DNA Content of Beta vulgaris Chloroplasts during Leaf Cell Expansion

During the growth of beet leaves from 2 to 3 to 25 to 30 centimeters, the leaf cells increase in size, the average number of chloroplasts per cell increases from 11 to 65 and the amount of chloroplast DNA per cell increases from 1100 to 1900 plastome copies. The average number of copies of the plastome per chloroplast decreases from 104 in 2 to 3-centimeter leaves to 29 in 25 to 30-centimeter leaves during a period when the chloroplasts undergo two to three rounds of division and increase diameter from 1.5 to 4.9 micrometers. This result is at variance with previously published studies of beet chloroplasts but agrees with the conclusions reached in more recent studies of pea and spinach and wheat leaf cell expansion.     

Changes in Chloroplast DNA Levels during Growth of Spinach Leaves

In young spinach leaves, 1–4 mm long, 7–10% of thetotal DNA of the leaf was chloroplast (pt) DNA. Growth in theseleaves was mainly by cell division with plastid division keepingpace with cell division and maintaining about 10 plastids percell. About 1% of the leaf cells were formed in 4.0 mm leaves.Both cell division and cell expansion contribute to the nextstage of leaf growth, which was quantitatively the major periodof new cell formation, nuclear DNA synthesis and ptDNA synthesis.Relative to the nuclear DNA level ptDNA levels rose to 21% ofthe total DNA and chloroplast.plastome copy numbers from 1500to 5000 per cell while chloroplast numbers rose from 10 to 30per cell. In the final period of leaf growth, cell expansionwas the main determinant of growth and chloroplast number percell rose to 180. In contrast to young leaves, newly emergedcotyledons contained 20% of their DNA as ptDNA and, during cellexpansion, cell number per cotyledon doubled. On average, thecells became octoploid, and chloroplast numbers and plastomecopy numbers rose to 500 and 22 000 per cell respectively. Similarlevels of nuclear ploidy, chloroplast number and plastome copynumber were induced in the first leaf pair of spinach followingdecapitation. When senescence was induced in mature leaves byshading, no loss of nuclear or ptDNA occurred. Following theonset of leaf yellowing and a form of senescence induced bynitrogen deficiency in leaves which had not fully expanded,there was preferential loss of ptDNA which fell from 8200 to3700 plastome copies per cell over an 11 d period. Key words: Spinach, Chloroplast, DNA, Ploidy     

Effects of nitrogen deprivation on cell division and expansion in leaves of Ricinus communis L.

The effects of nitrogen deprivation on leaf extension, cell numbers and epidermal cell size were followed in leaves of Ricinus communis L. The extent to which reductions in final cell number or final epidermal cell size contributed to the reduction in final leaf size depended on the developmental stage of the leaf at the time of N deprivation. In leaves which already had their full complement of cells (leaf 2), the reduction in final leaf size following nitrogen deprivation was associated with a reduction in final cell size. In leaves that were at earlier stages of development at the onset of N deprivation (leaves 3 and 4), the reduction in final leaf size was greater than in leaf 2. In these younger leaves, the final cell size was even smaller than in leaf 2, but the greatest contribution to reduced final leaf size was a reduction in the number of cells produced. This accounted for approximately 80% of the reduction in final leaf size in leaf 4. During leaf development, the contribution from different tissue layers to the total cell number changed. In the smallest leaf sizes, the contribution from upper and lower epidermis and spongy parenchyma was greater than that from palisade parenchyma. As the leaf size increased, cells in the palisade parenchyma continued to divide for longer than in the other layers. At final leaf size, the contribution from the different tissue layers to total cell number was the same for leaves 2, 3 and 4, irrespective of N treatment. In these final leaf structures, palisade parenchyma contributed 60% of the total cell number. Thus, although nitrogen deprivation affected leaf size variously through cell division and cell expansion, depending on leaf developmental stage at the time of nitrogen deprivation, the ratio of cell numbers and sizes in different tissue layers, at final leaf size, was unaffected.     

