Ethylene biosynthesis in Regnellidium diphyllum and Marsilea quadrifolia

The pathway of ethylene biosynthesis was examined in two lower plants, the semi-aquatic ferns Regnellidium diphyllum Lindm. and Marsilea quadrifolia L. As a positive control for the ethylene-biosynthetic pathway of higher plants, leaves of Arabidopsis thaliana (L.) Heynh. were included in each experiment. Ethylene production by Regnellidium and Marsilea was not increased by treatment of leaflets with 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene in higher plants. Similarly, ethylene production was not inhibited by application of aminoethoxyvinylglycine and -aminoisobutyric acid, inhibitors of the ethylene biosynthetic enzymes ACC synthase and ACC oxidase, respectively. However, ACC was present in both ferns, as was ACC synthase. Compared to leaves of Arabidopsis, leaflets of Regnellidium and Marsilea incorporated little [¹⁴C]ACC and [¹⁴C]methionine into [¹⁴C]ethylene. From these data, it appears that the formation of ethylene in both ferns occurs mainly, if not only, via an ACC-independent route, even though the capacity to synthesize ACC is present in these lower plants.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AdoMet S-adenosyl-l-methionine - AIB -aminoisobutyric acid - AVG aminoethoxyvinylglycine This research was supported by the U.S. Department of Energy through grant No. DE-FG02-91ER20021 and, in part, by a fellowship of the National Engineering and Research Council of Canada to Jacqueline Chernys.     

Giant Stomata in Regnellidium diphyllum Lind.

The occurrence of abnormally large stomata on the pinnae ofRegnellidium diphyllum is reported. The plants were grown underin vitro conditions in Knop's sugar medium. These stomata aredifferent from the normal ones, both in size as well as shape,but structurally similar to them and probably functioning inthe same way.     

Zeta potential measurements of cell wall preparations from Regnellidium diphyllum and Nymphoides peltata

Abstract. Cell wall particles were prepared from the semi-aquatic plants Regnellidium diphyllum and Nymphoides peltata with minimum disruption to the integrity of the cell wall. The behaviour of freshly-prepared and frozen-thawed particles in a D.C. electric field was monitored with a microscope attached to video recording apparatus. From the respective particles mobility in a well-defined electric field. it was possible to determine their electrostatic potential and consequently estimate the corresponding surface charge density. Experiments were performed in media of different pH and cation concentration (ie, K⁺ Ca²⁺). A significant electronegative potential was found in cell wall preparations of both plants. Freezing and thawing further reduced the electrostatic potential for both plant species in all the media utilized for electrophoresis. A reduction of pH or an increase of the cation concentration was found to neutralize the electrostatic potential in a sigmoidal fashion. Ca²⁺ was more than 10 times more effective than K⁺ at neutralizing the apparent electrostatic potential of the cell wall preparations. Regnellidium was found to have a lower electrostatic potential than Nymphoides , although both responded in a similar manner to the various treatments. The possible relevance of the cell wall electrostatic potential, pH and [Ca²⁺] and particularly their inter-relationship is discussed for the two species of plants in terms of their differing growth responses to the ionic environment of the plant.     

Cell Expansion and Wall pH in the Fern Regnellidium diphyllum, A Plant Lacking Acidinduced Growth

Petioles of the semi-aquatic fern Regnellidium diphyllum donot show acid growth but low wall pH is a necessary conditionfor maximum rates of IAA-induced cell expansion. Measurementsof wall pH by two indirect methods indicate an unusually lowvalue, in the range pH 4 to 5. This is one to two pH units belowthat estimated for petioles of the semi-aquatic dicotyledonNymphoides peltata, a species in which IAA aand ethylene causegrowth responses very similar to those in Regnellidium but whereacid growth occurs. Having shown previously that fusicoccinenhances proton secretion in both Regnellidium and Nymphoides,we now show that although it causes a reduction in the estimatedapoplast pH to below 4·0 in Regnellidium, cell expansionis not promoted. The FC-induced reduction in pH in Nymphoidesis less and occurs more slowly, but growth is promoted significantly;when IAA and fusicoccin are present together, growth promotionis approximately additive for Nymphoides A model is proposed for Regnellidium in which equilibrium wallpH is maintained at a low value that is optimal for acid growth,the availability of acid-labile sites in the wall being thechief limitation to cell extension. We suggest that this controlmechanism may be widespread for organs without a cuticle, includingroots and the gametophytes of lower plants growing in acidicconditions. Key words: Acid growth, wall pH, fusicoccin, Regnellidium diphyllum, Nymphoides peltata     

