Abscisic-acid-induced drought tolerance in Funaria hygrometrica Hedw.

Three-week-old protonemata of Funaria hygrometrica Hedw. cultivated in Petri dishes tolerate slow drying (24 h to complete dryness) but not rapid drying (1h to complete dryness). Slowly dried mosses show, on a dry-weight basis, a sixfold increase in abscisic-acid (ABA) contents during the drying process. Rehydrated, slowly dried protonemata have the ability to tolerate subsequent rapid drying. When ABA is added to three-week-old protonemata at a concentration of 10 M for 16 h, tolerance to rapid drying is induced. These data indicate that the induction of drought tolerance in Funaria hygrometrica is mediated by ABA. Mosses treated with ABA loose their water as fast as controls do; therefore, ABA does not act via reduced water loss. However, induction of synthesis of new proteins by ABA may form an important part of the drought tolerance because 10 M cycloheximide inhibits the ABA-mediated tolerance to rapid drying.Abbreviations ABA abscisic acid - CHI cycloheximide - DW dry weight - FW fresh weight - RWL relative water loss This work was supported by grants from the Deutsche Forschungs-gemeinschaft and by a NATO fellowship awarded to R.M. Ros Espin.     

Distribution of Potassium, Calcium and Magnesium in the Gametophyte and Sporophyte Generations of Funaria hygrometrica Hedw

Analyses have been made of the K, Ca and Mg in separated sporophytes,spores and gametophytes of the moss Funaria hygrometrica duringsporophyte development. Degeneration of the gametophyte wasaccompanied by loss of K and a gain in Ca while the K contentof the developing sporophyte increased more rapidly than thatof Ca. The presence of an air–gap in the expanding capsulesignificantly influenced the observed cellular location of ions.Relative to the sporophyte the spores were shown to have a higherK and a lower Ca concentration. The behaviour of Mg was intermediatebetween K and Ca throughout. Funaria hygrometrica, potassium, calcium, magnesium, cation distribution, gametophyte, sporophyte     

Epiphytic Bacteria Associated with the Bryophyte Funaria hygrometrica: Effects of Methylobacterium Strains on Protonema Development

Abstract: Bacteria represent a substantial fraction of the microorganisms that inhabit leaf surfaces. We collected samples of the moss Funaria hygrometrica (L.) in the field and analysed the epiphytes on the gametophyte by the agar impression method and scanning electron/fluorescence microscopy. On the phylloid surface numerous bacteria were detected, notably in the grooves between adjacent lamina cells. Methanol-ammonium salts agar surfaces impressed with isolated phylloids of green gametophytes resulted in the growth of methylotrophic colonies. Two Methylobacterium strains ( M. mesophilicum and M. sp., isolated from the Funaria phylloids) were found to simulate the well-known effect of cytokinin application on bud formation in Funaria protonemata. In addition, Methylobacterium inoculation promoted the growth of protonemal filaments. The significance of this novel Methylobacterium -land plant interaction is discussed.     

Germination of the Spores and Development of Primary and Secondary Protonema of Funaria hygrometrica

Abstract

1. The primary protonema of Funaria hygrometrica, cultivated on Knop's or Marchal's agar in the light, proved to consist of filaments with much chlorophyll, a hyaline membrane, perpendicular cross-walls and branches equal to the main filament (chloronema). These filaments grow on the surface of the agar, the branches may also grow vertically. Sometimes filaments with less chlorophyll occur immediately after the germination. The caulonema described by Sironval has not been observed. Thus the rhizoid-like forms mentioned in the literature should more likely be considered as a result of external conditions (see Schoene, Bauer, Heitz and Fitting). Therefore it remains doubtful if a distinction between rhizoids and chloronema on the primary protonema is of any importance as it is impossible to give a good definition of either form.

At the base of moss plants main filaments with brown membranes, oblique septa and without chlorophyll may develop (rhizoids). They grow on the surface or within the agar. In F. hygrometrica especially, the stem seems to influence the occurrence of these rhizoids. The main filaments form buds on the basal cell of the branches and thus serve for vegetative reproduction. The branches show the characteristics of the chloronema. This is contrary to the conclusion of Westerdijk that rhizoids would pass into chloronema only when they are damaged or when the growth of the end bud of the plant is inhibited. At the base of the plant, moreover, little ramified, short branches with oblique septa appear which do not produce buds.

2. Branches may develop in the first growth stages of the primary protonema at any point of the cells. One single cell of a main filament can produce none, one, or more than one branch. Later the branches appear immediately behind the acroscopic cross-wall except in a few cases. Each cell then produces one branch.

