REGULATION OF GAMETOGENESIS IN SCENEDESMUS OBLIQUUS (CHLOROPHYCEAE)1

The effects of nutrients, temperature and light on gametogenesis in Scenedesmus obliquus (Turp.) Kütz, were studied in culture. Concentrations of nitrogen in the medium employed showed a marked influence on gamete production. Gametogenesis is inhibited by N excess but is not a response to N starvation or depletion. A drop in N level from that of the growth medium is not required, nor does it per se trigger gametogenesis. The N concentration satisfying growth requirements insufficiently low to permit sexual differentiation. Nitrogen level in the growth medium has no effect on subsequent N to maintain a typical culture. Number of gametes present at maximum production time is inversely related to N concentration, but neither time of onset of gametogenesis, nor time of maximum gamete production is affected by N concentration. Cultures incubated at 15 C in medium lacking N take a minimum of 20—24 h to develop cells irreversibly committed to gamete formation. At the concentrations tested, no medium component other than the N-containing salt affected gametogenesis. Temperature influences both time of maximum production and numbers present at maximum production time. Time of maximum production is inversely related to incubation temperature; a 15 C incubation temperature yielded highest gamete production. Light enhances gametogenesis but gamete formation can occur in absence of light. Achievement of a light-saturated response is dependent upon illumination given at two critical periods: one occurs shortly after N withdrawal; the other occurs later, when cells are becoming irreversibly committed to gamete formation. Ability to produce gametes diminished with prolonged laboratory culture.     

GRAZER-INDUCED COENOBIAL FORMATION IN CLONAL CULTURES OF SCENENDESMUS OBLIQUUS (CHLOROCOCCALES, CHLOROPHYCEAE)

Five strains of the phenotypically plastic alga Scenedesmus obliquus (Turpin) Kützing were cultured in standard medium or in medium containing filtered water from a Daphnia culture to examine grazer-induced coenobia formation. Strains varied considerably in their response to Daphnia chemicals. A rapid formation of four- and eight-celled coenobia was observed in the presence of Daphnia water in three strains. One strain showed numerous irregular aggregates with more than eight cells per colony, and no colony formation occurred in a fifth strain. In standard medium, all cultures remained unicellular. Growth rates were affected by Daphnia water in four strains and were different among strains. The chl a concentration was significantly different among strains but was not influenced by Daphnia water. The specific absorption coefficient was reduced at larger colony sizes, suggesting a possible cost associated with colony formation. The phenomenon of grazer-induced coenobia formation seems widespread but not universal within the species S. obliquus.     

GRAZER-INDUCED COLONY FORMATION IN SCENEDESMUS ACUTUS (CHLOROPHYCEAE): ECOMORPH EXPRESSION AT DIFFERENT TEMPERATURES

Scenedesmus acutus Meyen was cultured at four temperatures (9.5°, 16.5°, 24°, and 29° C) in standard medium or in medium with filtered water from a Daphnia culture. Growth was significantly reduced at low temperatures. At 9.5° C it took more than a week before formation of eight-celled coenobia occurred in both the absence and presence of water from a Daphnia culture. At higher temperatures, formation of four- and eight-celled coenobia occurred more rapidly and was already observed in the presence of Daphnia water within 2 days. As cultures aged, also in the absence of Daphnia water, four-celled coenobia became dominant. At cold temperature, cell volume initially was significantly larger but declined after 3–4 weeks. Grazer-induced colony formation had occurred independent of the incubation temperature, but the number of cells per colony was increased with declining temperature. The morphological expression may be interpreted as a cyclomorphosis driven by nutrients, temperature, and chemical cues from grazers.     

CHANGES IN HYDROGENASE ACTIVITY DURING THE SYNCHRONOUS GROWTH OF SCENEDESMUS OBLIQUUS D3

1. A fairly good synchronization of Scenedesmus cells was obtainedby transferring the cells grown in a medium containing a lowconcentration of iron into a medium containing relatively highconcentration of iron.
2. During the synchronous culture in themineral medium, a goodparallelism between the average cellvolume and hydrogenaseactivity was observed.
3. Effect of glucoseon the development of the hydrogenase activitywas variabledepending on the stage of algal growth.
4. Iron is essentialfor the development of the hydrogenase activityand glucosesupplementary.

¹On leave from Laboratory of Applied Botany, Faculty of Agriculture,Kyoto University, Kyoto.     

