SHOOT APICAL MERISTEMS AND MUTATION: FIXATION OF SELECTIVELY NEUTRAL CELL GENOTYPES

Shoot apical meristems are interpreted as either structured, that is having a permanent set of apical initials, or stochastic, having apical initials which represent “... momentary representatives of the continuous meristematic residue at the apex of the relevant layer or zone” (Newman, 1965). The two main parameters of stochastic growth are the average number of apical initials (α) and the number of mitotic cycles (r) of the initials and their daughter cells prior to the random selection of subsequent initials. Mathematical analysis and computer simulation studies of stochastic growth have shown that if one starts with 1 mutant initial and α-1 nonmutant initials, eventually a mosaic plant results. The frequency of shoot apices composed of mutant cells is 1/α and the frequency of shoot apices composed of only nonmutant cells is (1 – α)/α. These asymptotics are only attained after considerable growth, thus mericlinal chimeras can persist for many nodes and give the appearance that a permanent set of initials is present.     

THE CONCEPT OF INCIPIENT VASCULAR TISSUE IN FERN APICES

A comparison of shoot apices of runners and rosettes of Nephrolepis with shoot apices of Adiantum indicates that a uniform concept of apical organization can be applied throughout. The region directly below the apical initials is designated a “central zone,” by analogy with that found in Lycopodium. The central zone is regarded as undifferentiated meristematic tissue rather than incipient vascular tissue.     

STUDIES ON THE ONTOGENY OF THE DWARF MISTLETOES,ARCEUTHOBIUM. II. HOMOLOGY OF THE ENDOPHYTIC SYSTEM

Cohen , L. I. (Washington State U., Pullman.) Studies on the ontogeny of the dwarf mistletoes, Arceuthobium. H. Homology of the endophytic system. Amor. Jour. Bot. 50(5): 409–417. Illus. 1963.—Development of the seedling in Arceuthobium, as well as the mode of penetration and infection, is described. The promeristem of the root apex of the seedling, as in the embryo, is composed of 4 zones: (1) a uniseriate layer of apical surface initials; (2) a subapical zone of central initials; (3) a peripheral zone; and (4) a zone of procambial initials. After germination, the seedling grows on the host surface until the root apex is confronted by a spur shoot or by a fragment of raised bark. The root apex becomes attached to the host by means of a massive holdfast. It is the procambial initials which actually invade the host tissue. After penetrating the host, the individual procambium initials proliferate in the cortex, each giving rise to a cortical strand. It was concluded that the endophytic system of Arceuthobium cannot be interpreted in terms of classical concepts of plant homology; it is, in fact, an organ sui generis.     

THE CYTOHISTOLOGICAL AND CYTOHISTOCHEMICAL ZONATION OF THE SHOOT APEX OF BOTRYCHIUM MULTIFIDUM

Cytohistologically, the shoot apex of Botrychium multifidum is composed of three zones—a zone of surface initials in which there is usually a centrally located apical cell, a zone of subsurface initials, and a cup-shaped zone that is subdivided into a pheripheral zone and a rib meristem. The results of cytohistochemical tests for total protein, RNA, total carbohydrate, histones, and DNA localization support this concept. Thus, the cytohistological zonation of the apical meristem of the Ophioglossales is essentially identical to that of the Filicales, and furthermore, is comparable to that of the seed plants.     

AN ELECTRON MICROSCOPIC EVALUATION OF CYTOHISTOLOGICAL ZONATION IN THE SHOOT APICAL MERISTEM OF PINUS BANKSIANA

Shoot apical meristems of jack pine (Pinus banksiana) were examined by light and electron microscopy. Cytohistological zonation was evident when meristems were fixed in Craf IV, embedded in paraffin, and stained with Chlorazol Black E. When meristems were fixed for electron microscopy the cytoplasm of the apical initials and central mother cells each contained numerous lipid bodies and their nuclei contained little, if any, heterochromatin. The cytoplasm of the peripheral zone was rich in ribosomes. The nuclei of the peripheral zone and rib meristem were heterochromatic. Thus, the lack of heterochromatin in the nuclei and the dissolution of lipids in the cytoplasm of the apical initials and central mother cells appeared to contribute most to the organization and appearance (cytohistological zonation) of the shoot apex when standard histological techniques are used.     