Devlopmental Physiology of Sugar-Beet: II. EFFECTS OF TEMPERATURE AND NITROGEN SUPPLY ON THE GROWTH, SOLUBLE CARBOHYDRATE CONTENT AND NITROGEN CONTENT OF LEAVES AND ROOTS

Plants were grown at temperatures of 15 and 25 ?C with two ratesof nitrogen supply. The changes in dry weight, leaf area, cellnumber, mean cell volume, soluble carbohydrate, and total nitrogenconcentration of the cotyledons, the first and second pair oftrue leaves, and the storage root were measured. Changes incell number and cell volume of the first pair of true leavesand storage root of plants were also measured at 11, 18, 25,and 32 ?C. Leaf growth before unfolding was chiefly by increase in cellnumber and after unfolding by increase in mean cell volume,while the growth of the storage root was almost entirely byincrease in cell number. The rates of cell division and cellexpansion were fastest at 25 ?C, but the initially high ratesof cell division in the terminal bud and in individual leavesdecreased rapidly and greater rates were maintained at the sub-optimaltemperatures, i.e. 15 and 18 ?C. After an initial period ofslow growth, the first-formed leaves grew faster and becamelarger at 15 than at 25 ?C. Leaves were produced, unfolded,grew faster, and became larger with increase in the externalconcentration of nitrogen, because cells divided and expandedfaster, so that nitrogen increased the number and size of cells. Sugar concentration was greater at 15 than at 25 ?C in leavesbut not in the storage root. Sugar concentration in the petiolesof the first and second pair of true leaves increased to 1.2and 2.0 per cent fresh weight respectively. Decreased nitrogensupply temporarily increased the sugar concentration of cotyledonpetioles and the seedling hypocotyl, but later decreased itin the leaves and storage root. Nitrogen concentration was greaterin the leaves and storage root at 15 than at 25 ?C with thelarger nitrogen supply. Nitrogen concentrations were similarin young leaves of all treatments but as the size of leavesincreased nitrogen concentrations decreased most rapidly at25 ?C with the smaller nitrogen supply. It is suggested that when increased leaf production and storage-rootgrowth occurs at temperatures below the growth optimum (25 ?C),they may be due to an effect of increased carbohydrate supplyon cell division and sugar storage.     

Changes in chloroplast number per cell during leaf development in spinach

Summary The amounts of chlorophyll and nitrogen and the numbers of cells per unit area change as the green leaves of spinach plants grow and increase in size in the light. The changes in the numbers of chloroplasts per cell were measured by a new method. A 5-fold increase in the numbers of chloroplasts per cell took place in both palisade and mesophyll cells over a growing period of 10 days during which time the area of the leaves increased from 1 to 50 cm². Proplastids were not present in the young green leaves but electron-microscope and phase-contrast observations showed the presence of grana-containing chloroplasts, many of which appeared to be undergoing division by constriction. It is suggested that the large increase in chloroplast numbers as leaf cells grow and expand in the light is from the division of differentiated chloroplasts containing grana.     

Response of cassava leaf area expansion to water deficit: cell proliferation, cell expansion and delayed development

BACKGROUND AND AIMS: Cassava (Manihot esculenta) is an important food crop in the tropics that has a high growth rate in optimal conditions, but also performs well in drought-prone climates. The objectives of this work were to determine the effects of water deficit and rewatering on the rate of expansion of leaves at different developmental stages and to evaluate the extent to which decreases in cell proliferation, expansion, and delay in development are responsible for reduced growth. METHODS: Glasshouse-grown cassava plants were subjected to 8 d of water deficit followed by rewatering. Leaves at 15 developmental stages from nearly full size to meristematic were sampled, and epidermal cell size and number were measured on leaves at four developmental stages. KEY RESULTS: Leaf expansion and development were nearly halted during stress but resumed vigorously after rewatering. In advanced-stage leaves (Group 1) in which development was solely by cell expansion, expansion resumed after rewatering, but not sufficiently for cell size to equal that of controls at maturity. In Group 2 (cell proliferation), relative expansion rate and cell proliferation were delayed until rewatering, but then recovered partially, so that loss of leaf area was due to decreased cell numbers per leaf. In Group 3 (early meristematic development) final leaf area was not affected by stress, but development was delayed by 4-6 d. On a plant basis, the proportion of loss of leaf area over 26 d attributed to leaves at each developmental stage was 29, 50 and 21 % in Group 1, 2 and 3, respectively. CONCLUSIONS: Although cell growth processes were sensitive to mild water deficit, they recovered to a large extent, and much of the reduction in leaf area was caused by developmental delay and a reduction in cell division in the youngest, meristematic leaves.     

