Time course of exudation from excised corn root segments of different stages of development

Xylem exudates were collected at hourly intervals from short segments which had been excised from two portions of the primary root of corn (Zea mays L.) seedlings and partially immersed in experimental salt solution containing ⁸⁶Rb. All segments showed variation in rates of output of both volume and ions for several hours, after which a steady state was attained which persisted for many hours. Apical segments produced little or no exudate for several hours and did not reach a steady state until 18 or more hours after excision. Basal segments produced exudate containing detectable quantities of isotope within an hour and they reached a steady state about 12 hours after excision. During their respective steady states, apical segments produced three times the volume per hour and translocated eight times as much Rb per hour as did basal segments.     

Selective effects of victorin on growth and the auxin response in Avena

Victorin, the pathotoxin from the host-specific pathogen, Helminthosporium victoriae, promotes the growth of coleoptile segments when given at concentrations that are high but which still show selective effects on susceptible and resistant tissue. The latent period in the growth response of both susceptible and resistant tissue is about 3.6 minutes compared to 11.0 minutes in the response of these tissues to auxin. The victorinpromoted rate of elongation of 8-millimeter segments is about 0.2 millimeter per hour in susceptible tissue and about 0.1 millimeter per hour in resistant tissue compared to about 0.4 millimeter per hour in response to auxin. At low concentrations, the toxin has no growth-promoting effect in either susceptible or resistant coleoptile segments. Over a wide range of concentrations, victorin inhibits the growth response of susceptible tissue to auxin completely while having no effect on the response of resistant tissue to auxin.     

TIME RELATIONS OF GROWTH : III. GROWTH CONSTANTS DURING THE SELF-ACCELERATING PHASE OF GROWTH.

Growth curves consist, in all cases, of two major segments. The first major segment is, in the case of higher animals and plants, made up in turn of several (probably five) shorter segments during each of which growth takes place at a constant percentage rate. The transitions between the successive stages are abrupt, the abruptness being of the order of metamorphosis in cold blooded animals. It has been made clear in the first paper of this series that the time rate of growth following the major inflection declines at a constant percentage rate. The junction between the two major segments occurs at puberty in animals and flowering in plants. The two major segments are not symmetrical about the major inflection. The slope of the segment following the inflection is always less than the slope of the segment preceding the inflection. The major inflection does not occur in the center of the growth curve. The instantaneous rate of growth at the beginning of growth is of the order of 100–200 per cent per day (i.e. the body weight is doubled in from 7 to 17 hours). It may be mentioned that 2 months after conception the rate of growth in man is only 8 per cent per day. This is contrary to all the published statements. Thus, Minot concluded that growth begins at 1000 per cent per day; Jackson concluded that in man, growth during the 1st month takes place at 57.5 million per cent per month; during the 2nd month 990 per cent per month; during the 3rd month 390 per cent per month (8 per cent per day is only 240 per cent per month). The reason for the discrepancy between the values derived, by the method adopted by the writer, and the values given in the literature is explained by Fig. 1.     

Flower Formation in Excised Tobacco Stem Segments: III. Deoxyribonucleic Acid Content in Stem Tissue of Vegetative and Flowering Tobacco Plants

A method has been developed that extracts DNA from stem tissue of flowering tobacco plants, Nicotiana tabacum cv. Wis. 38. The DNA content of stem tissue from a flowering tobacco plant is correlated with its capacity to flower in vitro. Stem segments known to form 100% floral buds contain 10 times more DNA per gram fresh weight than segments that form 5% floral buds and 95% vegetative buds, and in the uppermost 28 centimeters of flowering tobacco plant stems the DNA content decreases roughly in parallel with the floral gradient.     