The effect of ethylene and auxin on cell wall extensibility of the Semi-aquatic fern,Regnellidium diphyllum

Ethylene and auxin both enhance cell elongation growth in the rachis of the frond of Regnellidium diphyllum. Measurements of the stress relaxation modulus of the walls of methanol-killed rachis segments show that both auxin and ethylene cause an increase in cell wall extensibility, that the effects are additive, and that they occur in the presence of hypertonic solutions of mannitol that preclude cell elongation. The results are taken as evidence for the operation of two separate mechanisms for cell wall loosening.Abbreviation IAA indol-3yl-acetic acid     

Ethylene and auxin-induced cell growth in relation to auxin transport and metabolism and ethylene production in the semi-aquatic plant,Regnellidium diphyllum

Cell elongation in the rachis of the semiaquatic fern Regnellidium diphyllum is induced by the addition of ethylene or indoleacetic acid (IAA). Experiments with whole plants or rachis segments have shown that ethylene-induced growth requires the presence of auxin. Ethylene does not cause a modification in either endogenous auxin levels or in the extent of auxin metabolism but auxin transport is reduced. Rates of ethylene production in Regnellidium are not altered by either mechanical excitation or by the addition of auxin. A two-hormone control of cell expansion is proposed in which an initial, auxin-dependent growth event pre-conditions the cells to a further subsequent (or synchronous) ethylene-dependent growth event.Abbreviation IAA indole-3yl-acetic acid     

The Formation and Structure of the Cell Wall in Fibres and Tracheids

A study has been made of the changes in organization of theprimary wall of xylem fibres and tracheids during the phaseof surface growth using both direct electronmicroscopic observationsand replica techniques. It has been demonstrated that the transversemicrofibril orientation on the inner surface of the cell walldiffers from that on the outer surface. It has been concludedthat the observations made are consistent with some form ofmulti-net mechanism of growth in these cells. Autoradiographicstudies were consistent with this conclusion. Measurements of the variation in path difference in fibres andtracheids undergoing secondary wall formation showed a maximumnear the centre of the cells decreasing towards the cell tips.From these observations and parallel autoradaographic studiesit has been concluded that secondary thickening begins nearthe centre of the cells and progresses towards the cell tips.Since all layers of the secondary wall are lamellated theseobservations imply that more lamellae are present near the centreof the cells than at their tips, so that as secondary thickening proceeds the concentric lamellae of microfibrils grow progressivelytowards the ends of the cells. The lamellae of the secondarywall appear to develop in close association with the limitingcytoplasmic membrane (the plasmalemma). No evidence was obtainedof the inclusion of cytoplasmic components in the developingsecondary wall. At the conclusion of secondary thickening thecytoplasmic membranes retract on to the inner surface of thesecondary wall, forming the so-called ‘terminal lamella’which shows some fibrillar texture. It has been concluded thatthis appearance reflects some intrinsic texture of the lamellaitself, rather than the presence of cellulose microfibrils.     

Sulla cariologia di Gennaria diphylla (Link) Parl. (Coeloglossum diphyllum Fiori)

Abstract

OBSERVATIONS ON THE CARYOLOGY OF GENNARIA DIPHYLLA (LINK) PARL. (COELOGLOSSUM DIPHYLLUM FIORI) — Some caryological characters of Gennaria diphylla (Link) Parl, have been here investigated. Chromosome counts from somatic metaphases show a 34 chromosomes diploid set; the resting nuclei have been classified as half-reticulated with chromocenters (« noyaux semi-reticulé à chromocentres » according to Delay's classification); in prophases we have noticed that chromocenters often appear like prochromosomes. The chromosome number 17 (2n = 34) is new for the Ophrydinae.

The interest of this research lies in the fact that the species here dealt with has been differently classified by various Authors.     

