Abscisic-acid-induced turion formation in Spirodela polyrrhiza L. IV. Comparative ion flux characteristics of the turion and the vegetative frond and the effect of ABA during early turion development

Abstract Using the method of compartmental analysis, the ion fluxes and compartment concentrations of Ca²⁺, K⁺ and Cl^- have been compared in the untreated vegetative frond and the abscisic acid (ABA) induced turion of Spirodela polyrrhiza. The ABA-induced turion is characterized by reduced Ca²⁺ exchange across the tonoplast and low vacuolar Ca²⁺ concentration relative to the vegetative frond. In addition the turion exhibits a higher plasmalemma flux with a correspondingly high Ca²⁺ concentration in the cytoplasm. The concentration of K⁺ and Cl^- is much lower in the cytoplasm of the ABA-induced turion than in the vegetative frond with the influx/efflux ratio at both the plasmalemma and the tonoplast being less than 1, a finding exhibited also in dormant storage tissue. Treatment of vegetative fronds with ABA for 18 h resulted in a reduced K⁺ plasmalemma efflux relative to untreated vegetative fronds and a concomitant increase in the cytoplasmic concentration. There was no rapid effect of ABA on Ca²⁺, K⁺ or Cl^- fluxes through either membrane. These results are consistent with the notion that drastic changes in ion fluxes and concentrations in the turion are a secondary consequence of ABA-induced development, possibly due to prior regulation by ABA of enzymes inherent to processes involved in membrane transport.     

Abscisic-acid-induced turion formation in Spirodela polyrrhiza L. I. Production and development of the turion

Abstract The production, growth, and development of the abscisic-acid-induced turion (a small dormant bud) of Spirodela polyrrhiza were investigated. Addition of ABA to a culture of S. polyrrhiza resulted in growth inhibition at concentrations as low as 10⁻⁶molm⁻³, growth being completely arrested at 10⁻² mol m. Over a single order of magnitude range around I0⁻⁴molm⁻³, ABA also induced the production of turions. The range of turion-producing concentrations of ABA was found to be much narrower than previously reported, turion production having a clearly defined threshold, optimum, and upper limit. The possibility that growth inhibition and turion formation are integrally linked aspects of a single response is discussed. Only primordia ≤0.7 mm long at the time of ABA addition could be induced to develop into turions and the events leading to turion formation were found to be reversible up to 72 h in ABA . It is concluded that in terms of turion formation there is a sensitivity window to abscisic acid lasting some 4–20h in the normal developmental life of frond cells. Providing cells experience the appropriate signal in this sensitivity window they initiate a new programme which eventually leads to turion formation. Microscopical analysis showed that the cells within this sensitivity window were still actively dividing. It is suggested that the developmental switch-over to rapid cell expansion and separation marks the end of this ABA sensitivity window.     

Co‐action of temperature and phosphate in inducing turion formation in Spirodela polyrhiza (Great duckweed)*

Increased phosphate concentration, higher temperature and addition of glucose all increased the number of fronds and turions of the duckweed Spirodela polyrhiza formed under in vitro conditions. Increasing the number of turions by increasing the plant biomass does not mean that the developmental process (switch of the programme of the primordia from vegetative fronds toward resting turions) has been specifically influenced. The specific turion yield (STY; number of turions formed by one frond) and the time of onset of turion formation have been used as more specific measures of turion induction. At more than 30 µm initial phosphate the STY was increased by lower temperature (15 °C) and became independent of the phosphate concentration. Between 10 and 30 µm and at higher temperatures (25 °C) the STY was increased by lower phosphate levels. The stimulatory effect of lower temperature was more pronounced than that of lower phosphate concentrations. Decreased phosphate concentration highly accelerated the formation of the first turions. The influence of low temperature was small at lower phosphate concentration but became dominant at higher concentrations (especially in autotrophic cultures). Low phosphate levels (e.g. 10 µm ) and low temperatures (e.g. 15 °C) both represent specific turion‐inducing factors having significant interactive effects. In S. polyrhiza, these signals may replace the interactive effects of photoperiods and low temperature known from other hydrophytes in turion induction under natural conditions.     

