Regeneration of Triticum aestivum apical explants after microinjection of germ line progenitor cells with DNA

A highly efficient method of regenerating fertile, phenotypically normal plants from shoot apex cultures of T. aestivum was developed. The hypodermal layer (L2) of the vegetative apex containing germ line precursor cells could be located with bright field microscopy and targeted for microinjection. Fluorescently labelled dextrans were used as markers to develop a microinjection procedure which did not disrupt nuclear or cytoplasmic structure. This procedure was used to inject plasmid DNA into L2 cells. Capillary microinjection did not shear the plasmid DNA and delivery of DNA was confirmed by polymerase chain reaction analysis of DNA isolated from injected apices. The significance of these findings in relation to the development of cereal transformation systems will be discussed.     

Somatic and Dendritic Encoding of Spatial Variables in Retrosplenial Cortex Differs during 2D Navigation

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Nauclea diderrichii: rooting of stem cuttings,clonal variation in shoot dominance,and branch plagiotropism

Summary Nauclea diderrichii (De Wild, and Th. Dur.) Merill (Rubiaceae), an indigenous hardwood of West Africa, is increasingly being grown commercially. This study investigates the potential for vegetative propagation and clonal selection, and raises some fundamental questions about the physiology of apical dominance and of plagiotropism. Rooting ability was high, with up to 100% rooting in 2–4 weeks, when different Indole-3-butyric acid (IBA) concentrations and leaf areas were tested. Auxin applications greatly increased the numbers of roots per cutting. The decapitation of unbranched plants revealed clonal variation in apical dominance and also in the establishment of outright dominance by the two shoots formed from the outgrowth of the axillary buds of the opposite leaves at the top node. Regression analysis of the Dominance Ratio (length of dominant: length of the sub-dominant shoot at the time of achieving dominance) against overall lateral bud activity (r = 0.82), showed that when the two top shoots co-dominate they provide a more powerful source of Correlative Inhibition than when one of the top shoots dominates the other. The imposition of plagiotropism in the axillary bud occurred over a period of a few days as the terminal and axillary buds emerged from the stipule. Growth of accessory buds on intact plants and debranched cuttings was orthotropic. These results are discussed with regard to the role of the leaf in root formation and the understanding of dominance relationships, branching and crown development in trees.     

Compensatory growth in shoot populations of young white pine trees

Summary White pine (Pinus strobus L.) trees have shoot populations composed of subpopulations of terminal and lateral shoots. I tested whether the subpopulations would show compensatory (increased) growth when separated from each other. Ten-year-old white pine (Pinus strobus L.) trees growing under an oak (Quercus) overstory were untreated or treated in winter by removing either all terminal, or all lateral buds (10 trees per treatment). Growth was compared between control and treated shoot subpopulations. In the 1st year, shoot-length frequency distributions were similar between control and treated subpopulations. There was significant compensatory shoot elongation (mean of 1.5 cm per shoot) in both treated subpopulations. In the 2nd year each subpopulation produced both terminal and lateral shoots. Shoot-length frequency distributions were similar, but shifted toward longer shoots in treated populations. Shoot number, mean length and total shoot length were greater in treated populations. The increased growth in treated subpopulations was due both to differences in parent shoot length and to compensatory shoot production and elongation.     

Dual effect of light on flowering and sprouting of rose shoots

Shade, caused by a dense leaf canopy in the light conditions of a normal greenhouse, reduced sprouting of the third axillary bud (from the top) on decapitated rose branches ( Rosa hybrida cv. Marimba) in comparison to less shaded buds on branches protruding above the canopy and sparsely spaced. Flowering of the third young shoot on shaded branches bearing 3 lateral shoots was totally inhibited. Mixed fluorescent and incandescent light in a growth chamber reduced sprouting of the third bud on decapitated rose branches in comparison to decapitated branches on rose plants held in fluorescent light of similar photon flux density. This was attributed to the higher R:FR ratio in fluorescent vs mixed light that reached the third bud, and in exposed vs shaded branches. Flowering of the third shoot was promoted by several factors: high photon flux density, 0.5 m M gibberellic acid (GA) or 0.2 m M benzyladenine (BA). BA was the most effective treatment. Treatments promoting flowering of the third shoot did not reduce growth or flowering of the upper shoots. However, spraying the uppermost shoot with BA suppressed the growth of the shoots below. It is concluded that light affects flowering in two ways. The effect on bud sprouting is related mainly to R:FR ratios, while the effect on flower development is related mainly to photon flux density. Cytokinins may substitute for the light effect on flower development.     

