ON THE MECHANISM OF DECUSSATE PHYLLOTAXIS: BIOPHYSICAL STUDIES ON THE TUNICA LAYER OF VINCA MAJOR

Phyllotaxis theory typically assumes that an acropetal influence from recently formed leaves acts on the apical dome to initiate new leaves. Biophysical theory postulates that established plant organs elongate because their primary walls, particularly those in the organ surface layer, are transversely reinforced by cellulose to give the organ overall hoop reinforcement. These two postulates are combined here in a biophysical theory for phyllotaxis. The essential acropetal influence from young leaves is proposed to be the stretching of the adjacent dome tissue by the growth of leaf bases. Cytoskeletal responses on the dome produce reinforcement patterns which initiate new hoop reinforced leaves. Growth of these leaves remodels the dome for the next round of organs. Data pertinent to this theory are presented here for Vinca major. The surface (tunica) layer of the apical dome was isolated by paradermal cuts. Using polarized light, the cellulose alignment in this surface layer was determined, cell by cell, for various stages of the plastochron. The growing dome is typically elliptical, with the major axis shifting by 90° during each plastochron. The periphery of the dome always has cellulose oriented parallel to its margin; the central region, when the major axis is pronounced, has reinforcement normal to this axis. During the plastochron this reinforcement pattern is modified, by plausible biophysical mechanisms, to account for the three major activities of the dome: 1) production of a hoop-reinforced leaf at each end of the ellipse, 2) formation of a hoop-reinforced stem segment, 3) revision of dome structure to produce the same initial reinforcement pattern as at the start of the plastochron, but at 90°.     

A Sequential Study of Cell Divisions and Expansion Patterns on a Single Developing Shoot Apex of Vinca major

During the growth of a single developing vegetative apex ofVinca major, both the orientation and frequency of cell divisions,and the pattern of cell expansion, were observed using a non-destructivereplica technique. Micrographs taken at daily intervals illustratethat the central region of the apical dome remains relativelyinactive, except for a phase of cell division which occurs after2 d of growth. The majority of growth takes place at the proximalregions of the dome from which develop the successive pairsof leaves. The developing leaf primordia are initiated by aseries of divisions which occur at the periphery of the centraldome and are oriented parallel to the axis of the subsequentleaves. The cells which develop into the outer leaf surfaceof the new leaves undergo expansion and these cells divide allowingfor the formation of the new leaf. This paper describes thefirst high-resolution sequential study of cell patterns in asingle developing plant apex. Sequential development, cell division, expansion patterns, SEM, Vinca major, apical dome, leaf primordium, leaf initiation     

A theory for inflorescence development and flower formation based on morphological and biophysical analysis in Echeveria

Floral development is generally viewed as involving interactions between recently made organs and generative activity on the apical dome; one set of floral organs is thought to induce the next. To investigate such interactions, flowering in Echeveria derenbergii (J. Purpus) was studied at two levels of structure. At the larger, morphological, level the inflorescence apex is shown to have simple cyclic development. Seen from above, it elongates horizontally, then forms a transverse cleft to demarcate a flower primordium in one of two rows. The meristem then elongates at 90° to its previous axis, also horizontally, and demarcates a flower in the other row. Activity on the apical surface correlates well with the nature and activity of adjacent sub-apical organs. For example, the 90° shifts in elongation of the meristem correlate with that tissue's being attached, laterally, to successive large growing bracts whose bases lie at 90°. Also, on the flower primordium, the five sepals arise in a spiral sequence which correlates with one of increasing age, since formation by the cleft, of the edges of the primordium.The second level of study was to test whether the developmental correlations could have a biophysical explanation. By biophysical theory, organs arise where the dome surface is structurally predisposed to bulge. This is a function of the cellulose reinforcement pattern in the surface. Successive patterns of cellulose reinforcement in isolated surface layers from floral organs were determined using polarized light. This was done for the cyclic activity of the inflorescence meristem and the development of the flower. The results indicate that patterns of cellulose reinforcement on the apical dome surface could lead to the production of organs, through local promotion of bulging of the tunica. Subsequent growth of the base of each organ stretches the adjacent dome tissue in a directional fashion. Cytoskeletal responses of these stretched cells lead to new cellulose alignments on the dome which generate the reinforcement pattern for the next round of organs.Abbreviations F floral meristem tissue which will directly produce a flower, starting with sepals - I inflorescence meristem tissue, generally oval in top view and bounded by two bracts, that produces both floral tissue (F) and additional I meristem tissue - I-max the maximal size of I tissue before it bifurcates into F tissue and I tissue (I-min) - I-min the minimal size of I tissue just after it has bifurcated to produce F tissue and I tissue     

