大型绿藻浒苔藻段及组织块的生长和发育特征

通过将浒苔叶状体分为基部、中部和顶端三部分分别进行切段和切碎处理,在实验室条件下,用液体浅层培养的方法,系统地研究了其组织和细胞的生长和发育特性。显微观察的结果显示：切段培养条件下,基部和中部的藻段均可在其形态学下端形成假根,在形态学上端产生类似叶状体的突起。藻段的发育具极性,但是其极性并不绝对的,在1.0 mm的基部藻段两端都观察到了假根的形成。虽然顶端的藻段和组织块全都形成和释放了孢子,未见明显的营养生长,但是在培养早期,其下端仍然具有形成假根的能力。浒苔各部位藻段和组织块释放的和滞留于孢子囊内的孢子都可以立即萌发成苗。快速生长的中基部藻段形成了气囊,致使其漂浮于培养基上。有很多藻段和组织块形成和释放了生殖细胞,释放到外界以及滞留于孢子囊内的孢子均可立即附着萌发。数据分析表明：藻段的生长具有极性,不同部位相同长度的藻段生长率差异明显,基部藻段的生长率高于中部藻段,顶部藻段无明显的营养生长。藻段的生长与其原始长度和在藻体中所处的位置有密切关系;藻段和组织块的再生与藻体的完整性及其在藻体中所处的位置有关。     

Some observations on polarity and regeneration in Enteromorpha

The regeneration of two types of Enteromorpha has been investigated. Small tubular sections cut from the thallus develop rhizoids along the basal cut edge and papillae from the apical cut edge. The capacity for regeneration is greatest in segments from the middle and base of the thallus and least in apical sections. Regeneration in various liquid culture media at different light intensities and temperatures and on solid agar media has been tested. The addition of growth substances and extracts from Enteromorpha thalli always stimulate regeneration but in no way alter the polarity. The results are compared with previous conflicting accounts of regeneration of Enteromorpha.     

Inheritance of potential for in vitro androgenesis in spring barley

Inheritance of the ability to produce androgenic structures and regenerate plants in an in vitro anther culture with doubled-haploid spring barley lines derived from F₁ reciprocal hybrids from crosses of cultivars with contrast (high × low), high (high × high), and low (low × low) growth capacity is studied. A general trend towards transgressive segregation in terms of the indices, “number of anthers with new growths” and “rate of regeneration of plants,” was found. The data attest to a major role of nonallelic interaction of genes in genetic control of in vitro androgenesis and their variance distribution in the parent forms of the hybrids.     

Structure and Protein Composition of a Basal‐Body Scaffold (“Cage”) in the Hypotrich Ciliate Euplotes

Cilia on the ventral surface of the hypotrich ciliate Euplotes are clustered into polykinetids or compound ciliary organelles, such as cirri or oral membranelles, used in locomotion and prey capture. A single polykinetid may contain more than 150 individual cilia; these emerge from basal bodies held in a closely spaced array within a scaffold or framework structure that has been referred to as a basal‐body “cage”. Cage structures were isolated free of cilia and basal bodies; the predominant component of such cages was found on polyacrylamide gels to be a 45‐kDa polypeptide. Antisera were raised against this protein band and used for immunolocalizations at the light and electron microscope levels. Indirect immunofluorescence revealed the 45‐kDa polypeptide to be localized exclusively to the bases of the ventral polykinetids. Immunogold staining of thin sections of intact cells further localized this reactivity to filaments of a double‐layered dense lattice that appears to link adjoining basal bodies into ordered arrays within each polykinetid. Scanning electron microscopy of isolated cages reveals the lower or “basal” cage layer to be a fine lacey meshwork supporting the basal bodies at their proximal ends; adjoining basal bodies are held at their characteristic spacing by filaments of an upper or “medial” cage layer. The isolated cage thus resembles a miniature test‐tube rack, able to accommodate varying arrangements of basal‐body rows, depending on the particular type of polykinetid. Because of its clear and specific localization to the basal‐body cages in Euplotes, we have termed this novel 45‐kDa protein “cagein”.     

