The cellular parameters of leaf development in tobacco: a clonal analysis

The cellular parameters of leaf development in tobacco (Nicotiana tabacum L.) have been characterized using clonal analysis, an approach that provides unequivocal evidence of cell lineage. Our results indicate that the tobacco leaf arises from a group of around 100 cells in the shoot apical meristem. Each of these cells contributes to a unique longitudinal section of the axis and transverse section of the lamina. This pattern of cell lincage indicates that primordial cells contribute more or less equally to the growth of the axis, in contrast to the more traditional view of leaf development in which the leaf is pictured as arising from a group of apical initials. Clones induced prior to the initiation of the lamina demonstrate that the subepidermal layer of the lamina arises from at least six files of cells. Submarginal cells usually divide with their spindles parallel to the margin, and therefore contribute relatively little to the transverse expansion of the lamina. During the expansion of the lamina the orientation and frequency of cell division are highly regulated, as is the duration of meristematic growth. Initially, cell division is polarized so as to produce lineages that are at an oblique angle to the midrib; later cell division is in alternating perpendicular planes. The distribution of clones generated by irradiation at various stages of development indicates that cell division ceases at the tip of the leaf when the leaf is about one tenth its final size, and then ceases in progressively more basal regions of the lamina. Variation in the mutation frequency within the lamina reflects variation in the frequency of mitosis. Prior to the mergence of the leaf the frequency of mutation is maximal near the tip of the leaf and extremely low at its base; after emergence, the frequency of mutation increases at the base of the leaf. In any given region of the lamina the frequency of mutation is highest in interveinal regions, and is relatively low near the margin. Thus, both the orientation and frequency of cell division at the leaf margin indicate that this region plays a minor role in the growth of the lamina.Abbreviation MF mutation frequency     

The role of the sheath tube in the development of expanding leaves in perennial ryegrass

A possible morphogenic effect of leaf sheaths on subsequent leaf development was investigated by varying sheath tube (pseudostem) length in plants of perennial ryegrass (Lolium perenne) cv. Talbot by either incising longitudinally or excising the distal portion of the sheath tube, while leaving the basal length of the tube intact. The tube was maintained at predetermined lengths by incising and excising new growth daily. The youngest rapidly expanding leaf was allowed to grow through the tube and was measured when fully expanded. Reducing tube length by excision or by incision from 60 mm to just above the cowl leaf on the apex reduced lamina length by 87% and 77% respectively. Over all tube lengths, laminae in incised treatments were almost twice as long as those in excised treatments. Sheath length followed a similar pattern. The effect on developing leaves of artificially extending sheath tubes (previously excised to 15 mm) to 30 or 45 mm with foil was similar to that of initially excising sheath tubes to 30 or 45 mm. The shorter the sheath tube (reduced by incision) through which the leaves had to grow, the shorter the cells, especially in the laminae. The estimated cell number per row along the length of the laminae ranged from 190 in tillers (shoots) with a very short tube (just above the cowl leaf) to 454 in intact control tillers. It is concluded that the sheath tube has a morphogenic influence on the development of subsequent leaves due to the change in environment of the leaf lamina on appearance, affecting both cell elongation and cell division.     

DNA SYNTHESIS,CELL DIVISION,AND CELL DIFFERENTIATION DURING LEAF DEVELOPMENT OF XANTHIUM PENNSYLVANICUM

Leaf growth consists of two basic processes, cell division and cell enlargement. DNA synthesis is an integral part of cell division and can be studied with autoradiographic techniques and incorporation of some labeled precursor. Studies were made on the synthesis of nuclear DNA through incorporation of ³H-thymidine in various parts of the lamina during the entire course of leaf development of Xanthium pennsylvanicum. The time course analysis of DNA synthesis was correlated with cell division and rates of cell enlargement. Significant differences in ³H-thymidine incorporation were found in various parts of the lamina. Cell division and DNA synthesis were highest in the early stages of development. Since no ³H-thymidine was incorporated after cessation of cell division (LPI 2.8) in the leaf lamina, it appears that DNA synthesis is not needed for enlargement and differentiation of Xanthium cells. Rates of cell enlargement were negligible in the early development and reached their maximum after cessation of mitoses, between plastochron ages (LPI) 3 and 4. Cells matured between LPI's 5 and 6. Enzymatic activity was correlated with cell division and cell differentiation at various stages of leaf development.     

