PHYSIOLOGICAL INVESTIGATIONS ON ALARIA ESCULENTA (LAMINARIALES,PHAEOPHYCEAE). II. ROLE OF TRANSLOCATION IN BLADE GROWTH1,2

Laboratory studies on blade growth in Alaria esculenta (L.) Grev. showed 3 periods of rapid blade elongation during the year: October–November, February–April and late June. The first two periods are characteristic of many Laminariales; the unique June peak may reflect local nutrient conditions. While the distal blade functions as a source, supplying organic matter to the blade meristem, the stipe can be a source during periods of rapid growth or a sink during late summer when blade growth is slow. Maximum enhancement of elongation rate of blade meristems was observed in 40–50 cm blades; longer blades showed no further increase in growth rate. This blade length-growth promotion relationship may be independent of seasonal variations in meristematic activity. ¹⁴C tracer experiments suggested that separate growth promotion effects by distal blade, sporophylls and stipe were not additive in the intact thallus. The preferential source of assimilate for blade meristem growth was the distal blade. Secondary sources: sporophylls, which were activated following excision of the primary source; and stipe, which began to translocate assimilate when both sources were removed. The role of secondary sources in nature is discussed. Profiles of radioactivity in alcohol-soluble organic matter in blades are evaluated in relation to tracer profiles in higher plants and mechanisms of translocation.     

PHYSIOLOGICAL INVESTIGATIONS ON ALARIA ESCULENTA (LAMINARIALES,PHAEOPHYCEAE). III. EXUDATION BY THE BLADE1,2,3

Comparison of exudation rate of medullary conducting cells in the midrib of Alaria esculenta (L.) Grev. showed lowest rates for the blade meristem and highest rates in the non-growing region, 300–500 mm from the meristem. Holding plants under continuous darkness or severing the wings from the midrib reduced exudation rate by 26 and 37%, respectively. Osmotic pressure of exudate in sink (meristem) and source (non-growing region, up to 500 mm from the meristem) were similar (34.6–36.1 · 10²kpa). Pressure flow mechanism of translocation is evaluated in Alaria.     

SOURCE-SINK RELATIONSHIPS IN THE BLADE OF ALARIA ESCULENTA (LAMINARIALES,PHAEOPHYCEAE)1

The translocation of ¹⁴C-labelled photoassimilate was studied in blades of the kelp, Alaria esculenta (L.) Grev., which had been surgically modified to produce source and sink regions of various sizes. Thirty cm above the blade base a 2.5 cm dia. section of the blade received a 1 h pulse of H¹⁴CO₃^?. Efflux of ¹⁴C-labelled photoassimilate from this “source disc” was monitored over the next 8 days using a Geiger-Müller detector probe. Accumulation of ¹⁴C-labelled solutes by a “sink disc” of similar diameter, 20 cm below the source disc, was also monitored. Rate of ^l4C efflux from the source disc was governed by two factors: (1) total sink size and (2) feed-back from competing sources. In the latter case, source export was depressed if the portion of the blade, just distal to the source disc, was present. While the initial ¹⁴C influx rate into the sink disc was greater when competing sinks were present, the total accumulation of ¹⁴C-photoassimilates was 2–3 times higher in the sink disc when competing sources were not present. Basipetal translocation velocity (1.3–1.7 cm h^?1) was unaffected by competing sources and sinks.     

THE RATE OF TRANSLOCATION IN ALARIA ESCULENTA (LAMINARIALES,PHAEOPHYCEAE)12

The rate of translocation of organic carbon in blades of Alaria esculenta (L.) Grev. (Laminariales) was calculated from blade growth data. The cross sectional area of sieve filaments in the midrib medulla was estimated from light microscopic examination of fresh material. Files of these filaments form a perimedullar ring occupying ca. 2/3 of the medulla. Values computed for specific mass transfer of carbon into the blade meristem ranged from 36.2 to 60.8 mg C.wk^?1.0.1 mm^?2 sieve filaments.     

