Comparison of Leaf Plastochron Index and Allometric Analyses of Tooth Development in Arabidopsis thaliana

Abstract Two methods of analyses were used to investigate tooth development in serrate (se) mutant and wild-type Columbia-1 (Col-1) Arabidopsis thaliana leaves. There were almost twice as many teeth with deeper sinuses and two orders of toothing on the margins of serrate compared with Columbia-1 leaves. The main objective of this study was to test three hypotheses relative to the source of polymorphism in tooth development: (i) Teeth share similar growth rates and initial sizes, but the deeper teeth are initiated earlier in leaf development. (ii) Teeth share similar timing of initiation and growth rates, but the deeper teeth have a larger initial size. (iii) Teeth share similar timing of initiation and initial sizes, but the deeper teeth have a faster growth rate. Leaf plastochron index (LPI) was used as the time variable for leaf development. Results showed teeth in se were initiated at −27 LPI, 15 plastochrons earlier than those of Col-1. Serrate leaf expansion was biphasic, with the early phase expanding at half the relative plastochron rate of the later phase, which equaled the constant relative expansion rate of Col-1 leaves. Allometric analyses of tooth development obscured the interactions between time of tooth and leaf initiation and the early phase of leaf expansion characteristic of serrate leaves and teeth. Timing of developmental events that allometric analysis obscured can be readily detected with the LPI as a developmental index. Received 25 January 2000; accepted 17 March 2000     

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.     

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     

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.     

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.     

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.     

SPECIAL PAPER: THE PLASTOCHRON INDEX: A REVIEW AFTER TWO DECADES OF USE

The plastochron index provides a morphological time scale which has proved more reliable than chronological age in studies relating morphological and physiological development of a whole plant or plant organ. Since its inception in 1957, the index has been utilized in a variety of investigations from leaf ontogeny in cottonwood trees to rhizoid cluster initiation in algae. The plastochron index has been extensively used in studies involving source and sink relationships, leaf anatomy, cell differentiation, and primary vascularization. It has been used in investigations of hormonal regulation of plant growth and in studies of the effects of various environmental factors on developmental processes in crops. This paper reviews some of the literature from 1957 to present concerning the development and use of the plastochron index.     

14CO2 Fixation, Translocation, and Carbon Metabolism in Rapidly Expanding Leaves ofPopulus deltoides

To examine ¹⁴CO₂ fixation, potential translocation, and carbonflow among leaf chemical fractions of young developing leaves,the shoot tip of 24-leaf cottonwood (Populus deltoides Bartr.ex. Marsh) plants were cut off under water, placed in artificialxylem sap, and treated with ¹⁴CO₂ in continuous and pulse-chaseexperiments. Additional leaves on whole plants were spot treatedon the lamina tip to follow export from the tip only. The analysedleaves ranged in age from leaf plastochron index(LPI) –5to 3, the spot treated leaves from LPI 2 to 5. After 30 minfixation, the specific activity in the lamina tip increasedlinearly with leaf age from LPI –5 to 1 (0.5 to 4.5 kBqmg^–1). Specific activity in the lower lamina increasedslowly with leaf age and did not reach 500 kBq mg^–1 untilLPI –1. Total ¹⁴CO₂ fixed in the lower lamina exceededthat fixed in the tip by LPI –2 because of the large amountof tissue present in the lower lamina. Although the lamina tipfixed high levels of ¹⁴CO₂, pulse-chase studies coupled withautoradiography indicated no vein loading or translocation fromthe tip until about LPI 4 or 5. The ¹⁴C fixed in both tip andlower lamina was incorporated at the site of fixation and wasnot distributed to younger tissue or translocated from the lamina.Although the percentage distribution (¹⁴C in each chemical fractioncompared with the total in all fractions) of ¹⁴C among certainchemical fractions, e.g. sugars, amino acids and proteins, indicatedthat the mesophyll of the tip was more mature than the lowerlamina, physiologically both leaf sectors were immature basedon the expected ¹⁴C distribution in mature tissue. Informationfrom this and other studies indicates that the extreme tip ofa developing cottonwood leaf first begins to export photosynthateabout LPI 4 or 5 on a 24-leaf plant. The first photosynthatetranslocated may be incorporated into the vascular tissues andmesophyll directly below the tip. However, as the tip continuesto mature photosynthate is translocated past the immature lowerlamina into the petiole and out of the leaf. Populus deltoides Bartr. ex. Marsh, eastern cottonwood, translocation, leaf development, ¹⁴C fixation, carbon metabolism     

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.     

