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.     

THE EFFECTS ON CHRONIC GAMMA RADIATION ON THE INTERNAL APICAL CONFIGURATIONS OF THE VEGETATIVE SHOOT APEX OF COLEUS BLUMEI

Clonal plants of a bushy variety ‘Trailing Queen’ of Coleus blumei Benth. were exposed to 600–700 R per day for 28 days. Shoot apices of control and irradiated plants were studied microscopically. The normal vegetative shoot apex has two tunica layers and a corpus which is partly layered and partly composed of randomly divided cells. Five cytohistological zones were observed, including a cup-shaped zone, imposed upon the tunica-corpus relationships. Response to gamma radiation resulted in abnormal internal apical configurations. All irradiated shoots was found to have the same internal configurations. Only a single tunica layer broken by occasional periclinal divisions was observed. The cytohistological organization of irradiated apices was different from that seen in the normal shoot tips in that two zones were found. Evidence of a radiosensitivity gradient within the vegetative shoot apex was also observed. The possible significance of the results is discussed.     

EARLY HISTOGENESIS AND SEMIQUANTITATIVE HISTOCHEMISTRY OF LEAF INITIATION IN TRITICUM AESTIVUM

The shoot apex of Triticum aestivum cv. Ramona 50 was investigated histologically to describe cell lineages and events during leaf initiation. During histogenesis three periclinal divisions occurred in the first apical layer, with one or two divisions in the second apical layer. This sequence of cell divisions initially occurred in one region and spread laterally in both directions to encircle the meristem. Cells of the third apical layer were not involved in leaf histogenesis. Initially, young leaf primordia were produced from daughter cells of periclinal divisions in the two outer apical layers. Nuclear contents of protein, histone, and RNA in the shoot apex were evaluated as ratios to DNA by means of semiquantitative histochemistry. Daughter cells of periclinal divisions in the outer apical layer which produced the leaf primordia had higher histone/DNA ratios than cells of the remaining meristem. However, protein/DNA and RNA/DNA ratios were similar in both regions. Leaf initial cells had a higher ³H-thymidine labeling index, a higher RNA synthesis rate, and smaller nuclear volumes than cells of the residual apical meristem.     

FLEXIBILITY IN ONTOGENY AS SHOWN BY THE CONTRIBUTION OF THE SHOOT APICAL LAYERS TO LEAVES OF PERICLINAL CHIMERAS

The development of leaves on apically stable, periclinal chimeras was studied in a number of dicot genera. The mutant cell layers of the shoot apex and the tissues derived from them were as active developmentally as the normal layers. Ontogeny was the same in these chimeras as in nonchimeras, and growth of their leaves can be outlined as follows. Formation of the buttress, the axis, and the lamina of simple dicot leaves were independent events. In each the first growth included derivatives of the apical layers, usually three in number, found in the apex of the shoot and the lateral buds. Most cell divisions in the outer layers (L-I and L-II) were anticlinal relative to the new structures. Therefore, in the proximal regions of the buttress, axis (petiole and midrib), and lamina, the derivative cells of L-I and L-II were usually present in single layers. The rest of the internal tissue was from L-III. As formation of the axis and the lamina proceeded, derivatives of L-II replaced L-III internally in the distal and marginal regions leaving cells of L-III behind. Both the determinate growth of leaves and the pattern of cell divisions at and near the leading edges of growth meant that no cells in the leaf were comparable to the initial cells of the shoot apex. As the lamina extended, there were extensive intercalary cell divisions, both anticlinal and periclinal, so that in any given region of a leaf the layers of internal cells were from either L-II or L-III. At any point along the axis, L-III participated or did not participate in laminar extension. At any given stage in laminar growth either of two sister cells in any internal layer divided either a few times or extensively. The extreme variability in direction and frequency of cell division during leaf development was under an overriding genetic control, which resulted in the normal or typical size, shape and thickness of leaves.     