ULTRASTRUCTURE OF SWARMERS IN THE LAMINARIALES (PHAEOPHYCEAE). II. SPERM1

The ultrastructure of sperm from 13 species in 11 genera of Laminariales collected in the northeast Pacific Ocean is unique in the brown algae. The sperm are elongate, and possess a nucleus, several mitochondria and two or three chloroplasts, but no eyespot. The anterior flagellum bears mastigonemes on the proximal half of its length; a distal “whiplash” portion lacks mastigonemes and is an extension of only the two central singlet microtubules of the axoneme. A peculiar feature of these sperm is the posterior flagellum, which is longer than the anterior flagellum and tapers distally as the doublet microtubules become singlets and decrease in number. This feature contrasts with the laminarialean zoospore, which possesses a short posterior flagellum with the usual “9 + 2” axoneme. The structure of these sperm differs from that reported for Chorda, the sperm of which resembles a primitive brown algal zoospore. The facts support the concept that Chorda is the most primitive member of the Laminariales.     

ULTRASTRUCTURE OF SWARMERS IN THE LAMINARIALES (PHAEOPHYCEAE). I. ZOOSPORES1

Zoospores of 17 species in 14 genera of Laminariales, collected in the northeast Pacific Ocean, were studied by electron microscopy. These zoospores are unique in the brown algae in lacking both an eyespot in the single chloroplast and any associated swelling at the base of the shorter, posterior flagellum. Spores of all species examined possess a distal whiplash portion on the longer, mastigoneme-bearing anterior flagellum. This appendage may sometimes be as long as the mastigoneme-bearing portion of the flagellum, but it is only seldom preserved in the preparations for electron microscopy. A microtubular cytoskeleton is probably responsible for maintaining the shape of the spore. It consists of a short band of about 10 microtubules between the two basal bodies, scattered tubules converging at the anterior of the spore, a band of 7–9 tubules directed anteriorly from the anterior basal body, and a band directed posteriorly from the posterior basal body. These anterior and posterior bands may form one continuous band looping around the periphery of the spore. Variation with possible taxonomic significance was found in the ultrastructure of vesicles which apparently contain adhesive material, and which are extruded through the plasmalemma when the zoospores settle.     

CRYOPRESERVATION OF THE GAMETOPHYTIC CELLS OF LAMINARIALES (PHAEOPHYTA) IN LIQUID NITROGEN1

The gametophytic cells of six species of Laminariales, Laminaria japonica Areschoug, L. longissima Miyabe, Kjellmaniella crassifolia Miyabe, Ecklonia stolonifera Okamura, E. kurome Okamura, and Undaria pinnatifida (Harvey) Suringar, were subjected to cryopreservation in liquid nitrogen. The cells were suspended in various cryoprotective solutions and slowly cooled to –40°C over a period of 4 h. After this slow cooling step, the suspensions were immediately immersed in liquid nitrogen. All the species of Laminariaceae used in the present study survived maximally in a mixture of ethylene glycol and proline. On the other hand, the gametophytic cells of Undaria pinnatifida, a member of the Alariaceae, survived maximally in the mixture of glycerol and proline. The viability of the thawed gametophytic cells decreased during postthawing incubation. The decrease in viability continued for 4–6 days, and the minimum levels ranged from 36.2% to 67.2%. After 4–6 days of incubation, the percentage viability of all strains began to increase due to the renewal of cell division.     

PHYSIOLOGICAL INVESTIGATIONS ON ALARIA ESCULENTA (L.) GREV. (LAMINARIALES). I. ELONGATION OF THE BLADE1,2,3

A seasonal rhythm in blade elongation rate was observed in Alaria esculenta. Maximum (3.0–3.6 cm/week) and minimum (0.5–1.0 cm/week) rates were recorded in June and September, respectively. Translocation of growth-promoting metabolites occurs within the main blade and between the sporophylls and blade. Bidirectional flow of growth-promoting metabolities within the stipe was demonstrated. Seasonal effects of nutrients (NO₃- and PO₄^3-) and photoperiod on blade elongation were shown in long-term laboratory experiments. Control of seasonal blade growth by exogenous and endogenous factors is discussed.     