Asymmetric auxin response precedes asymmetric growth and differentiation of asymmetric leaf1 and asymmetric leaf2 Arabidopsis leaves

We have analyzed the development of leaf shape and vascular pattern in leaves mutant for ASYMMETRIC LEAVES1 (AS1) or AS2 and compared the timing of developmental landmarks to cellular response to auxin, as measured by expression of the DR5:beta-glucuronidase (GUS) transgene and to cell division, as measured by expression of the cycB1:GUS transgene. We found that the earliest visible defect in both as1 and as2 first leaves is the asymmetric placement of auxin response at the distal leaf tip. This precedes visible changes in leaf morphology, asymmetric placement of the distal margin gap, formation of margin gaps along the leaf border, asymmetric distribution of marginal auxin, and asymmetry in cell division patterns. Moreover, treatment of developing leaves with either exogenous auxin or an auxin transport inhibitor eliminates asymmetric auxin response and subsequent asymmetric leaf development. We propose that the initial asymmetric placement of auxin at the leaf tip gives rise to later asymmetries in the internal auxin sources, which subsequently result in asymmetrical cell differentiation and division patterns.     

Polyploidy and cellular mechanisms changing leaf size: comparison of diploid and autotetraploid populations in two species of Lolium

BACKGROUND AND AIMS: Growth and development of plant organs, including leaves, depend on cell division and expansion. Leaf size is increased by greater cell ploidy, but the mechanism of this effect is poorly understood. Therefore, in this study, the role of cell division and expansion in the increase of leaf size caused by polyploidy was examined by comparing various cell parameters of the mesophyll layer of developing leaves of diploid and autotetraploid cultivars of two grass species, Lolium perenne and L. multiflorum. METHODS: Three cultivars of each ploidy level of both species were grown under pot conditions in a controlled growth chamber, and leaf elongation rate and the cell length profile at the leaf base were measured on six plants in each cultivar. Cell parameters related to division and elongation activities were calculated by a kinematic method. KEY RESULTS: Tetraploid cultivars had faster leaf elongation rates than did diploid cultivars in both species, resulting in longer leaves, mainly due to their longer mature cells. Epidermal and mesophyll cells differed 20-fold in length, but were both greater in the tetraploid cultivars of both species. The increase in cell length of the tetraploid cultivars was caused by a faster cell elongation rate, not by a longer period of cell elongation. There were no significant differences between cell division parameters, such as cell production rate and cell cycle time, in the diploid and tetraploid cultivars. CONCLUSION: The results demonstrated clearly that polyploidy increases leaf size mainly by increasing the cell elongation rate, but not the duration of the period of elongation, and thus increases final cell size.     

Division and differentiation during normal and liguleless-1 maize leaf development

The maize leaf is composed of a blade and a sheath, which are separated at the ligular region by a ligule and an auricle. Mutants homozygous for the recessive liguleless-1 (lg1) allele exhibit loss of normal ligule and auricle. The cellular events associated with development of these structures in both normal and liguleless plants are investigated with respect to the timing of cell division and differentiation. A new method is used to assess orientation of anticlinal division planes during development and to determine a division index based on recent epidermal cross-wall deposition. A normal leaf follows three stages of development: first is a preligule stage, in which the primordium is undifferentiated and dividing throughout its length. This stage ends when a row of cells in the preligule region divides more rapidly in both transverse and longitudinal anticlinal planes. During the second stage, ligule and auricle form, blade grows more rapidly than sheath, divisions in the blade become exclusively transverse in orientation, and differentiation begins. The third stage is marked by rapid increase in sheath length. The leaf does not have a distinct basal meristem. Instead, cell divisions are gradually restricted to the base of the leaf with localized sites of increased division at the preligule region. Divisions are not localized to the base of the sheath until near the end of development. The liguleless-1 homozygote shows no alteration in this overall pattern of growth, but does show distinct alteration in the anticlinal division pattern in the preligule region. Two abnormal patterns are observed: either the increase in division rate at the preligule site is absent or it exhibits loss of all longitudinal divisions so that only transverse (or cell-file producing) divisions are present. This pattern is particularly apparent in developing adult leaves on older lg1 plants, in which sporadic ligule vestiges form. From these and results previously published (Becraft et al. (1990) Devl Biol. 14), we conclude that the information carried by the Lg1+ gene product acts earlier in development than formation of the ligule proper. We hypothesize that Lg1+ may be effective at the stage when the blade-sheath boundary is first determined.     