A critical reappraisal of the status of Crataegus palmstruchii Lindman (Rosaceae)

C palmstruchii Lindman does not justify specific or subspecific rank, as it cannot be recognised by an association of distinct characters. Its habitat is not separate from that of C. laevigata (Poiret) DC., and it does not occupy a distinct geographical area, hut occurs as part of a range of continuous variation in most populations of C. laevigata.     

Characterization of microsatellite loci for a barrenwort species (Epimedium diphyllum,Berberidaceae)

Microsatellites were identified and characterized from Epimedium diphyllum, a species of barrenworts, both attractive garden plants and valuable medicinal plants. Some Japanese species of Epimedium are threatened with extinction, and are listed in the Red Data Book of plants in Japan. Natural hybrid zones also have been reported among some taxa of Japanese Epimedium. We developed eight polymorphic microsatellite primers for population genetic analyses of E. diphyllum. The numbers of alleles per locus ranged from 10 to 20, with observed levels of heterozygosity between 0.85 and 1.00. These primer sets yielded amplification in the other three Japanese Epimedium. These markers will be valuable for conservation genetics, evolutionary biology, pharmacognostic study, and horticultural study of Epimedium.     

Nucleic acid and protein synthesis during lateral root initiation in Marsilea quadrifolia (Marsileaceae)

The pattern of DNA, RNA, and protein synthesis during lateral root initiation in Marsilea quadrifolia L. was monitored by autoradiography of incorporated of 3H-thymidine, 3H-uridine, and 3H-leucine, respectively. DNA synthesis was associated with the enlargement of the lateral root initial prior to its division. Consistent with histological studies, derivatives of the lateral root initial as well as the cells of the adjacent inner cortex and pericycle of the parent root also continued to synthesize DNA. RNA and protein synthetic activities were found to be higher in the lateral root initials than in the endodermal initials of the same longitudinal layer. The data suggest a role for nucleic acid and protein synthesis during cytodifferentiation of a potential endodermal cell into a lateral root initial.     

Vessels in Nelumbo (Nelumbonaceae)

Vessels have been previously reported in the roots (but not rhizomes) of Nelumbo on the basis of light microscopy. We have reinvestigated vessel occurrence in Nelumbo by means of scanning electron microscopy (SEM). On the basis of these studies, vessels are characteristic of root metaxylem. Pores in primary walls of pits of end walls of these vessels are various in size, but feature incomplete lysis of pit membranes, often with residual webs or threads of primary wall material, much as in vessels of primitive woody dicotyledons. In addition, we newly report occurrence of vessels in metaxylem of rhizomes; pores in these vessels are smaller and more confined than in those of roots. The present study offers not only data of possible use in determining the phylogenetic position of Nelumbonaceae, but also contributes evidence that vessel origin in Nelumbonaceae relates to habit and ecology. We conclude that organographic distribution of vessels in Nelumbo follows the patterns seen in monocotyledons, which, like Nymphaeaceae and other aquatics, have sympodial architecture.     

植物管状分子三维重建的树脂复型法

介绍了用于观察植物管状分子内部三维结构的树脂复型法及其具体的操作步骤和适用范围     

Vessels in the roots of Barclaya rotundifolia (Nymphaeaceae)

Scanning electron microscope (SEM) studies of paraffin-sectioned material of stems and roots of Barclaya rotundifolia Hotta revealed perforations on tracheary elements of roots, but not on those of stems. End walls of vessels are identical with lateral walls except for the presence of perforations. Perforations can only be clearly revealed with SEM, and this method is advocated for further study of tracheary elements of Nymphaeaceae in particular, and primary xylem of ∗∗∗angiosperms in general. Vessel presence may be related to the habitat of this species, which unlike other members of the Nymphaeaceae (sensu stricto) has only aerial leaves and a rhizomatous system that is not inundated for prolonged periods.     