Microspectrofluorometric Measurement of Chloroplast DNA in Dividing and Expanding Leaf Cells of Spinacia oleracea

Absolute DNA amounts of individual chloroplasts from mesophyll and epidermal cells of developing spinach leaves were measured by microspectrofluorometry using the DNA-specific stain, 4,6-diamidino-2-phenyl indole, and the bacterium, Pediococcus damnosus, as an internal standard. Values obtained by this method showed that DNA amounts of individual chloroplasts from mesophyll cells fell within a normal distribution curve, although mean DNA amounts changed during leaf development and also differed from the levels in epidermal chloroplasts. There was no evidence in the data of plastids containing either the high or low levels of DNA which would be indicative of discontinuous polyploidy of plastids, or of division occurring in only a small subpopulation of chloroplasts. By contrast, the distribution of nuclear DNA amounts in the same leaf tissues in which cell division was known to be occurring showed a clear bimodal distribution. We consider that the distribution of chloroplast DNA in the plastid population shows that there is no S-phase of chloroplast DNA synthesis, all chloroplasts in the population in young leaf cells synthesize DNA, and all chloroplasts divide.     

Profiles of photosynthetic oxygen-evolution within leaves of Spinacia oleracea

Oxygen evolution was measured from mesophyll tissues in spinach leaves using a photoacoustic technique. The photosynthetic capacity of individual cell layers was measured by directing microscopic beams of light, 40 μm wide, to cells exposed within a leaf cross section. The resulting profile for oxygen-evolution potential was relatively flat, indicating a uniform capacity for photosynthesis in leaf mesophyll tissues. Two experimental approaches were used to estimate the photosynthetic performance of individual mesophyll cell layers when white light was applied to the adaxial leaf surface. These experiments indicated that oxygen was produced relatively uniformly across the mesophyll and that oxygen evolution increased with irradiance of the white light applied to the leaf surface. The measured profiles for oxygen evolution and capacity are flatter than previous measurements of profiles of fixed carbon and estimates of profiles for absorbed light within spinach leaves.     

Leaf initiation and development in soybean under phosphorus stress

Experiments investigated changes in leaf development in young soybean plants progressing into P stress. The apical meristem and leaf structure were examined anatomically to evaluate the involvement of cell division and cell expansion in the restriction of leaf number and individual leaf size. Seedlings were deprived of P for 32 d following germination. Leaf initiation rates declined noticeably after about 2 weeks, even though the apical dome was of similar size and had a similar number of cells as controls. Primordia appeared morphologically similar also. Expansion of primary and the first three trifoliolate leaves of -P plants was severely reduced, and expansion of each leaf ceased, uniformly, when an area of about 40 cm(2) was obtained. Leaf epidermal cell size in the lateral plane was unaffected. The results indicate that expansion of leaves under P stress was limited by the number of cell divisions, which would imply control of cell division by a common regulatory factor within the leaf canopy.     

Partitioning of (13)C-photosynthate from spur leaves during fruit growth of three Japanese pear (Pyrus pyrifolia) cultivars differing in maturation date

BACKGROUND AND AIMS: In fruit crops, fruit size at harvest is an important aspect of quality. With Japanese pears (Pyrus pyrifolia), later maturing cultivars usually have larger fruits than earlier maturing cultivars. It is considered that the supply of photosynthate during fruit development is a critical determinant of size. To assess the interaction of assimilate supply and early/late maturity of cultivars and its effect on final fruit size, the pattern of carbon assimilate partitioning from spur leaves (source) to fruit and other organs (sinks) during fruit growth was investigated using three genotypes differing in maturation date. METHODS: Partitioning of photosynthate from spur leaves during fruit growth was investigated by exposure of spurs to (13)CO(2) and measurement of the change in (13)C abundance in dry matter with time. Leaf number and leaf area per spur, fresh fruit weight, cell number and cell size of the mesocarp were measured and used to model the development of the spur leaf and fruit. KEY RESULTS: Compared with the earlier-maturing cultivars 'Shinsui' and 'Kousui', the larger-fruited, later-maturing cultivar 'Shinsetsu' had a greater total leaf area per spur, greater source strength (source weight x source specific activity), with more (13)C assimilated per spur and allocated to fruit, smaller loss of (13)C in respiration and export over the season, and longer duration of cell division and enlargement. Histology shows that cultivar differences in final fruit size were mainly attributable to the number of cells in the mesocarp. CONCLUSIONS: Assimilate availability during the period of cell division was crucial for early fruit growth and closely correlated with final fruit size. Early fruit growth of the earlier-maturing cultivars, but not the later-maturing ones, was severely restrained by assimilate supply rather than by sink limitation.     