Mechanisms of leaf tooth formation in Arabidopsis

Serration found along leaf margins shows species‐specific characters. Whereas compound leaf development is well studied, the process of serration formation is largely unknown. To understand mechanisms of serration development, we investigated distinctive features of cells that could give rise to tooth protrusion in the simple‐leaf plant Arabidopsis. After the emergence of a tooth, marginal cells, except for cells at the sinuses and tips, started to elongate rapidly. Localized cell division seemed to keep cells at the sinus smaller, rather than halt cell elongation. As leaves matured, the marginal cell number between teeth became similar in any given tooth. These results suggest that teeth are formed by repetition of an unknown mechanism that spatially monitors cell number and regulates cell division. We then examined the role of CUP‐SHAPED COTYLEDON 2 (CUC2) in serration development. cuc2‐3 forms fewer hydathodes and auxin maxima, visualized by DR5rev::GFP, at the leaf margin, suggesting that CUC2 patterns serration through the regulation of auxin. In contrast to a previous interpretation, comparison of leaf outlines revealed that CUC2 promotes outgrowth of teeth rather than suppression of growth at the sinuses. We found that mutants with increased CUC2 expression form ectopic tissues and mis‐express SHOOT MERISTEMLESS (STM) at the sinus between the enhanced teeth. Similar but infrequent STM expression was found in the wild type, indicating STM involvement in the serration of simple leaves. Our study provides insights into the morphological and molecular mechanisms for leaf development and tooth formation, and highlights similarities between serration and compound leaf development.     

Epidermal Cell Division and the Coordination of Leaf and Tiller Development

Initiation and development of grass leaves and tillers are oftendescribed individually with little attention to possible interrelationshipsamong organs. In order to better understand these interrelationships,this research examined epidermal cell division during developmentaltransitions at the apical meristem of tall fescue (Festuca arundinaceaSchreb.). Ten seedlings were harvested each day for a 9-d period,and lengths of main shoot leaves and primary tillers were measured.In addition, numbers and lengths of epidermal cells were determinedfor 0·5 mm segments along the basal 3 mm of each leafand tiller. Primordia development and onset of rapid leaf elongationwere characterized by an increase in the number of cells perepidermal file with mean cell length remaining near 20 µmper cell. After the leaf had lengthened to 1-1·5 mm,cells near the leaf tip ceased dividing and increased in length,at which time leaf elongation rate increased rapidly. Liguleformation, marking the boundary between blade and sheath cells,occurred prior to leaf tip emergence above the whorl of oldersheaths, while the earliest differentiation between blade andsheath cells probably began when leaves were < 1 mm long.Major transitions in leaf and tiller development appeared tobe synchronized among at least three adjacent nodes. At theoldest node, cessation of cell division in the leaf sheath wasaccompanied by initiation of cell division and elongation inthe associated tiller bud. At the next younger node the ligulewas being initiated, while at the youngest node cell divisioncommenced in the leaf primordium, as elongation of a new leafblade began. This synchronization of events suggests a key rolefor the cell division process in regulating leaf and tillerdevelopment.Copyright 1994, 1999 Academic Press Festuca arundinacea Schreb., tall fescue, cell division, leaf initiation, tillering, ligule development     

Manipulation of leaf shape by modulation of cell division

The role of cell division as a causal element in plant morphogenesis is debatable, with accumulating evidence supporting the action of cell division-independent mechanisms. To directly test the morphogenic function of cell division, we have utilised a microinduction technique to locally and transiently manipulate the expression in transgenic plants of two genes encoding putative effectors of the cell cycle, a tobacco A-type cyclin and a yeast cdc25. The results show that local expression of these genes leads to modulation of cell division patterns. Moreover, whereas altered cell division in the apical meristem had no influence on organogenesis, local induction of cell proliferation on the flanks of young leaf primordia led to a dramatic change in lamina development and, thus, leaf shape. These data indicate that the role of cell division in plant morphogenesis is context dependent and identify cell division in the leaf primordium as a potential target for factors regulating leaf shape.     