Growth and turion formation of Potamogeton crispus in response to different phosphorus concentrations in water

The vegetative growth and turion formation of Potamogeton crispus, a submersed aquatic macrophyte, was investigated under a range of phosphorus (P) concentrations (0.025, 0.25, 2.5 and 25 mg P L^?1) in the ambient water free of algae, aiming to identify the responses of submersed aquatic macrophytes to nutrient enrichment, a common eutrophication problem in China and worldwide. Plant growth was not affected by different P concentrations in terms of biomass accumulation of stems and leaves. However, the contents of chlorophyll a and starch in plants decreased with increasing water P levels, whereas chlorophyll b and carotenoids declined with P level ranging from 0.025 to 2.5 mg P L^?1. The soluble sugar content decreased when water P concentration increased up to 2.5 mg L^?1. The P content in plants increased with increasing water P levels, whereas plant N content decreased and soluble protein increased when water P concentration increased over 0.25 mg L^?1, implying that P. crispus may have modified its metabolism to adapt to water P availability. When P concentration increased to 25 mg L^?1, the number and dry matter production of turions per plant decreased significantly. Meanwhile, there was a significant reduction in turion weight and the accumulations of soluble sugar and starch in turion, when water P concentration was over 0.25 mg L^?1. The results suggest that turion formation in P. crispus is sensitive to P concentration in the ambient water, and high P levels may lead to decreases in P. crispus populations due to the decline in turion production.     

Abscisic-acid-induced turion formation in Spirodela polyrrhiza L III. Specific changes in protein synthesis and translatable RNA during turion development

Abstract The developmental process leading to the formation of the abscisic acid (ABA) induced turion of Spirodela polyrrhiza was accompanied by a repression of nucleic acid and protein synthesis. DNA synthesis in the developing lurion (induced by 10⁻⁴mol m⁻³ ABA) was inhibited within 3h of ABA addition, followed by a repression of protein synthesis after 24 h, while RNA synthesis was not inhibited until 3 d. The inhibitory effect of ABA on protein synthesis was found to be selective and the synthesis of several novel proteins appeared to be induced. These effects were specific to ABA-sensitive tissue. The relationship between the changes in the protein and mRNA profiles during the development of the turion was investigated. The rapid general inhibition of protein synthesis at early stages of lurion formation could not be accounted for by the level of translatable mRNA, indicating an effect of ABA at the translational level. The specific alteration to the pattern of in vivo labelled proteins could have resulted, however, from control of the level of specific mRNAs for those particular proteins. Only after 3 d in ABA, when the developing primordium is committed to the turion developmental pathway, is there a total inhibition in the production of mRNA leading to the shutdown of all primary processes and the onset of the irreversible events leading to the dormant state.     

Clonal Differences in the Formation of Turions are Independent of the Specific Turion-inducing Signal in Spirodela polyrhiza (Great Duckweed)

Abstract: Turion (survival organ) formation in Spirodela polyrhiza includes a switch in the programming of the primordia from the formation of vegetative fronds toward resting turions. The specific turion yield (SY; number of turions formed by one frond) is used to evaluate the effect of three turion-inducing signals: low phosphate concentration (depleted due to frond growth), low temperature (15 °C) and exogenously applied abscisic acid (1 μM). The formation of turions was observed in the presence of any of the turion-inducing factors in all three clones of S. polyrhiza investigated (clones 9256 from Finland, SJ from Germany and SC from Cuba). The clone SC showed no specific induction by low temperature or phosphate limitation in one nutrient medium. Regardless of the specific signal applied, the SYs were highest in clone 9256 and lowest in clone SC, demonstrating signal-independent clonal differences. Clonal differences are therefore located in the developmental-specific common phase of the transduction chains leading to turion formation. We intend to use clonal differences in the molecular analysis of turion formation, e.g., by cDNA-based amplified fragment length polymorphism, to distinguish signal-specific and developmental-specific gene expression. In contrast, the total turion yield is useful in an ecological context to evaluate the number of turions available to support the survival of a population of plants but gives little information about the physiological process.     

Influence of salinity and high temperature on turion formation in the duckweed Spirodela polyrhiza

The influence of NaCl (salinity; 0-5 ‰) and higher temperature (heat stress; 32 °C) on yield of turion formation has been tested in the duckweed Spirodela polyrhiza for the first time. Turion formation was more sensitive to both stressors than the growth of the vegetative fronds: (1) the concentration of NaCl which produces half-maximal inhibition was lower for turion formation than for growth by the factor of five. (2) At 32 °C turion formation was completely blocked whereas growth rates decreased by only 20% as compared with 28 °C.     

