Bud Activity Research Articles

Feather buds exert a polarizing activity when transplanted to chick limb buds.

Homeoproteins have been shown to be expressed in a position-specific manner along the anterior-posterior axis in the developing chick feather bud, as seen also in the developing limb bud. These facts raise the possibility that there may be common mechanistic features in the establishment of the anterior-posterior polarity between both organs. In order to investigate this possibility, feather bud tissues were transplanted into the anterior region of limb buds to determine whether feather bud tissues possess properties such as the zone of polarizing activity of the limb bud. The manipulated limb bud formed a mirror image duplication of the skeletal elements, mainly (2)2234 digit pattern or sometimes 3(2)234. Both the anterior and posterior halves of feather bud tissue exhibited almost equal activity in inducing ectopic skeletal elements. Hoxd-12 and Hoxa-13 were expressed coordinately around the transplanted site of the operated limb bud. This secondary axis-inducing activity of the feather bud was enhanced when grafts were pretreated with trypsin. In contrast, the presumptive feather bud tissue and inter-feather bud tissue did not induce a secondary axis of the limb bud. These results suggest that the feather bud contains a region that exerts polarizing activity and that this region may play key roles in the formation of the anterior-posterior and, if it exists, proximal-distal axis of the feather bud, possibly via the regulation of region specific expression of Hox genes.

Population-level compensation after loss of vegetative buds: interactions among damaged and undamaged cotton neighbours.

Population-level compensation occurs "when herbivore attack on one individual allows another individual to grow more rapidly". This form of compensation was investigated in high and low density cotton crops subjected to three treatments: (i) undisturbed controls, (ii) uniformly damaged, in which all plants were damaged, and (iii) non-uniformly damaged, in which every second plant was damaged. Damaged plants had their vegetative buds manually removed to simulate damage by Helicoverpa spp. (Lepidoptera). Removal of vegetative buds did not reduce seed cotton production per unit ground area. In uniformly damaged crops, compensation was essentially the result of profuse branching after release of apical dominance and activation of axilary buds. In non-uniformly damaged crops, population level mechanisms acted that involved strong plant-plant interactions. At both plant densities, undamaged plants grown alongside damaged neighbours accumulated more root and shoot biomass and produced more seed cotton than undamaged plants in uniform crops. Different degrees of symmetry in the relationship between damaged and undamaged neighbours lead to different degrees of compensation, viz. seed cotton production of non-uniformly damaged crops ranged from 98 to 125% of that in controls. At high plant density, neighbour status also affected flowerbud initiation and/or retention. Changes in competitive relationships as well as early detection of and response to neighbour status were likely involved in these responses.

Retinoic acid signaling is required during early chick limb development

In the chick limb bud, the zone of polarizing activity controls limb patterning along the anteroposterior and proximodistal axes. Since retinoic acid can induce ectopic polarizing activity, we examined whether this molecule plays a role in the establishment of the endogenous zone of polarizing activity. Grafts of wing bud mesenchyme treated with physiologic doses of retinoic acid had weak polarizing activity but inclusion of a retinoic acid-exposed apical ectodermal ridge or of prospective wing bud ectoderm evoked strong polarizing activity. Likewise, polarizing activity of prospective wing mesenchyme was markedly enhanced by co-grafting either a retinoic acid-exposed apical ectodermal ridge or ectoderm from the wing region. This equivalence of ectoderm-mesenchyme interactions required for the establishment of polarizing activity in retinoic acid-treated wing buds and in prospective wing tissue, suggests a role of retinoic acid in the establishment of the zone of polarizing activity. We found that prospective wing bud tissue is a high-point of retinoic acid synthesis. Furthermore, retinoid receptor-specific antagonists blocked limb morphogenesis and down-regulated a polarizing signal, sonic hedgehog. Limb agenesis was reversed when antagonist-exposed wing buds were treated with retinoic acid. Our results demonstrate a role of retinoic acid in the establishment of the endogenous zone of polarizing activity.