THE RELEASE,SETTLEMENT AND GERMINATION OF ZOOSPORES IN CHORDA TOMENTOSA (PHAEOPHYCEAE,LAMINARIALES)1,2

Details of zoospore germination in Chorda tomentosa Lyngb. are outlined. Uninucleate zoospores, when released are embedded in a mucilaginous mass of carbohydrate which dissolves and the biflagellate zoospores become motile. The long anterior flagellum is composed of a highly coiled terminal region and a rigid lower section bearing mastigonemes. The rigid, short posterior flagellum lacks mastigonemes. After initial contact by the tightly coiled region of the anterior flagellum, the zoospore draws itself to the substrate by flagellar resorbtion. After deposition of 3 wall layers the germling produces a germ tube. During this time the disc-shaped chloroplast enlarges undergoing changes in shape. As the germ tubes reach ca. 15 μm they cease forward growth and swell at their tips. The majority of cytoplasm of the original zoospore moves into the tube. Just before the nucleus enters the tube, centriole replication occurs. Mitosis is presumed to take place somewhere in the germ tube so that at 24 h, 2-celled gametophytes are produced.     

EFFECTS OF EXTERNAL INORGANIC NITROGEN CONCENTRATION ON METABOLISM,GROWTH AND ACTIVITIES OF KEY CARBON AND NITROGEN ASSIMILATORY ENZYMES OF LAMINARIA SACCHARINA (PHAEOPHYCEAE) IN CULTURE1

The growth rate of Laminaria saccharina (L.) Lamour. is dependent on inorganic nitrogen in culture. Growth rates were saturated between 5 and 10 μmol · L^?1 nitrate. The activities of ribulose-1,5 bisphosphate carboxylase, phosphoenolpyruvate carboxykinase, mannitol-1-phosphate dehydrogenase, nitrate reductase and glutamine synthetase also varied with the concentration of inorganic nitrogen in the medium. All enzyme activities were lowest at 2.5 μmol · L^?1 nitrate (the lowest concentration used) increasing to a maximum activity between 10 and 30 μmol · L^?1 nitrate. Most enzyme activities followed a hyperbolic curve resembling those described by the Michaelis-Menten equation, with different half-saturation constants.     

LIFE HISTORY AND SYSTEMATIC POSITION OF PSEUDOCHORDA GRACILIS SP. NOV. (LAMINARIALES,PHAEOPHYCEAE)1

Pseudochorda gracilis sp. Nov. (Pseudochordacease, Laminariales) is described from the Japan Sea coast of Hokkaido/ the species is subtidal, epilithic and annual, appearing in spring and maturing in winter. Erect thalli grow solitary or in tufts on a small discoid holdfast. They are simple, cord-shaped and hollow, with inner hyphal filaments, cylindrical medullary cells and paraphyses consisting of 3–6 cells. Hair tufts are observed only in young thalli. Unilocular sporangia are sessile and narrowly ovate. In culture, P. gracilis shows a heteromorphic life history with oogamy, characteristic of the order Laminariales. Gametophytes are dioecious and dimorphic. Gametophytes mature under lower temperature conditions (usually below 10°C), and sporophytes mature under low temperature and short-day conditions (5°C, SD). The seasonal growth pattern of the species results from the photoperiod-temperature conditions controlling saprophyte maturation.     

EFFECTS OF NITRATE CONCENTRATION ON THE GROWTH AND PHYSIOLOGY OF LAMINARIA SACCHARINA (PHAEOPHYTA) IN CULTURE1,2

Laminaria saccharina Lamour. sporophytes were grown in enriched and synthetic media through a range of nitrate concentrations, There was an approximately linear relationship between growth and nutrient concentration up to 10 μ substrate concentration. The half-saturation constant (K_{2) was ca. 1.4 μ NO3-. The internal levels of NO3- increased at substrate concentrations above 10 μM b>3- and reached levels several thousand times higher than the surrounding medium. Thus there is evidence for luxury consumption of NOsb>3}^-. The chlorophyll content and photosynthetic capacities of plants also increased with increasing external NO₃^- The ecological implications of this work are considered.     