The clonal dependence of turion formation in the duckweed Spirodela polyrhiza—an ecogeographical approach

Formation of turions, the vegetative perennation organs, plays an important role in the survival strategy of Spirodela polyrhiza (L.) Schleiden. Turion formation [quantified as number of turions formed per frond; specific turion yield (SY)] was investigated in 27 clones collected from a wide geographical range. The Pearson correlation was tested with (1) duration of growing season (monthly average temperature of ≥10°C), (2) relative growth rate of the fronds, (3) longitude and latitude, and (4) several climatic parameters, in all possible single and multiple regressions. All single coefficients of determination were below 0.10. The highest correlation (R² = 0.61; adjusted for the number of explaining variables 0.54) was found in a multiple linear regression with the following five parameters: average temperatures over the year and during the growing season, duration of the growing season and precipitation over the year and during the growth period. All these parameters were shown to have significant contributions. This equation was used successfully to predict the SY of five newly isolated clones. Finally, on the basis of all 32 clones the following conclusions were drawn: The mean annual temperature has the highest impact. It is suggested that lower temperatures decrease the survival rate of turions and that adaptation refers to increasing SY. The different levels of SY in the clones (ranging from SY = 0.22 to 5.9) were detected even after several years of in vitro cultivation. It is therefore assumed that these adaptations to the climatic conditions are genetically determined.     

Environmental and hormonal control of turion formation in Myriophyllum verticillatum

The turions of Myriophyllum verticillatum, an aquatic vascularplant, develop in the fall and function in propagation and dispersalas well as in over-wintering. Experiments with controlled evnironmentsindicate that both temperature and photoperiod regulate turionformation. Turions can be induced at 15°C or lower, butnot at 20°C. At 15°C, turions form in both 8- and 12-hrdays, but not in 16-hr days. Plants collected in early springdo not form turions readily in response to short days unlesspreviously exposed to long days; thus, turion formation is along-day-short-day response. This combination of photoperiodand temperature requirements probably prevents turion developmentin early spring when the temperature and photoperiod are similarto those in the fall. Treatment of plants with ABA (10^–5M) enhances turion development under marginally inductive conditions(12-hr days at 15°C) but cannot induce it under long days.On the other hand, the cytokinin benzyladenine (10^–5 M)blocks turion formation. GA3 (10^–5 M) and AMO-1618 (10^–5M) exert only small qualitative effects on turion development,while IAA (10^–5 M) retards it. During turion development,the level of ABAlike activity and of one or two unidentifiedinhibitors increases. Cytokinin activity decreases at the startof turion formation, increases during development, then decreasesat abscission. Thus two lines of evidence suggest that a decreasein cytokinin activity and an increase in acidic inhibitor activityplay important roles in turion induction. ¹Present address: Biological Station, University of Michigan,Ann Arbor, Michigan 48109, U. S. A. (Received December 1, 1975; )     

Low‐molecular weight carbohydrates modulate dormancy and are required for post‐germination growth in turions of Spirodela polyrhiza

The aquatic duckweed Spirodela polyrhiza propagates itself vegetatively by forming turions – bud‐like perennation organs – in the autumn, which spend the winter on the bottom of ponds and then germinate in the following spring and proliferate on the water surface. Newly formed turions usually require a period of cold after‐ripening and light to germinate effectively, but an ample supply of exogenous sugar can lead to germination even in the dark and independent of after‐ripening. The results of the present study indicate that the availability of readily metabolised carbohydrates is a determining factor for turion germination. Freshly harvested turions do not contain soluble, low‐molecular weight carbohydrates at a level sufficient to allow germination to take place, but after‐ripened turions do. Augmentation of the soluble carbohydrate content during after‐ripening derives from gradual breakdown of reserve starch of the turions. The long time required for any germination to be observed in turions incubated in darkness and the limited frequency of germination in the dark (about 50% of turion population), even with an ample external sugar, supply emphasise that both after‐ripening and light are essential for ensuring rapid germination and subsequent frond proliferation at an ecologically appropriate time. The carbohydrate supply required for rapid proliferation of the fronds produced at germination is provided by the rapid light‐induced breakdown of turion reserve starch.     