DEVELOPMENT OF FLORAL ORGANS IN THE SITES OF LEAF PRIMORDIA IN PINGUICULA VULGARIS

Plants of Pinguicula vulgaris L. have either clockwise or counterclockwise spiral phyllotaxy. The inception of floral primordia occurs in leaf sites as a normal sequence of development. Only two leaf primordia initiated late in the season develop into floral primordia in the following year. They do not represent a direct modification of the apical meristem nor of the detached meristem. The apical meristem continues to produce leaves in the vegetative phase and flowers in the reproductive phase, and thus the plants show a monopodial growth. Axillary buds are not developed in this perennial species and instead additional buds of adventitious ontogeny appear. Such buds are produced on the older leaves of larger plants, and they are extremely useful in the vegetative propagation of the species.     

Learning to recognize objects

Several aspects of systems for learning pattern or object recognition rules are discussed. First, how are recognition rules developed and to what extent is structural pattern information embedded into these recognition rules. Second, how are these rules applied to the recognition of complex patterns such as objects embedded in scenes and how is evidence from different rules combined into a single evidence vector. Third, how can learned rules be improved through performance evaluation and feedback to rule generation stages.     

Comparing alternative models to empirical data: cognitive models of western scrub-jay foraging behavior

Animals often select one item from a set of candidates, as when choosing a foraging site or mate, and are expected to possess accurate and efficient rules for acquiring information and making decisions. Little is known, however, about the decision rules animals use. We compare patterns of information sampling by western scrub-jays (Aphelocoma californica) when choosing a nut with three decision rules: best of n (BN), flexible threshold (FT), and comparative Bayes (CB). First, we use a null hypothesis testing approach and find that the CB decision rule, in which individuals use past experiences to make nonrandom assessment and choice decisions, produces patterns of behavior that more closely correspond to observed patterns of nut sampling in scrub-jays than the other two rules. This approach does not allow us to quantify how much better CB is at predicting scrub-jay behavior than the other decision rules. Second, we use a model selection approach that uses Akaike Information Criteria to quantify how well alternative models approximate observed data. We find that the CB rule is much more likely to produce the observed patterns of scrub-jay behavior than the other rules. This result provides some of the best empirical evidence of the use of Bayesian information updating by a nonhuman animal.     

Microtubule patterning in apical epidermal cells ofVinca minor preceding leaf emergence

Summary Serial thin sectioning for electron microscopy was carried out on the cortical cytoplasm of surface cells of the apical dome ofVinca minor. The cellulose reinforcement pattern in the outer epidermal walls forming this surface is known to correlate well with the decussate phyllotaxis pattern. The purpose of this study was to determine the location of microtubules immediately under these epidermal walls as a first step toward finding out how the cellulose pattern arises. First, correspondence between the patterns of microtubules and cellulose was checked. Second, the role of potential orienting cues for the alignment of microtubule arrays in specific cells was evaluated.Microtubule arrays which were well or moderately ordered (70% of the total interphase cells) generally had alignment parallel to the adjacent leaf base, as has been seen for cellulose. The aligned features or cues potentially correlating with a given array were: (1) orientation and length of the previous anticlinal cross-wall, (2) alignment of microtubules in adjacent cells, and (3) direction of inferred stretch, parallel to the nearby leaf bases. All three features were found to agree with the microtubule alignment in 17 of 34 cells with ordered arrays. At least two features agreed in 33 of the 34 cases. All 34 cells with ordered arrays had at least one feature parallel to the array. Random association between microtubule orientation and these features would lead to such correlations less than 0.01% of the time. Thirty percent of the interphase cells showed no obvious order. Most of these cells were located in the central linear corridor region of the apex. The unordered cells were more likely than the ordered cells to have more than one orientation specified by the potential cues; i.e., no single orientation parallel to all of the cues existed. This indicates that uniformity of the orientation cues may be as important as their direction.     

