CYTOKININ- AND GIBBERELLIC ACID-INDUCED EFFECTS ON THE STRUCTURE AND METABOLISM OF SHOOT APICAL MERISTEMS IN OPUNTIA POLYACANTHA (CACTACEAE)

The dormant axillary buds of Opuntia polyacantha can be activated by either cytokinins or gibberellic acid. Under the influence of benzylaminopurine (BAP), the axillary bud meristem increases greatly in size and becomes mitotically active. The primordia produced by the meristem develop as normal photosynthetic leaves. Gibberellic acid (GA) also causes the meristem to become mitotically active, but the meristem does not increase in size. The primordia produced under the influence of GA develop as normal cactus spines. Leaf-producing meristems and spine-producing meristems have the same zonation, despite the differences in size. The meristems are composed of a uniseriate tunica, a central mother cell zone, peripheral zone, and a pith rib meristem. The mitotic activity of each of the zones in the leaf-producing meristem differs significantly from the mitotic activity of the corresponding zones in the spine-producing meristem.     

Activities of DNA polymerases and RNA polymerases detected in situ in growing and differentiating cells of root cortex

Summary Activities of DNA polymerases and RNA polymerases were studied by autoradiographic methods in growing and differentiating root cortex cells ofZea mays — a species in which endomitosis occurs — andTulipa kaufmanniana — in which this process does not occur. InTulipa kaufmanniana, the highest activity of DNA polymerase appears in the nuclei of meristematic zone during the S phase of the cell cycle. InZea mays, endomitotic replication of DNA occurs in all growth and differentiation zones and the activity of DNA polymerase in the nuclei is similar to that in the meristematic zone. In both species, nuclear RNA synthesis, measured with³H uridine incorporation, is highest in the meristematic zone and declines steadily with development. Activity of nuclear RNA polymerase is present in all developmental zones in both species and is similar to that in the meristematic zone.³H uridine incorporation into nucleoli decreases markedly in both species, whereas the activity of nucleolar RNA polymerase remains at a high level in all root segments inZea mays and decreases slightly inTulipa kaufmanniana.It is argued that the differences between the incorporation of³H uridine and that or³H UMP may be caused by a reduction of the pool of endogenous ribonucleoside triphosphates. Marked activities of DNA polymerase and RNA polymerase in cytoplasm are possibly related to the growth and division of plastids and mitochondria.     

Structure and seasonal study of the shoot apex of two species ofAraucaria

Summary and Conclusions The shoot apices ofAraucaria columnaris andA. bidwillii exhibit a tunica-corpus organization. In the former the tunica is one-layered during spring and summer months and becomes more stratified during winter months. InA. bidwillii it is usually two-layered, with a tendency to further stratification.The corpus is divisible into three zones, the corpus initial zone, the flanking zone and the rib meristem zone. The corpus initial zone is the seat of formation of the rest two. The flanking zone is concerned with the production of leaf primordia, in which the covering tunica layer has no part.The rib meristem zone varies in depth according to the season of the year and the pith cells mature gradually in these species.InA. columnaris, where a seasonal study was undertaken, no period of complete dormancy was observed.     

Apical reactions to gibberellic acid application according to the genotype inSilene armeria

Two selected strains ofSilene armeria L. were used: S_1-2 (GA-line, not induced to flowering by GA₃ in SD of 8 h) and S_2-1 (GA+ line which reacts to GA₃ with flowering in non-inductive photoperiod). Moreover S_1-2 and S_2-1 differ in their critical daylength 14.5 and 8.0 h, respectively. Changes in the mitotic index and DNA content of cells in the various zones of the apical meristem during GA₃ treatment were described. At the start of the experiment, the functioning of the apex was characterized by a predominance of G₁ phase in the two strains. Therefore S_2-1 differed by a higher mitotic activity specially in the central zone. In S_1-2, GA₃ acted mainly on the rib-meristem cells, which resulted in stem elongation. Lack of response in the cells of the central zone explains why GA₃ fails to induce flowering in this strain. In S_2-1, GA₃ acted mainly on the central zone where a gradual increase in mitotic acitivity and in the percentages of nuclei at the (S + G₂) level was recorded. This stimulation brought the meristems into the prefloral stage. The differences between the two strains are discussed according to the status of control meristems. The reactions induced by GA₃ were compared with the pattern of changes during induction, in LD. This work was supported by grants from the “Centre National de la Recherche Scientifique” (UA 572).     