氯氰菊酯对斜生栅藻的毒性研究

研究了农药氯氰菊酯对斜生栅藻的毒性效应,结果显示：以丙酮为溶剂量,其96h EC50为112.45mg/L;以乙酸乙酯为溶剂时,其96hEC50为112.81mg/L。同时也研究了氯氰菊酯对斜生栅藻的光合色素含量、色素光谱、可溶性蛋白质含量和超氧物歧化酶活性的影响。     

PHOTOBIOTREATMENT: INFLUENCE OF NITROGEN AND PHOSPHORUS RATIO IN WASTEWATER ON GROWTH KINETICS OF SCENEDESMUS OBLIQUUS

Nitrogen and phosphorus concentration in the effluent of a wastewater treatment plant can vary significantly, which could affect the growth kinetic and chemical composition of microalgae when cultivated in this medium. The aim of this work was to study the rate of growth, nutrient removal and carbon dioxide biofixation as well as biomass composition of Scenedesmus obliquus (S. obliquus) when it is cultivated in wastewater at different nitrogen and phosphorus ratio, from 1:1 to 35:1. A more homogeneous method for calculating productivities in batch reactors was proposed. The proper N:P ratio for achieving optimum batch biomass productivity ranged between 9 and 13 (263 and 322 mg L^?1 d^?1 respectively). This was also the ratio range for achieving a total N and P removal. Above and below this range (9–13) the maximum biomass concentration changed, instead of the specific growth rate.The maximum carbon dioxide biofixation rate was achieved at N:P ratio between 13 and 22 (553 and 557 mg CO2 L^?1 d^?1 respectively). Lipid and crude protein content, both depend on the aging culture, reaching the maximum lipid content (34%) at the lowest N:P (1:1) and the maximum crude protein content (34.2%) at the highest N:P (35:1).     

酚类对栅列藻细胞的影响

作者测定了120余种化学药物,其中30余种为酚类化合物对斜生栅列藻的毒性影响。所用方法为在培养基内混入溶解后的各种药物,接种藻类后进行培养,观察效果。实验证明邻羟基苯甲醛和5,5’-二氯-2,2’-二羟基二苯甲烷及5种胺类化合物对藻类具有毒性。50ppm剂量的邻羟基苯甲醛和DDM足以杀死藻类,使藻类由绿色变成白色,叶绿体破坏,叶绿素消失。扫描电镜和透射电镜照片表明,两种药物的作用主要在破坏叶绿体和贮藏物淀粉粒。     

DISTRIBUTION OF DEFENSE NECTARIES IN IPOMOEA (CONVOLVULACEAE)

The structure and distribution of defense nectaries in the genus Ipomoea (Convolvulaceae) were investigated. These nectaries do not reward pollinators and probably contribute to antiherbivore defense. Of 22 species sectioned, 15 had defense nectaries on the sepals. Of 12 other species observed, ten had sepal nectaries and two did not. Structurally, 14 of the species sectioned had crypt sepal nectaries and one had a basin nectary. Of the 14 species with crypt nectaries, two had invaginated spaces adding greatly to the internal area of these nectaries, and forming the most complex nectaries that have been reported. We term these labyrinthine crypt nectaries. All three types of nectaries are lined with secretory trichomes along the proximal surfaces of the crypts. Species with defense nectaries on the sepals tend to have petiolar defense nectaries as well, but the two locations may have different nectary types; e.g., basins on the petiole and crypts on the sepals. Since most reports of the function of these nectaries have shown antiherbivore defense by nectar feeders, the distributions of defense nectaries with respect to region and life history of the species were sought. Plants without sepal nectaries were found to have significantly smaller seeds than plants with sepal nectaries; they were also more frequently annuals. No significant relationship was found between region or breeding system and defense nectaries.     

SECONDARY CAROTENOID ACCUMULATION IN SCENEDESMUS KOMAREKII (CHLOROPHYCEAE, CHLOROPHYTA)

As Scenedesmus komarekii Hegewald was cultured under high light intensity and nitrogen limitation, the color of cells progressed from green to brown, and finally through orange to brick red. The secondary carotenoids astaxanthin and canthaxanthin, as well as apolar carotenoids, were detected in the brown and orange cells. These carotenoids were contained in lipoidal globules that were first formed at the periphery of the cell and progressively propagated toward its inside, eventually filling most of it. The chloroplast was single and parietal in the green cells. As the cells turned brown, the chloroplast divided into several small lobes and was pushed toward the interior by the accumulating lipoidal globules. Sometimes the outer layer of the wall of the brown cell developed one or two diametrically opposed swellings. Once the cells became orange or red, neither lipoidal globules nor any major organelles were distinguishable. The cell wall in the orange cells became thick because of the formation of electron-dense granules between its outer and inner layers. The mode of the secondary carotenoid accumulation in S. komarekii differs from that of Haematococcus, an alga well known for its ability to accumulate secondary carotenoids, but resembles that of " Chlorella " zofingiensis (= Mychonastes zofingiensis (Dönz) Kalina et Punčochářová).     