SHOOT APEX ORGANIZATION AND ORIGIN OF THE RHIZOME-BORNE ROOTS AND THEIR ASSOCIATED GAPS IN DENNSTAEDTIA CICUTARIA

The shoot apex of Dennstaedtia cicutaria consists of three zones—a zone of surface initials, a zone of subsurface initials, and a cup-shaped zone that is subdivided into a peripheral region and central region. A diffuse primary thickening meristem, which is continuous with the peripheral region of the cup-shaped zone, gives rise to a broad cortex. The roots occurring on the rhizomes are initiated very near the shoot apex in the outer derivatives of the primary thickening meristem. The roots that occur on the leaf bases also differentiate from cortical cells. Eventually, those cortical cells situated between the newly formed root apical cell and the rhizome procambium (or leaf trace) differentiate into the procambium of the root trace, thus establishing procambial continuity with that of the rhizome or leaf trace. Parenchymatous root gaps are formed in the rhizome stele and leaf traces when a few of their procambial cells located directly above the juncture of the root trace procambium differentiate into parenchyma. As the rhizome procambium or leaf trace continues to elongate, the parenchyma cells of the gap randomly divide and enlarge, thus extending the gap.     

ONTOGENY OF THE INFLORESCENCE IN CHENOPODIUM ALBUM

Gifford , Ernest M., Jr ., and Herbert B. Tepper . (U. California, Davis.) Ontogeny of the inflorescence in Chenopodium album. Amer. Jour. Bot. 48(8): 657–667. Illus. 1961.—Chenopodium album, a short-day plant, was induced to flower by subjecting it to successive cycles of 7 hr light and 17 hr darkness. After 4 inductive days, the first macroscopic change is evident in the appearance of precocious axillary bud primordia. After 5–6 days, a primordial inflorescence is visible, and after 7–8 days a terminal flower appears on the main inflorescence axis. The vegetative apex has a biseriate tunica, the cells of which are larger than those of the corpus. The cells of the tunica stain lighter, possess larger nucleoli, and are more vacuolate than cells of the subjacent corpus. After photoinduction, the tunica-corpus organization is maintained, and after 4 short-days, the shoot apex possesses a mantle of 3–4 layers of cells because there are few periclinal divisions in the cells of the outer corpus. The cells of the mantle stain uniformly and are more chromatic than those of the underlying tissue. Mitotic activity was recorded in the upper 40-μ segment of the apex. In the vegetative apex, mitotic activity is greater in the lower portion of the segment. Following photoinduction, mitoses increase throughout the apex until a maximum is reached on the 4th day. Also, the plastochronic interval decreases after photoinduction. Nucleoli of cells of the corpus enlarge following induction until all nucleoli of the apex are nearly equal. Included in the paper are discussions of the general morphological differences between vegetative and flowering shoots.     

HISTOCHEMICAL AND AUTORADIOGRAPHIC STUDIES OF FLORAL INDUCTION IN CHENOPODIUM ALBUM

Gifford , Ernest M., Jr. , and Herbert B. Tepper . (U. California, Davis.) Histochemical and autoradiographic studies of floral induction in Chenopodium album. Amer. Jour. Bot. 49(7): 706–714. Illus. 1962.—Chenopodium album was induced to flower using short-day photoperiods. Changes in the chondriome, starch, total protein, ribonucleic acid (RNA), deoxyribonucleic acid (DNA), and histone distribution in cells of vegetative and inflorescence shoot apices were studied. The distal cells of the vegetative apex (especially the axial tunica cells) possess larger nucleoli and vacuoles, less granular mitochondria, and more differentiated plastids than do other cells of the apex; the distal cells stain lightly with dyes that indicate the presence of DNA and histone. RNA is distributed relatively uniformly in the shoot apex; the cells at sites of leaf initiation and young leaf primordia contain slightly higher concentrations of RNA than the axial cells of the shoot apex. Protein is uniformly distributed throughout the vegetative as well as the inflorescence apex. Upon induction, the chemical and morphological differences between cells in the shoot apex gradually disappear. RNA concentration of cells in the apex increases, reaching a maximum after 4 inductive cycles. Protein concentration of cells also increases, but this increase lags behind that of RNA.     