EFFECT OF LATRUNCULIN B AND BREFELDIN A ON CYTOKINESIS IN THE BROWN ALGA SCYTOSIPHON LOMENTARIA (SCYTOSIPHONALES,PHAEOPHYCEAE)1

In zygotes of the brown alga Scytosiphon lomentaria (Lyngb.) Link, cytokinesis proceeds by growth of membranous sacs, which are formed by fusion of Golgi vesicles and flat cisternae accumulated at the future cytokinetic plane. It has been reported that depolymerization of actin filaments by latrunculin B does not inhibit mitosis. However, this molecule prevents the formation of the actin plate, which appears at the region of intermingled microtubules from each centrosome just before and during cytokinesis. In this study, zygotes treated with latrunculin B were observed using EM. Remarkably, this reagent inhibited the formation of flat cisternae. Golgi vesicles gathered around the midzone between the two daughter nuclei and fused with the plasma membrane there. As a result, the plasma membrane invaginated, in a complicated manner, into the cytoplasm. However, these invaginations of the plasma membrane never produced a continuous partition membrane. The ultrastructure of zygotes treated with brefeldin A, which prevents Golgi‐mediated secretion, was also examined. Flat cisternae appeared at the future cytokinetic plane, and a new cell partition membrane was formed. However, the partition membrane became thick, because it was filled with amorphous material rather than the normal rigid fibrous material. These results suggested that actin is involved in the formation of flat cisternae, where it is necessary for completion of the new cell partition membrane, and that Golgi vesicles may play an important role in the deposition of cell wall material.     

ETIOLOGY AND DEVELOPMENT OF HYPERPLASIA INDUCED BY STREBLONEMA SP. (PHAEOPHYTA) ON MEMBERS OF THE LAMINARIALES (PHAEOPHYTA)1

Filaments of Streblonema sp. isolated from hyperplasia (galls) on Nereocystis luetkeana (Mert.) Post. et Rupr. induced similar gall growths when inoculated on young sporophytes of Nereocystis luetkeana, Macrocystis integrifolia Bory, and Laminaria japonica Aresch. Filaments were irregularly branched and uniseriate with cells 5–8 μm in diameter and 10–30 μm long. Growth under varied cultured conditions produced unilocular sporangia. Zoospores released from the sporangia germinated into identical filaments which also formed unilocular sporangia. Gall tissue originated from the innermost cells of the epidermal meristematic zone. When infected these cells divided in an irregular and unorganized manner. The resulting structure was a pronounced departure from normal morphology.     

大豆-根瘤菌共生体系光合与固氮关系研究

水培大豆和田间生长的大豆,接种根瘤菌 Rhizobium B16-11C 后植株全氮含量、叶片叶绿素含量和净光合速率及种子产量都明显增加。比较 Clark 大豆的结瘤品系和不结瘤品系获类似结果。摘除根瘤后3天内叶片净光合速率无明显变化。大豆植株遮阴、去叶或切掉地上部导致根瘤活性明显下降。但去豆荚不能提高根瘤固氮的比活性。根瘤活性的日变化不能用根瘤蔗糖、淀粉含量或周围温度的变化来解释,其控制因子尚待深入研究。     

VARIATION IN NITROGEN PHYSIOLOGY AND GROWTH AMONG GEOGRAPHICALLY ISOLATED POPULATIONS OF THE GIANT KELP,MACROCYSTIS PYRIFERA (PHAEOPHYTA)1

Three geographically isolated populations of the giant kelp, Macrocystis pyrifera (L.) C. Ag., were examined for responses to nitrate availability in batch culture experiments using juvenile sporophytes reared from spores in the laboratory. Although maximum rates of nitrate-saturated growth were similar among groups, there were significant quantitative differences in the response to nitrate limitation that can be related to natural patterns of nutrient availability at these sites. Plants from Santa Catalina Island (most oligotrophic) achieved maximum growth rates at ambient nitrate concentrations that were lower than those for plants from Monterey Bay, California (most eutrophic), or Refugio State Beach (near Santa Barbara, California). Tissue nitrogen and amino acid concentrations were highest in plants cultured from Santa Catalina Island populations at all external nitrate concentrations, suggesting that differences in nitrate requirements for growth may reflect the efficiency of nitrate uptake and assimilation at subsaturating nitrate concentrations. Given the different physical environments from which these plants came, the data suggest that geographically isolated populations of M. pyrifera have undergone genetic divergence that can be explained by ecotypic adaptation to unique habitat conditions at these sites.     