Multilocus genetic structure at contrasted spatial scales of the endangered water fern Marsilea strigosa Willd. (Marsileaceae, Pteridophyta)

Marsilea strigosa (Marsileaceae, Pterydophyta) is a rare water fern found in the Mediterranean basin, in temporary flooded habitats only. We analyzed the level and the distribution of genetic variation at seven microsatellite loci, both at the Mediterranean scale and at a narrower scale within a highly fragmented French metapopulation. Genetic diversity among individuals within each pond suggests that M. strigosa reproduces predominantly through selfing. The very high population differentiation at the Mediterranean scale indicates that gene flow (if any) is highly restricted. Similar differentiation is also found at the scale of a single metapopulation. The distribution of multilocus genotypes suggests that the genetic variation in this species is maintained mainly through the interplay of mutation and low recombination.     

水青树特殊管胞的分布位置及其形态特征的研究

针对水青树（Tetracentron sinense）中一类特殊管胞进行较为全面的观察研究,判断细胞种类并分析维管组织输导机理及树木进化过程中的细胞演化规律。通过切片和解离技术,借助光学显微镜和电子显微镜对34年生水青树特殊管胞的分布位置和形态特征进行观察。结果表明：(1)特殊管胞在树木水平方向自内向外径向呈串排列,并贯穿年轮界限,多为一列,少数两列,且较为稀见。每个特殊管胞弦向左右两侧或单侧均与木射线细胞相连通。纵向上,特殊管胞单独或数个上下端接相连。(2)特殊管胞主要有以下3种类型：无端壁的纺锤形,有一个倾斜端壁,以及有两个倾斜端壁。特殊管胞的平均长度为286.44μm;横切面为四边形,平均弦向宽度为55.22μm,其平均壁厚为1.53μm。(3)特殊管胞两端封闭,无穿孔。(4)特殊管胞侧面壁上的纹孔数量较多且纹孔膜明显可见,具体表现为：弦面壁上布满特殊管胞之间的具缘纹孔,呈对列、互列偶见梯状排列;径面壁上存在与射线细胞间的具狭缘单纹孔,呈大圆形至椭圆形,每区域多为2～10个纹孔,呈1～4排横列;径面壁上与正常管胞间几无纹孔。水青树特殊管胞分布有一定规律,其长度远小于水青树正常管胞...     

Collapse of Water-Stress Emboli in the Tracheids of Thuja occidentalis L

We report the kinetics of embolus formation and collapse in the tracheids of Thuja occidentalis L. stem segments. Radial wood sections were trimmed to 4 mm long paralleling the tracheids by 1 mm wide and 0.1 mm thick. They were observed under a dissecting microscope at 128x while sections were dehydrated and rehydrated. During dehydration, cavitations resulted in the formation of emboli in tracheids, but we concluded that the cavitated tracheids did not immediately fill with air at atmospheric pressure. This conclusion was based on the time required for the emboli to collapse after the rewetting of the dehydrated segment. By hypothesis, the time for the emboli to collapse should be proportional to the amount of air in the emboli. The time for all the emboli to collapse was a linear function of the dehydration time for times up to 15 min. For dehydrations greater than 80 min, the time for collapse after rewetting was constant, and we concluded that the tracheids have saturated with air by 80 min of dehydration. The kinetics of embolus formation is discussed in terms of the air-seeding hypothesis for cavitation, and collapse is discussed in terms of the physics of gas dissolution and diffusion. Embolus formation and dissolution in intact herbaceous and woody plants should follow the same physical laws.     

Insulin-like Growth Factor 1 (IGF-1) Stabilizes Nascent Blood Vessels

Here we report that VEGF-A and IGF-1 differ in their ability to stabilize newly formed blood vessels and endothelial cell tubes. Although VEGF-A failed to support an enduring vascular response, IGF-1 stabilized neovessels generated from primary endothelial cells derived from various vascular beds and mouse retinal explants. In these experimental systems, destabilization/regression was driven by lysophosphatidic acid (LPA). Because previous studies have established that Erk antagonizes LPA-mediated regression, we considered whether Erk was an essential component of IGF-dependent stabilization. Indeed, IGF-1 lost its ability to stabilize neovessels when the Erk pathway was inhibited pharmacologically. Furthermore, stabilization was associated with prolonged Erk activity. In the presence of IGF-1, Erk activity persisted longer than in the presence of VEGF or LPA alone. These studies reveal that VEGF and IGF-1 can have distinct inputs in the angiogenic process. In contrast to VEGF, IGF-1 stabilizes neovessels, which is dependent on Erk activity and associated with prolonged activation.