Consequences of herbivory in the mountain birch (Betula pubescens ssp tortuosa): importance of the functional organization of the tree

Summary Three types of experiments indicate that the functional organization of the mountain birch may influence the ways in which the tree responds to simulated or natural herbivory. The first experiment showed that herbivory to both short and long shoot leaves affects plant development but, because growth largely proceeds by resources of the previous year, is manifested only in the year following the damage. The second experiment showed that even partial damage to a single long shoot leaf caused the axillary bud of that leaf to produce a shorter shoot the next year. Therefore, the value of a leaf depends also on the organ which it is subtending. In the third experiment we manipulated the apical dominance of shoots in ramets and caused improvement to leaf quality in extant shoots. Ramets within a tree responded individually, probably mediated by disturbance of the hormonal control because removal of apical buds elicited the response although removal of the same number of basal buds did not. Induced amelioration is a different response to induced resistance. The two responses are triggered by different cues and may occur in the same plant. By altering hormonal balance of shoots it is potentially possible for herbivores to induce amelioration of food quality. The ways in which herbivory is simulated may explain variability of results obtained when herbivory-induced responses in plants have been studied.     

Ca-sensitive sodium absorption in the colon of Xenopus laevis

Summary Transepithelial electrogenic Na transport (I_Na) was investigated in the colon of the frog Xenopus laevis with electrophysiological methods in vitro. The short circuit current (I_sc) of the voltage-clamped tissue was 24.2±1.8 A·cm^-2 (n=10). About 60% of this current was generated by electrogenic Na transport. Removal of Ca²⁺ from the mucosal Ringer solution stimulated I_Na by about 120%. I_Na was not blockable by amiloride (0.1 mmol·l^-1), a specific Na-channel blocker in epithelia, but a fully and reversible inhibition was achieved by mucosal application of 1 mmol·l^-1 lanthanum (La^3-). No Na-self-inhibition was found, because I_Na increased linearly with the mucosal Na concentration. A stimulation of I_Na by antidiuretic hormones was not possible. The analysis of fluctuations in the short circuit current (noise analysis) indicated that Na ions pass the apical cell membrane via a Ca-sensitive ion channel. The results clearly demonstrate that in the colon of Xenopus laevis Na ions are absorbed through Ca-sensitive apical ion channels. They differ considerably in their properties and regulation from the amiloride-sensitive Na channel which is typically found in the colon of vertebrates.Abbreviations G _T transepithelial conductance - I _sc short circuit current - I _Na transepithelial Na-current - m mucosal - s serosal - PDS power density spectrum - f frequency - f _c corner frequency of the Lorentzian component of the PDS - S(f) power density of the Lorentzian component of the PDS - S_o plateau value of the Lorentzian component of the PDS     