THE RELATIVE IMPORTANCE OF WATER MOTION ON NITROGEN UPTAKE BY THE SUBTIDAL MACROALGA ADAMSIELLA CHAUVINII (RHODOPHYTA) IN WINTER AND SUMMER1

The influence of seawater velocity (1.5–12 cm · s^?1) on inorganic nitrogen (N) uptake by the soft‐sediment perennial macroalga Adamsiella chauvinii (Harv.) L. E. Phillips et W. A. Nelson (Rhodophyta) was determined seasonally by measuring uptake rate in a laboratory flume. Regardless of N tissue content, water velocity had no influence on NO₃^? uptake in either winter or summer, indicating that NO₃^?‐uptake rate was biologically limited. However, when thalli were N limited, increasing water velocity increased NH₄⁺ uptake, suggesting that mass‐transfer limitation of NH₄⁺ is likely during summer for natural populations. Uptake kinetics (V_max, K_s) were similar among three populations of A. chauvinii at sites with different mean flow speeds; however, uptake rates of NO₃^? and NH₄⁺ were lower in summer (when N status was generally low) than in winter. Our results highlight how N uptake can be affected by seasonal changes in the physiology of a macroalga and that further investigation of N uptake of different macroalgae (red, brown, and green) during different seasons is important in determining the relative influence of water velocity on nutrient uptake.     

THE EFFECTS OF TEMPERATURE,IRRADIANCE, AND NITROGEN ON THE ENCYSTMENT AND GROWTH OF THE FRESHWATER DINOFLAGELLATES PERIDINIUM CINCTUM AND P. WILLEI IN CULTURE (DINOPHYCEAE)1

Effects of temperature, irradiance, and nitrogen availability on the encystment and growth of the freshwater dinoflagellates Peridinium cinctum Ehrenberg and Peridinium willei Huitfeld-Kaas were studied in culture. Lack of nitrogen was the main trigger of encystment in both species. Irradiance had a secondary effect on the percentage of the population of each species that encysted. Temperature did not significantly affect encystment in either species. In both species, only a small percentage of the population underwent encystment. Low light had an inhibitory effect on the growth of P. willei growing in nitrogen-sufficient medium.     

THE EFFECT OF GROWTH IRRADIANCE ON THE COUPLING OF CARBON AND NITROGEN METABOLISM IN CHAETOMORPHA LINUM (CHLOROPHYTA)

The influence of growth irradiance on the non-steady-state relationship between photosynthesis and tissue carbon (C) and nitrogen (N) pools in Chaetomorpha linum (Muller) Kutzing in response to abrupt changes in external nitrogen (N) availability was determined in laboratory experiments. For a given thallus N content, algae acclimated to low irradiance consistently had a higher rate of light-saturated photosynthesis (P_max normalized to dry weight) than algae acclimated to saturating irradiance; for both treatments, P_max was correlated to thallus N. Both P_max and the photosynthetic efficiency (α_dw) were correlated in C. linum grown at either saturating or limiting irradiance over the range of experimental conditions, indicating that variations in electron transport were coupled to variations in C-fixation capacity despite the large range of tissue N content from 1.1% to 4.8%. Optimizing both α and P_max and thereby acclimating to an intermediate light level may be a general characteristic of thin-structured opportunistic algae that confers a competitive advantage in estuarine environments in which both light and nutrient conditions are highly variable. Nitrogen-saturated algae had the same photosynthesis–irradiance relationship regardless of light level. When deprived of an external N supply, photosynthetic rates did not change in C. linum acclimated to low irradiance despite a two-fold decrease in tissue N content, suggesting that the active pools of chlorophyll and Rubisco remained constant. Both α and P_max decreased immediately and continuously in algae acclimated to high irradiance on removal of the N supply even though tissue N content was relatively high during most of the N-starvation period, indicating a diversion of energy and reductant away from C fixation to support high growth rates. Carbon and nitrogen assimilation were equally balanced in algae in both light treatments throughout the N-saturation and -depletion phases, except when protein synthesis was limited by the depletion of internal N reserves in severely N-starved high-light algae and excess C accumulated as starch stores. This suggests that the ability for short-term adjustment of internal allocation to acquire N andC in almost constant proportions may be especially beneficial to macroalgae living in environments characterized by high variability in light levels and nutrient supply.