Cellular basis of developmental plasticity observed in heterophyllous leaf formation of Ludwigia arcuata (Onagraceae)

When heterophyllous plants of Ludwigia arcuata Walt. (Onagraceae) were transferred from aerial condition to submergence, young developing leaves were matured into leaves with intermediate shape between aerial-type and submerged-type, showing spatulate shape (spoon-shaped). This change was also induced by the exposure of plants to ethylene. On the other hand, when the plants were transferred from submergence to aerial conditions, young developing leaves were matured into intermediate-type leaves with elliptic shape (spearhead shape). Anatomical analysis revealed that the formation of spatulate leaf was caused by the reduction of the number of epidermal cells aligned in the leaf transverse direction in the basal region of the leaf while the tip regions remained as before and did not respond to this treatment. During development, the ethylene-induced spatulate leaves showed that three types of alterations in epidermal cell division were involved in this process. Changes in the distribution of cell divisions in leaf lamina were detected by the first day of ethylene exposure, and changes in the orientation of cell division planes were detected by the second day. However, changes in the number of cells aligned in the leaf transverse direction were not detected by this time. Three days after ethylene exposure, frequency of cell divisions changed, and by the time changes of cell numbers aligned in the leaf transverse direction were observed. Thus, the formation of intermediate-type leaves in L. arcuata was ascribed to the alterations of cell division patterns which was induced by ethylene.     

QUANTITATIVE ANALYSIS OF LEAF DEVELOPMENT IN XANTHIUM PENSYLVANICUM

Maksymowych , Roman . (Villanova U., Villanova, Pa.) Quantitative analysis of leaf development in Xanthium pensylvanicum. Amer. Jour. Bot. 46(9): 635–644. Illus. 1959.—An attempt was made to find a quantitative way of describing the development of the leaf and to correlate the developmental processes, designating precisely their sequence. The processes were presented in terms of the absolute and relative rates of leaf length, expansion of lamina in surface, increase in thickness, rates of cell division of leaf 9 and 13, and tissue differentiation of 3 portions of the lamina. All rates were estimated over the entire period of development, from initiation of a primordium to its maturity. The leaf plastochron index (L.P.I.) was used as a morphological time-scale. The relative plastochron rates were used for the purpose of correlation of the developmental processes. Leaf 9 elongates exponentially up to 3.0 L.P.I. with an average relative rate (dlnL/dpl) of about 0.78 pl^-1, and it stops growing around 8.0 L.P.I. The lamina stops elongating about 1.5 plastochrons before the petiole. The tip of the lamina expands its surface at a constantly lower relative rate than the middle and the basal portions of the blade. The average relative rate of expansion in area (dlnA/dpl) for the whole lamina is 1.7pl^-1 during the exponential stage. Differentiation of the laminar tissues proceeds basipetally, from the tip toward the base of the leaf. The relative rate of expansion of lamina in thickness (dlnT/dpl) is 0.55 pl^-1 at 1.5 L.P.I. and after 4.0 L.P.I. all cells cease elongating in a plane perpendicular to the leaf surface. The formation of cells proceeds exponentially up to 3.0 L.P.I. and about this time cell divisions stop in all parts of the lamina. The mean relative rate of cell formation (dlnC/dpl) at the exponential phase is 1.41 pl^-1, an increase of about 31% per day. At least 27 generations of cells are involved in the process of leaf formation and the generation time was calculated to be 0.5 plastochron or 2.2 days.     

Effect of drought stress on the cytological status in Ricinus communis

Growing leaves of dicots are characterized by the simultaneous development of cytological structure and physiological function. Cytological development of growing leaves of castor bean (Ricinus communis L.) and the impact of drought on this process was studied. Cell division was observed when the middle lobe of the leaf was below 8 cm length. Cell densities dropped when the middle leaflet had reached 4 cm. Identical relationships between leaf size (length of the middle lobe) and (I) exposed surface area of epidermal cells (ii) height of palisade cells, (iii) cell density and stomatal density were observed. During drought, areal growth decreased, but the relationships between the cytological parameters and leaf size did not change. The impact of drought on the cellular growth processes depended on the stage of cytological development at the onset of the drought. These results are the basis for an analysis of physiological and biochemical parameters in the forthcoming studies.     