Chlorophyll and light gradients in sun and shade leaves of Spinacia oleracea

Abstract. Light gradients were measured and correlated with chlorophyll concentration and anatomy of leaves in spinach (Spinacia oleracea L.). Light gradients were measured at 450, 550 and 680 nm within thin (455 μm) and thick (630 μm) leaves of spinach grown under sun and shade conditions. The light gradients were relatively steep in both types of leaves and 90% of the light at 450 and 680 nm was absorbed by the initial 140 μm of the palisade. In general, blue light was depleted faster than red light which, in turn was depleted faster than green light. Light penetrated further into the thicker palisade of sun leaves in comparison to the shade leaves. The distance that blue light at 450 nm travelled before it became 90% depleted was 120 μm in sun leaves versus 76 μm in shade leaves. Red light at 680 nm and green light at 550 nm travelled further but the trends were similar to that measured at 450nm. The steeper light gradients within the palisade-of shade leaves were caused by increased scattering of light within the intercellular air spaces and/or cells which were less compact than those in sun leaves. The decline in the amount of light within the leaf appeared to be balanced by a gradient in chlorophyll concentration measured in paradermal sections. Progressing from the adaxial epidermis, chlorophyll content increased through the palisade and then declined through the spongy mesophyll. Chlorophyll content was similar in the palisade of both sun and shade leaves. Chloroplast distribution within both sun and shade leaves was relatively uniform so that the chlorophyll gradient appeared to be caused by greater amounts of chlorophyll within chloroplasts located deeper within the leaf. These results indicate that the anatomy of the palisade may be of special importance for controlling the penetration of photo-synthetically active radiation into the leaf. Changing the structural characteristics of individual palisade cells or their arrangement may be an adaptation that maximizes the absorption of light in leaves with varying mesophyll thickness due to different ambient light regimes.     

Organ shape and size: a lesson from studies of leaf morphogenesis

Control of the shape and size of indeterminate organs, such as roots and stems, is directly related to the control of the shape and size of the cells in these organs, as predicted by orthodox cell theory. For example, the polarity-dependent growth of leaf cells directly affects the polar expansion of leaves. Thus, the control of leaf shape is related to the control of the shape of cells within the leaf, as suggested by cell theory. By contrast, in determinate organs, such as leaves, the number of cells does not necessarily reflect organ shape or size. Genetic evidence shows that a compensatory system(s) is involved in leaf morphogenesis, and that an increase in cell volume can be triggered by a decrease in cell number and vice versa. Studies of chimeric leaves also suggest interaction between leaf cells that coordinates the behaviour of these cells at the organ level. Moreover, leaf size also appears to be coordinated at the whole-plant level. The recently hypothesised neo cell theory describes how leaf shape- and size-control mechanisms control leaf shape at the organ-level via cell-cell interaction.     

SOME EFFECTS OF NITROGEN NUTRITION ON THE MORPHOLOGY AND ANATOMY OF MARROW-STEM KALE

Marrow-stem kale plants grown on plots receiving frequent additions of sulphate of ammonia showed a 40% increase in length of internode and a 25% increase in number of nodes per plant, and the leaf size was increased by between 50 and 70% over plants in plots receiving no N fertilizer. Leaves of kale continue to increase in area until they turn yellow, and the high N leaves showed a greater rate of increase in area at every stage in the life of the leaf.
Various features of leaf structure, such as stomatal index, and thickness of palisade and mesophyll, were unaffected by N treatment. The size of the epidermal cells of the leaves was very variable, and although the high N leaves showed a 12% increase in area per epidermal cell over the low N leaves, this difference is not statistically significant. The increased area of the high N leaves can therefore be attributed mainly to increased cell division during the life of the leaf. Only a very slight increase in rate of cell division is necessary to produce the observed effect.
The greater leaf area of the high N plants can be attributed mainly to increased size of individual leaves, but there was also a significantly greater number of living functional leaves per plant on the high N plants; at 23 weeks from sowing the high N plants had an average of 13.4 living leaves, while the low N plants had only 11.7 living leaves per plant.
There was an appreciable degree of N succulence in the high N kale leaves, which showed a 2% greater moisture content than the low N leaves.
A seasonal drift in epidermal cell size, palisade thickness, and total leaf thickness, is shown to be fully significant, statistically. Marked variations in stomatal frequency are barely significant at the 5% level.     

