CHANGES IN MITOTIC ACTIVITY AND CELL SIZE IN THE APICAL MERISTEM OF HELIANTHUS ANNUUS L. DURING THE TRANSITION TO FLOWERING

Cellular changes in the shoot apical meristem of Helianthus annuus L. have been investigated in relation to its progress towards flowering. During the strictly vegetative phase, lasting for 6–7 days from sowing, mitotic divisions were confined to the peripheral zone, while the central mother cells zone proper, together with the distal cells positioned above the mother zone and also the central portion of the tunica, were relatively quiescent. Mitotic activity increased in the distal cells zone on day 8 and reached the level of that in the peripheral zone by day 12. This was accompanied by an enlargement of this zone and the consequent recession of the mother zone away from the central tunica. At the same time there was a substantial increase in the amount of cytoplasm in the cells of the central tunica. Mitotic activity in the central tunica began on day 12 and reached a peak on day 16. This zone then lost its distinct entity and was replaced by a uniform dome-shaped meristematic layer that became apparent by day 16. The cells of the mother zone remained quiescent during the transition period.     

VARIATION OF NUCLEAR DNA CONTENT IN HELIANTHUS ANNUUS (ASTERACEAE)

Nuclear 2C DNA content was determined by laser flow cytometry for 13 diploid (2n = 34) lines (cultivated varieties and inbred lines) of Helianthus annuus. Mean DNA amount of second leaf nuclei varied from 6.01 to 7.95 pg (32%) among lines. Mean DNA content varied up to 19% within lines. Variability in mean DNA content exceeding 27% and 48% was detected among leaves from different nodes of plants of the open-pollinated variety, Californicus, and the inbred line, RHA 299, respectively. The root tip and shoot tip nuclei of H. annuus have been reported to consist of a mixture of aneuploid (17 to 33 chromosomes) and diploid (34 chromosomes) cells, a condition called aneusomaty. Chromosome counts of root tips and an analysis of the distribution of DNA content of large numbers of nuclei from leaves indicate that aneusomaty either does not occur, or is not common, among the lines investigated. The intraspecific, intraline, and intraplant variation in DNA content in H. annuus support the concept that a sizable portion of a plant genome is unstable and subject to rapid changes in DNA amount.     

HISTOCHEMICAL STUDY OF ENZYME ACTIVITY IN THE SHOOT APICAL MERISTEM OF BRASSICA CAMPESTRIS DURING TRANSITION TO FLOWERING. IV. GLUCOSE-6-PHOSPHATASE

Glucose-6-phosphatase (G6P) activity was determined in fresh-frozen, cryostat sections in the shoot apical meristem of Brassica campestris L. Enzymatic activity was differentially distributed in a zonate pattern in the vegetative meristem, but not in the transition and floral meristem. Vegetative apices showed a heterogenous localization with the highest activity in the central zone and the pith-rib meristem zone. At the early transition stage of development, G6P activity in the peripheral zone increased slightly. At the late transitional (prefloral) stage, G6P activity was not localized within the peripheral zone in island-like areas of activity. This is the first demonstration of G6P in shoot apical meristem at the vegetative, transition, and floral stage. The results indicate that G6P activity 1) is an accompanying event of evocation, but 2) does not mark incipient floral primordia. G6P may play an important role in the maintenance of glucose-6-phosphate homeostasis in an evoked shoot apical meristem.     

NUCLEAR DNA CONTENT VARIATION IN HELIANTHUS (ASTERACEAE)

Nuclear 2C DNA content was microspectrophotometrically determined for 19 diploid (2n=34) species of Helianthus L. DNA amount varied over a fourfold range. Helianthus neglectus had the lowest (53.40 ± 0.51 Feulgen absorbancy units [FAU]) and H. agrestis the highest (216.30 ± 2.30 FAU) DNA content. Chromosome size ranged from 1.1–1.7 μm long in H. neglectus to 3.2-5.0 μm long in H. agrestis. When the highly divergent H. agrestis was excluded from analyses, the mean of the annual species (70.05 FAU) was significantly different from the mean of the perennials (97.70 FAU). No correlations of DNA content with habitat, soil type, annual precipitation, or geographical location were apparent among species.     

VARIATION AND LOCALIZATION OF FLAVONOID AGLYCONES IN HELIANTHUS ANNUUS (COMPOSITAE)

Flavonoid aglycone variation within Helianthus annuus, a species widely distributed throughout North America, was analyzed. Flavonoid aglycones of H. annuus consist of two types, flavones and chalcones. The flavone aglycones are sequestered in glandular trichomes that occur on both leaf surfaces, whereas the chalcone aglycones appear to be incorporated in the waxy leaf cuticle. Considerable variation in flavonoid profile was observed with some plants exhibiting as few as one, and others as many as seven of the eight aglycones detected. No definable phytogeographic patterns were observed for this flavonoid variation. Flavonoid aglycone variation also did not differentiate the infraspecific taxa within H. annuus.     