3. Buds always develop at the basal cell of a primary branch of a green main filament or of a rhizoid derived from a moss plant.

4. In two ways the protonema may fall into pieces, which can develop into new main filaments:

(a) By forming brood cells; rounded cells which get detached by splitting of the septum. This phenomenon is very frequent. Contrary to Servettaz's opinion it seems to occur particularly under unfavourable conditions.

(b) By forming special cells, tmemata, whose walls are rent. These occur on the primary protonema contrary to the observations of Correns and Bauer, but they are much less frequent than the brood cells. No observations have been made on the circumstances of their occurrence.     

Polarity and growth of caulonema tip cells of the moss Funaria hygrometrica

In the caulonema tip cells of Funaria hygrometrica, chloroplasts, mitochondria, and dictyosomes have differences in structure which are determined by cell polarity. In contrast to the slowly growing chloronema tip cells the apical cell of the caulonema contains a tip body. Colchicine stops tip growth; it causes the formation of subapical cell protrusions, redistribution of the plastids, and a loss of their polar differentiation. Cytochalasin B inhibits growth and affects the position of cell organelles. After treatment with ionophore A23 187, growth is slower and shorter and wider cells are formed. D₂O causes a transient reversion of organelle distribution but premitotic nuclei are not dislocated. In some tip cells the reversion of polarity persists; they continue to grow with a new tip at their base. During centrifugation, colchicine has only a slight influence on the stability of organelle anchorage. The former polar organization of most cells is restored within a few hours after centrifugation, and the cells resume normal growth. In premitotic cells the nucleus and other organelles cannot be retransported, they often continue to grow with reversed polarity. Colchicine retards the redistribution of organelles generally and increases the number of cells that form a basal outgrowth. The interrelationship between the peripheral cytoplasm and the nucleus and the role of microtubules in maintaining and reestablishing cell polarity are discussed.Abbreviations DMSO dimethylsulfoxide - CB cytochalasin B Dedicated to Prof. Dr. A. Pirson on the occasion of his 70. birthday     

Lead uptake,localization and changes in cell ultrastructure ofFunaria hygrometrica protonemata

The main sites of lead entering the protonemata were spores with broken cell walls and apical cells of protonemata. The lead ions could enter the symplast probably by endocytosis but we can not exclude diffusion as the way of entering. Lead deposits inside a protoplast were observed after 2 h of lead application. After 12 h, Pb deposits were commonly present in all cell structures surrounded by a membrane. Some disturbances in ultrastructure of organelles were found after 8 h. The most frequent were: change of endoplasmatic reticulum configuration from linear to concentric one in many cases surrounding other organelles (e.g. mitochondria) and large and numerous plastoglobuli in chloroplasts. Longer treatment of protonemata with lead (24 h or 48 h) caused degeneration of organelles and even death of the cell. The present work was supported by a grant from the State Commitee for Scientific Research no. 1575 9101 for E.A.G.     

Tip-localised H+-fluxes and the applicability of the acid-growth hypothesis to tip-growing cells: Control of chloronemal extension in Funaria hygrometrica by auxin and light

The role of tip-localised H⁺ secretion in regulating chloronemal tip growth in the moss Funaria hygrometrica Hedw. was investigated. pH was monitored with pH microelectrodes placed close to the cell surface while the rate of extension growth was manipulated by illumination and by the application of indole-3-acetic acid. Growth stimulations were accompanied by acidification of the external solution; this acidification was most pronounced at the growing tip. The timing and extent of acification external to the tip correlated well with the magnitude and time course of growth stimulations. The maintenance of both growth and H⁺ efflux under CO₂-free conditions indicated that neither photosynthetic nor respiratory CO₂ metabolism were involved. Artificially acidifying the nutrient solution rapidly but transiently stimulated elongation in both white light and darkness. Furthermore, the stimulation of elongation caused by white light was inhibited if the nutrient solution was buffered strongly near neutrality. We conclude that the acid growth hypothesis is applicable to tip growth in Funaria and that light and exogenous indole-3-acetic acid act at least in part by stimulating localised H⁺-ion efflux.Abbreviations D darkness - IAA indole-3-acetic acid - WL white light     

Effect of Lead and Colchicine on Morphogenesis in Protonemata of the Moss Funaria hygrometrica