Effects of sediment anoxia and light on turion germination and early growth of Potamogeton crispus

Potamogeton crispus is a cosmopolitan aquatic species and is widely used as a pioneer species for vegetation restoration of eutrophic lakes. However, many restoration projects applying P. crispus turions have not been successful. Earlier studies focused on effects of light and temperature on turion germination. The purpose of this study was to determine whether sediment anoxia and light interactively affected the turion germination and early growth of P. crispus. Anoxic conditions in the experiment were produced by adding sucrose to the sediment. The germination rate of the turions was 68–73% lower in the highly anoxic condition treatment than in the control. Medium light intensity (10% of natural light at the water surface) was more favorable for germination under slightly anoxic conditions than either low or high light intensity. The growth of newly-formed sprouts was also significantly inhibited by sediment anoxia. Photosynthesis and shoot biomass were reduced under sediment anoxia, whereas total chlorophyll content, root biomass, and soluble protein content were highest in the low anoxic condition treatment. Medium light improved net photosynthesis and biomass production of the sprouts. We conclude that turion germination and sprout growth can be significantly inhibited by sediment anoxia. Medium light intensity may alleviate this inhibition by anoxia, but light has little effect when sediment anoxia is severe. For the purposes of vegetation restoration, more attention should be paid to the role of sediment anoxia, and it is necessary to improve sediment and light conditions for turion germination and early growth of P. crispus in eutrophic lakes. These results will contribute to a more complete understanding of turion germination dynamics of P. crispus and will be useful for future restoration programs. Handling editor: S. M. Thomaz     

Photophysiology of turion germination inSpirodela polyrhiza (L.)Schleiden. The cause of germination inhibition by overcrowding

Red-light-induced (via phytochrome) germination decreased with increasing numbers of turions per germination flask (overcrowding). Three hypotheses concerning the mechanism of this germination inhibition were tested, related to abscisic acid, ethylene, and oxygen deficiency: (i) Although abscisic acid is a powerful inhibitor of turion germination it had to be excluded as a cause, because abscisic acid was not secreted from turions into the nutrient solution, (ii) Ethylene (ethrel) strongly inhibited growth of newly formed sprouts, but germination response itself was not inhibited, (iii) Germination inhibition did not appear if short light pulses were substituted by continuous irradiation. It reappeared in the presence of the photosynthesis inhibitor 3-(3, 4-dichlorophenyl)-l, 1-dimethylurea, but it was not observed in aerated nutrient solutions, or when Petri dishes instead of Erlenmeyer flasks were used. Decreased oxygen concentrations in the nutrient solution were produced by turion respiration. Consequently, anaerobiosis within the nutrient solution caused by turion respiration was the reason for germination inhibition by overcrowding.     

No photoperiodoc control of the formation of turions in eight clones of Spirodela polyrhiza

The influence of daily photoperiod (8, 16, 24 h) on eight clones of Spirodela polyrhiza was tested in two different nutrient media. The number of vegetative fronds and resting turions formed after 50 days of cultivation were scored. The specific turion yield (STY; number of turions formed per vegetative frond) was used to evaluate the effectiveness of turion formation of the tested clones. All clones formed turions in both nutrient media. The STY varied substantially between the different clones, ranging from 0.22 +/- 0.03 (clone SC from Cuba) to 3.9 +/- 0.3 (clone 9256 from Finland) in continuous light. The STY increased with increasing duration of the photoperiod. This increase may have been due to the extended period of photosynthesis rather than that of a photoperiodic long-day response. Shorter photoperiods did not stimulate turion formation in any of the clones. S. polyrhiza is a day-neutral plant with respect to turion formation, as noted previously (Appenroth et al. 1990. Annals of Botany 66: 163-168). In accordance with this conclusion, no correlation was detected between the STY and the latitude at which the clones occur naturally. Environmental factors other than shortening of photoperiods seem to be effective in signalling seasonal changes of growth conditions in advance to S. polyrhiza.     

MORPHOLOGY OF THE SUBANTARCTIC RED ALGA CENACRUM SUBSUTUM GEN. ET SP. NOV. (RHODYMENIALES) FROM MACQUARIE ISLAND1

The red alga Cenacrum subsutum gen. et sp. nov. is described from material collected at Macquarie Island in the subantarctic between November 1977 and February 1978. The habit and carposporophyte development are similar to members of the family Rhodymeniaceae (Rhodymeniales), but certain vegetative features are unique. The frond is a variously incised or lobed foliose blade with hollow apices above and a medulla which becomes progressively filled basipetally with ingrowing rhizoidal filaments. Details of carpogonial branch, auxiliary cell, connecting cell and gonimoblast anatomy are given, as well as observations on the habitats and distribution of the species.     

