CHARACTERIZATION OF PIGMENT MUTANTS IN A BLUEGREEN ALGA,ANACYSTIS NIDULANS1

Characteristics of 2 types of pigment mutants of the bluegreen alga, Anacystis nidulans, are described. “Yellow-green” mutants (YG) which have normal chlorophyll but only half the phycocyanin of the parent are similar to the parent in number of reaction centers/cell, number of chlorophylls/reaction center, maximum turnover rate of the reaction centers, quantum yields at 620 and 686 nm and specific growth rate; they have a reduced action at 620 nm. “Blue” mutants (BL) with somewhat higher phycocyanin but only one-third the chlorophyll of the parent are dissimilar to the parent. BL's have fewer reaction centers/cell, a smaller number of chlorophylls/reaction center, a higher maximum turnover rate of reaction centers, and a lower specific growth rate. BL's show ca. half the quantum yield of the parent at 620 nm and at 686 nm show a “red rise” rather than a “red drop.” The consequences attending low chlorphyll in the BL's are more drastic than those attending the low phycocyanin of the YG's.     

MORPHOLOGY AND GENETICS OF A PERIODIC COLONIAL MUTANT OF NEUROSPORA CRASSA

A mutant of strain 69–1113a of Neurospora crassa, which shows periodic growth upon both complete and minimal media, was named “clock,” and some of the morphological and genetic differences between this mutant and the “patch” and “wild” strains were investigated. In contrast to the uniform growth of “wild,” the “clock” mutant produces a series of bands formed by cymelike aggregations of hyphae which become progressively more dense and finally mark the end (front) of a growth band. A new growth band is formed by a number of hyphae which grow out as in “wild” strains and dichotomize and form new cymes which again become progressively more dense and finally form a new front. It is shown that “clock” continues its rhythmic growth when cultured in continuous darkness. Some “wild” strains were induced to grow periodically on appropriate media. A medium containing equal quantities of sorbose and sucrose caused strain 65–811A (a “wild”) to produce the “patch” type growth. Random isolation, as well as ordered isolation, of ascospores following a cross between “wild” and “clock” show a 1: 1 segregation indicating that “clock” differs from “wild” by a single gene.     

PHOTOREGULATION OF PHYCOBILISOME STRUCTURE DURING COMPLEMENTARY CHROMATIC ADAPTATION IN THE MARINE CYANOPHYTE PHORMIDIUM SP. C861

Changes in the molecular structure of phycobilisomes during complementary chromatic adaptation were studied in the marine cyanophyte Phormidium sp. C86. This strain forms phycoerythrin (PE)-less phycobilisomes under red light but synthesizes PE-rich phycobilisomes under green light. Analysis of phycobiliprotein composition and electron microscopic examination of phycobilisomes in ultra-thin sections of cells and of isolated phycobilisomes were performed for cells acclimated to red and green light, respectively. The structure of phycobilisomes formed under red light conditions was typically hemidiscoidal. Phycobilisomes in cells acclimated to green light were twice as large in size as those in cells acclimated to red light. This increase in phycobilisome size was a result of the increase in the molar ratio of antenna pigment (PE and phycocyanin) to allophycocyanin, from 3.5 to 11.3. Pigment composition and fine structure of phycobilisomes formed under green light were similar to those of “nonhemidiscoidal” phycobilisomes reported in Phormidium persicinum. These results suggest that changes occur not only in the molecular species of peripheral rods but also in the structure of rods and probably of cores in relation to their connection with rods during chromatic adaptation of Phormidium sp. C86.     