"Pure" partial trisomy 4q25-qter owing to a de novo 4;22 translocation.

A girl with a new de novo translocation of 4q onto the short arm of acrocentric 22 is reported as a case of "pure" partial trisomy 4q. Her karyotype was 46, XX,-22, +mar, t(4;22)(q25-->qter;p11) identified by Giemsa staining and FISH. Comparison of the proband with previously reported cases of "pure" partial trisomy 4q showed the main clinical features to be growth retardation, psychomotor retardation, microcephaly, large, low set, malformed ears, prominent nasal bridge, ptosis and epicanthus.

The enamel knot as a signaling center in the developing mouse tooth

Mammalian tooth forms are produced during development by folding of the enamel epithelium but the molecular mechanisms involved in the formation and patterning of tooth cusps are not understood. We now report that several key signaling molecules found in well-known vertebrate signaling tissues such as the node, the notochord, the apical ectodermal ridge, and the zone of polarizing activity in the limb bud are specifically expressed in cells of the enamel knot, which is a transient cluster of dental epithelial cells. By comparing three-dimensional reconstructions of serial sections following in situ hydridization we localized Sonic hedgehog, Bone morphogenetic proteins-2, -4 and - 7, as well as Fibroblast growth factor-4 in nested domains within the enamel knot. We suggest that the enamel knot acts as a signaling or organizing center, which provides positional information for tooth morphogenesis and regulates the growth of tooth cusps.

Hensen's Node from Vitamin A-Deficient Quail Embryo Induces Chick Limb Bud Duplication and Retains Its Normal Asymmetric Expression ofSonic hedgehog(Shh)

Both Hensen's node, the organizer center in chick embryo, and exogenous retinoic acid are known to induce limb duplication when grafted or applied to the host chick limb bud. Retinoic acid is known to be present in the node and has been proposed as the putative morphogen for chick limb development. Here, we report that Hensen's node from vitamin A-deficient quail embryo induces limb duplication in the host chick embryo similar to that induced by the node from vitamin A-sufficient control embryos. We also demonstrate that the expression ofSonic hedgehog(Shh), recently shown to be the mediator of polarizing activity in the chick limb bud, is not affected by the endogenous vitamin A status of the embryo. Furthermore, whole-mountin situhybridization revealed asymmetry ofShhexpression in the Hensen's node of both vitamin A-sufficient and -deficient quail embryos. Retinoids were not detectable in the eggs from which vitamin A-deficient embryos were obtained. Extracts from normal embryos induced a level of expression of reporter gene equivalent to the presence of 3.4 pg of active retinoids per embryo, while those from vitamin A-deficient embryos induced a baseline level of reporter gene expression similar to that of the controls. Our studies suggest that endogenous retinoic acid is not involved inShhexpression nor in regulating its asymmetry during normal early avian embryogenesis and support the current view that endogenous retinoic acid may not be a direct morphogen for limb bud duplication.

Controlling rootstock sprouts of Agonis flexuosa, (Willd.) Sweet at ambient and elevated CO2 by multiple applications of low concentration NAA

A method for grafting variegated scion material to green leafed Agonis flexuosa (Willd.) Sweet stock was developed to overcome the difficulty of striking cuttings. However sprouting of both seedling and cutting-grown A. flexuosa rootstocks was a significant problem. Microwedge grafting of actively growing leafy scions and stocks in fog at 32/22�C gave 90-100% success and scion bud activity was stimulated within 2 weeks. Weekly or fortnightly spray applications of 100 mg NAA/L (napthaleneacetic acid), starting at the time of grafting, gave effective sprout control whereas a single pre-graft spray of 200 mg NAAL was not effective. CO2 enrichment of the fog environment was investigated as a means of enhancing scion growth. CO2 at 80 kPa increased scion dry weight (DW), leaf and branch numbers, but had no effect on rootstock sprout or stem DW or sprout numbers. Optimum NAA concentrations for rootstock sprout suppression under elevated CO2 with fog, were 50-100 mg/L, which were not deleterious to scion shoot length, when sprayed on the stock portion only. Stock sprout numbers, scion leaf and branch numbers were negatively correlated with NAA concentration. Sprout growth at 100 mg NAA/L was about 5% of control sprout growth. Concentrations >200 mg NAA/L caused leaf tip necrosis and excess stem callusing. Scion growth was inversely related to the degree of resprouting in control treatments.