Homeotic gene function in the muscles of Drosophila larvae

The segmental musculature of Drosophila melanogaster larvae consists of 24-30 muscles per segment. Unique patterns of muscles are found in the three thoracic segments and the first and last abdominal segments; the remaining abdominal segments share the same pattern. Mutations in Ultrabithorax (Ubx) cause partial transformation of the muscle pattern of larval abdominal segments towards metathorax. The muscles of the thorax are not affected. In the first two abdominal segments the changes include the loss of at least 11 `abdominal' muscles and the gain of 11 `thoracic' muscles. Less extensive transformations are seen in more posterior abdominal segments. Anterobithorax, bithorax, postbithorax and bithoraxoid mutations also induce transformations of the larval musculature. Each allelic group affects a domain that is a subset of the entire Ubx domain but these domains are not restricted to compartments or segments and may extend through as many as five segments. In the muscles the segmental distribution of Ubx antigen correlates with the segments affected by Ubx mutations. The different domains of Ubx in mesoderm and ectoderm argue that the segmental diversity of the muscle pattern is not simply induced by the overlying epidermis and that Ubx function in the mesoderm is required for the correct development of abdominal segments.     

Encapsulation, cold storage, and growth of Hibiscus moscheutos nodal segments

Nodal segments of Hibiscus moscheutos (hardy hibiscus) were excised from proliferating axillary shoot cultures and encapsulated in high density sodium alginate hardened by 50 mM CaCl₂. Nodal segments 4 mm long grew as well as and were easier to encapsulate than 8 mm long nodal segments. Although nodal segments grew regardless of the concentration of sodium alginate, 2.75% was determined to produce the highest quality encapsulated nodal segments beads (sufficient alginate coating and ease of use) because of the viscosity produced by the 2.75% sodium alginate solution. When encapsulated segments were stored at 5°C they did not grow in light or darkness. During the first month on fresh proliferation medium under normal incubation conditions following 5°C storage in the dark for up to 24 weeks, root number and root and shoot elongation were inhibited linearly as storage time increased. All encapsulated nodal segments survived 24 weeks of 5°C storage in two separate experiments. In fact, 80% of encapsulated hardy hibiscus nodal segments survived refrigerated storage for 1½ years (78 weeks) and after 3 months on proliferation medium, the nodal segments produced nearly the same length axillary shoots with the same number of axillary nodes per shoot as compared to encapsulated segments either not stored at 5°C or stored for 24 weeks at 5°C. Growth from encapsulated and cold-stored ‘Lord Baltimore’ nodal segments was more vigorous than from ‘Southern Belle’ nodal segments.     

Micropropagation of Mandevilla moricandiana (A.DC.) Woodson

A protocol was developed for micropropagation of Mandevilla moricandiana (A.DC.) Woodson, a native plant from Brazil. Shoots, obtained from in vitro plantlets were used as source of nodal segments for shoot production from axillary buds. The nodal segments were grown on Murashige and Skoog medium supplemented with different concentrations of 6-benzyladenine and/or indole-3-acetic acid to induce axillary bud elongation. After a 2-mo culture period, the medium supplemented with 1.0 mg?L^?1 6-benzyladenine gave the largest number of nodal segments per explant. The nodal segments obtained from plants developed under these conditions were grown on medium supplemented with different concentrations indole-3-acetic acid, ??-naphthaleneacetic acid, and indole-3-butyric acid. The use of the medium supplemented with indole-3-acetic acid and indole-3-buryric induced shoot elongation and shoot development, formation of basal callus, and/or indirect organogenesis of roots. Following transfer of shoots to soil, the plants with only basal callus showed 10% survival and developed roots from callus, while in vitro-rooted plants had a maximum 40% survival rate ex vitro. Regardless of the auxin added to the rooting medium, the acclimatization period allowed the plants rooted in vitro to develop their shoots fully. The protocol developed here is suitable for the production of shoots and rooted plantlets of M. moricandiana.     