BIOCHEMICAL AND MORPHOLOGICAL EFFECTS OF TEMPERATURE ON TYPHA LATIFOLIA L. (TYPHACEAE) ORIGINATING FROM DIFFERENT ENDS OF A THERMAL GRADIENT. I. CONTROLLED ENVIRONMENTAL STUDIES

T. latifolia grows along most of the shoreline of Par Pond, a cooling reservoir for the reactors at the Savannah River Plant in Aiken, South Carolina. A thermal gradient exists because of the movement of hot water from one end of the pond to the other. This study was undertaken to determine the biochemical and morphological effects of temperature on Typha latifolia clones originating from different ends of this thermal gradient. The data indicated that both “hot” and “cold” end plants react similarly to temperature. Forty-eight rhizome plants from both ends of Par Pond were transplanted into tanks at 20 C and 30 C. Total levels of malic dehydrogenase activity, proportions of mitochondrially bound MDH and growth rates were significantly different in plants grown in the two tanks, but no significant differences were observed within each tank between individuals from the “hot” and “cold” end. The relation of these results to hypotheses concerning quantitative strategies of enzyme adaptation are discussed in the paper.     

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.     

OBSERVATIONS ON THE LIFE HISTORY OF PAPENFUSSIELLA CALLITRICHA (PHAEOPHYCEAE,CHORDARIALES) IN CULTURE1

Papenfussiella callitricha (Rosenv.) Kylin from eastern Canada was studied in culture. Zoids from unilocular sporangia develop into microscopic, filamentous, dioecious gametophytes which produce isogametes in filament cells and few-chambered plurilocular gametangia. Unfused gametes germinate to reproduce the gametophytes. Fusion takes place between a settled (“female”) and a motile (“male”) gamete. The zygote gives rise to a filamentous plethysmothallus that reproduces asexually by zoids formed in thallus cells and in few-chambered plurilocular zoidangia. Erect macrothalli are produced on the plethysmothallus, beginning with the formation of upright filaments. Later on, these filaments become the terminal assimilators of the macrothalli. Further assimilatory filaments, rhizoids, and unilocular sporangia are produced in a branching region at the base of the terminal assimilator. Zoids from unilocular sporangia formed in culture germinate to reestablish the gametophyte phase. Chromosome counts yielded n = 19 ± 3 for the gametophytes, and 32 ± 6 for the sporophyte, both plethysmothallus and macrothallus.     

PRIMARY PHLOEM REGENERATION WITHOUT CONCOMITANT XYLEM REGENERATION: ITS HORMONE CONTROL IN COLEUS

Phloem regeneration in the absence of xylem regeneration was evoked in number 5 internodes of Coleus blumei Benth. by severing xylemless phloem bundles. Its quantitative extent was estimated. To determine whether phloem regeneration is directly affected by auxin, or whether it is a secondary consequence of the auxin-dependent xylem regeneration which usually accompanies it, phloem regeneration was measured in decapitated plants from which auxin-producing leaves and buds had been removed (i.e., in “plant stumps”). In these stumps, 1% IAA in lanolin completely restored phloem regeneration to the intact plant level. In such stumps from which roots had been excised, and in excised internodes, IAA failed to restore it to this level. However, zeatin or zeatin riboside in aqueous solution applied to the bases of excised internodes receiving IAA at their apical ends restored phloem regeneration to the level of that in whole plants. When similarly tested, other cytokinins (kinetin, kinetin riboside, 2iP, and 2iPA), gibberellic acid (GA₃), glutamine, proline, sucrose, and a mixture of mineral salts failed to promote phloem regeneration. Glutamic acid, tested only once, was slightly promotive of it.     