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.     

Gibberellins and the photoperiodic control of stem elongation in the long-day plant Agrostemma githago L.

Agrostemma githago is a long-day rosette plant in which transfer from short days (SD) to long days (LD) results in rapid stem elongation, following a lag phase of 7–8 d. Application of gibberellin A₂₀ (GA₂₀) stimulated stem elongation in plants under SD, while 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidine-carboxylate methyl chloride (AMO-1618, an inhibitor of GA biosynthesis) inhibited stem elongation in plants exposed to LD. This inhibition of stem elongation by AMO-1618 was overcome by simultaneous application of GA₂₀, indicating that GAs play a role in the photoperiodic control of stem elongation in this species. Endogenous GA-like substances were analyzed using reverse-phase high-performance liquid chromatography and the d-5 corn (Zea mays L.) assay. Three zones with GA-like activity were detected and designated, in order of decreasing polarity, as A, B, and C. A transient, 10-fold increase in the activity of zone B occurred after 8–10 LD, coincident with the transition from lag phase to the phase of rapid stem elongation. After 16 LD the activity in this zone had returned to a level similar to that under SD, even though the plants were elongating rapidly by this time. However, when AMO-1618 was applied to plants after 11 LD, there was a rapid reduction in the rate of stem elongation, indicating that continued GA biosynthesis was necessary following the transient increase in activity of zone B, if stem elongation was to continue under LD. It was concluded that control of stem elongation in A. githago involves more than a simple qualitative or quantitative change in the levels of endogenous GAs, and that photoperiodic induction alters both the sensitivity to GAs and the rate of turnover of endogenous GAs.Abbreviations AMO-1618 2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidine-carboxylate methyl chloride - GA(s) gibberellin(s) - LD long day(s) - LDP long-day plant(s) - SD short day(s)     

Location of cell cycle changes in relation to morphological changes in the shoot apex ofSilene coeli-rosa immediately before sepal initiation

Summary Changes in morphology, the mitotic index and the proportions of cells in G₁ and G₂ were measured in shoot meristems ofSilene coeli-rosa immediately before floral morphogenesis in order to determine whether the known changes to the cell cycle at this time are restricted to a particular region of the apex. Twenty-eight day-old plants were given either 7 long days (LD) plus 2 short days (SD) (day 8 of the LD treatment) or 9 SD [day 8 of the SD control (SDC) treatment]. Plants were sampled on day 8 every 2 h for 12 h and the various cell cycle measurements were performed on sections of the apical meristem. In the inductive LD treatment there was a peak in the mitotic index at 13.00 h and, possibly, the start of another at 19.00 h. At 21.00 h all meristems in this treatment initiated sepals. The mitotic activity at 13.00 and 19.00 h in the LD treatment was a result of significant increases in the mitotic index in the axial, lateral and central sub-axial areas of the apex compared with the corresponding zones in the SDC treatment. At 13.00 h of day 8, 80% of cells were in G₂ phase in the axial region in the LD treatment whilst 85% of cells were in G₁ in the axial zone in the SDC treatment. In the other zones significantly more cells were in G₂ in the LD compared with the SDC treatment as was the case at 19.00 h although not to the same extent as the axial zone at 13.00 h. Thus these data emphasize, for the first time, the mitotic activation and predominance of the G₂ population of cells particularly in the axial zone of shoot meristems in the LD treatment. These data are discussed in relation to the synchronisation of cell division which could occur in the prefloral shoot meristem at this time, affecting each shoot apical zone.Abbreviations LD long day - SD short day - SDC short day control     

Comparison of two serpentine species with different nickel tolerance strategies

The effects of nickel were studied in two serpentine species with different metal tolerance strategies:Silene italica L., which limits nickel uptake and translocation, andAlyssum bertolonii Desv., a serpentine endemic, which accumulates nickel mostly in the leaves. InS. italica, nickel 7.5 μM inhibited root growth and depressed mitotic activity in root tips. Peroxidase activity and phenol concentration both in roots and shoots were increased; under the same conditions nickel did not produce any relevant effect onA. bertolonii. InS. italica an adequate calcium concentration (25 mM) was able to reverse the effects of nickel on root growth and metabolism. InA. bertolonii the same calcium concentration reduced root growth, confirming this species adaptation also to low calcium concentrations, typical of serpentines.     