FLORAL BUD DIFFERENTIATION AND ACTIVITY OF ANNA APPLE TREES ON DIFFERENT ROOTSTOCKS

FLORAL BUD DIFFERENTIATION AND ACTIVITY OF ANNA APPLE TREES ON DIFFERENT ROOTSTOCKS

Anatomy and development of the fern sporophyte

Recent research on the developmental anatomy and morphology of the fern sporophyte is reviewed. Detailed histological and experimental studies of the organization of the fern shoot apical meristem have reconfirmed the recently controversial role of the shoot apical cell as the single apical initial of the meristem. The shoot apical meristem is nevertheless an anatomically and functionally complex structure with a strongly zoned cytohistological organization. Fern shoot apex organization can be compared with that of seed plants. The control of leaf initiation and phyllotaxy remains poorly understood. Studies differ as to whether leaf initiation in ferns involves one leaf mother cell or a multicellular region of the shoot apex. The concept of non-appendicular fronds is refuted for living ferns. The later developmental changes in the determinate leaf apical and marginal meristems of the leaf primordium form an area that is still largely unexplored but could be investigated by methods similar to those used to study shoot and root apices. Branching in ferns is morphologiclaly and developmentally diverse. There is apparently more than one developmental mode of dichotomous branching, and several modes of lateral bud formation have been described, including the phyllogenous initiation of branches at the base of leaf primordia. Developmental changes in bud meristems related to apical dominance, inhibition, and bud activation is another major area for continued study. The traditional concept of the role of the root apical cell has been reestablished by studies similar to those made of the shoot apex. Detailed ultrastructural investigations of the root ofAzolla have given a sophisticated new picture of developmental processes in that organ. Fern roots show remarkably precise patterns of histogenesis in relation to apical segmentation. The formation of secondary vascular tissue inBotrychium suggests that the Ophioglossales may be related to the seed plants. The causal relationship of leaf (and branch and root) formation and the initiation of vascular tissue in the shoot needs more study. Although still poorly understood, protoxylem systems in ferns are variable and may have morphological and systematic significance. Recent investigations of hydraulic conductance in fern stems have found possible correlations of conductance levels with growth forms. The anatomical diversity of ferns makes comparative functional anatomy a promising field for future study.

Ultrastructural Changes in Chloroplasts in Harvested Broccoli Flower Buds During Senescence

Green broccoli (Brassica oleracea L. Italica Group) flower heads were stored in perforated polyethylene bags at 20C. Green color of sepals in broccoli flower buds changed to yellow and individual flower buds wilted gradually during storage. Chlorophyllase activity in flower buds tended to increase and chlorophyll content decreased during senescence. Optical and scanning electron microscopic observations were conducted to elucidate the structural changes of chloroplasts in sepals of broccoli flower buds through the senescence. The chloroplasts observed with optical microscope were fine and green when the stage of broccoli flower buds was all green. However, at half-yellow stage, the shapes of chloroplasts obscured and the green color faded. After this stage, colored small particles appeared in the cells and the number of particles tended to increase as yellowing of the flower buds progressed through the senescence. Scanning electron micrographs indicated that the small particles were formed in the chloroplasts and come out from them with senescence followed by aggregation with each other.