The impact of internodal segmentation in biophysical nerve fiber models

Implementation of double cable models to simulate the behavior of myelinated peripheral nerve fibers requires defining a segmentation of the internode between successive nodes of Ranvier. The number of internodal segments is a model parameter that is not well agreed on, with values in the literature ranging from 1 to more than 500. Moreover, a lot of studies also lack a sensitivity study or a rationale behind the implementation used. In a model of a myelinated nerve fiber developed in our group, the segmentation scheme (i.e., the number of segments and their individual morphology) strongly influenced model outcomes such as action potential shape and velocity, stimulation threshold and absolute refractory period. In the present study these influences were investigated systematically in homogeneous neurons with different diameters. Uniformly segmented internodes were found to require several hundreds of segments (and associated computational power) to reach model outcomes differing by less than 1 % from the asymptotic value. In fact, in the majority of segmentation schemes the main determinant is not the number of segments, but the length λ of the internodal segments directly adjacent to the nodes of Ranvier. If λ is larger than approximately 10 μm, model outcomes for the tested fibers are almost independent of the total number of segments. Furthermore, λ can be optimized to enable models using just three segments per internode, to reach physiologically relevant model outcomes with limited computational resources. However, to study anatomical or physiological details of the internode itself, an appropriately detailed segmentation scheme is crucial.     

Effect of auxins and cytokinins on plant regeneration from hypocotyls and cotyledons in niger (Guizotia abyssinica Cass.)

Tissue cultures were established from hypocotyl and cotyledonary leaf segments ofGuizotia abyssinica Cass. on MS medium supplemented with various concentrations of auxins (IAA, NAA, IBA or 2,4-D) and cytokinins (KN or BA). Expiants cultured on media with cytokinins or in combination with auxins produced shoot buds. Maximum number of shoot buds (20–25 per culture) were differentiated from cotyledonary leaf segments on medium with 2 mg 1^-1 each of KN and IBA. Rooting of regenerated shoot buds was acheived on medium with NAA. The obtained plantlets were successfully transferred to soil.     

Effect of Ethylene Treatment on Polar IAA Transport, Net IAA Uptake and Specific Binding of N-1-Naphthylphthalamic Acid in Tissues and Microsomes Isolated from Etiolated Pea Epicotyls

The effect of ethylene treatment on polar indole-3-acetic acid (IAA) transport, net IAA uptake in the presence and absence of N-1-naphthylphthalamic acid (NPA) and [³H]NPA binding characteristics was investigated in tissue segments or microsomes isolated from etiolated pea (Pisum sativum L. cv Alaska) epicotyls. Basipetal IAA transport in 5 millimeter segments isolated from ethylene-treated seedlings was inhibited by ethylene in a dose-dependent manner. Threshold, half-maximal and saturating concentrations of ethylene were 0.01, 0.55, 10.0 microliters per liter, respectively. This inhibition became apparent after 6 to 8 hours of ethylene treatment. Transport velocity in both control and ethylene-treated tissues was estimated to be 5 millimeters per hour. Net IAA uptake was stimulated in ethylene-treated tissues and the relative ability of the phytotropin NPA to enhance net IAA uptake was reduced in treated tissues. Specific binding of [³H]NPA to microsomes prepared from both control and ethylene-treated tissues was saturable and consistent with the existence of a single class of binding sites with an apparent affinity (K_d) toward NPA of 8 to 9 nanomolar. The density of these binding sites (per milligram protein) was lower (36% of control) in ethylene-treated tissues. Direct application of ethylene to microsomal preparations isolated from untreated seedlings had no effect on the level of specific [³H]NPA binding.     

Spatial distributions of potassium, solutes, and their deposition rates in the growth zone of the primary corn root

Densities of osmoticum and potassium were measured as a function of distance from the tip of the primary root of Zea mays L. (cv WF9 × mo17). Millimeter segments were excised and analyzed for osmotic potential by a miniaturized freezing point depression technique, and for potassium by flame spectrophotometry. Local deposition rates were estimated from the continuity equation with values for density and growth velocity. Osmotic potential was uniform, −0.73 ± 0.05 megapascals, throughout the growth zone of well-watered roots. Osmoticum deposition rate was 260 μosmoles per gram fresh weight per hour. Potassium density fell from 117 micromoles per gram in the first mm region to 48 micromoles per gram at the base of the growth zone. Potassium deposition rates had a maximum of 29 micromoles per gram per hour at 3.5 millimeters from the tip and were positive (i.e. potassium was being added to the tissue) until 8 millimeters from the tip. The results are discussed in terms of ion relations of the growing zone and growth physics.     