Analysis of shoot apical organization in six species of the Cupressaceae based on chimeric behavior

Six species of the Cupressaceae, the variegated Leyland cypress (Cupressocyparis leylandii 'Silver Dust'), savin (Juniperus sabina variegata Laws), davurian juniper (Juniperus davurica 'expansa variegata'), California incense cedar (Calocedrus decurrens 'Aureovariegata'), the American arbor vitae (Thuja occidentalis 'lutae zebrina' Kent), and the sawara false cypress (Chamaecyparis pisifera 'nana aureovariegata') were examined for the behavior of albino-green shoot chimeras. The fate of the variegations in these six plants is the same in two important respects. First, the majority (89%) of sprays with an original sector become completely white. Second, sectorial branch sprays of the original sectorial sprays become either completely green or white in a 1?:?1 ratio. Based on the first finding it is concluded that there is one rather than the two to four apical initials in the shoot apex, as generally postulated. This single apical initial, actually an apical cell lineage, residing in the tunica layer can both form the leaf epidermis and by rare periclinal divisions form sectorial chimeras. The second finding is that there is no selection advantage of either type, a feature also postulated by others.     

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.     

ONTOGENY OF THE INFLORESCENCE OF SAURURUS CERNUUS (SAURURACEAE)

The spicate inflorescence of Saururus cernuus L. (Saururaceae) results from the activity of an inflorescence apical meristem which produces 200–300 primordia in acropetal succession. The inflorescence apex arises by conversion of the terminal vegetative apex. During transition the apical meristem increases greatly in height and width and changes its cellular configuration from one of tunica-corpus to one of mantle (with two tunica layers) and core. Primordia are initiated by periclinal divisions in the subsurface layer. These are “common” primordia, each of which subsequently divides to produce a floral apex above and a bract primordium below. The bract later elongates so that the flower appears borne on the bract. All common primordia are formed by the time the inflorescence is about 4.4 mm long; the apical meristem ceases activity at this stage. As cessation approaches, cell divisions become rare in the apical meristem, and height and width of the meristem above the primordia diminish, as primordia continue to be initiated on the flanks. Cell differentiation proceeds acropetally into the apical meristem and reaches the summital tunica layers last of all. Solitary bracts are initiated just before apical cessation, but no imperfect or ebracteate flowers are produced in Saururus. The final event of meristem activity is hair formation by individual cells of the tunica at the summit, a feature not previously reported for apical meristems.     

FOLIAR ONTOGENY AND HISTOGENESIS IN MAGNOLIA GRANDIFLORA L. I. APICAL ORGANIZATION AND EARLY DEVELOPMENT

Foliar ontogeny of Magnolia grandiflora was studied to elucidate possible unique features of evergreen leaves and their development. The apex of Magnolia grandiflora is composed of a biseriate or triseriate tunica overlying a central initial zone, a peripheral zone and a pith rib meristem. Leaf primordia are initiated by periclinal divisions on the apical flank of the tunica in its second layer. This initiation and expansion is seasonal just as in related deciduous magnolias. Following leaf initiation, a foliar buttress is formed and the leaf base gradually extends around the apex. As growth continues, separation of the leaf blade primordium from the stipule proceeds by intensified anticlinal divisions in the surface and subsurface layers near the base. Marginal growth begins in the blade primordium when it reaches approximately 200 μm in height and results in the formation of two wing-like extensions, the lamina. This young blade remains in a conduplicately folded position next to the stipule until bud break.     

FLORAL DEVELOPMENT OF POTAMOGETON RICHARDSONII

The inception and development of the sterile floral appendages of Potamogeton richardsonii have been re-investigated with a refined dissection technique (Sattler, 1968) and improved microtechnical methods (Feder and O'Brien, 1968). The results obtained by Sattler (1965) are confirmed, i.e., the sterile appendages are initiated at the flanks of the floral apex before the stamen primordia are formed. Consequently, they may be homologized with tepals or perianth members, although in the mature flower they are inserted at the stamen connective, due to growth between and at the base of each developing tepal and stamen. Each carpel arises as a radial primordium which becomes peltate immediately after its inception. One ovule primordium is initiated at the cross-zone. The stigma becomes bilobed. A slight outgrowth develops at the abaxial side of the style. The floral apex has a two-layered tunica. The primordia of the tepals, carpels, and ovules arise by periclinal divisions in the second tunica layer, whereas the stamen primordia are initiated by periclinal divisions in the corpus and second tunica layer. Variation in floral pattern, especially with regard to the number of appendages, has been observed in flowers near the tip of the inflorescence axis.     

Some Aspects of Floral Histogenesis in Bajra (Pennisetum typhoides S. & H.)