Far-red light-insensitive, phytochrome A-deficient mutants of tomato

We have selected two recessive mutants of tomato with slightly longer hypocotyls than the wild type, one under low fluence rate (3 mol/m²/s) red light (R) and the other under low fluence rate blue light. These two mutants were shown to be allelic and further analysis revealed that hypocotyl growth was totally insensitive to far-red light (FR). We propose the gene symbol fri (far-red light insensitive) for this locus and have mapped it on chromosome 10. Immunochemically detectable phytochrome A polypeptide is essentially absent in the fri mutants as is the bulk spectrophotometrically detectable labile phytochrome pool in etiolated seedlings. A phytochrome B-like polypeptide is present in normal amounts and a small stable phytochrome pool can be readily detected by spectrophotometry in the fri mutants. Inhibition of hypocotyl growth by a R pulse given every 4 h is quantitatively similar in the fri mutants and wild type and the effect is to a large extent reversible if R pulses are followed immediately by a FR pulse. After 7 days in darkness, both fri mutants and the wild type become green on transfer to white light, but after 7 days in FR, the wild-type seedlings that have expanded their cotyledons lose their capacity to green in white light, while the fri mutants de-etiolate. Adult plants of the fri mutants show retarded growth and are prone to wilting, but exhibit a normal elongation response to FR given at the end of the daily photoperiod. The inhibition of seed germination by continuous FR exhibited by the wild type is normal in the fri mutants. It is proposed that these fri mutants are putative phytochrome A mutants which have normal pools of other phytochromes.     

Pigment cell differentiation in the fire-bellied toad,Bombina orientalis. I. Structural,chemical, and physical aspects of the adult pigment pattern

Wild-collected adults of Bombina orientalis are bright green dorsally and red to red-orange ventrally. As a prelude to an analysis of the differentiation of pigment cells in developing B. orientalis, we describe structural and chemical aspects of the fully differentiated pigment pattern of the “normal” adult. Structurally, differences between dorsal green and ventral red skin are summarized as follows: (1) Dorsal green skin contains a “typical” dermal chromatophore unit comprised of melanophores, iridophores, and xanthophores. Red skin contains predominantly carotenoid-containing xanthophores (erythrophores), and skin from black spot areas contains only melanophores. (2) In ventral red skin, there is also a thin layer of deep-lying iridophores that presumably are not involved in the observed color pattern. (3) Xanthophores of red and green skin are morphologically distinguishable from each other. Dorsal skin xanthophores contain both pterinosomes and carotenoid vesicles; ventral skin xanthophores contain only carotenoid vesicles. Carotenoid vesicles in dorsal xanthophores are much larger but less electron dense than comparable structures in ventral xanthophores. The presence of carotenes in ventral skin accounts for the bright red-orange color of the belly of this frog. Similar pigments are also present in green skin, but in smaller quantities and in conjunction with both colored (yellow) and colorless pteridines. From spectral data obtained for xanthophore pigments and structural data obtained from the size and arrangement of reflecting platelets in the iridophore layer, we attempt to explain the phenomenon of observed green color in B. orientalis.     

PROTOPLAST FUSION AND GENETIC COMPLEMENTATION OF PIGMENT MUTATIONS IN THE RED MICROALGA PORPHYRZDZUM SP.1

Protoplasts from two green pigment mutants of Porphyridium sp. (UTEX 637) containing a low phycoerythrin level were fused by exposure to polyethylene glycol (MW 6000) combined with a short heat shock (45° C, 5 min). Following regeneration on agar plates, red colonies arose in which complementation of the phycoerythrin deficiency had occurred. The complementation frequency was estimated to be 0.2%. Eight progeny showing red pigmentation were isolated and purified by consecutive transfers on agar plates. Characterization of the fusion progeny revealed that their phycobiliprotein and chlorophyll contents per cell were higher than those of their parental mutant strains and, in most strains, similar to that of the wild type. The fusion products proved to be stable over many growth cycles. The DNA content of the wild type and of the parental mutant strains was about 0.05 pg-cell^?1. Fusion progeny strains showed a variable DNA content: a few fusants contained the same amount of DNA as the wild type and the parental strains, while others had about 50% more DNA per cell. The DNA content of one of the progeny strains (CF1c) was double that of the wild type (0.1 pg. cell^?1). Cells of this fusion progeny contained one nucleus per cell, which suggests that nuclear fusion and the formation of a stable diploid followed cell fusion. Analysis of phycobilisome components of CF1c revealed complementation of linker polypeptides associated with phycoerythrin (γ subunits). CF1c contained, like the wild-type strain, four linker polypeptides; all of these were absent in one parental strain and one was absent in the second. To the best of our knowledge, this is the first report of protoplast fusion, formation of somatic hybrids, and the apparent completion of a parasexual cycle in a red microalga.     

