A new method to measure leaf age: Leaf measuring-interval index

We propose a new method, the leaf measuring-interval index (LMI), to estimate leaf age in morphological and physiological studies of leaves. When the plastochron, the interval between the initiation of successive leaves, is constant, the well-known leaf plastochron index (LPI) provides a robust measure of leaf age. When the duration of the plastochron is not uniform, however, we show that the LPI can (in simulations) and does (with actual data) turn variation in duration of the plastochron into variance about the regression estimates of leaf growth curves. The method we present in this paper, the LMI, is plastochron independent. This new method is particularly suited, therefore, for studies of plants growing in natural environments rather than in controlled growth facilities where the assumptions of the LPI method can be met.     

LEAF AND APICAL GROWTH CHARACTERISTICS IN TRITICUM

Leaf initiation rate, leaf primordium growth rates, and apical volume growth rates were determined for seedlings of Triticum aestivum cv. Ramona 50 under controlled environmental conditions. Three leaf primordia are present in the caryopsis, and three more leaves are initiated within the first two weeks after germination with a mean plastochron length of 95.5 hr. Volume growth rates of the apical region were determined on six apices which had six primordia each. The mean radial expansion rate was 0.467/plastochron, and the vertical expansion rate was 0.457/plastochron. The volume expansion rate was 1.393/plastochron. The mean volume doubling time was 0.498 plastochrons or 47.1 hr.     

STRUCTURAL CHANGES IN POPULUS DELTOIDES TERMINAL BUDS AND IN THE VASCULAR TRANSITION ZONE OF THE STEMS DURING DORMANCY INDUCTION

Morphological and anatomical changes in shoots of vigorously growing cottonwood plants (Populus deltoides Bartr.) were studied during dormancy induction in 8-hr short days (SD) and in control plants grown in 18-hr long days (LD). Pronounced structural changes occurred in terminal buds after 4 wk and full dormancy was achieved in 7 wk of SD. Leaf expansion ceased after 5 wk of SD as foliage leaves matured to the terminal bud base at leaf plastochron index 0 (LPI 0). Within the bud, total leaf length (lamina + petiole) decreased and stipule length increased progressively each week; thus, the ratio total leaf length/stipule length decreased rapidly, especially at the position of incipient bud-scale leaves LPI - 1 and LPI - 2. These bud-scale leaves were fully developed by wk 6 and were derived from enlarged stipules and aborted laminae. The full complement of primordia within the bud at the start of SD eventually matured as foliage leaves and the first bud-scale leaf (LPI - 1) was initiated immediately following transfer to SD. Acropetal advance of the primary-secondary vascular transition zone (TZ) was associated with leaf maturation. However, it did not advance throughout the entire vascular cylinder as in LD, but only in those leaf traces serving mature leaves beneath the terminal bud. In both LD and SD treatments the same linear relationship was maintained between LPI of the TZ and LPI of the most recently matured leaf; both parameters simultaneously increased in LD and decreased in SD. Thus, the relationship between leaf maturation and advance of the TZ was maintained irrespective of environment.     

14C fixation,metabolic labeling patterns,and translocation profiles during leaf development in Populus deltoides