THE ROLE OF UNICELLS IN THE POLYMORPHIC SCENEDESMUS ARMATUS (CHLOROPHYCEAE)1

The UTEX 2193 strain of Scenedesmus armatus (Chod.) Chod, when cultured in any of several media (whether natural or artificial, concentrated or dilute) produced a variety of colonial morphologies as well as a unicell population. Morphological expression was related to culture ape. When the initial cell density was just a feu1 hundred cells per mL. the culture first produced a unicell population, then spiny colonies, and as stationary phase was approached, spine-less colonies. Two classes of spiny colonies were detected. Type I colonies had elongate cells with the terminal cells shorter than median cells. Spines were longer than cell length. The wider, oval, grainy cells of Type II colonies were uniform m length. Spines were shorter and thicker than those on Type I colonies. Only Type I colonies produced unicells: the latter appeared as two morphs. The smaller unicell was obovoid with four delicate spines: the larger had ovate cells bearing four thicker spines. Control of unicell development in all media was achieved by carefully monitoring colony type and cell number used for the inoculum. A unicellular population developed in batch culture in defined media, both concentrated and dilute, when the initial cell density (either Type I or Type II colonies) was low (below 1000 cells-mL^?1), as well as in synchronous cultures. With higher initial cell densities, e.g. 2 × 10⁴ cells·mL^?1, the inoculum had to contain Type I colonies to produce unicells. Unicells were also produced in water from Agronomy Pond, where the strain originated. We discuss the role of unicell populations in the distribution of Scenedesmus.     

INFLUENCE OF THE CLAY TYPES ILLITE AND MONTMORILLONITE ON THE UPTAKE OF65 Zn BY SCENEDESMUS OBLIQUUS.

SUMMARY

The stimulatory effect of the clays illite and montmorillonite on zinc uptake by Scenedesmus obliquus is discussed. The competitive effects of hydrogen, phosphate, magnesium and copper ions on zinc uptake are illustrated. This competition is reduced in the presence of illite and montmorillonite and zinc uptake is stimulated. Ferrous ions have no effect on zinc uptake, while calcium ions antagonize zinc uptake at high concentrations. The latter effect was remarkable in the presence of montmorillonite.     

FUNCTION OF MENTZELIA NUDA (LOASACEAE) POSTFLORAL NECTARIES IN SEED DEFENSE

Mentzelia nuda is a coarse, short-lived perennial of the High Plains of North America. The flowers secrete nectar which serves as an attractant for pollinating bees. Postfloral nectar secretion attracts ants whose presence significantly enhances seed set. This is the first demonstration of advantage to postflowering activity of a floral nectary.     

EFFECT OF GRAZING-ASSOCIATED INFOCHEMICALS ON GROWTH AND MORPHOLOGICAL DEVELOPMENT IN SCENEDESMUS ACUTUS (CHLOROPHYCEAE)

Populations of the phenotypically plastic alga Scenedesmus acutus Meyen were cultured in standard medium or in medium with filtered water from a Daphnia culture to examine the expression of ecomorphs in both water types. A rapid formation of four- and eight-celled coenobia was observed in the presence of Daphnia water, but not in standard medium. Moreover, cell dimensions were increased in the Daphnia water containing medium. Population growth rates were comparable in both water types; however, the carrying capacity appeared significantly lower in Daphnia water. Daphnia needed to feed on digestible food to produce the colony-inducing chemicals; medium from starved animals and Daphnia fed polystyrene beads appeared inactive. Neither suspensions of homogenate of Scenedesmus and Cryptomonas nor auxins affected colony formation or growth rate in S. acutus. The colony-inducing infochemicals are probably not constituents of the algae themselves; however, modification of algal constituents in the digestive system of daphniids may be involved.     

CYCLOMORPHOSIS IN SCENEDESMUS ARMATUS (CHLOROPHYTA): AN ORDERED SEQUENCE OF ECOMORPH DEVELOPMENT1

Scenedesmus armatus (Chod.) Chod, growth and morphology were monitored in medium 7 (oligotrophic) and Bristol's medium (eutrophic); cultures in both media were incubated at 10 and 22° C. Growth rate at 10° C was reduced, i.e. only one doubling in 7 days in medium 7 and 2.3 doublings in Bristol's, compared to 4.3 and 6 doublings at 22° C over the same period. Unicells as well as cells of colonies were larger at the cold temperature. The lengths of cells were not significantly different regardless of temperature or medium, but cell width was markedly increased at the lower temperature. Additionally, an arcuate, eight-celled, multispined ecomorph, which resembled several previously described taxa, was produced at 10° C. It becomes a component of a previously published ordered sequence of ecomorph development for this species. Based on data now accumulated in both the laboratory and the field, these temporal changes are interpreted to be a cyclomorphosis, driven by a coupling of nutrient availability and temperature. Within the addition of new cold temperature (spring) ecomorphs, the ordered sequence of ecomorphs for S. armatus is a succession from unicells to multispined eight-celled colonies to quadricaudate colonies, ending with acaudate four-celled ecomorphs.     