Blue–green opponency and trichromatic vision in the greenhouse whitefly (Trialeurodes vaporariorum) explored using light emitting diodes

Visual orientation in the greenhouse whitefly (Trialeurodes vaporariorum Westwood, Hemiptera: Aleyrodidae) is the result of “wavelength‐specific behaviours.” Green–yellow elicits “settling behaviour” while ultraviolet (UV) radiation initiates “migratory behaviour.” The only available physiological study of the photoreceptors' spectral efficiency showed peaks in the green and the UV range and whitefly vision was said to be dichromatic so far. In order to study the visual behaviour of T. vaporariorum, 19 narrow‐bandwidth light emitting diodes (LEDs) covering the UV‐A and visible range were used in combination with light scattering acrylic glass screens in a small‐scale choice arena under greenhouse conditions. Multiple‐choice and dual‐choice assays were performed, resulting in LED‐based behavioural action spectra of settling (green) and migratory behaviour (UV). A potential inhibitory blue–green chromatic mechanism was studied by combining yellow with different bluish LEDs. Intensity dependencies were illustrated by changing LED intensities. Regarding the “settling response,” highest attraction was achieved by a green LED with a centroid wavelength of 550 nm, while a blue LED with 469 nm proved to be most inhibitory. Besides this inhibitory interaction, an intensity dependence was observed within the action spectrum in the green–yellow range. “Migratory behaviour” was elicited the most by the UV LED with the shortest available wavelength of 373 nm. The results provide compelling behavioural evidence for the presence of a green and a yet undescribed blue sensitive photoreceptor and a blue–green opponent mechanism. Furthermore, empirical colour choice models were built and receptor peaks were estimated around 510–520 nm (green), 480–490 nm (blue) and 340–370 nm (UV). Consequently, a trichromatic receptor setup is suggested for T. vaporariorum.     

VARIATION IN THE STRENGTH AND SOFTNESS OF SELECTION ON DELETERIOUS MUTATIONS

Deleterious alleles constantly enter populations through mutation. Understanding the nature of selection against such alleles is required to assess their impact on populations. In a subdivided population, two distinct aspects of selection are important: the strength and softness of selection. Using Drosophila melanogaster, we estimated both aspects of selection for each of eight loci across two environments. These data allow us to test conflicting predictions about the factors affecting the softness of selection. First, we show that the softness of selection is not determined by ecological conditions alone. Second, we find that resource limitation makes selection stronger but does not make it softer. Third, we find that wild‐type individuals tend to benefit more than mutants from being reared with competitors of low genetic quality. This means that selection is effectively “harder” on mutants than wild types. A model is presented showing that the sensitivities of mutants and wild types to local competitors differentially affect equilibrium mutation frequency and measures of load.     

N-methyl-N′-nitro-N-nitrosoguanidine and UV induced mutants of the dinoflagellate Crypthecodinium cohnii*

SYNOPSIS. Mutant strains were chemically induced by treatment with N-methyl-N′-nitro-N-nitrosoguanidine (NTG) and UV irradiation. UV and NTG mutation rates were obtained that were both consistent with the organism being haploid. Three types of mutants were produced: (a) strains deficient in both β- and γ-carotene, the only carotenoids found in the wild type; phenotypes include albinos (translucent, dull white, “snow white”) and cream-colored on agar as compared to the yellow-orange color of wild type colonies; (b) strains requiring adenine, guanine or cytosine in addition to the minimal medium for growth; (c) mutants that grow at a rate less than 40% of the wild type in minimal medium.     