The incorporation of photosynthetically fixed ¹⁴CO₂ and the distribution of ¹⁴C among the main chemical constituents of laminae and petioles were examined in cottonwood (Populus deltoides Bartr. ex Marsh.) leaves ranging in age from Leaf Plastochron Index (LPI) 3 (about one-quarter to one-third expanded) to LPI 30 (beginning of senescence). In addition, carbon flow among chemical fractions and translocation from leaves of LPI 7 and 14 were examined periodically up to 24 h after labeling. Specific activity of ¹⁴C (on dry-weight basis) increased in developing laminae to full leaf expansion, decreased in the mature leaves to LPI 16, then remained constant to LPI 30. In developing leaves (LPI 3-5), after 2 h, most of the ¹⁴C was found in protein, pigments, lipids, and other structural and metabolic components necessary for cell development; only 28% was in the sugar fraction of the lamina. In fully expanded leaves (LPI 6-8), after 2 h, the sugar fraction contained 50–60% and about 90% of fixed ¹⁴C in the lamina and the petiole, respectively. In a pulsechase kinetic series with recently mature leaves, 60% of the ¹⁴C was found in the sugar fraction after 15 min of ¹⁴CO₂ fixation. Over the 24-h translocation period, ¹⁴C decreased in sugars to 23% and increased in the combined residue fraction (protein, starch, and structural carbohydrates) to about 60% of the total activity left in the lamina. Within 24 h after labeling, the turnover of ¹⁴C-organic acids,-sugar, and-amino acids (either metabolzed or translocated from the leaf) was 30, 70 and 80%, respectively, of that initially incorporated into these fractions by a leaf at LPI 7 (turnover was 55% of ¹⁴C-organic acids, 80% of ¹⁴C-sugar, and 95% of ¹⁴C-amino acids at LPI 14). Anatomical maturity in cottonwood leaves is closely correlated with physiological maturity and with production of translocatable sugar.Abbreviations LPI leaf plastochron index - PI plastochron index Research Plant Physiologist and Chief Plant Physiologist, respectively     

LEAF INITIATION RATES AND VOLUME GROWTH RATES IN THE SHOOT APEX OF CHRYSANTHEMUM

The leaf initiation rate and apical volume growth rate of lateral shoots of Chrysanthemum morifolium ‘Improved Albatross III’ were determined for plants growing under controlled environment conditions. After removal of the terminal bud, the top two lateral buds produced leaves at a rate of 1.4 to 1.6 leaves/day for the first 9 or 10 days. At about the 10th day there appeared to be an abrupt shift to a lower initiation rate of 0.7 to 0.8 leaves/day. Defoliation by periodic removal of leaves larger than 1 cm caused a statistically significant increase in initiation rate, but not to as high a rate as that of the first 10 days of bud growth. Volume growth rates of the apical region were determined from transverse sections of five apices, according to the method of Richards (1951). The apical region was found to have a radial expansion rate of 0.1623/plastochron and a vertical expansion rate of 0.0494/plastochron. The volume growth rate was calculated to be 0.3740/plastochron and the volume doubling time to be 1.85 plastochrons or 56 to 63 hr.     

PETIOLE DEVELOPMENT AND XYLEM DIFFERENTIATION IN XANTHIUM REPRESENTED BY THE PLASTOCHRON INDEX

Petiole development and formation of xylem vessels have been investigated in Xanthium leaves from early ontogeny to maturity. Kinetics of growth was presented in terms of absolute and relative elemental rates of elongation. The process of vascularization was assessed by the number of differentiated xylem vessels. The leaf plastochron index (LPI) developed by Erickson and Michelini (1957) was used for designating the various stages of development. An exponential increase in petiole length was observed between the LPIs –3 and +4 indicating a constant relative rate of 0.20 or 20% increase per day. After cessation of lamina elongation at LPI 8, petiole elongation continued for an additional 5 day period, to LPI 9.5. Relative elemental rate analysis revealed that the basipetal pattern of elongation was maintained throughout the leaf development. At a specific plastochron age, the only growth was due to the petiole elongation. Leaves which ceased elongating had not completed their internal development, since the process of xylem formation continued for several plastochrons, or about 8 days. The highest rate of xylem formation was ten vessels per day at LPI 5. On the average, about five xylem vessels differentiated per day in the middle portion of a Xanthium petiole. Mature petioles contained an average of 218 xylem vessels. About 12 canals of schizogenous origin preceeded the development of the vascular tissue.     