The Relationship Between the Distribution of Periclinal Cell Divisions in the Shoot Apex and Leaf Initiation

Periclinal cell divisions in vegetative shoot apices of Pisumand Silene were recorded from serial thin sections by mappingall the periclinal cell walls formed less than one cell cyclepreviously. The distribution of periclinal divisions in theapical domes corresponded to the distributions subsequentlyoccurring in the apices when the young leaf primordia were forming.In Pisum, periclinal divisions were almost entirely absent fromthe I₁ region of the apical dome for half a plastochron justafter the formation of a leaf primordium and appeared, simultaneouslyover the whole of the next potential leaf site, about half aplastochron before the primordium formed. In Silene periclinaldivisions seemed to always present in the apical dome at thepotential leaf sites and also round the sides of the dome wherethe ensheathing leaf bases were to form. Periclinal divisionstherefore anticipated the formation of leaf primordia by occuring,in Pisum about one cell cycle and in Silene two or more cellcycles, before the change in the direction of growth or deformationof the surface associated with primordial initiation. Pisum, Silene, planes of cell division, orientation of cell walls, leaf primordia, shoot apical meristem, plastochron     

Life-history patterns in British Umbelliferae: a review

British Umbelliferae form a natural cohesive taxon, having patterns of habitat distribution, reproductive biology and life-history that distinguish them from other British dicotyledons. They are usually found in open sites, and are local or rare in distribution. Almost half of British species have the andromonoecious sex habit. Perfect flowers are usually protandrous. The floral composition of umbels, and the pattern of opening of the inflorescence combine to produce a protogynous effect. These have been interpreted as mechanisms which decrease competition for resources between male and female functions in the plant, such that separate maternal and paternal fitness can become optimal. Seeds often require a chilling treatment, alternating temperatures, or a period of after-ripening before they are capable of germination. Schizocarps are of low weight. This seems to relate to the relative openness of habitat wherein unbellifers are found. Patterns of dry matter distribution reflect the life-history strategies of various umbellifer species. Agromyzid leaf miners parasitize the more widespread species, and those which are found in a greater number of different habitats.     

Life-history patterns in British Umbelliferae: a review

British Umbelliferae form a natural cohesive taxon, having patterns of habitat distribution, reproductive biology and life-history that distinguish them from other British dicotyledons. They are usually found in open sites, and are local or rare in distribution.
Almost half of British species have the andromonoecious sex habit. Perfect flowers are usually protandrous. The floral composition of umbels, and the pattern of opening of the inflorescence combine to produce a protogynous effect. These have been interpreted as mechanisms which decrease competition for resources between male and female functions in the plant, such that separate maternal and paternal fitness can become optimal.
Seeds often require a chilling treatment, alternating temperatures, or a period of after-ripening before they are capable of germination. Schizocarps are of low weight. This seems to relate to the relative openness of habitat wherein unbellifers are found. Patterns of dry matter distribution reflect the life-history strategies of various umbellifer species. Agromyzid leaf miners parasitize the more widespread species, and those which arc found in a greater number of different habitats.     

Floral morphogenesis in Anagallis: Scanning-electron-micrograph sequences from individual growing meristems before,during, and after the transition to flowering

Non-destructive scanning electron microscopy allows one to visualize changing patterns of individual cells during epidermal development in single meristems. Cell growth and division can be followed in parallel with morphogenesis. The method is applied here to the shoot apex of Anagallis arvensis L. before, during, and after floral transition. Phyllotaxis is decussate; photoperiodic induction of the plant leads to the production of a flower in the axil of each leaf. As seen from above, the recently formed oval vegetative dome is bounded on its slightly longer sides by creases of adjacent leaf bases. The rounded ends of the dome are bounded by connecting tissue, horizontal bands of node cells between the opposed leaf bases. The major growth axis runs parallel to the leaf bases. While slow-growing at the dome center, this axis extends at its periphery to form a new leaf above each band of connecting tissue. Connecting tissue then forms between the new leaves and a new dome is defined at 90° to the former. The growth axis then changes by 90°. This is the vegetative cycle. The first observed departure from vegetative growth is that the connecting tissue becomes longer relative to the leaf creases. Presumably because of this, the major growth axis does not change in the usual way. Extension on the dome continues between the older leaves until the axis typically buckles a second time, on each side, to form a second crease parallel to the new leaf-base crease. The tissue between these two creases becomes the flower primordium. The second crease also delimits the side of a new apical dome with the major axis and growth direction altered by 90°. During this inflorescence cycle the connecting tissue is relatively longer than before. Much activity is common to both cycles. It is concluded that the complex geometrical features of the inflorescence cycle may result from a change in a biophysical boundary condition involving dome geometry, rather than a comprehensive revision of apical morphogenesis.Abbreviation SEM scanning electron microscopy, micrograph Use of the SEM facility of Professor G. Goffinet, Institute of Zoology, University of Liège, is greatly appreciated. We thank Dr. R. Jacques, C.N.R.S., Le Phytotron, Gif-sur-Yvette, France, for providing the experimental material, and Mr. Philippe Ongena for expert photography. Support was from grants from the U.S. Department of Agriculture and National Science Foundation as well as from the Fonds National de la Recherche Scientifique, Fonds de la Recherche Fondamentale et Collective, and the Action de Recherche Concertée of Belgium.     