The young reproductive apex in Bajra (Pennisetum typhoides S.& H.) possesses a biseriate tunica and a massive corpus.The cells of three or four peripheral layers and six to eightlayers at the summit of the apex are eumeristematic. The centralregion consists of elongated, highly vacuolated, and lightlystained cells arranged in files. The initiation of the spikeletbud is by periclinal divisions first in the corpus and laterin T₂ cells. Similarly the longer bristle or the extension ofthe fascicular axis develops from the corpus and T₂ cells. Theother bristles develop from the tunica layers. The chaff membersare initiated and develop like a leaf. The development of thestamen resembles that of a spikelet or an axillary bud. Thedevelopment of the carpel is similar to that of the leaf primordium.The origin and development of the male flower is like that ofan axillary bud.     

SOMATIC GENETIC ANALYSIS OF THE APICAL LAYERS OF CHIMERAL SPORTS IN CHRYSANTHEMUM BY EXPERIMENTAL PRODUCTION OF ADVENTITIOUS SHOOTS

Many commercial chrysanthemum cultivars display unusual somatic variability. The ‘Indianapolis’ family of chrysanthemum sports was analyzed for the genetic potential for color of each of the three layers in the apical meristem of their shoots. Populations of each cultivar were grown and sectors and off-color plants recorded. The location of the pigment within cells and between tissues was determined by microscopic examination of free-hand sections of fresh petals. Adventitious buds were forced from the stems of each cultivar by excising all normal lateral buds. These observations, showed 12 of the 16 ‘Indianapolis’ cultivars to be periclinal chimeras. Adventitious shoots often originated from two or more cells, derived from at least two different apical layers, and thus were themselves periclinal chimeras. While somatic mutation is the ultimate source of the variability in ‘Indianapolis’ chrysanthemums, the most frequent type of sporting resulted from the loss in mitosis of a chromosome carrying a supressor for the formation of yellow chromoplasts, giving a yellow sector or shoot. Sectors resulting from rearrangement of layers in the periclinal chimeras were less frequent than the sectors from chromosome loss.     

Arrangement of cortical microtubules in the shoot apex of Vinca major L.

The arrangements of cortical microtubules (MTs) and of cellulose microfibrils in the median longitudinal cryosections of the vegetative shoot apex of Vinca major L., were examined by immunofluorescence microscopy and polarizing microscopy, respectively. The arrangement of MTs was different in the various regions of the apex: the MTs tended to be arranged anticlinally in tunica cells, randomly in corpus cells, and transversely in cells of the rib meristem. However, in the inner layers of the tunica in the flank region of the apex, cells with periclinal, oblique or random arrangements of MTs were also observed. In leaf primordia, MTs were arranged anticlinally in cells of the superficial layers and almost randomly in the inner cells. Polarizing microscopy of cell walls showed that the arrangement of cellulose microfibrils was anticlinal in tunica cells, random in corpus cells, and transverse in cells of the rib meristem; thus, the patterns of arrangement of microfibrils were the same as those of MTs in the respective regions. These results indicate that the different patterns of arrangement of MTs and microfibrils result in specific patterns of expansion in the three regions. These differences may be necessary to maintain the organization of the tissues in the shoot apex.Abbreviations MT(s) microtubule(s) - lp length of the youngest leaf primordium     

Marking cell layers with spectinomycin provides a new tool for monitoring cell fate during leaf development

Spectinomycin, an inhibitor of plastid protein synthesis, can be used to mark specific cell layers in the shoot meristem of Brassica napus. Pale yellow-green (YG) plants resulting from spectinomycin-treatment can be propagated indefinitely in vitro. Microscopic examination showed that YG-plants result from inactivation of plastids in the L2 and L3 layers and are composed of a pale green epidermis covering a white mesophyll layer. Epidermal cells of YG and normal green plants are similar and contain 10-20 small pale green plastids. YG plants are equivalent to periclinal chimeras with the important distinction that there is no genotypic difference between the white and green cell layers. Periclinal divisions of epidermal cells take place at all stages of leaf development to produce invaginations of green mesophyll located in sectors of widely varying sizes. A periclinal division rate of 1 in 3000-4000 anticlinal divisions for the adaxial epidermis, was 2-3-fold higher than that estimated for the abaxial epidermis. Analysis of white and green mesophyll showed that chloroplasts are essential for palisade cell differentiation and this requirement is cell-autonomous. Stable marking of cell lineages with spectinomycin is simple, rapid and reveals the requirement for functional plastids in cellular differentiation.     

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.     

