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.     

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.     

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。     

Chara tomentosa Antheridial Plasmodesmata at Various Stages of Spermatogenesis

Chara tomentosa antheridial plasmodesmata are described during proliferation and spermiogenesis. In antheridial filament cells which are cycling completely synchronously, unplugged plasmodesmata are filled with light cytoplasm. The same plasmodesmata are observed after cessation of mitotic division followed by the onset of synchronous spermiogenesis. Walls separating cells at different cell cycle stages and dividing antheridial filaments into asynchronous domains are plugged with a dense osmophilic substance. Similarly plugged plasmodesmata are present between antheridial cells of different types, e.g., capitular cells and antheridial filaments. In mid spermiogenesis when abundant endoplasmic reticulum (ER) appears temporarily it penetrates into plasmodesmata enabling cell-to-cell transport via ER cisternae. In late spermiogenesis there are no cisternae in plasmodesmata. This revised version was published online in July 2006 with corrections to the Cover Date.     

中国及欧亚大陆新记录种—美洲葫芦藓

本文报道了河北省太行山小五台山脉所采集的中国及欧亚大陆藓类新记录种－美洲葫芦藓ＦｕｒｎａｒｉａａｍｅｒｉｃａｎａＬｉｎｄｂ。ｅｘＳｕｌｌ。     

Intercellular communication in Chara: factors affecting transnodal electrical resistance and solute fluxes

The permeability of plasmodesmata in the nodal complex of branch cells of Chara corallina was examined by measuring both the transnodal electrical resistance and transnodal fluxes of ³⁶CI and ¹⁴C-buty-rate. Under normal circumstances, the resistance across the node was low, but increased rapidly in response to metabolic inhibition, pressure gradients across the node or excision of one of the cells. For each of these treatments, there was a substantial reduction in solute transport between the cells. Acidification of the cytoplasm by weak acids or alkalinization by amines did not affect either the electrical resistance or the flux of solutes through the node between whorl cells. The transnodal resistance was significantly higher in older cell pairs, but was unaffected by large transnodal voltage differences or by the passage of action potentials. There was no evidence that short-term increases in cytoplasmic calcium have any effect on plasmodesmatal permeability.     

仙鹤藓属(Atrichum)藓类植物组织培养再生体系的建立

报道了仙鹤藓(Atrichum undulatum(Hedw.)P.Beauv.)和仙鹤藓小形变种(Atrichum undulatum var.minus(Hedw.)Par.)的组织培养再生体系的建立.为研究仙鹤藓属(Atrichum)藓类愈伤组织的诱导和再分化,将仙鹤鲜和仙鹤藓小形变种原丝体接种在含有4%葡萄糖和0.2～2.0 mg/L 6-BA的MS培养基上,培养一个月后,成功地诱导出疏松、易碎的绿色愈伤组织.愈伤组织诱导和常规继代培养较适合的培养基为含4%葡萄糖和1～2 mg/L6-BA的MS培养基.当将继代培养5次的脱分化藓类愈伤组织转移到含4%葡萄糖但无任何激素的MS培养基上时,能再分化形成原丝体,而在无任何碳源的Benecke培养基上培养时,能再分化形成经原丝体阶段发育来的直立配子体.     

垂蒴真藓原丝体发育特征的研究

在室内培养垂蒴真藓(Bryum uliginosum)孢子的基础上,对其孢子萌发、原丝体发育及配子体发生的全过程进行了跟踪观察。结果表明：该种孢子萌发后直接产生绿丝体。而轴丝体和假根仅在绿丝体上产生;配子体原始细胞产生于绿丝体基部细胞或轴丝体上。由此可以看出：垂蒴真藓属于真藓型孢子萌发型(Bryum-type)。     