Protonemata of Funaria hygrometrica grown in artificial mediacontaining different lead concentrations grow more slowly thancontrols and show a disturbance of polar growth, changed arrangementof chloroplasts, alterations of nucleus and septa position.Morphological effects are dose-dependent. At the lowest leadconcentration (10^-6 M), only a delay in development was observed,but no cellular alterations, At 10^-5 M Pb nuclear migration,cellular shape, size and position of plastids, were alteredand a variety of aberrant forms were present. At 10^-4 M, besidesthese alterations, a drastic reduction of the protonemal system,high vacuolation and the growth of protonemal filaments fromleaves were evident. The highest concentration, (10^-3 M), causeddeath. Patterns of protonemal development and cellular arrangementin lead-treated samples showed similarities as well as differences,if compared to alterations induced by colchicine. Indirect immunofluorescencedemonstrated a correlation between lead concentration and alterationof cytoskeletal organization (alterations similar to those inducedby colchicine). Hypotheses are raised to account for effects of lead on microtubulestructure, arrangement and cytoplasm organization.Copyright1995, 1999 Academic Press Funaria hygrometrica, lead, protonemal development, cytomorphogenesis, microtubules     

Observations on dividing plastids in the protonema of the moss Funaria hygrometrica Sibth.

The process of division was investigated in the different types of plastids found in the tip cell of the protonema of Funaria hygrometrica Sibth. There were no structural changes in the envelope membranes of any of the plastid types during the initial stage of division. As the process of constriction advanced, thylakoids were locally disintegrated and sometimes starch grains in the isthmus were locally dissolved. In the isthmus, tightly constricted plastids were characterized by an undulating envelope and an increasing number of vesicles. After three-dimensional reconstruction of electronmicrographs a distinct filamentous structure was observed in the plane of division outside the plastid but close to the envelope. At different stages of division the constricted regions were partly surrounded by one or a few filaments. The roundish plastids in the apical zone were accompanied by single microtubule bundles, and the spindle-shaped plastids in the cell base were surrounded by single microtubules and microtubule bundles. A model of co-operation between microtubules and the filamentous structure in the division process is discussed.A preliminary report was presented at the Tagung der Deutschen Botanischen Gesellschaft und der Vereinigung für Angewandte Botanik, Hamburg, September 1986     

细胞壁在植物重金属耐性中的作用

植物细胞壁主要是由多糖、蛋白质和木质素等组成的一个复合体,广泛参与植物生长发育及对各种逆境胁迫的响应,是重金属离子进入细胞质的第一道屏障。本文主要综述了植物细胞壁主要成分,包括细胞壁多糖、细胞壁蛋白质和木质素,在响应重金属胁迫反应中的作用及其参与重金属耐性的机制,以期能对植物细胞壁在重金属耐性中的作用有更深入的了解。     

Structure and Function of Transfer Cells in the Sporophyte Haustorium of Funaria hygrometrica Hedw: I. THE DEVELOPMENT AND ULTRASTRUCTURE OF THE AUSTORIUM

The development of the sporophyte-gametophyte interface in themoss, Funaria hygrometrica Hedw., is described with the aidof light- and electron-microscopy. The outer walls of the cellsthat abut the haustorial cavity in both generations developlabyrinths typical of transfer cells. This feature is more apparentin the epidermal cells of the sporophyte foot (haustorium),where development can be split into three main stages. The primarygrowth stage, which is complete at about the time the calyptradetaches from the ripened archegonium, involves the formationof transfer cells. The secondary stage is characterized by thedeposition of amorphous inclusions in the wall labyrinth ofthe transfer cells. The tertiary stage, which commences as thesporophyte capsule ripens, entails de-differentiation of thetransfer cell wall labyrinth to form a thick, heavily encrusted,outer cell wall. The pattern of development of these cells iscorrelated with changes in gametophyte- sporophyte translocationcapabilities.     

葫芦藓精子发生过程中胞间连丝与胞质桥的超微结构

从超微结构水平上对葫芦藓(Funaria hygrometrica Hedw.)精子发生过程中胞间连接系统的结构及其变化动态进行了研究。结果表明,同一区中的相邻生精细胞由大量胞质桥相连,而不同区的细胞之间则不存在胞质桥。胞间连丝存在于套细胞之间以及套细胞与生精细胞之间, 但它在生精细胞间不存在。在精子器发生的后期,当精子细胞壁开始降解时,同一个精子器中所有的精子细胞似乎都由扩大的胞质桥相互连接。胞质桥一直保持到精子分化的后期,最终精子细胞同步分化成精子。胞间连丝与胞质桥具有不同的内部结、分布以及生物发生机制,这表明它们在精子器的发育过程中可能扮演着不同的角色。     