Porphyra yezoensis bei Helgoland — eine entwicklungsgeschichtliche Studie

APorphyropsis-like epiphytic specimen found in the harbour of Helgoland was grown in culture and proved to be identical with the JapanesePorphyra yezoensis. Life history studies on this economically important alga resulted in some interesting and hitherto unknown details. The variability of the adult frond is fundamentally determined by the pattern of spore germination. Settled onChaetomorpha filaments, monospores elongate within 20 minutes; the epiphytic germlings are attached to the substrate by a typical basal cell and give rise exclusively to elongated fronds provided with a cuneate base. Unattached spores, however, germinate into buds with rhizoids; they develop into elongated elliptical to oval fronds provided with round or cordate bases. Only plants with male areas were observed in the cultures, butConchocelis was abundantly produced from cells of aged thalli. Grown in mussel-shells, the filamentous phase liberated conchospores for a long time.

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Herrn Paul-Heinz Sahling zum 75. Geburtstag gewidmet, mit Anerkennung und Dank für 50jährige Zusammenarbeit.     

Involvement of auxin and a homeodomain-leucine zipper I gene in rhizoid development of the moss Physcomitrella patens

Differentiation of epidermal cells is important for plants because they are in direct contact with the environment. Rhizoids are multicellular filaments that develop from the epidermis in a wide range of plants, including pteridophytes, bryophytes, and green algae; they have similar functions to root hairs in vascular plants in that they support the plant body and are involved in water and nutrient absorption. In this study, we examined mechanisms underlying rhizoid development in the moss, Physcomitrella patens, which is the only land plant in which high-frequency gene targeting is possible. We found that rhizoid development can be split into two processes: determination and differentiation. Two types of rhizoids with distinct developmental patterns (basal and mid-stem rhizoids) were recognized. The development of basal rhizoids from epidermal cells was induced by exogenous auxin, while that of mid-stem rhizoids required an unknown factor in addition to exogenous auxin. Once an epidermal cell had acquired a rhizoid initial cell fate, expression of the homeodomain-leucine zipper I gene Pphb7 was induced. Analysis of Pphb7 disruptant lines showed that Pphb7 affects the induction of pigmentation and the increase in the number and size of chloroplasts, but not the position or number of rhizoids. This is the first report on the involvement of a homeodomain-leucine zipper I gene in epidermal cell differentiation.     

DEVELOPMENTAL CHANGES IN THE ALGAL COENOCYTE CAULERPA PROLIFERA (SIPHONALES) AFTER INVERSION WITH RESPECT TO GRAVITY

The algal coenocyte Caulerpa prolifera produces three types of organs, each with a different orientation. The effect of gravity in controlling the development of these organs was investigated. We inverted plants by rotating them 180° around the horizontal rhizome axis, then compared development of the inverted plants with plants matched by size and differentiation. Quantitative data were obtained from photographic records. After inversion the site of organ differentiation was changed with no change in the timing: the next rhizoid (and all subsequent ones) differentiated on the “now-under side” of the inverted rhizome, the next leaf formed on the “now-upper side.” Despite the fast change in the site of organ differentiation, other parameters were not changed by inversion (e.g., rate of elongation of rhizomes or leaves, rate of formation of leaves or rhizoids). Because such changes also occurred in plants with balanced lighting from two sides, it is clear that gravity alone can control these changes without reinforcement from top illumination. After leaves were initiated, they did not change their orientation, showing neither positive phototropism (at our light levels) nor negative geotropism, even when elongating substantially. Torsion of the rhizome tip did not precede the change in site of rhizoid initiation.     

Development of the siphonous green alga Penicillus and the Espera state*

The thallus of Penicillus is composed of two filament types: axial filaments of indeterminate growth and laterals of determinate growth. In vegetative reproduction new plants arise from horizontal rhizoids. Four stages can be distinguished in development. In the primordium stage the tip of a rhizoid swells and forms a primordium. In the germling stage ascending and descending axial filaments arise from the primordium, the former grow into a fascicle and give rise to lateral saccate branches, each of which forms an ascending and a descending arm and branches further into rhizoid-like branchlets. Together these structures constitute the foundation of the stipe. At the same time the descending axial filaments elongate and become main rhizoidal filaments with lateral rhizoidal branchlets. In the early juvenile stage the stipe is formed. The elongating ascending axial filaments form a medulla while their laterals produce a cortex. In the late juvenile stage the axial filaments form the capitulum. The Espera state of Penicillus lacks a stipe since the ascending axial filaments do not join in a fascicle. Espera has been grown in laboratory cultures from Penicillus plants collected in the Caribbean region and also been found uncommonly in nature in this area. This state may be a response to environmental stress. The flattening of thalli and their orientation perpendicular to the direction of waves are discussed. A comparison of Penicillus and Codium indicates that at least two types of development exist in multiaxial Eusiphoniidae.     