Formation of branched plasmodesmata in regenerating Solanum nigrum-protoplasts

During the early stages of culture, discontinuous branched half-plasmodesmata were found randomly scattered in the newly formed outer cell walls of regenerating Solanum nigrum L. protoplasts. During later culture stages, most of these outer-wall plasmodesmata, which had been exposed to the culture medium, disappeared, except for those near the periphery of division walls between daughter cells and those near non-division walls between secondarily associated unrelated cells. Moreover, in the peripheral parts of older division walls, there were continuous branched plasmodesmata which showed the typical morphological characteristics of secondary cell connections: several cytoplasmic strands joined in the median plane of the cell wall and were often linked by so-called median cavities. Evidence is presented that this type of continuous plasmodesma originates from the fusion of the half-plasmodesmata which persisted in the outer walls adjacent to the division wall. Due to growth of the cells after division, opposite parts of the outer walls of the daughter cells come into close contact and fuse, elongating the original division wall peripherally. Opposite half-plasmodesmata remaining in these parts of the outer wall may thereby also be brought into contact and fuse to form a continuous secondary cell connection in the secondarily coalesced wall part. Our assumption is supported by further experiments: (i) longterm video observations of living cells showed differences in the development of the shapes of regenerating cells and (ii) electron-microscopical investigations showed differences in the frequency of the, presumably secondary, cell connections in the peripheral parts of the division walls — both related to the firmness of the embedding medium. In the central parts of division walls, unbranched primary cell connections were found as well as a second type of continuous branched plasmodesma showing an entirely different branching pattern: the region of the middle lamella was always traversed by straight, unbranched parts of these plasmodesmata and the branches occurred exclusively within the younger wall layers. Evidence is given that these branches are modifications of originally unbranched primary plasmodesmata, developing during subsequent thickening of the division wall.The authors are indebted to Prof. H. Binding, Botanisches Institut, Universität Kiel, for making his cell-culture laboratory available to us and to Dr. F. Grundler, Institut für Phytopathologie, Universität Kiel, for placing the video equipment at our disposal. The work was supported by the Deutsche Forschungsgemeinschaft.     

不同培养基及酶对立碗藓原丝体的作用研究

采用含不同成分的MS培养基处理立碗藓的原丝体,发现随蔗糖浓度的提高原丝体的生长相对趋于老化;当培养基中附加不同浓度的2,4-D和6-BA 时,10日龄的原丝体出现了明显变化,当二者浓度均为0.5 mg*L-1,幼嫩的原丝体上能长出疏松易碎的白绿色愈伤组织.利用1%纤维素酶和0.8%的果胶酶分别处理10日龄、20日龄的原丝体和10日龄的茎叶体的叶片2～10 h,结果发现10日龄的原丝体在酶解2 h后能游离出大量的原生质体;而20日龄的原丝体效果较差,只能游离出极少量的原生质体;对于10日龄的茎叶体酶解10 h仍看不到原生质体游离出来.这表明立碗藓原丝体对不同的外界条件可产生不同的反应,但原丝体的生理状态不同对激素和酶类的反应具有非常显著的差别.     

仙鹤藓属（Atrichum）藓类植物组织培养再生体系的建立

报道了仙鹤藓（Atrichum undulatum(Hedw.)P.Beauv.)和仙鹤藓小形变种（Atrichum undulatum var.minus(Hedw.)Par.)的组织培养再生体系的建立。为研究仙鹤藓属（Atrichum)藓类愈伤组织的诱导和再分化,将仙鹤藓和仙鹤藓小形变种原丝体接种在含有4%葡萄糖和0.2-2.0mg/L 6-BA的MS培养基上,培养一个月后,成功地诱导出疏松、易碎的绿色愈伤组织。愈伤组织诱导和常规继代培养较适合的培养基为含4%葡萄糖和1-2mg/L 6-BA的MS培养基。当将继代培养5次的脱分化藓类愈伤组织转移到含4%葡萄糖但无任何激素的MS培养基上时,能再分化形成原丝体,而在无任何碳源的Benecke培养基土培养时,能再分化形成经原丝体阶段发育来的直立配子体。     