SIMILAR UNSTABLE MUTATIONS IN THREE SPECIES OF GRACILARIA (RHODOPHYTA)1

Unstable mutants with similar variegated pigmentation were genetically characterized in the red algae. Gracilaria tikvahiae (McLachlan), G. foliifera (Forsk.) Børg. and. G. sjoestedtii (Kylin). All three mutants were green plants with flecks of red tissue where cells had reverted to wild type. The mutant green phenotypes were all recessive, and their genetic behavior in crosses indicated that each was the result of a single, unstable, nuclear gene. Wild-type revertant tissue was stable one it arose. Revertant plants obtained from spores and revertant fronds taken from variegated plants could not be distinguished from the normal wild type, either phenotypically or genetically. Reversion to wild type occurred during all phases of the life cycle. In crosses between the mutants and wild type, most of the F₁ tetrasporophytes were heterozygous wild-type plants, an observation consistent with the recessive nature of the mutations; however, a low frequency of homozygous unstable-green F₁ tetrasporophytes was also obttained from these crosses. The molecular basis of neither the mutant instability, i.e. the reversion to wild type, nor of the process producing the unstable green F₁ tetrasporophytes can yet be deduced, but the phenotype of the plants and genetic results suggest the involvement of transposable genetic elements.     

Evaluation of colors in green mutants isolated from purple bacteria as a host for colorimetric whole-cell biosensors

The change in carotenoid-based bacterial color from yellow to red can be applied to whole-cell biosensors. We generated several green mutants to emphasize the color change in such biosensors. The blue-green crtI-deleted mutant, Rhodopseudomonas palustris no.711, accumulated the colorless carotenoid precursor, phytoene. Green Rhodovulum sulfidophilum M31 accumulated neurosporene, a downstream product of phytoene. Another green mutant, Rhodobacter sphaeroides Ga, accumulated neurosporene and chloroxanthin, which are both downstream products of phytoene. All green mutants accumulated bacteriochlorophyll a. Photosynthetic membrane obtained from the green mutants all exhibited decreased absorption of wavelength range at 510–570 nm. Therefore, these indicate that the greenish bacterial colors were mainly caused by the existence of bacteriochlorophyll a and the changes in carotenoid composition in photosynthetic membrane. The colors of the green mutants and their wild-type strains were plotted in the CIE-L*a*b* color space, and the color difference (ΔE*ab) values between a green mutant and its wild type were calculated. ΔE*ab values were higher in the green mutants than in Rdv. sulfidophilum CDM2, the yellowish host strain of reported biosensors. These data indicate that change in bacterial color from green to red is more distinguishable than that from yellow to red as a reporter signal of carotenoid-based whole-cell biosensors.     

GROWTH AND CELL CYCLE OF TWO CLOSELY RELATED RED TIDE-FORMING DINOFLAGELLATES: GYMNODINIUM NAGASAKIENSE AND G. CF. NAGASAKIENSE1

Small-sized vegetative cells were found to co-occur with normal-sized cells in populations of the European bloom-forming dinoflagellate Gymnodinium cf. nagasakiense Takayama et Adachi, currently known as Gyrodinium aureolum Hulburt, but not in populations of the closely related Japanese species Gymnodiniumn agasakiense. We examined how cell size differentiation may influence growth and cell cycle progression under a 12:12-h light: dark cycle in the European taxon, as compared to the Japanese one. Cell number and volume and chlorophyll red fluorescence in both species varied widely during the photocycle. These variations generally appeared to be related lo the division period, which occurred at night, as indicated by the variations of the fraction of binucleated cells (mitotic index) as well as the distribution of cellular DNA content. “Small” cells of G. cf. nagasakiense divided mainly during the first part of the dark period, although a second minor peak of dividing cells could occur shortly before light onset. In contrast, “large” cells displayed a sharp division peak that occurred 9 h after the beginning of the dark period. The lower degree of synchrony of “small” cells could be a consequence of their faster growth. Alternatively, these data may suggest that cell division is lightly controlled by an endogenous clock in “large” cells and much more loosely controlled in “small” cells. Cells of the Japanese species, which were morphologically similar to “large” cells of the European taxon, displayed an intermediate growth pattern between the two cell types of G. cf. nagasakiense, with a division period that extended to most of the dark period.     