ACTIVITY OF MARGINAL AND PLATE MERISTEMS DURING LEAF DEVELOPMENT OF XANTHIUM PENNSYLVANICUM

The mitotic and biosynthetic activities of the marginal and plate meristems were studied during the entire course of leaf development of Xanthium pennsylvanicum. In contrast to statements in the literature, marginal meristem activity is long in duration, as assayed by the mitotic counts and H³-thymidine incorporation. This me istem is active 23 days. The plate meristem is active for an additional 3 days after cessation of cell division in the marginal meristem, but the total duration of its mitotic activity is also approximately 23 days. Numerous periclinal cell divisions of the plate meristem form additional cell layers and contribute to the growth of the lamina in thickness. Incorporation of H³-thymidine increased during the course of leaf development. Cells between plastochronic ages 0 and 2.0 incorporated more of the radioisotopic precursor than those of younger leaf primordia. The uptake and incorporation of H³-thymidine into nuclear DNA was more sluggish during the early stages of development than in the more expanded leaves. No DNA synthesis was demonstrated after cessation of cell division in the leaf lamina. Metabolic or endomitotic DNA synthesis after leaf plastochron index (LPI) 3.0 seems improbable. No significant differences in the incorporation of H³-thymidine could be demonstrated between the marginal and plate meristems. This would indicate no distinct biosynthetic differences between the two meristems. The definitions of the marginal and plate meristems of Xanthium leaves were formulated in view of the above findings.     

ANATOMICAL CHANGES DURING LEAF ONTOGENY IN POPULUS DELTOIDES

The leaf plastochron index (LPI) was used to interpret the anatomical changes during leaf ontogeny in the developing leaf zone of young cottonwood trees and to relate leaf anatomical structure to physiological function. The lamina tip matured precociously with respect to both structure and function. Below the lamina tip the intercellular spaces, stomates, and minor veins matured basipetally, while the major veins developed acropetally. Ontogenetically, maturation progressed from LPI –1.0, which was anatomically immature except for its lamina tip, to the first fully expanded leaf at LPI 6.0, which was anatomically mature. Physiological maturity also occurred at LPI 6.0, thus signifying a transition with respect to both structure and function. By evaluating the anatomical observations in conjunction with physiological data collected at comparable LPI's in other experiments, it could be demonstrated that anatomical development was a limiting factor in photosynthesis and translocation of assimilates.     

A METHOD FOR DETERMINING PLASTOCHRON INDICES DURING HETEROBLASTIC SHOOT GROWTH

A plastochron is defined as the time interval between two successive recurring events during the growth of plant shoots, such as leaf initiation. The plastochron index (PI) formulated by Erickson and Michelini (1957, American Journal of Botany 44: 297–305) provides a method for determining 1) morphological equivalence in a developmentally variable sample of shoots and 2) rates of development in microscopic tissues and organs, by expressing shoot age as a function of plastochron number. The PI assumes that homologous organs at successive nodes grow exponentially, at equal rates, and the plastochron remains constant. These three conditions are not met in many shoots that exhibit heteroblasty in their plastochron and the growth rate of organs at successive nodes. An alternative computational method for the PI is presented that uses two measurements taken at different times from the same organ during its exponential growth phase. The method does not assume that the PI is a linear function of time. Results of an analysis of cyme internode growth in two races of Arenaria uniflora (Caryophyllaceae) demonstrate that the method proposed is in good agreement with Erickson and Michelini's (1957) method when shoot growth is not markedly heteroblastic. The current method is also used to determine the nonlinear relation between PI and time in a race of A. uniflora that has heteroblastic cyme growth. The results generalize the PI for use in studies of heteroblasty, and for shoots where the relative plastochron rate cannot be directly determined.     

PLASTOCHRON2 regulates leaf initiation and maturation in rice

In higher plants, leaves initiate in constant spatial and temporal patterns. Although the pattern of leaf initiation is a key element of plant shoot architecture, little is known about how the time interval between initiation events, termed plastochron, is regulated. Here, we present a detailed analysis of plastochron2 (pla2), a rice (Oryza sativa) mutant that exhibits shortened plastochron and precocious maturation of leaves during the vegetative phase and ectopic shoot formation during the reproductive phase. The corresponding PLA2 gene is revealed to be an orthologue of terminal ear1, a maize (Zea mays) gene that encodes a MEI2-like RNA binding protein. PLA2 is expressed predominantly in young leaf primordia. We show that PLA2 normally acts to retard the rate of leaf maturation but does so independently of PLA1, which encodes a member of the P450 family. Based on these analyses, we propose a model in which plastochron is determined by signals from immature leaves that act non-cell-autonomously in the shoot apical meristem to inhibit the initiation of new leaves.     