Study of CO2 exchange processes,resistances to carbon dioxide and chlorophyll content during leaf ontogenesis in poplar

Net photosynthetic (P _N) and dark respiration (R _D) rate, stomatal (rs′) and internal (ri′) resistances to carbon dioxide were measured by gas exchange methods on leaves of different ages, expressed in leaf plastochron index units (LPI) for a fast growing poplar cultivar Unal 2. Although the optimal leaf age differs slightly for the different gas exchange parameters, leaf ontogeny is reflected in the same way in these different parameters. MaximalP _N and minimalrs′ and ri′ values were found at LPI between 6 and 10. Chlorophyll concentrations were lowest at LPI lower than 10 although an increase in two steps was found, when leaf age increases up to maturity.     

Developmental process of sun and shade leaves in Chenopodium album L.

The authors’ previous study of Chenopodium album L. revealed that the light signal for anatomical differentiation of sun and shade leaves is sensed by mature leaves, not by developing leaves. They suggested that the two‐cell‐layered palisade tissue of the sun leaves would be formed without a change in the total palisade tissue cell number. To verify that suggestion, a detailed study was made of the developmental processes of the sun and shade leaves of C. album with respect to the division of palisade tissue cells (PCs) and the data was expressed against developmental time (leaf plastochron index, LPI). The total number of PCs per leaf did not differ between the sun and shade leaves throughout leaf development (from LPI ?1 to 10). In both sun and shade leaves, anticlinal cell division of PCs occurred most frequently from LPI ?1 to 2. In sun leaves, periclinal division of PCs occurred synchronously with anticlinal division. The constancy of the total number of PCs indicates that periclinal divisions occur at the expense of anticlinal divisions. These results support the above suggestion that two‐cell‐layered palisade tissue is formed by a change of cell division direction without a change in the total number of PCs. PCs would be able to recognize the polarity or axis that is perpendicular to the leaf plane and thereby change the direction of their cell divisions in response to the light signal from mature leaves.     

FLORAL INDUCTION DOES NOT ALTER THE GROWTH PARAMETERS OF FUCHSIA

Floral induction by night interruption of Fuchsia hybrida cv. Lord Byron, a quantitative long-day plant with decussate phyllotaxis and an indeterminate flowering habit, altered neither the rate of leaf initiation nor the rate of leaf expansion; nor did flower initiation and development change the vegetative growth of the plants. This was diagnosed using plastochron duration and plastochron ratio measurements before, during, and after a 10-day induction period. A comparison between indeterminate and determinate flowering is made using these two parameters.     

Changes in photosynthetic characteristics during leaf development in apple

A comprehensive developmental survey of leaf area, chlorophyll, photosynthetic rate, leaf resistance, transpiration ratio, CO₂ compensation point and photorespiration was conducted in apple. The largest changes in each of the photosynthetic characteristics studied took place during the earliest stages of leaf development, coinciding with the period of greatest leaf expansion and chlorophyll synthesis. During early development, photosynthesis increased 5-fold, reaching a maximum rate of 40 mg CO₂ dm^-2 hr^-1 at a leaf plastochron index (LPI) of 10. During this same period, leaf resistance, transpiration ratio, CO₂ compensation point and mesophyll resistance decreased, while carboxylation efficiency increased. Two especially interesting aspects of the data discussed are simultaneous changes that occur at a LPI of 10 and 12 in all of the photosynthetic characteristics examined and an apparent decrease in photorespiration as leaves age. From our results it is clear that stage of leaf development is an important factor affecting the rate of photosynthesis and photorespiration.Scientific Paper No. 5687, College of Agriculture, Washington State University, Pullman. This work is supported by the National Science Foundation Grant 80-10958 and the Columbia River Orchards Foundation.     