Genetic analysis of instability inPetunia hybrida

Summary InPetunia hybrida frequent mutations of unstable alleles give rise to different types of periclinal chimeras. If genes expressed in the epidermis, such as the geneAn1 for flower colour, are concerned, mutations in the dermal layer of the shoot apex will result in changes in the phenotype but not in the offspring. Mutations in the subdermal layer will not lead to an altered phenotype, but to changes in the sporogenous tissues and, thus, to deviating segregations in progenies. Therefore, in crossing experiments with such an unstable mutant, it is always necessary to take the possibility into account that the plant may be a chimera, so as to prevent an incorrect interpretation of the recorded segregational ratios. Mutations of unstable alleles expressed in the mesophyll, such as geneYg3 for leaf colour, also give rise to chimeras. In such instances, however, a change in phenotype always involves a change in segregational ratios as well, since both the mesophyll and the sporogenous tissues are derived from the subdermal layer of the shoot apex.     

THE SHOOT APEX AND EARLY LEAF DEVELOPMENT IN CLEMATIS

Tepfer , Sanford S. (U. Oregon, Eugene.) The shoot apex and early leaf development in Clematis . Amer. Jour. Bot. 47 (8): 655–664. Illus. 1960.—The high-domed shoot apex comprises a 2-layered tunica and shallow corpus. The rib meristem at times extends to within 5 cells of the summit. The cells of tunica and corpus are uniform cytologically, distinguishable only by the orientation of division planes. No zonation is visible within the corpus. No evidence was found of the existence of a méristème d'attente; mitotic figures appear frequently in the central region of the tunica and corpus. Decussately arranged leaf primordia arise high on the flanks of the apex. Periclinal divisions in the inner tunica and outermost corpus layers mark the site of initiation. Details of the growth and early differentiation of the leaf primordia follow the usual pattern of buttress formation, growth through apical and subapical initials. Apical growth continues beyond the early stages of leaf ontogeny; the blade-forming marginal meristems do not appear until after leaflet primordia are formed. There are 5 primary leaflets, pinnately arranged. Each leaflet is 3- to 5-lobed. In primordium P₃ expansion of the adaxial-lateral margins occurs at the base, but not above. This marks the upper limits of the basal pair of lateral leaflets. In P₄ the upper limits of the upper lateral leaflets become demarcated in similar fashion.     

INFLORESCENCE AND FLOWER DEVELOPMENT IN THE PIPERACEAE. I. PEPEROMIA

Flowers of Peperomia species are the simplest structurally of any of the members of the Piperaceae. The spicate inflorescences form terminally and in axillary position; in each, the apex first is zonate in configuration with a two-layered tunica while 3-4 leaves are initiated. Later, when the inflorescence apical meristem begins bract initiation, the biseriate tunica persists, but zonal distinctions diminish and the apex can be described in terms of a simple tunicacorpus configuration. The inflorescence apex aborts after producing 30-40 bracts in acropetal succession an abscission layer forms across the base of the apex, and the meristem dries and drops off. Bracts are produced by periclinal divisions in T₂ (and occasionally also in the third layer as well); the later-formed floral apices arise by periclinal divisions in T₂ and the third layer. Each floral apex is at first a long transverse ridge in the axil, perpendicular to the long axis of the inflorescence. This establishes bilateral symmetry in the flower, which persists throughout subsequent growth. The floral meristem becomes saddle-shaped, and two stamen primordia are delimited, one at either end and lower than the central floral apex. A solitary carpel is initiated abaxially, and soon forms a circular rim which heightens as a tube with an apical pore. Within the open carpel, a solitary ovule is initiated from the entire remains of the floral apical meristem; it, hence, is terminal in the flower, and its placentation is basal. Carpellary closure in P. metallica results from accelerated growth of the abaxial lip, and the two margins become appressed. Species differ greatly as to whether the abaxial or the adaxial lobe predominates in late stages of carpel development. In P. metallica, the receptive portion of the stigma forms from the shorter lobe which is overtopped. Stigmatoid tissue forms internal to the receptive stigma. The prevailing bilateral floral symmetry, absence of a perianth, and the spicate inflorescence are features which distinguish Peperomia (and Piperaceae) from the magnolialian line of angiosperms.     