FLEXIBILITY IN ONTOGENY AS SHOWN BY THE CONTRIBUTION OF THE SHOOT APICAL LAYERS TO LEAVES OF PERICLINAL CHIMERAS

The development of leaves on apically stable, periclinal chimeras was studied in a number of dicot genera. The mutant cell layers of the shoot apex and the tissues derived from them were as active developmentally as the normal layers. Ontogeny was the same in these chimeras as in nonchimeras, and growth of their leaves can be outlined as follows. Formation of the buttress, the axis, and the lamina of simple dicot leaves were independent events. In each the first growth included derivatives of the apical layers, usually three in number, found in the apex of the shoot and the lateral buds. Most cell divisions in the outer layers (L-I and L-II) were anticlinal relative to the new structures. Therefore, in the proximal regions of the buttress, axis (petiole and midrib), and lamina, the derivative cells of L-I and L-II were usually present in single layers. The rest of the internal tissue was from L-III. As formation of the axis and the lamina proceeded, derivatives of L-II replaced L-III internally in the distal and marginal regions leaving cells of L-III behind. Both the determinate growth of leaves and the pattern of cell divisions at and near the leading edges of growth meant that no cells in the leaf were comparable to the initial cells of the shoot apex. As the lamina extended, there were extensive intercalary cell divisions, both anticlinal and periclinal, so that in any given region of a leaf the layers of internal cells were from either L-II or L-III. At any point along the axis, L-III participated or did not participate in laminar extension. At any given stage in laminar growth either of two sister cells in any internal layer divided either a few times or extensively. The extreme variability in direction and frequency of cell division during leaf development was under an overriding genetic control, which resulted in the normal or typical size, shape and thickness of leaves.     

CELL DIVISION AND CELL ELONGATION IN LEAF DEVELOPMENT OF XANTHIUM PENSYLVANICUM

Maksymowych, Roman. (Villanova U., Villanova, Pa.) Cell division and cell elongation in leaf development of Xanthium pensylvanicum. Amer. Jour. Bot. 50(9) : 891–901. Illus. 1963.—Cell division in different parts of the lamina and cell enlargement of the upper epidermis and palisade mesophyll were studied in vertical and horizontal planes during the entire period of growth. The leaf plastochron index (L.P.I.) was used for designation of developmental stages of the leaf. From cell-length data and measurements of cell area the absolute rates of elongation (dX/dpl) and relative rates of elongation (dlnX/dpl) were calculated. The increase in number of cells in the early plastochrons is exponential and cell division stops at about L.P.I. 3.0. Divisions cease first at the tip and last in the basal lobes of the leaf, indicating a basipetal trend of this process. Cells are elongating while division is in progress, though this elongation proceeds at low rates and for a limited time. Palisade cells elongate in the vertical plane at higher rates and at least 1 plastochron sooner than the upper epidermis. The latter cells, however, expand in area with higher absolute and relative rates, and about 2 plastochrons in advance of the palisade mesophyll. The rates are not constant during the whole period of development but are represented by the bell-shaped curves with maximal peaks around L.P.I. 3.0 for the middle portion of the lamina. The increase in volume of the 2 types of cells stops around L.P.I. 5.0, or shortly after. In addition to unequal durations of cellular enlargement, both tissues expand at differential rates, which for the upper epidermis is high in the horizontal plane but low in the vertical plane, while the opposite is true for the palisade mesophyll. It is suggested that palisades and spongy mesophyll are separated and intercellular spaces formed during the course of development because of the greater rate of expansion in area of the upper epidermis.     

Studies on the Growth of Spinach Leaves (Spinacea oleracea)

The growth of spinach leaves has been studied from approximately1 cm long to full size. Over-all growth was measured in termsof area and total number of cells. The differential growth ofleaves was measured by the changes in the shape of squares drawnon the leaf surface. Growth differentials in terms of numbersof cells and number displaying mitotic figures were measuredin leaf discs taken from different positions within leaves. It was found that cell division in spinach leaves continueduntil the leaves reach from one-third to one-half full size.Cell division within the lamina of the leaves was not uniformbut ceased at an early stage of development in the leaf tipregion and continued for an extended period at the base.     

<Emphasis Type="Italic">ASYMMETRIC LEAVES1</Emphasis>, an <Emphasis Type="Italic">Arabidopsis</Emphasis> gene that is involved in the control of cell differentiation in leaves

During leaf development, the formation of dorsal-ventral and proximal-distal axes is central to leaf morphogenesis. To investigate the genetic basis of dorsoventrality and proximodistality in the leaf, we screened for mutants of Arabidopsis thaliana (L.) Heynh. with defects in leaf morphogenesis. We describe here the phenotypic analysis of three mutant alleles that we have isolated. These mutants show varying degrees of abnormality including dwarfism, broad leaf lamina, and aberrant floral organs and fruits. Genetic analysis revealed that these mutations are alleles of the previously isolated mutant asymmetric leaves1 ( as1). In addition to the leaf phenotypes described previously, these alleles display other phenotypes that were not observed. These include: (i) some rosette leaves with petiole growth underneath the leaf lamina; (ii) leaf vein branching in the petiole; and (iii) a leaf lamina with an epidermis similar to that on the petiole. The mutant phenotypes suggest that the ASYMMETRIC LEAVES1 ( AS1) gene is involved in the control of cell differentiation in leaves. As the first step in determining a molecular function for AS1, we have identified the AS1 gene using map-based cloning. The AS1 gene encodes a MYB-domain protein that is homologous to the Antirrhinum PHANTASTICA ( PHAN) and maize ROUGH SHEATH2 ( RS2) genes. AS1 is expressed nearly ubiquitously, consistent with the pleiotropic mutant phenotypes. High levels of AS1 expression were found in tissues with highly proliferative cells, which further suggests a role in cell division and early cell differentiation.     