Rapidly Induced Wound Ethylene from Excised Segments of Etiolated Pisum sativum L., cv. Alaska: I. Characterization of the Response

A rapidly induced, transitory increase in the rate of ethylene synthesis occurred in wounded tissue excised from actively growing regions of etiolated barley, cucumber, maize, oat, pea, tomato, and wheat seedlings. Cutting intact stems or excising 9-mm segments of tissue from near the apex of 7-day-old etiolated Pisum sativum L., cv. Alaska seedlings induced a remarkably consistent pattern of ethylene production. At 25 C, wound-induced ethylene production by segments excised 9 mm below the apical hook increased linearly after a lag of 26 minutes from 2.7 nanoliters per g per hour to the first maxium of 11.3 nanoliters per g per hour at 56 minutes. The rate of production then decreased to a minimum at 90 minutes, increased to a lower second maximum at 131 minutes, and subsequently declined over a period of about 100 minutes to about 4 nanoliters per g per hour. Removal of endogenous ethylene, before the wound response commenced, had no effect on the kinetics of ethylene production. Tissue containing large amounts of dissolved ethylene released it as an exponential decay with no lag period. Rapidly induced wound ethylene is synthesized by the tissue and is not merely the result of facilitated diffusion of ethylene already present in the tissue through the newly exposed cut surfaces. Previously wounded apical sections did not exhibit a second response when rewounded. No significant correlation was found between wound-induced ethylene synthesis and either CO₂ or ethane production.     

Age and orientation of the cotyledonary leaf explants determine the efficiency of de novo plant regeneration and Agrobacterium tumefaciens-mediated transformation in Jatropha curcas L.

Effects of age and orientation of the explant on callus induction and de novo shoot regeneration from cotyledonary leaf segments of Jatropha curcas were studied. The callus induction and shoot regeneration capacity of cotyledonary leaf segments were found significantly related to the age of the explants and their orientation in culture medium. The youngest explant, derived from the cotyledonary leaf of germinated seed induced the highest regeneration response as compared to one- and two-week-old explants. A gradient response with age of the explant was observed in percentage of callus induction, shoot regeneration from callus and the number of shoots per regenerating callus. The explants cultured with their abaxial side in medium showed significantly higher regeneration response. The youngest explant was found to be most amenable to Agrobacterium-mediated transformation as compared to older explants. The fact that callus induced from the edges of the explant followed by de novo shoot induction, and strong transient gus expression observed in the edges of the explant are significant for routine Agrobacterium-mediated transformation and generation of stable transgenic plants in J. curcas.     

Inhibition of stem elongation in cucumis seedlings by blue light requires calcium

The effects of blue light and calcium on elongation of hypocotyl segments of Cucumber (Cucumis sativa L. cv Burpee's Pickler) were studied. Cucumber seedlings grown in dim red light showed a rapid decline in the rate of hypocotyl elongation when irradiated with high intensity (100 micromoles per square meter per second) blue light. In intact, 4-day-old seedlings the inhibition began within 2 minutes after the onset of blue-light irradiation and reached a maximum of approximately 55% within 4 minutes. Hypocotyl segments cut from 4-day-old seedlings also showed an inhibition of elongation in response to blue light when segments were floated on aqueous buffer and exposed to blue light for 3 hours. In the presence of 2 micromolar indole-3-acetic acid, blue light caused a 50% inhibition of elongation. Buffering free calcium in the incubation medium with 0.1 millimolar ethylene glycol bis(-aminoethyl ether)- N,N,N′,N′-tetraacetic acid eliminated the blue-light inhibition of segment elongation. Several experiments confirmed a specific requirement for calcium for the blue-light-induced inhibition of segment elongation. Treating segments with 0.2 micromolar fusicoccin abolished the inhibition of elongation by blue light as did buffering the medium at pH 4. Adding 1 millimolar ascorbate to incubation medium also eliminated the inhibition of segment elongation caused by blue light. Several compounds implicated in cell-wall redox reactions alter the magnitude of the blue-light-induced inhibition. The activity of peroxidase isolated from the cell-wall free space of cucumber hypocotyls was inhibited by ascorbate and low pH. The results are consistent with the hypothesis that blue light inhibits elongation by inducing an increase in cell-wall peroxidase activity and implicate calcium ions in the response to blue light.     