TIMING APPLICATION OF HYDROGEN CYANAMIDE (DORMEX) TO PEACH TREES USING A MODIFIED CHILLING MODEL

This multifaceted study was conducted over the past 6 years in Alabama to determine the efficacy of using hydrogen cyanamide to replace lack of chilling in peaches and to develop a working chilling model to allow proper timing of application. Several timings (0%, 25%, 50%, and 75% chilling accumulation) for each chilling level and rates (0%, 0.5%, 1.0%, and 2.0% v/v of 50% hydrogen cyanamide) were evaluated in commercial orchards using replicated studies. It was determined that for Dormex to be effective, 60% to 65% of chilling for the cultivar involved must be accumulated, accompanied by no bud activity beyond bud swell. Rates of 0.5% and 1% v/v of 50% work well with the latter preferred. A computer chilling model was developed to assist growers with proper timing of application.

Expression of genes encoding bone morphogenetic proteins and sonic hedgehog in talpid (ta3) limb buds: Their relationships in the signalling cascade involved in limb patterning

The chicken mutant talpid3 (ta3) has polydactylous limbs with up to 7-8 morphologically similar digits. This lack of antero-posterior polarity in digit pattern is correlated with symmetrical expression of genes of the HoxD complex. We determined the distribution of polarizing activity in limb buds of the chick mutant ta3 by assessing the ability of mesenchyme from various positions along the antero-posterior axis to induce digit duplications when grafted anteriorly into a normal limb. Cells with highest polarizing activity were found at the posterior margin of the wing as in the polarizing region of normal limb buds. However, in contrast to normal limb buds, ta3 anterior mesenchyme also had low polarizing activity. Application of retinoic acid or a polarizing region graft to the anterior of ta3 limb buds changed digit morphology but did not induce digit duplications or digits with any characteristic a-p pattern. To determine which genes are associated with polarizing activity and which are associated with patterning of the digits, we examined expression of the genes Sonic hedgehog (shh), Bmp-2, and Bmp-7, whose expression is normally confined to the posterior margin of the early wing bud and is associated with the polarizing region. In addition, we determined the distribution of Fgf-4 transcripts which in normal limb buds are restricted to the posterior part of the apical ectodermal ridge. In ta3 limb buds, shh expression is restricted to the posterior limb mesenchyme, which has high polarizing activity, but is not expressed in regions which have low polarizing activity. In contrast, Bmp-2 and Bmp-7 are expressed uniformly along the a-p axis. Fgf-4 transcripts are present throughout the apical ectodermal ridge in ta3 limb buds. In the ta3 mutant, there is both an abnormal distribution of signalling activity and response to polarizing signals. In addition, the dissociation between the expression of shh and Bmps suggests distinct roles for the encoded molecules in signalling and response in a-p patterning of limb buds.

Plant Growth and Soil Processes in Temperate Grassland Communities at Elevated CO 2

Turves of a Mollic Psammaquent soil were used in controlled environment rooms to examine the response of managed temperate pasture communities to 350, 525 or 700 p.p.m. CO2. Yield of herbage (regrowth over 3-week intervals) increased only slightly with higher C02; however, the botan- ical composition was markedly different. At elevated CO2 Paspalum dilatatum (C4) and Lolium perenne (C3) declined as a proportion of harvested yield despite a stimulation of single leaf photosynthesis that was comparable to that found in Trifolium repens, a species that increased in abundance. Changes in species composition were largely a consequence of C02-induced differences in axillary bud activity. Net primary productivity below-ground was stimulated by CO2. Soil C02-C production was greater in elevated CO2 treatments, and was consistent with a greater input of herbage and root mass and/or metabolites and of more readily decom- posable material. Levels of microbial biomass were un- changed, but enchytraeids were more abundant at elevated CO2. Tracking of 14CO2 into the various C pools also indicated a more rapid turnover of C at elevated CO2 but no change in pool sizes. No consistent effects on net mineralization of N were observed.

Seasonal changes in the transient expression of a 35S CaMV-GUS gene construct introduced into Scots pine buds.

