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.     

Microculture and Electrofusion of Defined Protoplasts of the Moss Funaria hygrometrica

Protoplasts isolated from protonemata of Funaria hygrometrica wild type and developmentally abnormal mutants were individually selected and cultured in microdroplets of defined, unconditioned culture medium. Up to 60 % of the microcultured wild type protoplasts regenerated whole plants after 4 weeks. Electrofusion in different combinations of defined protoplast pairs of wild type and mutated forms was performed. The products derived from one-to-one electric field induced fusion were efficiently microcultured and somatic hybrids were regenerated.     

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     

Partial purification and characterization of cyclic AMP phosphodiesterases from Funaria hygrometrica

Natural-abundance ¹³C NMR spectroscopy at 15.04 MHz has been used to examine the effects of pH, calcium, and lanthanide ions on the polypeptide cardiac stimulant Anthopleurin-A in aqueous solution. The carboxyl resonance from the aspartic acid residue not observed in a previous study (R. S. Norton and T. R. Norton, 1979, J. Biol. Chem.254, 10220–10226) has been identified and an apparent pK_a of 3.4 obtained. More accurate estimates have been derived for the apparent pK_a values of the two histidine residues. Binding of Ca²⁺ ions has been found by equilibrium dialysis and ¹³C NMR to be weak (K_d > 0.1 M). The interaction with lanthanide ions is slightly stronger, but binding occurs at the C terminus as well as at a site involving one or both of the aspartate carboxylate groups. These results suggest that possible Anthopleurin-A-induced calcium translocation in the myocardial cell is a secondary effect. The interaction of Anthopleurin-A with lipid monolayers has also been examined. Binding occurs to neutral and zwitterionic lipids, but is stronger with negatively charged lipids, particularly cardiolipin. This interaction is also influenced by the presence of Ca²⁺ ions. The implications of these results for the mechanism of action of this polypeptide are discussed.     

苔藓植物孢子萌发与原丝体发育研究进展

孢子萌发与原丝体发育对于研究苔藓植物以及其他植物类群的系统发育与系统演化有着十分重要的作用。本文综述苔藓植物孢子萌发类型、原丝体发育特征以及环境影响因子等,还介绍了20世纪90年代以来该领域研究的新进展,如人们以苔藓植物原丝体为实验材料进一步揭示Ca2 ．CAM信号系统、植物激素、光信号系统等在植物体内的作用机制。     

苔藓植物孢子萌发与原丝体发育研究进展

孢子萌发与原丝体发育对于研究苔藓植物以及其他植物类群的系统发育与系统演化有着十分重要的作用。本文综述苔藓植物孢子萌发类型、原丝体发育特征以及环境影响因子等,还介绍了20世纪90年代以来该领域研究的新进展,如人们以苔藓植物原丝体为实验材料进一步揭示Ca2+·CAM信号系统、、植物激素、光信号系统等在植物体内的作用机制。     

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。     

五种藓类植物的孢子萌发与原丝体发育

本文对中华缩叶藓(Ptychomitrium sinense)、狭叶缩叶藓(P.linearifolium)、威氏缩叶藓(P.wilsonii)、葫芦藓(Funaria hygrometrica)和立碗藓(Physcomitrium sphaericum)5种国产藓类植物的孢子萌发和原丝体发育进行研究,结果表明不同藓类孢子萌发和原丝体发育类型不同,孢子萌发和原丝体发育方式与藓类植物的生态环境有关.     

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     

五种藓类植物的孢子萌发与原丝体发育

本文对中华缩叶藓(Ptychomitrium sinense)、狭叶缩叶藓(P. linearifolium)、威氏缩叶藓(P．wilsonii)、葫芦藓(Funaria hygrometrica )和立碗藓(Physcomitrium sphaericum ) 5种国产藓类植物的孢子萌发和原丝体发育进行研究, 结果表明不同藓类孢子萌发和原丝体发育类型不同, 孢子萌发和原丝体发育方式与藓类植物的生态环境有关。     

The impact of forest disturbance on the genetic diversity and population structure of a late-successional moss

Abstract

Human-induced disturbances threaten the genetic variation of wild plant populations. The genetic diversity and spatial population structure of the moss Isothecium myosuroides, a late-successional forest species, was investigated in subtropical cloud forests (La Gomera, Canary Islands) using inter-simple sequence repeat (ISSR) markers. Inter- and intrapopulation genetic variability was assessed in two ancient and four disturbed forest stands, which were classified according to their vegetation, forest age and type of disturbance. ISSR analysis of 144 epiphyte colonies with eight primers resulted in 211 reliably amplified bands. Our findings show that in disturbed forest stands, the population structure is increased, and the genetic diversity decreased compared with the levels observed in ancient forests. Although ancient and disturbed stands were located relatively close to each other, the (re-) established epiphyte populations did not reach their original genetic condition, 40 years following disturbance. Strong differentiation among populations of I. myosuroides at several spatial scales and differences in genetic diversity are mainly related to the local environmental conditions and the availability of suitable microhabitats in anthropogenically disturbed forest stands.     