Real-time lineage analysis reveals oriented cell divisions associated with morphogenesis at the shoot apex of Arabidopsis thaliana

Precise knowledge of spatial and temporal patterns of cell division, including number and orientation of divisions, and knowledge of cell expansion, is central to understanding morphogenesis. Our current knowledge of cell division patterns during plant and animal morphogenesis is largely deduced from analysis of clonal shapes and sizes. But such an analysis can reveal only the number, not the orientation or exact rate, of cell divisions. In this study, we have analyzed growth in real time by monitoring individual cell divisions in the shoot apical meristems (SAMs) of Arabidopsis thaliana. The live imaging technique has led to the development of a spatial and temporal map of cell division patterns. We have integrated cell behavior over time to visualize growth. Our analysis reveals temporal variation in mitotic activity and the cell division is coordinated across clonally distinct layers of cells. Temporal variation in mitotic activity is not correlated to the estimated plastochron length and diurnal rhythms. Cell division rates vary across the SAM surface. Cells in the peripheral zone (PZ) divide at a faster rate than in the central zone (CZ). Cell division rates in the CZ are relatively heterogeneous when compared with PZ cells. We have analyzed the cell behavior associated with flower primordium development starting from a stage at which the future flower comprises four cells in the L1 epidermal layer. Primordium development is a sequential process linked to distinct cellular behavior. Oriented cell divisions, in primordial progenitors and in cells located proximal to them, are associated with initial primordial outgrowth. The oriented cell divisions are followed by a rapid burst of cell expansion and cell division, which transforms a flower primordium into a three-dimensional flower bud. Distinct lack of cell expansion is seen in a narrow band of cells, which forms the boundary region between developing flower bud and the SAM. We discuss these results in the context of SAM morphogenesis.     

CINCINNATA controls both cell differentiation and growth in petal lobes and leaves of Antirrhinum

To understand how differentiation and growth may be coordinated during development, we have studied the action of the CINCINNATA (CIN) gene of Antirrhinum. We show that in addition to affecting leaf lamina growth, CIN affects epidermal cell differentiation and growth of petal lobes. Strong alleles of cin give smaller petal lobes with flat instead of conical cells, correlating with lobe-specific expression of CIN in the wild type. Moreover, conical cells at the leaf margins are replaced by flatter cells, indicating that CIN has a role in cell differentiation of leaves as well as petals. A weak semidominant cin allele affects cell types mainly in the petal but does not affect leaf development, indicating these two effects can be separated. Expression of CIN correlates with expression of cell division markers, suggesting that CIN may influence petal growth, directly or indirectly, through effects on cell proliferation. For both leaves and petals, CIN affects growth and differentiation of the more distal and broadly extended domains (leaf lamina and petal lobe). However, while CIN promotes growth in petals, it promotes growth arrest in leaves, possibly because of different patterns of growth control in these systems.     

Spatial and temporal analysis of non-steady elongation of rice leaves

A precise knowledge of the temporal and spatial distributions of cell division and tissue expansion is essential for appropriate leaf sampling in omics studies and for analyses of plant–environment relations. Elongating leaves of rice were studied during their whole development for elongation rate, distribution of cell length, cell production rate and spatial distribution of growth in the leaf. In seven genotypes, the pattern of leaf elongation rate followed three phases: (1) an exponential increase before leaf appearance; (2) a short phase (2–4 d at 20 °C) with a stable leaf elongation rate around leaf appearance; and (3) a phase of 8–10 d with a progressive decrease in elongation rate. The profile of cell length along the leaf changed with time during the first and last phases, but was time invariant around appearance. We propose a method adapted to non-steady elongation based on anatomical measurements, which was successfully tested by comparing it with the pricking method. It allowed analysis of the change with time in the spatial distribution of growth from initiation to end of leaf growth. The length of leaf zones with cell division and tissue elongation varied with time, with maximums of 21 and 60 mm respectively around leaf appearance.