Intestinal remodelling in mink fed with reduced protein content

Low protein intake occurs in humans in relation to diseases, starvation and post-operatively. Low-protein diets may affect the gastrointestinal structure and mechanical function. The aim was to study the passive biomechanical properties and tissue remodelling of the intestine in minks on reduced protein diets. Twenty-seven male minks were divided into three groups receiving different protein level in the diet for 6 weeks: High protein level (group H, 55% energy from protein), moderate protein level (group M, 30% energy from protein) and low protein level (group L, 15% energy from protein) (n=9 for each group). Ten centimetre long segments from duodenum, jejunum and ileum were excised at the end of the study period. The mechanical test was performed as a ramp distension experiment. The intestinal diameter and length, wall thickness, wall area and opening angle were obtained from digitized images of the intestinal segments at pre-selected pressures, no-load and zero-stress states, respectively. Circumferential and longitudinal stresses (force per area) and strains (deformation) were computed. The layer thickness was measured from intestinal histological images. No difference in body weight was found between groups at the start of the experiment. However, at the end of the experiment the body weight was smallest in group L (P=0.0003 and 0.0004 compared with groups H and M). Similarly, the wet weight per unit length, wall thickness and area were smallest in group L (P<0.05, P<0.01). The lowest wall thickness was found in the jejunum and ileum in group L (P<0.05), mainly due to decreased mucosa and submucosa thickness. The smallest opening angle and absolute values of residual strain were found in the jejunal segment in group L (P<0.05). No difference was observed for duodenal and ileal segments among the three groups. Feeding the low-protein diet shifted the stress–strain curves to the right for the circumferential direction, indicating the wall become softer in the circumferential direction. However, no significant difference was observed in the longitudinal direction for any of the intestinal segments. In conclusion, this study demonstrated that low-protein diet in minks induce histomorphometric and biomechanical remodelling of the intestine.     

Shoot regeneration and cryopreservation of shoot tips of apple (Malus) by encapsulation–dehydration

Here, we report an efficient and widely applicable method for cryopreservation of Malus shoot tips by encapsulation–dehydration using adventitious shoots. Shoots were induced from leaf segments cultured on a shoot induction medium containing 2–3 mg L^?1 thidiazuron, depending on genotype, and 0.5 mg L^?1 indole-3-butyric acid. Shoot tips (3 mm in length) containing six leaf primordia excised from 11-wk-old adventitious shoots were encapsulated and precultured with 0.5 M sucrose for 5 d, followed by air-drying for 6 h prior to direct immersion in liquid nitrogen. With our protocol, we obtained a mean organogenesis rate of 100%, a mean of 4.5 adventitious shoots per explant (leaf segment), and a mean shoot recovery of 57.0% from cryopreserved shoot tips in four Malus species. Inter-simple sequence repeat (ISSR) analysis did not reveal any polymorphic bands in regenerants recovered from either leaf segments or cryopreserved shoot tips of ‘Gala’. To the best of our knowledge, this is the first report on cryopreservation of Malus shoot tips using adventitious shoots derived from leaf segments and is the most widely applicable protocol so far reported for cryopreservation of Malus. Establishment of this protocol provides an alternative means for cryopreservation of Malus.     