EFFECT OF SPECTRAL QUALITY ON GROWTH AND PIGMENTATION OF PICOCYANOBACTERIA1

The influence of spectral quality on growth and pigmentation was compared among five strains of marine and freshwater picocyanobacteria grown under the same photon flux density (28 μE · m^?2·s^?1). Growth and phycoerythrin (PE) concentration per unit carbon increased when marine Synechococcus WH7803 was grown under green light as compared to red light, but no change in phycocyanin concentration occurred. Marine Synechococcus strain 48B66 also showed greater levels of PE when grown under green light than under red light, but no concomitant growth increase occurred. Both strains thus exhibited Group II chromatic adaptation. Additionally, strain 48B66 increased the relative level of phycourobilin compared to phycoerythrobilin when grown under red light. In contrast, both marine and freshwater Synechococcus strains containing no PE showed decreased growth under green light. Chlorophyll a concentrations were greatest or among the greatest in all strains grown under green light. These results suggest that light quality, through its effects on growth rate, may be an important factor controlling the distribution and abundance of the various pigment types of Synechococcus.     

Cell Surface Carbohydrate Differences in Wild and Mutant Strains of Crithidia fasciculata1

Wild type Crithidia fasciculata and three drug-resistant mutant strains that have shown “flagellar adherence” were studied as to their ability to agglutinate with lectins specific for receptor molecules containing N-acetyl glucosamine, N-acetyl galactosamine, galactose, mannose-like residues, fucose, and sialic acid. Escherichia coli with mannose-sensitive fimbriae was also used as an agglutination probe. The presence of D-GalNAc, D-Gal, and mannose-like residues was detected in the wild strain. Generally, in the mutants, residues of these sugar units were present in greater concentrations when compared to the wild type strain. β-Galactosidase treatment showed that β-D-Galp units are exposed on the cell membrane. All types of cell agglutination including flagellum-flagellum (F-F), flagellum-soma (F-S), and soma-soma (S-S) were observed when lectins were used; however, with E. coli only the F-F type of cell agglutination was observed with the wild type strain and the TFR^R1 mutant. All types of agglutination were observed with the other two mutants.     

Isolation and Characterization of a Xanthophyll Aberrant Mutant of the Green Alga Nannochloropsis oculata

Novel mutants (xan1 and xan2) of the unicellular green alga Nannochloropsis oculata are impaired in xanthophyll biosynthesis, thereby producing aberrant levels of xanthophylls. High-performance liquid chromatography (HPLC) analysis revealed that the xan1 and xan2 mutants have double the violaxanthin (V) content, but have significantly decreased lutein content in their cells compared to the wild type. Furthermore, these mutants contain two to three times more zeaxanthin than the wild type under low light (LL) growth conditions. However, this xanthophyll aberration in N. oculata did not affect the normal growth and the major cellular chemical composition of the xan1 strain. The xanthophyll pool size of the LL-grown mutant was 1.8-fold greater than that of the wild type. Under high light (HL) growth conditions, V content was substantially decreased in both the mutant and wild types because of the epoxidation state of the xanthophylls. Under LL growth conditions, the deepoxidation states of the xanthophyll pool sizes were 0.1 and 1.2 in the wild type and the mutant, respectively. However, the deepoxidation states of the xanthophyll pool sizes were 0.78 in the wild type and 0.87 in the mutant under HL growth conditions. We observed that the level of one of the commercially important xanthophylls, zeaxanthin, was higher in the mutant than in the wild type under all culture conditions. This mutant is discussed in terms of its commercial value and potential utilization by the algal biotechnology industry for the production of zeaxanthin.     