ETHEPHON (2-CHLOROETHYLPHOSPHONIC ACID) EFFECTS ON SCENEDESMUS QUADRICAUDA (CHLOROPHYCEAE)1,2

Growth of Scenedesmus quadricauda (Turp.) Bréb. was significantly increased when the cultures were grown in Ethephon 68–241 (2-chloroethylphosphonic acid) at concentrations of 0.001–1.0 μg · ml^?1. At 0.1 μg · ml^?1, Ethephon caused a significant increase in RNA levels on a dry weight basis. Total protein and DNA levels increased but not significantly. These data indicate that ethylene, the decomposition product of Ethephon at physiological pH, has some effect on the metabolism of algae.     

SCENEDESMUS MORPHOGENESIS. COLONY CONTROL IN DILUTE MEDIA1

Morphological stability, with the elimination of the unicellular singe, was achieved by culturing several strains of Scenedesmus in media with, low inorganic salt levels. Organisms stabilize and form only colonies at specific nutrient levels. One became colonial only when the salt level was dropped to 3.07 mg inorganic salts per liter, with N and P at levels found in nature.     

RESPONSE OF GRACILARIA CONFERTA (RHODOPHYTA) TO OLIGOAGARS RESULTS IN DEFENSE AGAINST AGAR-DEGRADING EPIPHYTES

Elicitation of Gracilaria conferta (Schousboe ex Montagne) J. et G. Feldmann with oligoagars resulted in a defense response that was strong enough to kill epiphytic bacteria associated with the alga. Up to 60% of the resident bacterial flora of healthy plants was eliminated within 60 min after addition of neoagarohexaose to the algal medium. Single isolates of agar-degrading bacteria that had been isolated previously from healthy or decaying algal tissues proved to be more sensitive. Some of them were generally unable to survive on healthy G. conferta. Others survived on unelicited plants. Approximately 90% of these more resistant agar degraders were eliminated within 15 min after elicitation. The bacterial degradation of dead tissue of G. conferta resulted in a release of elicitors. The elicitors accumulated in the medium and reached high enough concentrations within 24 h to induce a hypersensitive response in healthy algae. The eliciting agent could be destroyed with β-agarase and was thus probably oligoagar. Application of antibiotics prevented the accumulation of the elicitor, which indicated that bacteria were responsible for its release from the algal biomass. The hypersensitive response of G. conferta after contact with oligoagars is thus a true defense response, because it enables the plant to protect itself efficiently from enzymatic attacks on its cell wall.     

A STUDY OF WALL ORNAMENTATION IN CULTURES OF SCENEDESMUS

In a comparative study, six strains of Scenedesmus isolated from one small pond were examined in culture. Three of the isolates, S. denticulatus, S. dispar, and S. abundans, proved to be morphologically quite stable when studied in the laboratory. In an axenic culture of any one of the three pleomorphic strains one could find coenobia which resemble S. longispina or S. armatus. High percentages of four-celled coenobia with several ridges but with only two spines (the S. semipulcher type) were found typically in older cultures of two of the pleomorphic cultures. Long, delicate appendages (bristles) were easily detected on most coenobia in desiccated preparations of all but the S. dispar culture. The use of cultures in identifying Scenedesmus coenobia is discussed.     

阴离子通道参与了harpin_(Pss)诱导的烟草细胞早期防卫反应的信号传导

无论在harpin_(Pss)之前、同时、还是之后向烟草植株或悬浮培养系加阴离子通道的抑制剂DIDS(4,4’-diisothiocyanatostilbene-2,2’-disulfonic acid)或AgC(anthracene-9-carboxylic acid),都可以抑制harpin_(Pss)诱导的烟草植株过敏反应和悬浮细胞的活性氧的释放及胞外碱性化。DIDS和A9C还可以抑制harpin_(Pss)诱导的Ca~(2 )内流。而且DIDS的抑制效率比A9C高。推测质膜上的阴离子通道对钙离子通道有着正调节作用,harpin_(Pss)通过阴离子通道和钙离子通道介导的信号传导途径,调节胞内Ca~(2 )浓度,从而启动这些防卫反应。