Proteomic analysis reveals the spatial heterogeneity of immobilized Taxus cuspidata cells in support matrices

A proteomic approach was used to study the responses of Taxus cuspidata cells to local microenvironments in different zones of immobilized support matrices. Analysis of protein spots by 2-DE revealed significant differences in the abundance of 31 spots, 28 spots, and 23 spots in outer, middle, and central zone cells between the immobilized and suspended cells. Six of these proteins, identified by MALDI-TOF-MS, were involved in the regulation of carbohydrate, nitrogen, and sulfur metabolisms. Immobilization triggered an increase in taxol production of the immobilized cells in the middle and central zones compared to that of the suspended cells. A negative relation between taxol production and the mitotic index was observed in the cells in the immobilization support matrix. Cells in the outer zone had high mitotic index and low taxol production, while cells in the middle and central zones showed low mitotic index and high taxol production. The abundance of S-adenosylmethionine synthetase, which was identified as one of the differentially expressed proteins, was positively correlated to the cell division activity in the immobilized cell cultures.     

Comparative karyotype analysis ofCeratozamia mexicana andMicrocycas calocoma (Zamiaceae) using fluorochrome banding (CMA/DAPI) and fluorescence in situ hybridization of ribosomal DNA

Orcein staining, differential staining with CMA and DAPI, and FISH with an rDNA probe were used to compare somatic chromosomes ofCeratozamia mexicana andMicrocycas calocoma. CMA-positive dots and hybridization signals appeared on chromosomes at early interphase and mitotic prophase, but in significantly different number in the two species. InCeratozamia mexicana, the CMA-positive and DAPI-negative bands and the hybridization signal were located at the terminal region of the long arm of three median-centromeric chromosomes, the terminal region of the short arm of two median-centromeric chromosomes and the terminal region of the long arm of two subterminal-centromeric chromosomes. InMicrocycas calocoma, they were located at the pericentric region of two median-centromeric chromosomes. These chromosome data suggested thatMicrocycas has no simple Robertsonian relationship toCeratozamia.     

Internal dimorphism of chloroplasts in the leaves ofPeperomia pellucida (L.) H.B.K.

Summary The large chloroplasts found at the abaxial pole of palisade cells in the leaf ofPeperomia pellucida (L.) H.B.K. exhibit a peculiar ultrastructure, having a marginal and a central zones clearly distinguished from each other. This internal dimorphism may correspond to a functional specialization because starch grains concentrate at the central region, and grana show a much larger number of thylakoids at the marginal zone.     

EVIDENCE FOR POST-TRANSLATIONAL MODIFICATION OF TRIOSE PHOSPHATE ISOMERASE (TPI) IN ISOËTES (ISOËTACEAE)

As part of an electrophoretic study on Isoëtes, a number of Neotropical and North American species were examined for allozyme variation in TPI. Three of these species—I. storkii, I. flaccida, and I. mexicana—exhibit three distinct zones of TPI activity. The two most anodally migrating zones are comparable to the two zones found in most angiosperms and in several other species of Isoëtes. The single or three-banded phenotypes produced at these loci correspond, respectively, to the homozygous and heterozygous patterns typical of a dimeric enzyme. The most cathodal zone (zone III) differs in producing either single or two-banded phenotypes. Analyses of these three zones indicate a nearly perfect correlation between zones II and III in putative allelic constitution and relative allelic mobility. Explanations involving TPI gene duplications and/or null alleles fail to account for the peculiar banding characteristics and origin of activity zone III. An alternative hypothesis involving a protease duplication and differential post-translational modification is postulated. This hypothesis adequately explains the zone III phenotypes and fixation of the third activity zone in the species examined. Amino acid sequencing is suggested as the most direct test of this hypothesis. The taxonomic distribution of TPI III generally supports a previous, morphologically-based, hypothesis on species relationships in Isoëtes. The presence of this zone is regarded as an independent synapomorphy for a major clade of Neotropical Isoëtes.     

Synthesis of actinomycin-insensitive RNA during the first post-irradiation mitotic cycle,in the synchronously mitotic plasmodia of Physarum polycephalum

A sucrose density gradient analysis of³H-uridine pulse-labelled RNA from the first postirradiation mitotic cycle ofPhysarum polycephalum shows that all the density classes of RNA synthesized during this period are resistant to the peptide-antibiotic, actinomycin D. In fact, the synthesis is found to be greater in the presence of the drug. The heterogenously sedimenting synthetic activity here may represent a single species of RNA and its precursors or more than one kind of RNA. Further characterization of this RNA is meaningful in view of the actinomycin insensitivity of the postirradiation mitotic cycle itself to this antibiotic.     