Distribution of imported 14C in developing leaves of eastern cottonwood according to phyllotaxy

Summary Individual leaves of eastern cottonwood (Populus deltoides Bartr.), representing an ontogenetic series from leaf plastochron index (LPI) 3.0 to 8.0, were fed ¹⁴CO₂ and harvested after 2–24 h. Importing leaves from LPI-1.0 through 8.0 on each plant were sectioned into 9 parts, and each part was quantitatively assayed for ¹⁴C activity. The highest level of ¹⁴C import was by leaves from LPI 1.0 to 3.0, irrespective of source-leaf age. ¹⁴C was translocated preferentially to either the right or left lamina-half depending on the position of the importing leaf in the phyllotactic sequence and its stage of development. For example, import was high when the importing leaf and the source leaf had two vascular bundles in common, moderately high with one bundle in common, and low with no bundles in common. The distribution of ¹⁴C within young importing leaves was highest in the lamina tip and decreased toward the base. With increasing leaf age, incorporation declined in the lamina tip and increased in the base.It may be concluded that each cottonwood leaf progresses through a continuum of importing and exporting stages as its lamina expands. The photosynthate imported by a given leaf is compartmentalized, with different exporting leaves supplying photosynthate to rather restricted regions of the lamina. Such localization within the importing leaf depends on its vascular connections with each of the exporting leaves, and these are predictable from a knowledge of the phyllotaxy.Plant Physiologists.     

COMPARATIVE FOLIAR HISTOGENESIS IN ACORUS CALAMUS AND ITS BEARING ON THE PHYLLODE THEORY OF MONOCOTYLEDONOUS LEAVES

A comparative histogenetic investigation of the unifacial foliage leaves of Acorus calamus L. (Araceae; Pothoideae) was initiated for the purposes of: (1) re-evaluating the previous sympodial interpretation of unifacial leaf development; (2) comparing the mode of histogenesis with that of the phyllode of Acacia in a re-examination of the phyllode theory of monocotyledonous leaves; and (3) specifying the histogenetic mechanisms responsible for morphological divergence of the leaf of Acorus from dorsiventral leaves of other Araceae. Leaves in Acorus are initiated in an orthodistichous phyllotaxis from alternate positions on the bilaterally symmetrical apical meristem. During each plastochron the shoot apex proceeds through a regular rhythm of expansion and reduction related to leaf and axillary meristem initiation and regeneration. The shoot apex has a three- to four-layered tunica and subjacent corpus with a distinctive cytohistological zonation evident to varying degrees during all phases of the plastochron. Leaf initiation is by periclinal division in the second through fourth layers of the meristem. Following inception early growth of the leaf primordium is erect, involving apical and intercalary growth in length as well as marginal growth in circumference in the sheathing leaf base. Early maturation of the leaf apex into an attenuated tip marks the end of apical growth, and subsequent growth in length is largely basal and intercalary. Marked radial growth is evident early in development and initially is mediated by a very active adaxial meristem; the median flattening of this leaf is related to accentuated activity of this meristematic zone. Differentiation of the secondary midrib begins along the center of the leaf axis and proceeds in an acropetal direction. Correlated with this centralized zone of tissue specialization is the first appearance of procambium in the center of the leaf axis. Subsequent radial expansion of the flattened upper leaf zone is bidirectional, proceeding by intercalary meristematic activity at both sides of the central midrib. Procambial differentiation is continuous and acropetal, and provascular strands are initiated in pairs in both sides of the primordium from derivatives of intercalary meristems in the abaxial and adaxial wings of the leaf. Comparative investigation of foliar histogenesis in different populations of Acorus from Wisconsin and Iowa reveals different degrees of apical and adaxial meristematic activity in primordia of these two collections: leaves with marked adaxial growth exhibit delayed and reduced expression of apical growth, whereas primordia with marked apical growth show, correspondingly, reduced adaxial meristematic activity at equivalent stages of development. Such variations in leaf histogenesis are correlated with marked differences in adult leaf anatomy in the respective populations and explain the reasons for the sympodial interpretation of leaf morphogenesis in Acorus and unifacial organs of other genera by previous investigators. It is concluded that leaf development in Acorus resembles that of the Acacia phyllode, thereby confirming from a developmental viewpoint the homology of these organs. Comparison of development with leaves of other Araceae indicates that the modified form of the leaf of Acorus originates through the accentuation of adaxial and abaxial meristematic activity which is expressed only slightly in the more conventional dorsiventral leaf types in the family.     