梨蒴珠藓孢子萌发与原丝体发育特征研究

为了解梨蒴珠藓（Bartramia pomiformis）孢子萌发和原丝体发育特征,在显微镜下观察室内人工培养的梨蒴珠藓单倍配子体发育过程。结果表明,梨蒴珠藓孢子吸水膨胀5 d后,开始破壁萌发,原丝体系统以丝状绿丝体为主,轴丝体在绿丝体上分化产生。培养22 d后,配子体在轴丝体细胞上分化产生。参照Nishida的标准,梨蒴珠藓孢子萌发类型为真藓型（Bryum-type）。这为梨蒴珠藓的人工扩繁提供了发育学基础资料。     

Side branch formation and orientation in the caulonema of the moss,Funaria hygrometrica: Normal development and fine structure

Summary The regular branching of theFunaria caulonema filaments is partly related to rhythms in nuclear and cell division. The formation and development of the branches were studied by light and electron microscopy with particular attention directed to the distribution of microtubules and the polar organization of the cytoplasm. The new side branch breaks through the wall of the mother cell. The site of branch development is determined by the position of the nucleus of the mother cell. In protonemata which grow in vertically placed Petri dishes gravity influences the position of nuclei and side branches, and also the direction of oblique cross walls in the caulonema filaments to a certain extent.     

The quantitative response to cytokinin in the moss Funaria hygrometrica does not reflect differential sensitivity of initial target cells

Exposure to sufficient cytokinin induces the formation of buds from responsive cells in the protonema of Funaria hygrometrica. Initial perception of the phytohormone results in a Ca+2 cascade within minutes. A second cytokinin-mediated event occurs some days later, and converts incipient buds into stably committed buds. The concentration of exogenous cytokinin also regulates the total number of buds produced from a protonemal colony. This concentration-dependent production of buds has been thought to reflect differential sensitivity of target cells. Under that hypothesis, the regulation of bud number occurs during initial perception of hormone. This paper presents direct experimental evidence to the contrary and supports the alternate hypothesis that bud formation involves the gating of large numbers of responding cells by later events. Experiments transferring protonema between media with different levels of cytokinin show that the cytokinin concentration during the initial perception of cytokinin is unimportant in controlling bud number. Instead, bud number is found to be regulated by the concentration of exogenous cytokinin as incipient buds or bud initials become stably committed buds.     

Side branch formation and orientation in the caulonema of the moss,Funaria hygrometrica: Experiments with inhibitors and with centrifugation

Summary Colchicine treatment ofFunaria caulonemata, usually does not inhibit initiation of a side branch or its incipient elongation but does prevent movement of chloroplasts and the nucleus into the outgrowth. After colchicine and after cytochalasin B treatment side branches are formed about at the normal age of the cells; because of the inhibition of the apical cell they arise at an abnormal position,i.e., not in the third but in the second cell of a filament. After D₂O treatment the organelles are dislocated toward the basal cross wall. The site of side branch formation is then obviously determined by the position of the nucleus. Cells with an irreversibly reversed longitudinal polar axis can be found; by centrifugation in proximal direction the sites of side branch initiation likewise are displaced into the proximal region of the cell, especially if the remigration of the nucleus is inhibited by colchicine. High concentrations of Ca²⁺ ions induce the formation of side branch cells, without any outgrowth. The calcium ionophore A 23 187 influences the position of the nucleus and of the side branch only slightly. After these various treatments intercalary divisions frequently occur. The role and interrelationship of the nucleus and peripheral cytoplasm in establishing and maintaining the polar axes, and the role of microtubules are discussed.     

三种植物生长调节剂对密叶绢藓(Entodon challengeri)孢子萌发、原丝体发育及芽体发生的影响

研究了3种植物生长调节剂苯基噻二唑基脲(TDZ)、6-苄基腺嘌呤(6-BA)、萘乙酸(NAA)对密叶绢藓[Entodon challengeri(Paris)Cardot]孢子萌发、原丝体发育及芽体发生的影响,并对整个发育过程进行了显微观察和照相,结果表明:(1)3种植物生长调节剂对密叶绢藓孢子萌发影响不显著;(2)在原丝体发育阶段,1.0 mg/LNAA对原丝体初期的发育促进效果显著,0.4 mg/L TDZ对原丝体发育中期分枝的形成促进效果显著,6-BA处理效果不显著;(3)3种植物生长调节剂单独处理均促进芽体的发生,但0.4 mg/L TDZ效果最佳。而1.5 mg/L 6-BA+TDZ组合处理效果更加显著;(4)芽体的发生数量与芽体的长势无正相关性。