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.     

The role of auxin-binding protein 1 in the expansion of tobacco leaf cells

Tobacco leaf was used to investigate the mechanism of action of auxin-binding protein 1 (ABP1). The distributions of free auxin, ABP1, percentage of leaf nuclei in G2 and the amount of auxin-inducible growth were each determined in control tobacco leaves and leaves over-expressing Arabidopsis ABP1. These parameters were compared with growth of tobacco leaves, measured both spatially and temporally throughout the entire expansion phase. Within a defined window of leaf development, juvenile leaf cells that inducibly expressed Arabidopsis ABP1 prematurely advanced nuclei to the G2 phase. The ABP1-induced increase in cell expansion occured before the advance to the G2 phase, indicating that the ABP1-induced G2 phase advance is an indirect effect of cell expansion. The level of ABP1 was highest at the position of maximum cell expansion, maximum auxin-inducible growth and where the free auxin level was the lowest. In contrast, the position of maximum cell division correlated with higher auxin levels and lower ABP1 levels. Consistent with the correlations observed in leaves, tobacco cells (BY-2) in culture displayed two dose-dependent responses to auxin. At a low auxin concentration, cells expanded, while at a relatively higher concentration, cells divided and incorporated [3H]-thymidine. Antisense suppression of ABP1 in these cells dramatically reduced cell expansion with negligible effect on cell division. Taken together, the data suggest that ABP1 acts at a relatively low level of auxin to mediate cell expansion, whereas high auxin levels stimulate cell division via an unidentified receptor.     

Studies on the Expansion of the Leaf Surface: III. THE INFLUENCE OF RADIATION ON CELL DIVISION AND LEAF EXPANSION

The numbers of leaves and the areas and cell numbers of leavesfrom the first five nodes of the cucumber were determined throughouttheir development with three different amounts of daily radiationand two conditions of nutrient supply. The rate of leaf production was found to be constant with timefor any one amount of radiation and to increase with increasedradiation. The transition from the rate in darkness to thatin light was sharp and occurred more quickly the higher theradiation. Provided the nutrient supply was high the ultimate areas ofindividual leaves were greater the higher the radiation; ifmineral nutrients were depleted the maximum area of a leaf occurredwith an intermediate amount of light. This arose because thesefactors exercised a differential effect on the various phasesof growth. Each leaf commenced its life as a mass of dividing cells andthe mean rate of division remained constant until it unfoldedfrom the terminal bud. The mean rate of division was much greaterat high than at low levels of radiation and was interpretedas being regulated by carbohydrate supply. Although expansionof some cells was likely before unfolding, after this stagethere was a marked decrease in the proportion of cells proceedingto division. The duration of division in the leaf after unfoldingwas independent of radiation; although 70–98 per cent,of the final number of cells were formed after unfolding, theultimate number was effectively determined by the rate of divisionprior to unfolding. Low nutrient supply restricted the duration of division in theleaf to a significant extent and the expansion of cells to avery considerable degree. The smaller leaves on plants receivinghigh rather than medium radiation were due to their cells beingmuch smaller.     

Development of the Fifth Leaf is Indicative for Whole Plant Performance at Low Temperature in Tomato

In the search for early-detectable selection criteria for growthat low temperature conditions in tomato, first the initiationand growth of individual leaves was analysed. Scanning electronmicroscopy revealed that the first four primordia had alreadydeveloped during the germination period at 25°C. The primordiumof the fifth leaf, however, was initiated after the transferof seedlings to the experimental conditions. The increase inlength of the first three leaves, and to a lesser extent ofthe fourth leaf, was considerably smaller in comparison withthat of later formed leaves. Moreover, the morphology of thefirst three to four leaves was deviant, whereas the others showedthe normal compound leaf architecture. All these results indicatedthat the fifth leaf was the earliest formed leaf with growthcharacteristics that might reflect the growth potential of thewhole plant. Development of the fifth leaf was tested as a marker for wholeplant growth. At three temperature, 18, 15 and 12°C, growthresponses of the fifth leaf were similar to that of whole plantsin four tomato genotypes: Line A, Line B, Premier and MXXIV-13.Significant differences in relative growth rate of dry weightof whole plants and fifth leaves (RGR_W)and of leaf area of thefifth leaves (RGR_LA between two fast growing and two slow growinggenotypes were found. No genotype by temperature interactionfor RGR_W and RGR_LA was found, indicating that the effect oftemperature decrease was similar for the four genotypes. The structure of the mature fifth leaf of one fast and one slowgrowing genotype, Line A and MXXIV-13, was analysed. For bothgenotypes, leaves were small and thick at low temperature, 12°C.The total number of epidermis and palisade parenchyma cellsper leaf was smaller but the size of the cells developed at12°C was larger than at 18°C. Consequently, the slowgrowth at 12°C was due to a low rate of cell division. Atboth temperatures, the fifth leaf to MXXIV-13 was smaller comparedto that of line A. Since the size of the cells were similar,the smaller leaf size was due to lower number of leaf cells. The results confirm the suitability of the growth, especiallyexpressed as RGR_LA , of the fifth leaf as a nondestructive marketfor vegetative development of tomato at low temperature. Growthdifferences between genotypes were mainly reflected by differencesin cell number of leaves, which might be correlated with geneticallydetermined differences in cell number of leaf primordia.Copyright1993, 1999 Academic Press Lycopersicon esculentum Mill. genotypes, plant growth, selection criteria, low temperature, leaf initiation, leaf development, RGR, leaf structure, cell expansion     