Modellrechnung zur Datenverarbeitung beim Farbensehen des Menschen

Fig. 4 demonstrates the problem: How are the messages of the cones processed by the structures of the CNS before they are perceived in the form of colour impressions? The absorption curves of Fig. 2 are used as input functions of the three types of cones; they were adapted from the work of MacNichol et al. [2] and from similar results of other authors [1, 3, 4]. According to a well known hypothesis [8] the messages of the green receptors are supposed to be the antagonists to the messages of the “red” receptors, and the messages of the blue receptors the antagonists to the sum of the messages of the green and the “red” receptors (Fig. 11). But this hypothesis does not account for the red sensations present in the region of both the long and the short wave end of the visual spectrum (Fig. 14). Therefore another model was studied (Fig. 12): Blue (P 445, cyanolabe) being antagonistic to yellow (P 570, erythrolabe); and green (P 535, chlorolabe) being antagonistic to the combined action of blue and yellow (P 445 and P 570). This formalism explains the following experimental data: The distribution of colour sensations along the spectrum (Fig. 7, 13, 14); the wavelengths of antagonistic spectral colours (Fig. 5); the general form of the spectral hue discrimination function (Fig. 8); the number and the qualitative symptoms of colour deficiencies (Fig. 10, 18–24). A meaningful interpretation is given concerning the reduced colour vision of the distal parts of the retina.     

Biogenesis of mitochondria 23. The biochemical and genetic characteristics of two different oligomycin resistant mutants ofSaccharomyces cerevisiae under the influence of cytoplasmic genetic modification

Two classes ofSaccharomyces cerevisiae mutants resistant to oligomycin, an inhibitor of mitochondrial membrane bound ATPase are described. Biochemical analysis shows thatin vitro the mitochondrial ATPase of both types of mutant are sensitive to oligomycin.In vivo sensitivity of the mutants to oligomycin can be demonstrated following anaerobic growth of the cells, which grossly alters the mitochondrial membrane and renders the ATPase of the mutants sensitive to oligomycin. It is concluded that the mutation to oligomycin resistance in both mutant types is phenotypically expressed as a change in the mitochondrial membrane. The intact mitochondrial membrane in the wild type cell is freely permeable to oligomycin, whereas the resistant mutant is impermeable to oligomycin; alteration of the mitochondrial membrane during isolation of the organelle or physiological modification of the membranes of the mitochondria by anaerobic growth renders the membranes permeable.These mitochondrial membrane mutants differ in their cross-reference patterns and their genetics. One is resistant to oligomycin only, and behaves like previously reported cytoplasmic mutants. The other shows cross-resistance to inhibitors of mitochondrial protein synthesis as well as to oligomycin; although the mutant appears to arise from a single step mutation its genetic properties are complex and show part-nuclear and part-cytoplasmic characteristics. The implications of the observations are discussed.     

CELL SIZE DIFFERENTIATION IN THE BLOOM-FORMING DINOFLAGELLATE GYMNODINIUM CF. NAGASAKIENSE1

Two subpopulations differing essentially by their mean cell size were observed regularly in cultures and natural samples of the naked dinoflagellate Gymnodinium cf. nagasakiense Takayama et Adachi (currently known as Gyrodinium aureolum Hulburt), a species which frequently forms red tides in North European seas. “Large” cells represented the typical forms; they were morphologically similar to cells of the closely related Japanese species G. nagasakiense, which did not form any subpopulation of reduced size. “Small” and “large” cells of G. cf. nagasakiense had the same DNA content, but the nucleus of the former appeared to be much more condensed during interphase. Each cell type was able to divide and had its own growth dynamics; therefore, any intermediary between pure populations of “small” and of “large” cells were observed in culture. The “large” form generated a “small” cell by an atypical budding-like division, whereas the “small” form gave back a “large” form, once it ceased to divide, by simple enlargement of its cell body. Factory inducing cell size differentiation are yet unclear. Neither nitrogen nor phosphorus starvation induced a significant increase in the relative proportion of “small” and budding cells. Although cell size differentiation is associated with the formation of gametes in a variety of dinoflagellates, we demonstrated that “small” cells of G. cf. nagasakiense are able to divide asexually, in contrast to gametes of most other species. The high proliferative power of “small” cells as compared with normal cells suggests that they could play a significant role during red tides of G. cf. nagasakiense; in contrast, cells of the Japanese species G. Nagasakiense could sustain high growth rates with larger cell size because this species generally blooms in waters much warmer than those found in northern Europe.