SHOOT ORGANIZATION IN THE FILICALES: THE PROMERISTEM

The classical pteridophyte apical cell theory of meristem organization is not flexible or realistic enough to encompass the variation encountered in a comprehensive anatomical survey of fern meristems. The meristems of 28 taxa of ferns (both eu- and leptosporangiate) were studied. This analysis has led to the formulation of the concept of a promeristem composed of two zones: the surface and subsurface initials. This concept is flexible and sufficient to describe the variation encountered in the ferns as a group. No differences in promeristem organization were observed in plants with various rhizome morphology (e.g., upright and radial; prostrate and dorsiventral). Marking experiments, performed on living surface cells of fern promeristems, correlate with anatomical observations of division planes in the promeristem and indicate that the central, surface initials are not quiescent. Feulgen determinations indicate that Nephrolepis stolons have no endomitotically polyploid cells in the promeristem. Additional work is needed before a generalization can be made concerning ploidy levels in a more typical fern apex. Ferns, in general, have a zoned promeristem which is parallel to the zonation described for higher vascular plants.     

ONTOGENETIC AND HISTOCHEMICAL CHANGES IN THE VEGETATIVE SHOOT TIP OF CHENOPODIUM ALBUM

Gifford , Ernest M., Jr ., and Herbert B. Tepper . (U. California, Davis.) Ontogenetic and histochemical changes in the vegetative shoot tip of Chenopodium album. Amer. Jour. Bot. 49(8): 902–911. Illus. 1962.—A distinct central zone is present in apices of young, rapidly growing seedlings of Chenopodium album. This zone is clearly evident when the plane of section includes developing leaf buttresses and/or newly formed leaf primordia, but is not visible if the cells associated with these developing foliar organs are not in the plane of section. Cells in the central zone contain markedly lower concentrations of RNA than those of the peripheral zone. Differences in DNA, SH-protein, total protein, and histone concentration of cells in the apex parallel the zonal distribution of RNA, but are much less obvious. As the plant ages and the rate of leaf production decreases, the zonal distribution of the above-mentioned compounds becomes less and less obvious. Cells at sites of leaf initiation in older seedlings still have slightly higher RNA concentrations, but a central zone such as is visible in the young seedlings is no longer evident. The axial tunica cells do, however, contain noticeably lower DNA and histone concentrations than the remaining cells of the shoot apex. Changes occur in the size of nucleoli during ontogeny and the possible relationship between nucleolar size and modifying factors is discussed.     

ONTOGENY OF THE SHOOT APEX OF SEEDLINGS OF PINUS PONDEROSA

Dormant seeds of ponderosa pine (Pinus ponderosa) were stratified and then planted in the greenhouse. Changes occurring in the shoot apex during germination and growth of the seedlings were observed and are described. Cell divisions in germinating embryos are first noted on the flanks of the apex near the cotyledons. From these loci, mitotic reactivation proceeds up the flanks of the apex and inward toward cells of the rib meristem. Cells are dividing actively in all regions of the shoot apex of seedlings 12 days after planting, but no cytohistological zonation is evident due to differences in staining intensity and in cell and nuclear size. In 64-day-old seedlings the mitotic rate is reduced and the characteristic zonation of pine apices begins to appear. Zonation is even more evident in older seedlings. These observations are discussed in relation to the concept of initials and to theories concerning factors which regulate growth in the shoot apex.     

DETERMINATE BRANCH DEVELOPMENT IN ALSTONIA SCHOLARIS (APOCYNACEAE): THE PLAGIOTROPIC MODULE

The development of the vegetative lateral branches in Alstonia scholaris (L.) R. Br. was examined. The overall architecture conforms to Prévost's model and the branches are sympodial complexes of plagiotropic modules. Each module consists of two whorls of 6–11 foliage leaves and a whorl of four scale leaves. The apex is parenchymatized just distal to the scale leaves. Renewal branches grow from buds in the axils of the scale leaves. These large buds are initiated simultaneously with the scale leaves and “use up” a large portion of the original apex. Parenchymatization of the central region of the apex occurs after a period of lateral growth and development that separates and vascularizes the renewal branches. Branch extension occurs sympodially by substitution. More typical buds develop in the axils of the foliage leaves but grow out only in response to injury or pruning. They are smaller than the renewal buds and, unlike them, are delimited by a shell zone during early development.     