COMPARISON OF EARLY LATERAL VEIN FORMATION IN LINUM USITATISSIMUM LEAVES WITH THEORETICAL NETWORK MODELS

The plastochron age of the Linum leaf that first exhibited lateral leaf vein divergences, the divergent leaf, increased through shoot ontogeny, but the size of the divergent leaf remained constant. There were progressive decreases in the plastochron and relative plastochron rate of leaf elongation, but no significant change in relative chronological rate of leaf elongation, through ontogeny. Thus, divergent leaves of similar sizes occupied different relative positions in the array of leaves on stems of different plastochron ages. These observations are partially consistent with theoretical network model predictions on early leaf vein development. The empirical data of this study suggest additional features of leaf development that should be incorporated into future simulation models for leaf vein development.     

Rates of corolla growth in tobacco determined with the plastochron index

The growth of vegetative and reproductive shoots of Nicotiana tabacum L. cv. Xanthi is analyzed with the plastochron index to estimate the relationship between corolla growth and time. The plastochron of leaves 9 through 20 declines steadily at each successive node. The flower plastochron increases steadily during the growth of an individual cyme, with the most distal flower to open having the longest plastochron. Variation in the flower plastochron is the result of variation in the rate of flower initiation, not the growth rate of individual flowers. The corolla has an extended phase of approximately constant relative growth in length (between 0.2 · d^–1 and 0.3 · d^–1) until a peak of growth (0.5 · d^–1) 2–3 d before anthesis. Corollas also have periodic peaks and troughs of growth that are low in amplitude (0.1 · d^–1), but persist throughout most of corolla development. The pattern of corolla expansion contrasts strongly with earlier reports of the pattern of tobacco leaf growth.Abbreviations PI plastochron index - PR plastochron ratio - RGR relative growth rate in length The authors thank: Drs. T. Sage and E.G. Williams for the considerable time and space they invested; the members of Dr. R. Wyatt's laboratory for allowing us to use their computer facilities; A. Tull and M. Smith for their care taken in the green-house. This research was supported by U.S. Department of Agriculture grant GAM-89-01056 and by National Science Foundation grant DCB-87-15799.     

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.     

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.     

The function of the RNA-binding protein TEL1 in moss reveals ancient regulatory mechanisms of shoot development

The shoot represents the basic body plan in land plants. It consists of a repeated structure composed of stems and leaves. Whereas vascular plants generate a shoot in their diploid phase, non-vascular plants such as mosses form a shoot (called the gametophore) in their haploid generation. The evolution of regulatory mechanisms or genetic networks used in the development of these two kinds of shoots is unclear. TERMINAL EAR1-like genes have been involved in diploid shoot development in vascular plants. Here, we show that disruption of PpTEL1 from the moss Physcomitrella patens, causes reduced protonema growth and gametophore initiation, as well as defects in gametophore development. Leafy shoots formed on ΔTEL1 mutants exhibit shorter stems with more leaves per shoot, suggesting an accelerated leaf initiation (shortened plastochron), a phenotype shared with the Poaceae vascular plants TE1 and PLA2/LHD2 mutants. Moreover, the positive correlation between plastochron length and leaf size observed in ΔTEL1 mutants suggests a conserved compensatory mechanism correlating leaf growth and leaf initiation rate that would minimize overall changes in plant biomass. The RNA-binding protein encoded by PpTEL1 contains two N-terminus RNA-recognition motifs, and a third C-terminus non-canonical RRM, specific to TEL proteins. Removal of the PpTEL1 C-terminus (including this third RRM) or only 16–18 amino acids within it seriously impairs PpTEL1 function, suggesting a critical role for this third RRM. These results show a conserved function of the RNA-binding PpTEL1 protein in the regulation of shoot development, from early ancestors to vascular plants, that depends on the third TEL-specific RRM.