Sussex, 1985). Thus, we cannot divide the entire process into unit processes in accordance with the tenets of classical anatomy.Genetic approaches in Arabidopsis, a model plant (Meyerowitz and Pruitt, 1985), have provided a powerful tool for studies of mechanisms of leaf development in dicotyledonous plants, and various aspects of the mechanisms that control leaf development have been revealed in recent developmental and molecular genetic studies of Arabidopsis (for reviews, see Tsukaya, 1995 and 1998; Van Lijsebettens and Clarke, 1998; Sinha, 1999; Van Volkenburgh, 1999; Tsukaya, 2000; Byrne et al., 2001; Dengler and Kang, 2001; Dengler and Tsukaya, 2001; Tsukaya, 2001). In this review, we shall examine the information that is currently available about various mechanisms of leaf development in Arabidopsis. Vascular patterning is also an important factor in the determination of leaf shape, and this topic is reviewed in this resource by Turner (see also Dengler and Kang, 2001). The interested reader is also referred to work on the basic characterization of the vascular patterning in foliage leaves of Arabidopsis has been carried out by Candela et al. (1999) and Semiarti et al. (2001). For terminology, see (Fig. 1).

Local expression of expansin induces the entire process of leaf development and modifies leaf shape.

Expansins are a family of extracellular proteins proposed to play a key role in wall stress relaxation and, thus, in cell and tissue growth. To test the possible function of expansins in morphogenesis, we have developed a technique that allows transient local microinduction of gene expression in transgenic plants. We have used this system to manipulate expansin gene expression in various tissues. Our results indicate that local expansin expression within the meristem induces a developmental program that recapitulates the entire process of leaf formation. Moreover, local transient induction of expansin expression on the flank of developing primordia leads to the induction of ectopic lamina tissue and thus modulation of leaf shape. These data describe an approach for the local manipulation of gene expression and indicate a role for expansin in the control of both leaf initiation and shape. These results are consistent with the action of cell division-independent mechanisms in plant morphogenesis.

Restoration of cardenolide-synthesis in redifferentiated shoots from callus cultures of Digitalis purpurea

No cardenolide was detected in cultured cells and roots of root forming callus of Digitalis purpurea under such conditions as addition of auxins, varied sugar concentration of the medium and illumination. Cardenolides such as digitoxin and purpurea glycoside A were found in leaves from the regenerated shoot from callus cultures. The manifestation of cardenolide-synthesizing potentiality seems to be closely related to some process of leaf development.