Sensitivity of soybean leaf development to water deficits

Abstract. Drought effects on the final leaf area of individual leaves were hypothesized to depend on the leaf developmental stage at which drought occurred. To evaluate this hypothesis, final leaf area and cell number were measured for soybean ( Glycine max (L.) Merr.) leaves that were at different stages of development when single or cyclical drought treatment was imposed. Leaf emergence rate from the meristem, as depicted by changes in the plastochron index, was not as sensitive as leaf expansion to cyclical droughts. For leaf expansion, small leaves, once they emerged from the meristem, suffered larger decreases in growth than leaves undergoing rapid leaf area expansion. Decreases in final leaf area as a result of a cyclical drought were correlated with decreases in final cell number. Decreases resulting from a single 8-d drought were dependent on the age of the leaf at the time of drought, because small leaves were found to have proportionately larger decreases in final cell number and area than larger leaves. These results indicated that age-dependent leaf responses to drought are based on the relative activity of cell division and expansion at the time stress was imposed.     

Whole organ, venation and epidermal cell morphological variations are correlated in the leaves of Arabidopsis mutants

Despite the large number of genes known to affect leaf shape or size, we still have a relatively poor understanding of how leaf morphology is established. For example, little is known about how cell division and cell expansion are controlled and coordinated within a growing leaf to eventually develop into a laminar organ of a definite size. To obtain a global perspective of the cellular basis of variations in leaf morphology at the organ, tissue and cell levels, we studied a collection of 111 non-allelic mutants with abnormally shaped and/or sized leaves, which broadly represent the mutational variations in Arabidopsis thaliana leaf morphology not associated with lethality. We used image-processing techniques on these mutants to quantify morphological parameters running the gamut from the palisade mesophyll and epidermal cells to the venation, whole leaf and rosette levels. We found positive correlations between epidermal cell size and leaf area, which is consistent with long-standing Avery's hypothesis that the epidermis drives leaf growth. In addition, venation parameters were positively correlated with leaf area, suggesting that leaf growth and vein patterning share some genetic controls. Positional cloning of the genes affected by the studied mutations will eventually establish functional links between genotypes, molecular functions, cellular parameters and leaf phenotypes.     

Proteomic Analysis Reveals That Developing Leaves are More Sensitive to Nitrogen Fertilizer Than Mature Leaves

The size of rice leaves is tightly controlled by environmental and genetic factors. Several functional genes control leaf growth and development by regulating cell expansion and cell cycle activity. The regulation of leaf growth, particularly the effects of environmental conditions on leaf size, is still poorly understood. We examined the environmental control of leaf size in rice (Oryza sativa) by performing a comparative proteomic analysis, which showed that exposure to high-nitrogen levels produced enlarged leaves. The enhanced leaf growth occurred mainly as a result of an increased number of cell cycles. Two proteins related to cell division, FtsZ and ERBB3 binding protein, were increased by nitrogen treatment in the developing leaves. The expression of a type-A response regulator, OsRR2, was also elevated in developing leaves. OsRR2 acts as a negative regulator of cytokinin signaling and may reduce the cytokinin content in developing leaves; a low cytokinin level is necessary for leaf development. By analyzing the proteome response to nitrogen in both developing and mature leaves, we provide deeper insight into the mechanism by which nitrogen treatment affects the phenotype.