In Vitro Regeneration of Achillea millefolium L from Shoot-tips and Root Segments of Seedlings

This report describes, for the first time, an efficient plant regeneration system for Achillea millefolium L (yarrow), a medicinal plant, via shoot multiplication from shoot-tips and adventitious shoot regeneration from root segments. Higher numbers of shoots were obtained when shoot-tips were cultured on MSMO medium supplemented with 3.0 mg l^?1 BA and 0.5 mg l^?1 IAA, or 5.0 mg l^?1 KIN and 1.0 mg l^?1 IBA, producing 17.3 and 17.0 shoots per explant at 100% frequency, respectively. For adventitous shoot regeneration, only root segments developed shoots when cultured on medium containing a combination of 1 mg l^?1 TDZ, 0.5 mg l^?1 IAA and 0.5 mg l^?1 GA3 (18.9 shoots per explant at 100% frequency), while other types of explants (i.e., cotyledons, leaf lamina and petiole segments) or hormonal combinations tested were found ineffective. Regenerated shoots rooted readily on MSMO medium containing different concentrations of IAA, IBA, NAA or 2,4-D, however, NAA at 0.5 mg l^?1, or IBA at 0.5 or 1.0 mg l^?1 were found to be the most productive. Nearly all of the regenerated plants (98%) survived through the hardening process when the rooted plantlets were kept at 55–65% relative humidity for 2 weeks, which were then planted in pots containing potting soil and kept at 25–35% humidity.     

Micropropagation of Zeyheria montana Mart. (Bignoniaceae), an endangered endemic medicinal species from the Brazilian cerrado biome

Zeyheria montana Mart. has become endangered, primarily because of deforestation of its habitat, its use as a medicinal plant extract, and the strong endemism of the species. In this study, an efficient protocol was established for the micropropagation and conservation of Z. montana germplasm using isolated mature zygotic embryos as explants. Embryos germinated in vitro 4 d after isolation and inoculation on modified Murashige and Skoog (MS) medium containing 2.0 mg/l gibberellic acid (GA₃). The addition of GA₃ also improved the germination index and accelerated the process of germination. Nodal segments from seedlings were placed on modified ¼-strength MS medium containing 0.1 mg/l 6-benzyladenine and 0.5 mg/l GA₃. Nodal segments produced 7.3 shoots per explant within 60 d. Following transfer of shoots to a medium containing 1.5 mg/l indole-3-butyric acid, roots formed. All plantlets obtained were successfully acclimatized under greenhouse conditions, and approximately 68.5 acclimatized plants could be obtained per seed each year. This protocol provides a method to preserve this rare and endangered medicinal plant.     

Saturable uptake of indol-3yl-acetic Acid by maize roots

The uptake of 5-[³H]indol-3yl-acetic acid (IAA^*) by segments of Zea mays L. roots was measured in the presence of nonradioactive indol-3yl-acetic acid (IAA°) at different concentrations. IAA uptake was found to have a nonsaturable component and a saturable part with (at pH 5.0) an apparent K_m of 0.285 micromolar and apparent V_max 55.0 picomoles per gram fresh mass per minute. These results are consistent with those which might be expected for a saturable carrier capable of regulating IAA levels. High performance liquid chromatography analyses showed that very little metabolism of IAA^* took place during 4 minute uptake experiments. Whereas nonsaturable uptake was similar for all 2 millimeter long segments prepared within the 2 to 10 millimeter region, saturable uptake was greatest for the 2 to 4 millimeter region. High levels of uptake by stelar (as compared with cortical) segments are partly attributable to the saturable carrier, and also to a high level of uptake by nonsaturable processes. The carrier may play an essential role in controlling IAA levels in maize roots, especially the accumulation of IAA in the apical region. The increase in saturable uptake toward the root tip may also contribute to the acropetal polarity of auxin transport.