‘Seed’ production of Porphyra spp. by tissue culture

Differentiation of archeospores was observed from excised tissue of young thalli of various monoecious Porphyra species ( P. tenera, P. yezoensis, P. suborbiculata, P. okamurae) after 4–8 days in culture at temperatures of 20 and 25 °C. Excised tissue from adult thalli did not differentiate into archeospores, but rather regenerated directly into blades and rhizoids of foliose thalli. Tissues from young thalli of two dioecious Porphyra species ( P. dentata and P. pseudolinearis) also regenerated into blades and rhizoids after manipulation of the culture conditions. In addition, 1–2 celled tissue pieces of both monoecious and dioecious species were also seen to develop directly into blades. Polarity of regeneration of blades and rhizoids was observed in these species. These results suggest that ‘seed’ can be obtained through tissue culture instead of using conventional conchocelis culture for commercial nori aquaculture in Japan. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nutritional Study of Three Strains of Naegleria gruberi*

Naegleria gruberi strains cloned from amebas isolated from a Vero cell culture (“TS”), a sewer drainage ditch (“PD”), and an established laboratory line (“S”) were morphologically identical except for differences in size and flagellate transforming ability. Cultivation on a Trypticase-yeast extract-glucose medium (“TYG”) fortified with autoclaved E. coli resulted in increased cell size of 2 strains. Differences also were noted in growth rates and optimal growth temperatures. The autoclaved E. coli in TYG medium was replaceable with serum only for strains TS and PD. A basal salts medium + autoclaved E. coli supported growth of all 3 strains, but the basal salts medium + serum would not support growth of any of the strains.     

Plant Regeneration and Transformation of Sweet Papper (Capsicum frutescens)

Three strains of sweet pepper, (Capsicum frutescens) “Shuang Feng”, “Zhong Jiao No. 2” and “Zhong Jiao No.3” were screened out of six Chinese cuhivars for their high capacity of regeneration. The normal flowering and fertile regenerated plants have been obtained from cotyledons of seedlings from 10 to 16 days old by a four-step culture procedure; short induction, shoot elongation, rooting of excised shoot and transplanting into soil. MS was the basal medium in all steps, supplemented with different kind and different concentration Of phytohomores. Optimal shoot ioduction medium is MS +4–6mg/LBA+0.5mg/LIAA which gives rise a shoot regeneration frequency of 100%. 35% of the induced shoots elongated on the medium of MS+2mg/L Zeatin or 2mg/L. BA+l–3mg/L GA, and subsequently rooted on MS medium or in addition of 0.1–0.5mg/LNAA. The regenerants were transplanted into soil and developed into normal plants. In the transformation of sweet pepper using the leaf disc method, two kinds of wild type Agrobacterium tumefaciens, C58 and GV3111, have been screened out in regard to their high infection capacity. The transient expression of GUS gene was detected and Kanamycin-resistant shoots from infected cotyledons have been obtained. Further assay and transfering the TMV-resistant and CMV-resistant genes into sweet pepper are in progress.     

STUDIES ON SARGASSUM. I. A LIGHT MICROSCOPIC EXAMINATION OF THE WOUND REGENERATION PROCESS IN MATURE STIPES OF S. FILIPENDULA

Regrowth from wounded stipe explants of Sargassum can be divided into four stages based on cytological changes. The first stage involves changes associated with the wound reactions and the formation of a wound epidermis. The second stage includes the formation of a well defined medullary pit with meristematically active cells around its periphery. Several “bud primordia” are also formed which begin to grow by cell division towards the wound surface. The third stage involves a period of internal tissue differentiation in the “bud primordia” such that mitotic activity is localized in the bud tip and the basal cells grow by cell elongation. The fourth stage marks a major change in the morphology of the regeneration branch from a tubular structure to that of a flattened blade. This change in morphology is preceded by the formation of an apical pit around which the flattened growth appears to be organized.     