CHLOROPHYLL-DEFICIENT CELL LINES WHICH ARE GENETICALLY UNCHARACTERIZED CAN BE INAPPROPRIATE FOR USE AS PHENOTYPIC MARKERS IN DEVELOPMENTAL STUDIES

Periclinal chloroplast chimeras are genetic mosaics which possess shoot apices composed of one or more chlorophyll-deficient histogens and can exist as a series of arrangements of normal and mutant layers (A-B-B, A-B-A, etc.). Three periclinal chimeral cultivars of Sansevieria trifasciata L., each of which possesses normal green cell layer(s) but a genetically different chlorophyll-deficient cell layer(s), were utilized to study the effect of genotype on the ability of the cell layers of leaf cuttings and of cultured leaf tissue to regenerate shoots. The epidermis and LI derivatives were apparently incapable of shoot regeneration via leaf cutting, yet in two cultivars produced some shoots in vitro. In two of the cultivars, the chlorophyll-deficient cells never produced shoots. In the third, the capability of chlorophyll-deficient cell layers to produce shoots was less in vitro than in vivo, indicating that when determining morphogenic potential, direct comparisons between in vitro and in vivo systems may not be valid. Results also demonstrate that because genetically different albino cell layers can differ in their morphogenic response, utilizing a series of periclinal chimeras is useful only if the series is composed of the same two genotypes.     

Arrangement of cell layers in the shoot apical meristems of periclinal chimeras influences cell fate

Utilizing a complete set of six periclinal graft chimeras composed of Nicotiana tabacum and Nicotiana glauca (TGG, GTT, TTG, GGT, TGT, and GTG), the fate of the three apical cell layers in both vegetative and reproductive organs has been traced. An analysis of leaf phenotype indicated that only rarely did deviations from expected cell lineage occur and in only TTG did such deviations originate in the shoot apical meristem rather than during leaf development. In most plants that possess a stratified shoot apical meristem, gametes are derived from the second apical layer (L2). A phenotypic and/or DNA analysis of seed progeny following reciprocal crosses between all chimeras and their component species indicated that pollen and eggs were sometimes derived from non-L2 lineage in all but one periclinal chimera. There was no evidence for non-L2-derived gametes in 95 crosses where GTT was a parent whereas 40 of 104 crosses with TTG as a parent yielded some offspring that resulted from non-L2-derived gametes. Of these 40 cases, non-L2-derived pollen grains were responsible 39 times while non-L2-derived eggs were responsible just once. Therefore, the occurrence of non-L2-derived gametes was not random. The disruption of ‘normal’ lineage patterns was dependent on the specific arrangement of genetically dissimilar tissue layers in the shoot apices of the chimeras and was different for different organs.     

ANATOMY AND ASPECTS OF DEVELOPMENT OF THE STAMINATE INFLORESCENCES AND FLORETS OF SEVEN SPECIES OF ALLOCASUARINA (CASUARINACEAE)

The morphology, ontogeny, and vascular anatomy of the staminate inflorescences and florets of seven species of Allocasuarina are described. The generally terminal but open-ended inflorescences occur on monoecious or staminate dioecious trees and consist of whorls of bracts, each subtending a sessile axillary floret. Each floret consists of one terminal stamen with a bilobed, tetrasporangiate anther enclosed typically by cuculliform appendages, commonly considered bracteoles, an inner median pair and an outer lateral pair. The mature stamen is exerted, the anther is basifixed and is extrorsely dehiscent. In early development of a male inflorescence very little internodal elongation occurs and enclosing cataphylls appear. The inflorescence apex is a low dome with a uniseriate tunica and a small group of central corpus cells. Bract primordia are initiated by periclinal divisions of C₁ followed by further divisions of the corpus and anticlinal divisions in the tunica. The bracts are epinastic and become gamophyllous except apically by cell divisions in both sides of each primordium. Stomata are restricted to the axis furrows and the abaxial tips of the bracts. The axillary florets arise in acropetal succession initiated by periclinal divisions in C₁ accompanied by anticlinal divisions in the tunica. The lateral floral appendages are also initiated by C₁ followed by anticlinal divisions in the tunica. They become adnate basally later with the subtending bract. The median sterile appendages are initiated in a manner similar to the initiation of the outer appendages. The stamen is initiated by divisions in the outer layers of the corpus and in the tunica, and then develops first by apical growth followed by intercalary growth. The vascular system of the inflorescence is identical to that of the vegetative stem. Each floret is supplied by a single bundle that has its source in a branch from each of the two traces supplying a bract. Six bundles arise from the floral bundle; four of these terminate in the base of the stamen and two form an amphicribal bundle that supplies the anther. Pollen is binucleate, 3- to 7-porate. The exine is tegillate.