PLASTOCHRON INDICES FOR JUVENILE AND MATURE FORMS OF HEDERA HELIX L. (ARALIACEAE)

English ivy (Hedera helix) plants were assessed for the applicability of the plastochron index (PI). Juvenile ivy satisfied all requirements for the use of the PI and showed a plastochron of 4.23 days. Mature ivy grown under long day conditions flowered after 11–12 leaves. Two distinct groups of leaves were produced with different plastochrons (0.83 and 3.2 days, respectively) and leaf morphologies. Long-day-grown ivy did not satisfy the requirements for the use of the PI. Short-day-grown mature ivy continued production of leaves beyond the 12th leaf. Vegetative growth was perpetuated for at least 25 plastochrons. By 19 plastochrons (ca. 41 days after budbreak) a linear PI vs. time relationship was established with a plastochron of 3.16 days. This newly acquired ability to maintain vegetative growth in mature ivy plants may allow a direct comparison with the vegetative indeterminant juvenile in order to assess possible anatomical factors responsible for phase stability and phase change using the PI as a basis for comparison.     

Translocation pathways in the petioles and stem between source and sink leaves of Populus deltoides Bartr. ex Marsh.

Microautoradiography was used to follow the translocation pathways of ¹⁴C-labeled photosynthate from mature source leaves, through the stem, to immature sink leaves three nodes above. Translocation occurred in specific bundles of the midveins and petioles of both the source and sink leaves and in the interjacent internodes. When each of six major veins in the lamina of an exporting leaf was independently spot-fed ¹⁴CO₂, label was exported through specific bundles in the petiole associated with that vein. When the whole lamina of a mature source leaf was fed ¹⁴CO₂, export occurred through all bundles of the lamina, but acropetal export in the stem was confined to bundles serving certain immature sink leaves. Cross-transfer occurred within the stem via phloem bridges. Leaves approaching maturity translocated photosynthate bidirectionally in adjacent subsidiary bundles of the petiole. That is, petiolar bundles serving the lamina apex were exporting unlabeled photosynthate while those serving the lamina base were simultaneously importing labeled photosynthate. The petioles and midveins of maturing leaves were strong sinks for photosynthate, which was diverted from the export front to differentiating structural tissues. The data support the idea of bidirectional transport in adjacent bundles of the petiole and possibly in adjacent sieve tubes within an individual bundle.Abbreviations C central leaf trace - L left leaf trace - LPI leaf plastochron index - R right leaf trace     

COMPARATIVE FLOWER DEVELOPMENT IN THE CLEISTOGAMOUS SPECIES VIOLA ODORATA. I. A GROWTH RATE STUDY

In Viola odorata, chasmogamous (CH) or open flowers and small, short-petioled leaves are produced under 11 hr or less of daylight, cleistogamous (CL) or closed flowers and large, long-petioled leaves under 14 hr or more of daylight, and intermediate floral and leaf forms under transitional photoperiods. CL flowers are approximately four times smaller than CH flowers and differ morphologically in repressed growth of the anterior petal spur and staminal nectaries, and recurving of the style which remains enclosed within the cone formed by anther appendages. Both CH and CL shoot systems conform to a (2 + 3) phyllotaxis with minor differences in leaf divergence angles and phyllotactic indices. The larger CL leaf grows significantly faster than the CH leaf, and an increased rate of leaf initiation occurs in the CL apex represented by a plastochron of 3.4 days compared to 4.3 days in the CH apex. The plastochron index was used to indirectly age young floral primordia nondestructively until prophase of meiosis I within the anthers. This event occurs 8 days earlier in the CL than the CH flower. Time from meiosis until flower maturity, determined by direct observation, is about 14 days for the CL flower, versus 21 days for the CH flower.     

THE ELASTIC MODULI AND MECHANICS OF POPULUS TREMULOIDES (SALICACEAE) PETIOLES IN BENDING AND TORSION

Changes attending leaf development in the mechanical behavior and elastic moduli of the petioles of Populus tremuloides Michx. were examined in terms of total leaf weight (Wt), lamina and petiole weight (Wl and Wp), petiolar length (L), and lamina surface area (A). A primary concern was the extent to which two elastic moduli (Young's modulus E and the shear modulus G) of petioles changed over time and were correlated with one another. E and G were measured by means of multiple resonance frequency spectra, and together with the dimensions of cross sections through petioles, these two elastic moduli were used to estimate the stiffness of petioles in simple bending and torsion. Petiolar bending was predicted by means of a model incorporating expressions for both the bending stiffness (El) and torsional rigidity (GJ), where I is the second moment of area and J is the torsional constant. The predictions from these models were compared to observed petiolar bendings due to Wl. Additionally, the frequency of the oscillatory motion of leaves (placed in a wind tunnel with a 1 m sec^-1 ambient wind speed, directed normal to the blade of the leaf) was determined. Results indicate that L, A, Wl, Wp, and Wt were positively correlated with the age of leaves (crudely estimated as a function of leaf plastochron index, LPI); these morphometric parameters were also correlated with the magnitudes of E and G. Also, G was positively and linearly correlated with E, and was, on the average, an order of magnitude less than E. EI and GJ were positively correlated with LPI. The relationships among E, G, EI, C and Wl, Wp, Wt are discussed in terms of leaf allometries.     

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.