Seasonal changes in the transient expression of Beta-glucuronidase gene (GUS) driven by a constitutive 35S CaMV-promoter in Scots pine (Pinus sylvestris L.) buds were studied by the microprojectile DNA-delivery method. Buds were collected from 5-, 15- and 50-year-old trees. In buds from all age groups the amount of transient expression was dependent on the season; the highest values were found in March, and values were lowest both at the beginning and at the end of the growing season. Pretreatment with growth regulators increased both the amount of transient GUS expression and arginine decarboxylase (ADC) activity in buds indicating an increase in metabolic activity. These results confirm that the genetic transformation technique can be used to study seasonally dependent regulation in mature Scots pine tissues.

Effect of light quality (red: far-red ratio) and defoliation treatments applied at a single phytomer on axillary bud outgrowth in Trifolium repens L.

We studied the effects of light quality and defoliation on the rate of phytomer appearance and axillary bud outgrowth in white clover. The treatments were applied to one phytomer, a phytomer being defined as the structural unit comprising a node, internode, axillary bud, subtending leaf and two nodal root primordia. Light of a low red:far-red (R:FR) ratio (0.27) was applied to a "target" phytomer either (i) within the apical bud and then to the axillary bud after emergence of the phytomer from the apical bud, or (ii) to the axillary bud only after emergence. The light conditions were directed to these specific parts of the plant by collimating light from small FR light-emitting diodes; with this technique we were able to change the light quality without any change in the level of photosynthetically active radiation. The subtending leaf of the target phytomer was retained or defoliated when it had emerged from the apical bud. FR light applied from the time the phytomer was within the apical bud caused a delay in branch appearance at the target phytomer. In contrast, direct treatment of the axillary bud with FR light after it had emerged from the apical bud did not result in any delay in branch appearance. As the light treatment of the apical bud may have changed the light environment of any of the organs contained in the bud we were unable to ascribe the delay in branch appearance to light perception by any particular organ. However, indirect evidence leads to the conclusion that the likely site of light perception was the developing leaf subtending the axillary bud while it was the outermost phytomer within the apical bud. These results do not support the hypothesis that the R:FR ratio of light incident at an axillary bud site is the environmental factor that controls bud development. Defoliation of the unfolding leaf reduced the rate of phytomer appearance on the main stolon but had no immediate effect on branch appearance. As a consequence there was a reduction in the number of phytomers between the stolon apical meristem and the first phytomer with a branch. This is frequently taken to indicate a relaxation of apical dominance, but in this case was found not to involve a direct effect on bud activity. A current model of white clover growth suggests that there is integration of activity between apical meristems but independence of activity and response to the local micro-environment by axillary buds. In contrast, we found that (i) defoliation reduced phytomer appearance only at the main stolon apical meristem and not at all the meristems in the plant and (ii) that a change in the local light environment of an axillary bud had no discernible effect on bud activity once the bud had emerged from the apical bud but could delay branching if applied before emergence. These results are at variance with the predictions of the model.

Frost tolerance and bud dormancy of container-grown yellow birch, red oak and sugar maple seedlings.

Container-grown seedlings of red oak (Quercus rubra L.), sugar maple (Acer saccharum Marsh.) and yellow birch (Betula alleghaniensis Britton) in their first year of growth were overwintered outdoors. Tolerance of roots and stems to freezing was compared from late summer to the following spring. Mitotic activity in the apical bud was related more closely to air temperature than to bud dormancy as defined by days to bud break. In all species, stem hardening was observed before days to bud break reached a maximum. Dormancy release (days to bud break equal to zero) of yellow birch coincided with loss of stem hardening in the spring. Roots hardened more slowly, had a lower frost tolerance than stems in fall and winter, and dehardened earlier than stems in the spring. There were differences in stem and root hardiness among the species, with yellow birch being the most tolerant, followed by sugar maple and red oak. Primarily because of root sensitivity to frost, winter was a critical period for all three species, but particularly for red oak.