The Effects of Photoperiod and Mineral Nutrient Supply on Growth and Primordia Production at the Stem Apex of Barley Seedlings

Growth and development of the shoot apex in seedlings of threebarley cultivars was examined in two daylengths (8, 16 h) andat two mineral nutrient levels (x 1, x 0.1). Production of primordiawas greater at the higher nutrient level and in the longer days.The rate of production varied with cultivar but in all casesthe plastochron shortened with transition to spike formation.Early flowering (cv. Clipper) was associated with a high rateof primordial production and early transition to spike formation,late flowering (cv. Proctor) with a low rate of production anda longer vegetative phase. The cultivar Akka showed intermediatecharacteristics. The volume of the apical dome increased linearlywith increasing numbers of primordia, the rate of increase varyingwith cultivar and treatment. Enlargement of the dome was duemainly to increase in cell number. The transition of the apexto produce spikelet primordia occurred with widely differingvolumes of the apical dome, thus invalidating the hypothesisthat transition is dependent upon attainment of a critical domesize. Although both the rate of production of primordia andenlargement of the dome were markedly affected by photoperiod,both were unaffected when the photoperiodic treatment was givendirectly to the shoot apex. It is considered that the fate of a primordium once initiatedis determined by competition for available metabolites betweenit, other primordia and the apical dome. Hordeum vulgare L, barley, apical dome, primordia, plastochron, cell division     

Cryopreservation for eradication of Jujube witches' broom phytoplasma from Chinese jujube (Ziziphus jujuba)

Here, we report efficient eradication of Jujube witches' broom phytoplasma (Candidatus Phytoplasma ziziphi) from Chinese jujube (Ziziphus jujuba) by cryopreservation. Shoot tips (1.0 mm in size) with 5–6 leaf primordia (LPs) excised from diseased in vitro stock shoots were subject to droplet‐vitrification cryopreservation. Shoot tips following cryopreservation were post‐cultured on a recovery medium for survival. Plantlet regeneration was obtained by micrografting of surviving shoot tips upon in vitro rootstocks. With this protocol, 85% of shoot tips survived following cryopreservation, among which 75% regenerated into whole plantlets and all of them were free of phytoplasma, regardless of the sizes used for cryopreservation. Ultrastructural studies demonstrated that phytoplasma was absent in the apical dome, and leaf primordia (LPs) 1 and 2, while abundance of phytoplasma was present in the lower parts of shoot tips, leaf primordium 3 and older tissues. Histological observations showed that much more damage was found in cells located in the lower part of apical dome, leaf primordium 3 and older tissues than in those at the upper part of apical dome and in the LPs 1 and 2. These cells were most likely to survive and regenerate into phytoplasma‐free plantlets following cryopreservation and micrografting. Ploidy levels analyzed by flow cytometry (FCM) were maintained in plantlets regenerated from cryopreservation followed by micrografting. Results reported here would provide technical support for production of phytoplasma‐free plants and for long‐term storage of germplasm of Chinese jujube.     

Rates of Cell Division in the Shoot Apical Meristem of Pisum

The relative rates of cell division in different regions ofthe pea shoot apical meristem were obtained by measuring theincrease in the numbers of metaphases following applicationof colchicine to the plants. Absolute values for the rates ofcell division could be calculated since the average rate ofcell division for the whole apex was known. Measurements ofthe rates of cell division were obtained at defined intervalsduring the course of a single plastochron. Within each regionof the apex the rate of cell division did not change more thanabout two-fold throughout the plastochron. There was very littleor no increase in the rate of cell division associated withleaf initiation. The formation of a leaf primordium and thesubsequent growth of the apical dome apparently result fromchanges in the direction of growth rather than changes in therates of growth. Three main regions were discernible withinthe apical meristem: a region with a slow rate of cell divisionin the apical dome, a region of a faster rate of cell divisionat the base of the apical dome and at the site of initiationof procambial strands, and a region of an intermediate rateof cell division in the newly initiated leaf primordium andthe adjacent part of the shoot axis.     