INDUCED APOSPORY IN THE LIVERWORT BLASIA PUSILLA

Immature sporophytes of Blasia pusilla L. collected in the field were excised from the protective gametophytic tissues and cultured on a slightly modified Knop's agar substrate in microphytotrons. Under the experimental conditions the setae elongated and after 33½ to 6 weeks many began to give rise to from 1 to 20 or more aposporous outgrowths. These subsequently developed into young gametophytes. The apices were then cut off, sterilized, and grown on glucose-mineral agar in aseptic culture. The resulting gametophytes were similar to haploid plants derived from spores in pattern and rate of growth, and in possession of rhizoids, ventral scales, lateral lobes, auricles, and stellate and discoid gemmae. They produced archegonia but no antheridia. The chromosome number of the aposporous plants was 18 in contrast to the normal haploid number of 9.     

Isolation of protoplasts from fern prothallia and their regeneration to gametophytes

Protoplasts were isolated enzymatically from prothallia ofLygodium japonicum. The protoplasts grown in a culture medium containing 0.6 M mannitol and 0.05 M sucrose began to divide within 8 days of culture, and after 30 days 10-cell clusters were present. When the cell-clusters were transferred into fresh media followed by sequential reduction of mannitol concentration, they developed rhizoids and protonemata. The reduction of mannitol concentration to 0.3 M resulted in the regeneration of a common gametophyte within 50 days of culture, and subsequently the regenerated gametophytes produced sporophytic leaves and roots.     

Induction of callus in the liverwort Marchantia palmata Nees

Abstract

In Marchantia palmata Nees, normal growth of thalli occurs on Nitsch's basal medium with 1% sucrose. With 4% sucrose, dark green callus is initiated by light intensities of 150–4,500 lux after 13–15 days of growth. In the dark only rhizoids develop from callus and at low light intensity (150 lux) rhizoids as well as knob-like green outgrowths differentiate. Increased light intensities induce differentiation of thalli and rhizoids from callus. The callus inducing effect of increased sucrose level can be replaced by higher concentrations of mannitol. Histological studies reveal that callus initially arises from the lower epidermis.     

Morpho-histological characterization and direct shoot organogenesis in two types of explants from <Emphasis Type="Italic">Bacopa monnieri</Emphasis> on unsupplemented basal medium

In the present study, high frequency regeneration has been obtained via de novo direct shoot organogenesis from leaf and internode explants in Murashige and Skoog (MS) basal medium without any phytohormone supplementation in Bacopa monnieri, an indigenous traditionally used medicinal herb. Leaves and internodes from different positions were excised from 4-weeks-old in vitro propagated B. monnieri plants and cultured on MS basal medium supplemented with 3% (w/v) sucrose and 0.75% (w/v) agar for 4 weeks. The induction of de novo shoot buds was observed at petiolar cut edges of leaf and both proximal and distal cut ends of internode explants within 10–15 days of culture. The first histological changes could be observed after 4–5 days, with meristematic activity of vascular bundles. Proliferation of epidermal cells gave rise to dome-shaped protuberances followed by shoot apical meristems formation and their vascular connections with explant tissues within 2 weeks of culture. However, a basipetal gradient of shoot regeneration from both types of explants collected along the branch axis was noticed after 4 weeks of culture. Leaf and internode explants near the basal region exhibited significantly higher number of shoot buds and micro shoots (8.8/leaf explant and 15/internode explant). Microshoots (7–12 micro shoots/leaf or internode explants) elongated (shoot length 8–9 cm) within 8 weeks on phytohormone free MS medium. Excised micro shoots rooted (100%) in hormone free MS medium within two weeks of culture. Rooted plants were then acclimatized and transferred to field with 95% survival. This protocol may be used for micropropagation, genetic transformation as well as a model system for evaluation of changes associated with acquisition of competence of differentiated cells in phytohormone free medium.