Production of clonal plantlets of Grevillea robusta in in vitro culture via axillary bud activation

A micropropagation method for Grevillea robusta A. Cunn. was developed using explants from mature trees cultured on Woody Plant Medium plus 4.4 μM benzyladenine and 0.27 μM naphthaleneactic acid (NAA) for shoot proliferation. Each nodal explant produced three to five shoots within 10 to 12 weeks in three successive transfers. Rooting was obtained by dipping the shoots in 0.54 mM NAA solution. Rooted shoots were established well in soil.

Cholinesterases and peanut agglutinin binding related to cell proliferation and axonal growth in embryonic chick limbs

Embryonic cholinesterases are assigned important functions during morphogenesis. Here we describe the expression of butyrylcholinesterase and acetylcholinesterase, and the binding of peanut agglutinin, and relate the results to mitotic activity in chick wing and leg buds from embryonic day 4 to embryonic day 9. During early stages, butyrylcholinesterase is elevated in cells under the apical ectodermal ridge and around invading motoraxons, while acetylcholinesterase is found in the chondrogenic core, on motoraxons and along the ectoderm. Peanut agglutinin binds to the apical ectodermal ridge and most prominently to the chondrogenic core. Measurements of thymidine incorporation and enzyme activities were consistent with our histological findings. Butyrylcholinesterase is concentrated near proliferative zones and periods, while acetylcholinesterase is associated with low proliferative activity. At late stages of limb development, acetylcholinesterase is concentrated in muscles and nonexistent within bones, while butyrylcholinesterase shows an inverse pattern. Thus, as in other systems, in limb formation butyrylcholinesterase is a transmitotic marker preceding differentiation, acetylcholinesterase is found on navigating axons, while peanut agglutinin appears in non-invaded regions. These data suggest roles for cholinesterases as positive regulators and peanut-agglutinin-binding proteins as negative regulators of neural differentiation.

Fungitoxic effect of some organic volatile substances against fungi causing otomycosis.

The fungitoxic effect of seven volatile substances (ammonia, carbon disulphide, petroleum benzene, carbon dioxide, methanol, glacial acetic acid and hydrogen peroxide) against five fungi, i.e. Aspergillus niger, A. flavus, Absidia corymbifera, Penicillium nigricans and Candida albicans, was determined on the basis of their dry mycelial weight and sporulation or budding activity. These organisms were isolated from patients suffering from fungal infections of the ear. All the volatile substances tested were found to inhibit mycelial growth and sporulation or budding.

Plant Compensatory Responses: Bud Dormancy as an Adaptation to Herbivory

Some plants can compensate, and even overcompensate, for the loss of productivity caused by herbivory. The presence of latent meristems, or dormant buds, is one of the basic prerequisites of such compensation mechanisms. We present a mathematical model in order to analyze compensation responses in relation to the intensity of herbivory. The model generates a number of qualitatively different kinds of compensation curves when seed production is plotted against the proportion of active meristems lost per grazed plant. The shape of the curves depends on the proportion of dormant buds and their activation sensitivity in relation to meristem loss. Overcompensation is most probable when dormant buds are easily activated. When plants are grazed only once, as assumed in our model, selection favors high bud sensitivity. However, we expect that repeated damage may select for a more restrained pattern of bud activation. When relatively few buds remain dormant, plants can overcompensate for low levels of damage only. On the other hand, when most buds remain dormant, they can overcompensate even for high levels of damage. We consider compensation capacity a potential benefit of bud dormancy when plants are subject to damage. However, bud dormancy may also imply costs on plant productivity and fecundity in the absence of herbivory. Still, intense herbivory may favor bud dormancy in spite of the potential costs. Selection for bud dormancy requires both that the risk of herbivory is high and that herbivores remove a large fraction of active meristems per plant. Consequently, overcompensation is a theoretically plausible possibility, and intense herbivory is a potential selective force that favors bud dormancy. None of these results, however, imply that herbivory is beneficial to plants. In our case, plants with bud dormancy never have higher seed production than plants that have no dormant buds and that are not grazed.