Plasmodesmata as a Modulator of Osmotic Water Fluxes in Plants

Solutions to some key problems in the relationships between the structure and functions of plasmodesmata, a component of the plant intercellular communication system, are proposed on the basis of the theory of osmotic flows through porous membranes. The theory accounts for structural characteristics of plasmodesmata, such as their dimension, shape, and length. It considers the steric and adsorption potentials of the solution–cell wall interaction and estimates water and solute (e.g., sucrose) flows under the sustained difference of osmotic pressures at the ends of plasmodesmata. The theory predicts that the water flow through plasmodesmata increases with the widening of the neck constriction and reaches its peak when its size is equal to the diameter of the solute molecule. The water-flow direction was found to depend on the opening of the annulus in neck constrictions at negative adsorption potentials of the plasmodesmata channel walls. Taking into account the presence of sphincters in the neck constrictions, our data suggest the role of plasmodesmata as a modulator of osmotic water fluxes in plants.     

植物病毒在细胞间转运的机理探讨

植物病毒在寄主体内的移动包括细胞间转运和系统性转运两个部分。在这两个过程中,如何有效地利用和修饰胞间连丝,是病毒成功侵染的关键。病毒通过编码运动蛋白与寄主因子互作靶定于细胞质膜,然后通过一系列复杂机制修饰胞间连丝从而顺利完成细胞间转运。综述了植物病毒在细胞间转运过程中与寄主发生的一系列互作,着重阐述了病毒与胞间连丝之间互作的机制,旨在为相关研究工作提供参考。     

Novel conducting tissues in Lower Devonian plants

Elongate cells presumed to comprise water-conducting tissues are described from the central regions of short lengths of two naked, stomatiferous, coalified, axial fossils from Lochkovian (Lower Devonian) fluvial rocks in the Welsh Borderland. In one, a discrete central strand is predominantly composed of uniformly thickened cells that are compared with central tissues in coeval plants, e.g. Aglaophyton , and the hydroids of extant mosses. The other has at least two types of cells with pits of plasmodesmata dimensions that perforate only the inner layer of a bilayered wall. These are compared with liverwort and Takakia hydroids and the coeval S-type tracheids that characterize the Rhyniopsida. The affinities of the two axes remain equivocal. The relevance of plasmodesmata-derived pits to the evolution of diversity in water-conducting elements in early cmbryophytes is discussed.     

Fine structure of protonemal apical cells of the mossPhyscomitrium turbinatum

Summary Elongating caulonemal apical cells of the mossPhyscomitrium turbinatum were cultivatedin vitro and observed during successive stages of cell elongation and division. Actively-growing cells which had completed approximately half of their growth in length were examined by electron microscopy. The distribution of many organelles changes progressively from the cell tip to the distal edge of the large basal vacuole, establishing an apical-basal gradient in organization. Whereas the vacuoles become progressively more extensive in more mature parts of the cell, the dictyosomes, chloroplasts and smooth endoplasmic reticulum are more numerous in younger regions. Some mitochondria in the younger regions of the cell contain localized areas of membrane invagination. Attempts were made to clarify the origin and growth of vacuoles, which become increasingly prominent as the apical cell elongates.Morphological evidence suggests that vacuoles arise in close association with endoplasmic reticulum and dictyosomes as a result of ER dilation and/or cytoplasmic sequestration. The number of vacuolar profiles is highest at the cell tip, decreasing progressively toward the base of the cell, Conversely, the mean area of vacuolar profiles increases progressively toward more basal regions of the cell. These features, along with the increasing number of closely grouped vacuolar profiles along an apical-basal gradient are compatible with the concept of vacuolar growth by coalescence, culminating in their union with the basal vacuole.     

The mechanism of auxin uptake and accumulation in moss protonemata

Protonemata of the moss Funaria hygrometrica Sibth. (L.) show a strong pH dependence for auxin accumulation. IAA uptake is enhanced when the pH of the incubation medium is lowered from 7.6 to 4. In low light intensity grown protonemata (0.56 W m⁻²) a component of IAA uptake could be saturated by IAA; efflux of IAA could be inhibited by 2,3,5-triiodobenzoic acid. This is explained by the presence of influx and efflux carriers for IAA. In protonemata grown at high light intensity (2.00-2.30 W m⁻²) these carriers could not be shown to be present. These results are discussed with regard to the different physiological behavior of moss protonemata grown under different light conditions.