Effects of heavy metals on ultrastructure and HSP70s induction in the aquatic moss Leptodictyum riparium Hedw

The effects of heavy metals, both toxic (Pb, Cd) and essential (Cu, Zn) on the ultrastructure and the induction of Heat Shock Protein 70 (HSP70) have been studied in the aquatic moss Leptodictyum riparium Hedw. In vitro cultured L. riparium was treated with different heavy metals, both toxic, as cadmium or lead; and essential microelements such as Copper or Zinc concentrations ranging from 10(-3) to 10(-6) M to investigate both ultrastructural damage and HSP induction. TEM observations showed that sub-lethal concentrations of heavy metals caused only slight changes, largely localized in the chloroplasts. Among all the heavy metals tested, cadmium caused the most severe modifications. Heavy metals caused the decrease of the soluble protein content and the enhancement of proteins reacting versus HSP70 antibodies, suggesting that molecular chaperons might be involved in the resistance to toxic effects of lead, cadmium, copper and zinc. Therefore, the induction of HSP70 in L. riparium would confer a higher resistance to pollutants under stressful conditions lethal for other mosses and higher plant species. These results suggest that the moss L. riparium can tolerate heavy metals stress without incurring severe cellular/subcellular damage. Therefore it can be used as a useful indicator of heavy metals accumulation.     

The role of the cell wall in the mechanism of lead and zinc tolerance in Anthoxanthum odoratum L

The respiration rate and viability of cultured cells and protoplasts isolated from two clones of Anthoxanthum odoratum tolerant to both zinc and lead were unaffected by the presence of zinc. Although intact cells were largely unaffected by the presence of lead, protoplasts isolated from cultured cells were susceptible, showing a reduced respiration rate and a high mortality. In contrast cultured cells and protoplasts of non-tolerant clones of A. odoratum were susceptible to both zinc and lead. The results provide direct evidence that in A. odoratum the cell wall is part of the mechanism of tolerance to lead, but not to zinc.     

Phenylurea cytokinins assayed for induction of shoot buds in the moss Funaria hygrometrica

The induction of shoot buds from the filamentous protonema of moss is a classic bioassay for cytokinin. While a large literature documents this response in many species of moss and for a wide range of natural and synthetic cytokinins, to date only substituted adenine cytokinins have been examined in detail. This paper shows that at least some of the novel phenylurea cytokinins will induce bud formation in mosses. Funaria responds to thidiazuron much as it responds to benzyladenine. Exposure to either substance results in log-linear dose-dependent increases in bud number that reach similar maximal numbers of buds at the optimal concentration of compound. The related compound chloro-pyridyl-phenylurea (CPPU) is slightly less active, but induces buds over a wider range of concentration. Carbanilide (diphenylurea or DPU), an active cytokinin in other systems, induces very few buds in Funaria, but does so over a wide range of concentration. Bioassay of mixtures of benzyladenine and DPU finds no evidence of competition for cytokinin receptors. That result could support suggestions that the phenylurea cytokinins act indirectly, by altering endogenous cytokinin metabolism, but we favor another interpretation. Unlike other cytokinin-responsive systems, the induction of buds from moss protonema involves two cytokinin-mediated events. The number of buds is controlled by the second cytokinin-mediated event. If DPU has little or no affinity for the receptor triggering this second event, DPU treatments will produce few to no buds, and kinetic analysis using bud number would find no evidence for competition with benzyladenine. Our results would support the hypothesis that bud induction in Funaria involves two chemically distinct cytokinin receptors.     

Calcium pectate chemistry causes growth to be stored in Chara corallina: a test of the pectate cycle

Calcium pectate chemistry was reported to control the growth rate of cells of Chara corallina , and required turgor pressure ( P ) to do so. Accordingly, this chemistry should account for other aspects of growth, particularly the ability of plants to compensate for brief exposure to low P , that is, to 'store' growth. Live Chara cells or isolated walls were attached to a pressure probe, and P was varied. Low P caused growth to be inhibited in live cells, but when P returned to normal (0.5 MPa), a flush of growth completely compensated for that lost at low P for as long as 23–53 min. This growth storage was absent in isolated walls, mature cells and live cells exposed to cold, indicating that the cytoplasm delivered a metabolically derived growth factor needing P for its action. Because the cytoplasm delivered pectate needing P for its action, pectate was supplied to isolated walls at low P as though the cytoplasm had done so. Growth was stored while otherwise none occurred. It was concluded that a P -dependent cycle of calcium pectate chemistry not only controlled growth rate and new wall deposition, but also accounted for stored growth.