THE VASCULAR TRANSITION REGION OF HELIANTHUS ANNUUS. I. BILATERAL AND UNILATERAL PATTERNS OF DIFFERENTIATION

The vascular transition region of Helianthus annuus L. may exhibit either a unilateral or a bilateral differentiation of primary phloem groups. The unilateral type of transition has been previously described but the bilateral type with its two variations is described here for the first time. The three patterns of transition are compared, and the significance of the variability of transition regions is discussed in terms of basic transition types and phylogenetic importance.     

CHANGES IN GLYCERALDEHYDE 3-PHOSPHATE DEHYDROGENASE ACTIVITY IN SHOOT APICAL MERISTEMS OF BRASSICA CAMPESTRIS DURING TRANSITION TO FLOWERING

An assay system has been developed for the histochemical determination of glyceraldehyde 3-phosphate dehydrogenase (G3PD) activity to indicate glycolytic pathway capacity during evocation in the shoot apical meristem of Brassica campestris L. G3PD activity was differentially distributed in a zonate pattern within the meristems at the vegetative, the transition, and the floral stages. The activity of G3PD changed in all apical zones of evoked apices, but especially in the central and the peripheral zones of apices at the prefloral stage. In the prefloral stage of development heavy enzyme activity was localized in island-like areas within the peripheral zone. These results indicate that 1) the capacity of glycolysis fluctuates during evocation, 2) during floral evocation the capacity of the glycolytic pathway parallels the capacity of the citric acid cycle and the electron transport system only at the prefloral stage, and 3) G3PD activity marks incipient floral primordia. It is proposed that the enzymic marking of an incipient floral primordium indicates the end of evocation in Brassica.     

A QUANTITATIVE STUDY OF CELLULAR EVENTS IN THE SHOOT APICAL MERISTEM OF BRASSICA CAMPESTRIS (CRUCIFERAE) DURING TRANSITION FROM VEGETATIVE TO REPRODUCTIVE CONDITION

Vegetative plants were induced to flower by 16-hr-long days. Apical buds were collected at intervals during several developmental phases up to 63 hr. A stereologic analysis and mitotic index study was conducted on median longitudinal sections of shoot apical meristems. A rise in the mitotic index occurred between 12 and 24 hr within central, peripheral and pithrib meristem zones. Preceding the floral stage a second increase in the mitotic index was observed in peripheral and central zones, but not in the pith-rib meristem zone. A significant rise in apical volume, cell number, height, and width began in the transitional stage and continued to the floral stage. Significant correlation coefficients were observed between these apical parameters. Relative volume and cell population of each zone remained constant from the vegetative to the reproductive stage. Volume fraction occupied by the nucleus and nucleolus remained constant within each zone during the same time period. In each zone the volume of the nucleus was significantly correlated to volume of the nucleolus. It appears a pre-inflorescence apex, while larger, is structurally similar to a vegetative apex.     

向日葵未受精胚珠培养时胚状体发生的显微观察

由开花前1—4天的向日葵子房中取出胚珠,在液体培养基上进行漂浮培养,诱导了未受精的卵细胞发育为单倍体的胚状体.亦诱导了珠被绒毡层产生胚状体。对两种胚状体的发生和发育过程及其形态发生特点作了显微观察与描述。     

THE VASCULAR TRANSITION REGION OF HELIANTHUS ANNUUS (COMPOSITAE). II. EFFECT OF IONIZING RADIATION

The vascular pattern in the transition region of Helianthus annuus L. may be altered by ionizing radiation in dosages ranging from 650–19,200 R. The alteration of the vascular cylinder is expressed by (1) the failure of normal tracheary elements to mature and (2) by the induction of accessory xylem and/or phloem strands. The anatomical alterations in the vasculature of the transition region are presented and their significance discussed in terms of seedling survival and the mechanisms controlling vascular differentiation.     

CELL DIVISION KINETICS IN THE SHOOT APICAL MERISTEM OF CERATOPTERIS THALICTROIDES BRONG. WITH SPECIAL REFERENCE TO THE APICAL CELL

The duration of mitosis and the cell cycle were determined for defined cell populations of the shoot apical meristem of Ceratopteris thalictroides Brong. by using the colchicine-induced metaphase accumulation technique. The results indicate that the apical cell is mitotically active and cycles at an apparently greater frequency than the cells of subjacent populations. Duration of mitosis was similar for all cells of the meristem. These results are correlated with mitotic indices of control apices, the geometry of the apex, and the mean number of cells in the meristem. Shoot apices from adult plants were examined to determine mitotic indices within the meristem; mitotic activity was again noted for the apical cell. These results contradict recent proposals that the pteridophyte apical cell serves as a unicellular quiescent center which lacks histogenic potential and offer experimental support for the classical concept of apical cell function in those fern shoot meristems which terminate in a single apical cell.     