Evidence that cytokinin controls bud size and branch form in Norway spruce

Shoot elongation in many coniferous species is predetermined during bud formation the year before the shoot extends. This implies that formation of the primordial shoot within the bud is the primary event in annual shoot growth. Hormonal factors regulating bud formation are consequently of utmost importance. We followed the levels of the endogenous cytokinins zeatin riboside (ZR) and isopentenyladenosine (iPA) in terminal buds, whorl buds and lower lateral buds of the uppermost current-year whorl shoots of 15- to 20-year-old trees of Norway spruce [ Picea abies (L.) Karst.] from June to September. Cytokinins were isolated with affinity chromatography columns, purified by high performance liquid chromatography, and quantified by ELISA. The level of ZR was low in June but increased gradually in all buds until September. Throughout the measurement period, the ZR level was highest in terminal buds and lowest in the scattered lateral, buds, with the whorl buds intermediate. The level of iPA peaked in July and decreased later without any consistent differences among the three classes of buds. The development of different kinds of buds was followed by scanning electron microscopy. We found that bud growth was greatest during August and September. The final size of primordial shoots within the buds varied considerably and the weight of the terminal bud was three times that of the whorl buds and more than five times that of the other lateral buds.
We conclude that the increase in ZR level during the period of active bud development is indicative of the importance of cytokinin for this process. Furthermore, the positive correlation between the level of ZR and bud growth during the period of predetermination of next year's branch growth suggests that this hormone indirectly controls the form of single branches in the spruce tree.     

Mutations associated with floral organ number in rice

How floral organ number is specified is an interesting subject and has been intensively studied in Arabidopsis thaliana. In rice (Oryza sativa L.), mutations associated with floral organ number have been identified. In three mutants of rice, floral organ number 1 (fon1) and the two alleles, floral organ number 2-1 (fon2-1) and floral organ number 2-2 (fon2-2), the floral organs were increased in number centripetally. Lodicules, homologous to petals, were rarely affected, and stamens were frequently increased from six to seven or eight. Of all the floral organs the number of pistils was the most frequently increased. Among the mutants, fon1 showed a different spectrum of organ number from fon2 -1 and fon2 -2. Lodicules were the most frequently affected in fon1, but pistils of more than half of fon1 flowers were unaffected; in contrast, the pistils of most flowers were increased in fon2 -1 and fon2-2. Homeotic conversion of organ identity was also detected at a low frequency in ectopically formed lodicules and stamens. Lodicules and stamens were partially converted into anthers and stigmas, respectively. Concomitant with the increased number of floral organs, each mutant had an enlarged apical meristem. Although meristem size was comparable among the three mutants and wild type in the early phase of flower development, a significant difference became apparent after the lemma primordium had differentiated. In these mutants, the size of the shoot apical meristem in the embryo and in the vegetative phase was not affected, and no phenotypic abnormalities were detected. These results do not coincide with those for Arabidopsis in which clavatal affects the sizes of both shoot and floral meristems, leading to abnormal phyllotaxis, inflorescence fasciation and increased floral organs. Accordingly, it is considered that FON1 and FON2 function exclusively in the regulation of the floral meristem, not of the vegetative meristem.Abbreviation DIC differential interference contrast This work was supported in part by Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Science and Culture of Japan.     

Maxi K+ channels on human vas deferens epithelial cells

The vas deferens forms part of the male reproductive tract and extends from the cauda epididymis to the prostate. Using the patch clamp technique, we have identified a Ca²⁺-activated, voltage-dependent, maxi K⁺ channel on the apical membrane of epithelial cells cultured from human fetal vas deferens. The channel had a conductance of 250 pS in symmetrical 140 mm K⁺ solutions, and was highly selective for K⁺ over Na⁺. Channel activity was increased by depolarization and by an elevation of bath (cytoplasmic) Ca²⁺ concentration, and reduced by cytoplasmic Ba²⁺ (5 mm) but not by cytoplasmic TEA (10 mm). Channel activity was also dependent on the cation bathing the cytoplasmic face of the membrane, being higher in a Na⁺-rich compared to a K⁺-rich solution. We estimated that up to 600 maxi K⁺ channels were present on the apical membrane of a vas cell, and that their density was 1–2 per ² of membrane. Activity of the channel was low on intact cells, suggesting that it does not contribute to a resting K⁺ conductance. However, fluid in the lumen of the human vas deferens has a high K⁺ concentration and we speculate that the maxi K⁺ channel could play a role in transepithelial K⁺ secretion.Funded by grants from the Cystic Fibrosis Trust and the Medical Research Council (UK). We thank Mr. David Stephenson for excellent technical assistance.