Shoot apex,leaf development and unifacial tip inAgave wightii drumm. et prain (agavaceae)

The shoot apex has one tunica layer enclosing a mass of corpus which is differentiated cytohistologically into central mother cell zone, flank zone, rib zone and a ‘cambium-like’ zone. Occurrence of ‘cambium-like’ zone during minimal phase is considered as an expression of nodal region. Agave wightii shows spirodistichous arrangement of leaves which have an expanded photosynthetic surface with a reduced unifacial tip. Leaves are initiated by periclinal divisions in the second layer. Vertical growth in the leaves is by subapical initials and lateral growth is by marginal and submarginal initials in their early stages of development. The unifacial tip is formed by the extension of adaxial meristematic activity. The derivatives thus formed are pushed to the abaxial side of the primordiuj. Hence the unifacial part of the leaf is regarded as equivalent to a phyllode.     

THE DEVELOPMENTAL ANATOMY OF LONG-BRANCH TERMINAL BUDS OF PINUS BANKSIANA

A study of the composition of long-branch terminal buds (LBTB) of Pinus banksiana Lamb. and the yearly periodicity associated with their formation, development, and elongation was undertaken. Each LBTB has lateral bud zones and zones of cataphylls lacking axillary buds. When present, staminate cone primordia differentiate from the lowest lateral buds in the lowest lateral bud zone of the LBTB. Ovulate cone primordia and lateral long-branch buds can differentiate from the upper lateral buds in any lateral bud zone. When both types of buds are present, lateral long-branch buds are uppermost. Dwarf-branch buds occur in all lateral bud zones. During spring LBTB internodes elongate, new cataphylls are initiated, dwarf branches elongate, needles form and elongate, pollen forms and is released, and ovulate cones are pollinated. During summer buds form in the axils of the newly formed cataphylls. By early fall the new LBTB are in overwintering condition and the four types of lateral buds are discernable. The cytohistological zonation of the LBTB shoot apex is similar to that of more than 20 other conifer species. Cells in shoot apices of pine are usually arranged in distinct zones: apical initials, subapical initials, central meristem, and peripheral meristem. Periclinal divisions occur in the surface cells of the apex; therefore no tunica is present. At any given time, shoot apex volume and shape vary among LBTB in various positions on a tree. In any one LBTB on a tree, shoot apex shape changes from a low dome during spring to a high dome during summer to an intermediate shape through fall and winter.     

THE ROOT APEX OF LINUM

Several developmental stages of primary roots of 13 species of Linum were studied. The basic pattern of root apex organization in three species consisted of a single tier of cortical initials. Ten species had a basic pattern of two tiers of cortical initials. Variations, manifested by either an increase or a decrease in the tiers of cortical initials, were observed in some roots in those species that had a basic pattern of two tiers of cortical initials. Although these variations were interpreted as being ontogenetic, there was no total reorganization of the root apex. There was anatomical and cytological evidence that a quiescent center is established in Linum roots.     

Occurrence of a Putative Precursor to the Amyloplast,the “Amyloplast Initial” in Developing Stolons of the Potato

Cells at the apical part of developing stolons of the potato (Solatium tuberosum L. cv. Norin 1) were analyzed for the occurrence of putative precursors to amyloplasts, designated “amyloplast initial.” Ultrastructural studies showed that the cells contained the expected novel organelle. It was about 1 μm in diameter, devoid of thylakoid membranes, and was stained to a similar extent as the stroma of amyloplasts by uranyl acetate and lead citrate. Formation of thylakoid membranes and starch granules takes place at an early stage of development of these initials when they are just a few μm in diameter. At this stage, proliferation of the initials takes places by division at random sites.     

Structure and Cytogenesis of the Shoot Apex of Syringa oblata var. affinis Lingelsh