THE QUANTITATIVE ULTRASTRUCTURE OF THE MERISTEMATIC CELLS OF XANTHIUM STRUMARIUM DURING THE TRANSITION TO FLOWERING

Ultrastructural changes occurring in the central part of the apical meristem of the SDP Xanthium strumarium, induced to flower by a single 16-hr long night, were quantitatively investigated using stereological methods and compared to the changes previously reported in other species, particularly the LDP Sinapis alba. Changes detected in Xanthium, which are also found in other species, included: increase in cellular, cytoplasmic, cytoplasmic matrix and nucleolar sizes, change in nucleolar structure; increase in mitochondrial number and chondriome size, increase in dictyosome number. These changes are believed to be essential for floral evocation because of their universality. Other changes were specific to Xanthium and not detected in Sinapis. Accordingly, they were thought to be accompanying nonessential events of floral evocation in Xanthium. These changes included an increase in the number of plastid profiles and in plastidome size. The size of the nucleus, chromatin and vacuolar apparatus, as well as the number of vacuolar profiles, did not change in Xanthium, contrary to what was observed in other plants.     

STABILITY OF NUCLEAR DNA CONTENT DURING ADVENTITIOUS SHOOT FORMATION IN PINUS TAEDA L. TISSUE CULTURE

Attempts to preserve selected genotypes through clonal propagation may be hampered by genetic instability produced during the tissue culture phase of the propagation procedure. To investigate the genetic condition of loblolly pine (Pinus taeda L.) tissue cultures, nuclear DNA content in embryos grown in vitro and in regenerated buds were examined using two-wavelength microspectrophotometry. Embryos were grown on chemically defined media containing cytokinin as the sole growth regulator. Nuclear DNA content remained at or between 2 C and 4 C with no evidence of polyploidy during nearly 3 months of continuous culture. Benzylaminopurine, even at concentrations which were supraoptimal for shoot regeneration, had no effect on nuclear DNA content. Regenerated shoots also had stable nuclear DNA contents which correspond to a diploid condition. These results are of significance for the development of mass propagation techniques for this commercially valuable species.     

MITOTIC ACTIVITY IN THE ROOT APICAL MERISTEM OF AZOLLA FILICULOIDES LAM., WITH SPECIAL REFERENCE TO THE APICAL CELL

The meristematic activity of the apical cell and its derivatives (merophytes) in the unbranched, determinate roots of Azolla filiculoides Lam. was investigated. The plane of division of the apical cell indicates that it is the initial of each merophyte. The division plane of each newly formed merophyte is strictly periclinal to the root surface and provides confirmation that the immediate derivatives of the apical cell cannot be the ultimate root initials. The frequency of cell division as determined by the mitotic index, and by the duration of the cell cycle as determined by the colchicine method, confirmed the meristematic activity of the apical cell. As roots increase in length, the duration of the cell cycle in the total meristem increases, with the apical cell possessing the longest cell cycle, whereas the immediate derivatives maintain approximately the same cycle duration as in shorter roots. In determinate Azolla roots, cell division appears to play a major role up to a certain root length, then increase in length is produced mainly by cell elongation.     

CELL DIVISION STUDIES OF THE SHOOT APEX OF DATURA STRAMONIUM DURING TRANSITION TO FLOWERING

Seedlings of Datura stramonium L., although not photoperiodically sensitive, are useful for floral transition studies when raised in a growth chamber at a constant temperature of 25 C with a photoperiod of 8 hr of light (1,600-2,000 ft-c) and 16 hr of darkness. A terminal flower is formed after the seventh or eighth leaf primordium is produced. A constant rate of leaf initiation up to the time of flowering enables specific apical stages to be obtained and studied. Changes in the mitotic index, substantiated with calculated rates of cell division (measured by the accumulation of metaphases following treatment with colchicine) were studied in shoot apical zones during transition to flowering. Fluctuations in the mitotic index of each zone in the vegetative and transition apex with respect to apical stage as well as time of day were not statistically significant. The mitotic index of the summit zone of the vegetative apex was significantly lower than in the other zones whose mitotic indices were not significantly different from one another. During floral transition the mitotic index of the summit zone as well as the central zone (just below the summit zone) significantly increased while no significant changes were detected in the flank zones. It was shown that the mitotic index could be considered representative of the rates of cell division in Datura.