SHOOT HISTOGENESIS: THE EARLY EFFECTS OF GIBBERELLIN UPON STEM ELONGATION IN TWO ROSETTE PLANTS

Sachs , Roy M., Charles F. Bretz , and Anton Lang . (U. California, Los Angeles.) Shoot histogenesis: The early effects of gibberellin upon stem elongation in two rosette plants. Amer. Jour. Bot. 46(5): 376–384. Illus. 1959.—Within 24 hr. after the application of gibberellic acid (GA) to vegetative plants of biennial Hyoscyamus and of the long-day plant Samolus, a considerable increase in mitotic activity was observed in the pith, cortical, and vascular tissues of the rosette axis immediately below the apical meristem. As the treatment continued, the zone of cell division increased commensurate with the increase in length of the stem; the new cell divisions formed transverse walls predominantly and thus contributed to stem elongation. The cell contribution from the apical meristem was but a small fraction of the total produced by the subapical tissues, suggesting that the induced subapical mitotic activity is the main site of tissue development in the shoot. There was no evidence for cell elongation for at least 72 hr. after application of GA, and, hence, the initial increase in stem length was due solely to an increase in cell number. With regard to the general problem of shoot histogenesis, our data for the rosette plants and those for Xanthium and Chrysanthemum showing extensive cell division far below the apical meristem, are in full agreement with the studies by Bindloss (1942) with tomato, and support her conclusion that “. . . it is no longer possible to think that the chief center of cell division is in a relatively short zone 60 to 100 microns from the stem tip . . . and that cell division activity in the promeristem is not solely responsible for stem length.” On the contrary, the mitotic activity in the subapical regions is undoubtedly responsible for the major part of the cells found in the stem.     

Morphological variations of the cranium within the genusLeontopithecus

Multivariate analysis methods were applied to the cranial measurements ofLeontopithecus. InLeontopithecus chrysomelas, the face is generally narrow and the cranial shape is relatively unique. Especially, the male has extremely narrow face and quite unique cranial shape amongLeontopithecus. Leontopithecus rosalia has the broad face compared with the other species. The cranial size ofL. rosalia is as large as that ofL. chrysomelas. Male ofLeontopithecus chrysopygus is the largest in overall size of the cranium, and has the widest braincase.     

Patterns of vegetation surrounding springs in Goshen Bay,Utah County,Utah U.S.A.

Saline meadows in Goshen Bay, Utah County, Utah, USA, occur interspersed among large areas of salt playa. Springs irrigate these meadows which show concentric vegetational zones surrounding each spring. Each vegetational zone demonstrates its own unique dominant vascular plant species. The central zone is dominated primarily by Scirpus americanus, the middle zone by Eleocharis palustris and the outer zone by Juncus balticus, Distichlis spicata and Muhlenbergia asperifolia. As distance increases from the springs, pH and soluble salt concentration increase, while percent organic matter, percent moisture and phosphorus decrease. With the increase in salt levels and decrease in water levels, halophytic plant species generally increase with distance from the springs.     

THE EARLY ACTION OF THE FLORAL STIMULUS ON MITOTIC ACTIVITY AND DNA SYNTHESIS IN THE APICAL MERISTEM OF XANTHIUM STRUMARIUM

Vegetative plants of Xanthium strumarium (a short-day species) were induced to flower by exposure to a single 16-hr long night. By cutting off the induced leaf (half-expanded leaf) at various times, it was established that, by 8 hr after the end of the long night, a sufficient amount of floral stimulus had reached the meristem to induce a flowering response. The following sequence of events occurred in both the peripheral and central zones of the apical meristem of induced plants: 1) a rise in the mitotic index beginning at 28 hr after the end of the long night and culminating at 36 and 56 hr; 2) a stimulation of DNA synthesis starting at 32–36 hr and reaching a maximum at 60 hr; 3) an increase in nucleolus diameter starting at 32 hr. The cell population in the meristems of both vegetative and induced plants displayed a similar distribution, with about 80 % of the nuclei with the 2C amount of DNA. The comparison of the kinetic data concerning the mitotic index and DNA synthesis indicated that one of the early effects of the floral stimulus in the peripheral and central zones was the release in mitosis of cells whose nuclei were in the postsynthetic (G₂) phase of the mitotic cycle. In the pith-rib meristem, the following events were recorded: 1) a stimulation of DNA synthesis starting at 20 hr; 2) a rise of the mitotic index beginning at 28 hr; 3) the vacuolation and elongation of cells starting at 48 hr. All these events occurred well before the initiation of bract and flower primordia, which began at 96 and 136 hr, respectively. Neither stimulation of mitotic activity nor flowering occurred in the meristems of plants subjected to a long night interrupted at its midpoint by a 5-min light break. The results are discussed in relation to the early events which are known to occur in the meristems of other photoperiodic species in transition to flowering.     