The structure, growth and mitotic activity of 211 shoot apices of developing sprouts of Syringa oblata var. affinis Lingelsh. in longitudinal sections and 67 in transverse sections have been studied with the view to understanding the nature of zonation patterns and cytogenesis of the apical meristems during a double plastochron. The external morphology and the anatomical structure of the apices in 4 plastochronic stage-early, middle, late Ⅰ and late Ⅱ stages are described. In the shoot apices examined, especially those at late plastochronic stage, the following zones may be delimited: Zone of tunica initials, zone of corpus initials, peripheral zone and zone of rib meristem. The location and orientation of mitotic figures observed in longisections of the apices in 4 plastochronic stages are plotted in diagrams and the mitotic frequency has been calculated. Information obtained from these investigations reveals that the tunica and corpus inititals constitute an active region of the apex, but their mitotic activity changes periodically within the double plastochron. In the middle plastochronic stage when the apex is at its minimal area and the cells of peripheral zone and rib meristem zone have been completely transformed into constituent parts of foliar primordia and the subjacent tissues of the stem and the pith mother cells respectively, the mitotic frequency of the initials is at its maximium and its intensity of mitotic activity is not much lower than that of any other meristematic zone at any stage. When the apical dome is reformed by the activity of these initials in late plastochronic stages, the mitotic frequency of the initials gradually drops and the region of high mitotic frequency shifts to the flank of the apex, the peripheral zone. Anticlinal divisions are predominant in this zone. On the other hand, those cells directly left behind by the corpus initials, which constitute the rib meristem, are vacuolated and marked by the pre- dominance of transverse divisions. Thus the entire zonation pattern reappears. In the next early plastochronic stage, the mitotic frequency of the tunica and corpus initials drops to its mimimium, but other regions of the apex still maintain a high mitotic frequency. It may be concluded that the tunica and corpus initials form a cytogenerative center of the shoot, and the cytohistological zonation is actually a result of the fact that different regions of apical meristems are different in mitotic activety, different in state of cell differentiation and different in their function in morphogenesis.     

INFLORESCENCE DEVELOPMENT IN BRASSICA CAMPESTRIS L.

Vegetative seedlings of the Ceres strain Brassica campestris L., a quantitative, long-day plant, were induced to flower by exposure to a 16-hr, long-day cycle. Cytohistological and cytohistochemical changes associated with inflorescence development were examined. Developing shoot apices were classified in vegetative, transitional, and reproductive stages. The vegetative apex possessed a biseriate tunica, central zone, peripheral zone and pith-rib meristem. The transitional stage at 48 hr was marked by an increase in size and by a stratification of the upper cell layers of the shoot apex with a concurrent decrease of apical cytohistochemical zonation. The reproductive stage was initiated at 58 hr by periclinal cell divisions in the 3rd and 4th cell layers of the flank region. Cytohistochemical zonation in the vegetative apical meristem was restored in the floral apex. An “intermediate developmental” phase was not observed between the vegetative and reproductive stage.     

ROOT APEX STRUCTURE IN EPHEDRA MONOSPERMA AND EPHEDRA CHILENSIS (EPHEDRACEAE)

The root meristem of E. monosperma and E. chilensis possesses a central group of distinctive, large cells. These cells have large nuclei with scattered heterochromatin, proplastids with no starch, small vacuoles, mitochondria, few dictyosomes and endoplasmic reticulum cisternae, and lipid deposits. Over a 24 hr labelling period, the large cells fail to incorporate ³H-thymidine, whereas cells both distal and proximal to this region do. A quiescent center which includes these large cells is present therefore. Both species have an extensive root cap, the length being contributed by mitoses in many tiers of cells distal to the quiescent center. The root cap consists of a columella and peripheral regions. Distinctive amyloplasts, an increase in the number of endoplasmic reticulum cisternae and dictyosomes, large vacuoles, and lipid deposits are characteristic of differentiated columella cells. Peripheral cells elongate, lose most of their starch, and are eventually sloughed from the root.     

DEVELOPMENTAL ANATOMY IN ROOTS OF IPOMOEA PURPUREA. I. RADICLE AND PRIMARY ROOT

The developmental anatomy of the primary root of Ipomoea purpurea was studied at several growth stages, beginning with the radicle. The radicle is generally composed of three superimposed tiers of initials, which produce the vascular cylinder, cortex, and columella; and a peripheral band of lateral rootcap-epidermal initials. The radicular cortex contains 16–19 immature laticifers; none of the tissue regions in the radicle contains mature cells. Following germination and during the first 2–3 cm growth of the primary root the apical meristem and its derivative tissues undergo a series of modifications. Root apical diameter decreases as cells in lateral portions of the rootcap elongate; meanwhile, the columella enlarges vertically. The relationship between cortical and columellar initials changes as fewer mitoses occur in the former while the latter remain active. In longer roots the columellar initials are directly in contact with the vascular initials. Cortical size diminishes during early root growth as cortical laticifers and their associated cells cease to be produced by the outer cortical initials and ground meristem. Early procambium, at the level of vascular pattern initiation, decreases in diameter by cellular reorientation, and the vascular cylinder decreases in overall diameter although the tetrarch pattern remains unchanged.