Hermodactylus tuberosus L. (Iridaceae) pollen organisation before and after anther dehiscence

ABSTRACT

The morphology, cytology and viability of Hermodactylus tuberosus L. (Iridaceae) pollen were examined from the first mitosis until maturation and after anther opening. During maturation, the pollen coat becomes modified, and the vegetative cell cytoplasm accumulates several types of reserve substances. In the vegetative cell cytoplasm, starch is quickly utilised whereas lipid inclusions of different dimensions, shape and composition occur during pollen maturation. Pollen from opened anthers have a thin pollen coat; the cytoplasm has mostly lipid reserves, and many small vesicles and vacuoles. It is similar in size or larger than pollen located inside the anther, and its viability does not decrease until one day after anther dehiscence. Large osmiophilic bodies, different from those of the vegetative cell cytoplasm, are present in the generative cell cytoplasm starting from the first stage of pollen development. The poorly developed pollen coat in pollen from opened anthers suggests that it plays a minor role in attracting insects for pollination. The size and structural and ultrastructural features of mature pollen indicate that it does not undergo dehydration and possesses sufficient vigour for immediate germination.     

Germination of Spirodela polyrhiza turions: the role of culture conditions during turion development

The rapidly germinating "old" turions of Spirodela polyrhizawere shown to derive mainly from the slowly germinating "young"turions. This modification to "old" turions could occur evenin isolated "young" turions, and was accelerated by sucrose.It is suggested that this modification is a form of turion senescenceand that turion initiation and maturation are strongly influencedby exogenous carbon and nitrogen sources. (Received November 19, 1979; )     

A new aerial alga, Stichococcus ampulliformis sp. nov. (Trebouxiophyceae, Chlorophyta) from Japan

A new aerial alga, Stichococcus ampulliformis S. Handa sp. nov. (Trebouxiophyceae, Chlorophyta) is described based on a clone isolated from the bark of Cephalotaxus harringtonia (Knight ex Forbes) K. Koch collected from Taishaku‐kyo Gorge, Hiroshima Prefecture, south‐west Japan. This alga was examined by light microscopy and transmission electron microscopy and subjected to molecular phylogenetic analysis. Based on its morphological features and life‐cycle, especially short filament formation, the alga was assigned to the genus Stichococcus Nägeli. However, this alga differs from other described Stichococcus species in that it reproduces by a form of ‘budding’, producing two daughter cells of different sizes. The larger cell, enclosed within the mother cell wall, soon reaches the size of a normal vegetative cell. The smaller cell is extruded and takes longer to reach full vegetative size. A phylogenetic tree constructed using 18S rRNA sequences indicated that, within the Trebouxiophyceae, S. ampulliformis is closely related to S. bacillaris Nägeli and some species of Prasiola Meneghini.     

梁子湖苦草繁殖体的分布及其萌发初步研究

苦草鳞茎(冬芽)在梁子湖的垂直分布深度与其重量呈显著正相关,而重量也与鳞茎芽数显著相关 (P<0.05).埋藏深度、水分状况和鳞茎本身的大小都显著影响鳞茎的出土能力.去除第一位芽和切碎鳞茎后促进了其余芽的萌发;上年产生的未萌发的鳞茎在条件适宜的时候也可以很好的萌发,这些可能成为苦草在鳞茎被牧食后,维持种群数量的有效对策.沙质底比泥质底更有利于苦草种子的萌发,但在没有扰动的情况下,幼苗的定植能力在两种基质中没有显著差异.       

TURION FORMATION AND GERMINATION IN SPIRODELA POLYRHIZA

Three clones of Spirodela polyrhiza L. (Schleid.) formed dormant bodies called turions. A clone from Puerto Rico did not form turions under all conditions tried. In those clones producing turions, formation was stimulated by the addition of sucrose (10–50 mM) to the nutrient solution. Increased levels of Ca(NO₃)₂ plus sucrose stimulated turion production. In the absence of NO₃^–, Ca⁺⁺ was more effective than K⁺ in stimulating turion formation. Turion buoyancy was not light dependent, nor was it promoted by sucrose. Normal turions required light for germination, whereas sucrose-induced turions germinated in the dark. Dark germination was not promoted by either Ca⁺⁺ or K⁺. Sucrose stimulation of turion formation and subsequent promotion of dark germination was attributed to metabolic rather than osmotic effects. One hundred mM sucrose concentrations inhibited turion buoyancy and germination. Turions formed one primary abscission layer which separated them from the stolon and the mother frond. Subepidermal idioblasts appeared to seal the stolon stump after separation.