Spontaneous chromosome inversions of guatemalan teosintes (Zea mexicana)

Two paracentric inversions,In3 andIn9, were found in the F1 hybrids of maize and Florida teosinte and these inversions were contributed by the teosinte parent. The length ofIn3 was equivalent to about 35 percent of the length of the long arm of chromosome 3, while that ofIn9, about 60 percent of the length of the short arm of chromosome 9.There were also two paracentric inversions,In1 andIn9, in the F1 hybrids of maize and Jutiapa teosinte and these inversions were inherited from the teosinte parent. The length of theIn1 occupied 22 percent of the total length of the long arm of chromosome 1, while that ofIn9, 60 percent of the total length of the short arm of chromosome 9.Only one paracentric inversion,In9, was identified in the F1 hybrids of maize and Lake Retana teosinte, and this inversion was also from the teosinte parent. As length and location are considered, thisIn9 is the same as theIn9's of Florida teosinte and Jutiapa teosinte.At anaphases I and II of the microsporocyte divisions of the F1 hybrids, evidences of crossovers within the inverted segments, such as bridges and fragments, were obtained for all of these inversions. The interchromosome effect ofIn3 of Florida teosinte, and that ofIn1 of Jutiapa teosinte on the frequency of crossovers within the inverted segment ofIn9's are discussed.Chromosome inversions have probably accompanied the divergence of geographical races of teosinte. This might also be true for the race diversities of maize. The absence ofIn9 in certain teosinte races of southern Mexico and northern Guatemala is accounted for by the substitution of maize chromosome for this inversion.     

Cell cycle events during the development of the silk glands in the mulberry silkworm<Emphasis Type="Italic"> Bombyx mori</Emphasis>

Silk glands of the mulberry silkworm Bombyx mori are long and paired structures originating from the labial region and are anatomically and physiologically divided into three major compartments, the anterior, middle and posterior silk glands. The silk gland morphogenesis is complete by 8 days post egg laying. Extensive growth of silk glands during the larval stages is due to increase in tissue mass and not cell number. The cells in a completely formed silk gland pursue an endoreplicative cell cycle, and the genome undergoes multiple rounds of replication without mitosis or nuclear division. The expression patterns of cyclin B (mitotic cyclin) and cyclin E (G1 cyclin, essential for G1/S transition in both mitotic and endoreplicative cell cycles) in the course of silk gland development revealed that mitotic cell divisions take place only in the apex of the growing silk gland. However, the persistence of another mitotic focus in the middle silk gland even when the growing apex has moved well past this zone suggested the continued operation of mitosis for a while in this restricted region. The lack of cyclin B expression and abundance of cyclin E in the rest of the areas confirmed an alternation of the G1 and S phases of the cell cycle without an intervening mitotic phase. No expression of cyclin B was noticed anywhere in the silk glands after stage 25 of embryogenesis, indicating a complete switch over to the endomitotic mode of the cell cycle. The onset of expression of various genes encoding different silk proteins correlated with the onset of endomitotic events.Edited by D. Tautz     

Mitotic cycle time and DNA content in annual and perennialMicroseridinae (Compositae,Cichoriaceae)

Within eight annual and perennialMicroseridinae species studied, the duration of the mitotic cycle is positively correlated with the nuclear DNA content, cycle time (hrs) = 7.3 + 0.32 × pg DNA/nucleus. Within the generaAgoseris andMicroseris, the annuals have lower DNA contents and more rapid mitotic cycle times than do the perennials. This relationship is predicted by the nucleotypic theory ofBennett. Annual species ofPyrrhopappus have relatively high DNA contents and a proportionately longer mitotic cycle time, but contrary to that expected by the nucleotypic theory as originally proposed have the fastest growth rate and shortest generation time observed in theMicroseridinae. This rapid developmental rate is discussed, nucleotypically, however, by analyzing relationships between DNA content, mitotic cycle time, and cell size.