Shoot tip culture and cryopreservation for eradication of Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) from apple rootstocks ‘M9’ and ‘M26’

This study attempted to eradicate Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) from virus‐infected in vitro shoots of apple rootstocks ‘M9’ and ‘M26’ using shoot tip culture and cryopreservation. In shoot tip culture, shoot tips (0.2 mm in length) containing two leaf primordia failed to show shoot regrowth. Although shoot regrowth rate was the highest in the largest shoot tips (1.0 mm in length) containing four leaf primordia, none of the regenerated shoots was virus‐free. Shoot tips (0.5 mm in length) containing two and three leaf primordia produced 100% and 10% of ASPV‐free shoots, respectively, while those (1.0 mm) containing four leaf primordia were not able to eradicate ASPV. ASGV could not be eradicated by shoot tip culture, regardless of the size of the shoot tips tested. In cryopreservation, shoot tips (0.5 mm in length) containing two leaf primordia did not resume shoot growth. Although 1.0‐mm and 1.5‐mm shoot tips gave similarly high ASPV‐free frequencies, the latter had much higher shoot regrowth rate than the former. Very similar results of shoot regrowth and virus eradication by shoot tip culture and cryopreservation were observed in both ‘M9’ and ‘M26’. Histological observations showed that only cells in upper part of apical dome and in leaf primordia 1–3 survived, while other cells were damaged or killed, in shoot tips following cryopreservation. Virus immunolocalization found ASPV was not detected in upper part of apical dome and leaf primordia 1 and 2, but was present in lower part of apical dome, and in leaf primordium 4 and more developed tissues in all samples tested. ASPV was also detected in leaf primordium 3 in about 16.7% and 13.3% samples tested in ‘M9’ and ‘M26’. ASGV was observed in apical dome and leaf primordia 1–6, leaving only a few top layers of cells in apical dome free of the virus. Different abilities of ASPV and ASGV to invade leaf petioles and shoot tips were also noted.     

Arrangement of cortical microtubules at the surface of the shoot apex inVinca major L.: Observations by immunofluorescence microscopy

Arrangements of cortical microtubules (MTs) and of cellulose microfibrils at the surface of the vegetative shoot apex ofVinca major L. were examined by immunofluorescence microscopy and polarizing microscopy, respectively. Cortical MTs adjacent to the outermost walls of the apex were arranged more or less randomly in individual cells: especially in cells in the central region of the apex the arrangement was almost completely random. However, in the peripheral region MTs tended to show parallel alignment in individual cells, and an overall pattern that was roughly concentric around the apical dome was discerned. Observations of birefringence of cell walls indicated that cellulose microfibrils in the peripheral region of the apex were also arranged in a pattern which was roughly concentric around the apical dome. These patterns of arrangements of MTs and microfibrils are understood to be perpendicular to the radial cell files observed in the peripheral region of the apex, and can be related to the radial expansion of the surface of the apex.     

Effects of Shading the First Leaf of Barley Plants on Growth and Carbon Nutrition of the Stem Apex

Effects of shading the first leaf on development of the apicalregion were investigated by examining the growth of leaf primordiaand the apical dome in the early seedling stages. Shade treatment affects the size of the dome; it was shown thatvalues for height, width, and volume of the dome of 12-day controlplants were always higher than for shaded plants. Primordialgrowth, in terms of length and dry weight, was reduced by shadeand growth in dry weight of the second, third, and fourth leaveswas shown to be dependent on photosynthetic production by thefirst leaf. Incorporation of ¹⁴C in the apical region was detected by autoradiographyon day 6 and increased with age. Transfer of assimilated carbonfrom the first leaf to the apex occurred during the first 3h after exposure to ¹⁴CO₂. On a unit dry-weight basis it isshown that the third and fourth leaves and apex incorporatedproportionately more labelled carbon than the larger older organssuch as the, second leaf. Shade treatment reduced incorporationinto the apical region and this is associated with the failureof the apex to grow over the period up to day 15. Evidence isprovided to show that in control plants the second leaf suppliescarbon to the apex from about day 12. The crucial importanceof the contribution of the first leaf to plant development isdiscussed.     

LONG-TERM DEVELOPMENTAL CHANGES IN XANTHIUM INDUCED BY GIBBERELLIC ACID

One application of gibberellic acid (GA₃) to Xanthium shoots resulted in an initial large stimulation, followed by inhibition, of internode elongation. After presumed translocation of the hormone from the locus of its application to the stem apex several morphological changes were observed. There was a significant increase in number of mitotic figures in the apical meristem and a twofold increase in volume of the apical dome. With time, the rate of leaf production was accelerated about 1.8 times. The phyllotaxis of leaf primordia initiated under the influence of GA:, changed from a (2, 3) contact parastichy pattern in control shoot to a (3, 5) pattern. Final petiole length was smaller than the control, and the absolute rate of lamina expansion decreased under prolonged treatment. Gibberellic acid had a pronounced effect on leaf morphology. GA_a induced the development of lanceolate leaves instead of typical deltoid leaves. The reduction in leaf area coincided with a 32% reduction in the average area of epidermal cells. Plastochron changes were correlated with anatomical and morphological changes during the course of leaf development.