The effect of caffeine and light on killing of the blue-green alga Anabaena doliolum by ultraviolet radiation

Summary Pretreatment of spores of the blue-green alga Anabaena doliolum with caffeine is antagonistic to UV lethality and posttreatment with caffeine is synergistic to UV lethality and mutagenicity. The results of photoreactivation experiments suggest that photoreactivation is independent of photosynthesis in blue-green algae.     

Developmental mutants of Anabaena doliolum defective in repair of UV-damage

Summary Nitrosoguanidine induced blue pigment mutants of the blue-green alga Anabaena doliolum were isolated. The blue-mutants on further characterization were grouped into three developmental phenotypes — (i) those forming doli-form blue-spores of heterogenous size i.e., Ad 011, (ii) those forming spheroidal cells in the stationary phase, some of which behave like spores on transfer to fresh medium i.e., Ad 012, and (iii) those showing no sporulation and conditionally producing abnormal cells in the presence of combined nitrogen only i.e., Ad 007. The former two classes of mutants showed the formation of abnormal cells irrespective of the presence or absence of combined nitrogen sources in the medium. The formation of abnormal cells in the filaments of the above mutants were distinguished by their larger size and irregular mode of division leading to true-branch formation. The comparative characterization of these mutant strains with the parental one showed sluggish growth, increased UV-sensitivity, almost unchanged photorepair capacity, a marked change in the pigment composition and relative resistance to nitrosoguanidine. Irregular cell division in both space and time in the mutant strains and their increased sensitivity to ultraviolet irradiation indicate the possible involvement of dark repair system in maintaining the precision of cell cycle in this alga.     

Nutritional control of the frequency of MNNG-induced true branching habit in Anabaena doliolum

Summary MNNG survival and mutagenesis were studied in Anabaena doliolum Bharadwaja. The survival curves of germinating spores were exponential while those of resting spores and filament-fragments were sigmoidal. Blue mutants with much higher phycocyanin contents than the parent were recovered in varying frequencies; the frequency (1.1%) was the highest with 2 mg/ml MNNG for 15 min. Some of the blue mutants sporulated normally; while others did not. A nonsporulating blue mutant (M16) showed branched filaments in liquid cultures. In nitrogen free medium. M16 had a high frequency (70%) of branched filaments during the early phase of growth. In some filaments, the branch arose from a heterocyst showing three polar nodules. There was no other difference between the parent and the mutant in cell morphology and cellular differentiation. High light intensity adversely affected phycocyanin and chlorophyll a pigments and nitrogen fixation in the mutant. Frequency of mutant branched character appears to beta factor of inorganic nitrogen nutrition.Dedicated to the memory of my revered teacher the late Prof. R. N. Singh     

Factors modulating differentiation of spores: Characterization of mutants defective in sporulation in the cyanobacterium Anabaena doliolum (AdS strain)

Summary Mutants of Anabaena doliolum (AdS strain) altered with respect to the time of initiation and degree of sporulation were isolated following mutagenesis with N-methyl-N-nitro-N-nitrosoguanidine and hydroxylamine. The non-sporulating mutant showed a high phycocyanin (Pc): chlorophyll a (chl a) ratio (ca. 7.2) as compared to sporulating strains (Pc:chl a, 4.7–5.3). Also this strain seemed to have higher RNA pools per unit of genomic material as reflected in a higher RNA:DNA ratio. The data suggest that degradaton of phycocyanin and controlled RNA synthesis are prerequisites for sporulation. Mutants exhibiting non-sporulation and delayed initiation of sporulation accumulated more nitrogen through nitrogen fixation, probably indicating nitrogenase function over an extended vegetative phase.     

Genetic analysis of the non-sporulating phenotype of brewer's yeasts

The ploidies and sporulation abilities of six brewer's yeasts were examined. One (YB11-1) out of the six was triploid and sporulating, another (IFO2031) was haploid, and the others (IFO1167, IFO2003, S341 and YB3-7) were diploid and non-sporulating. The five non-sporulating strains did not have the premeiotic DNA synthesis. Their non-sporulating phenotypes were genetically analyzed by examining the sporulation abilities of hybrids between brewer's yeasts and standard genetic strains of Saccharomyces cerevisiae. All non-sporulating brewer's yeasts complemented 32 sporulation-deficient mutations (spoT–spoT23, spo1–spo5, spo7, spo8, spo10, and spo11). Hybrids between brewer's yeasts and haploid or diploid strains homozygous for the mating-type locus had poor or no sporualtion. On the contrary, hybrids between brewer's yeasts and diploid strains heterozygous for the mating-type locus sporulated at a high frequency. These results indicated that the non-sporulating phenotype of brewer's yeasts was caused by a deficiency of the mating-type genes rather than by mutations of sporulation genes. The Southern hybridization probed with the MATa gene showed polymorphisms in mating-type genes of brewer's yeasts.     

Pigment variations in ultraviolet-treated strains of the blue-green alga Anabaena doliolum

Summary Two kinds of cultures were raised from clones of Anabaena doliolum surviving on selective medium following exposure of spores to ultraviolet radiation. The pigments of these cultures have been characterized with respect to those of controls.     

Carbohydrate Metabolism During Ascospore Development in Yeast

Carbohydrate metabolism, under sporulation conditions, was compared in sporulating and non-sporulating diploids of Saccharomyces cerevisiae. Total carbohydrate was fractionated into trehalose, glycogen, mannan, and an alkali-insoluble fraction composed of glucan and insoluble glycogen. The behavior of three fractions was essentially the same in both sporulating and non-sporulating strains; trehalose, mannan, and the insoluble fraction were all synthesized to about the same extent regardless of a strain's ability to undergo meiosis or sporulation. In contrast, aspects of soluble glycogen metabolism depended on sporulation. Although glycogen synthesis took place in both sporulating and non-sporulating strains, only sporulating strains exhibited a period of glycogen degradation, which coincided with the final maturation of ascospores. We also determined the carbohydrate composition of spores isolated from mature asci. Spores contained all components present in vegetative cells, but in different proportions. In cells, the most abundant carbohydrate was mannan, followed by glycogen, then trehalose, and finally the alkali-insoluble fraction; in spores, trehalose was most abundant, followed by the alkali-insoluble fraction, glycogen, and mannan in that order.     

On a new species of Hapalosiphon naegeli (Myxophyceae)

Hapalosiphon fertilissima sp. nov. has been isolated from enrichment cultures of paddy field soils, Karjat (India). Morphology and reproduction of the alga have been studied in detail. The new species is distinct in producing entire sporogenous filaments with characteristic barrel-shaped spores which remain undissociated and may germinate in situ.     

Correlation between alkaline activation of Bacillus thuringiensis var. kurstaki spores and crystal production

The spores of crystal-forming (Cry⁺) and non-crystal-forming (Cry^-) strains of Bacillus thuringiensis var. kurstaki and Bacillus cereus were tested for the ability to be activated by 0.1 m K₂CO₃ (pH 10). Only the spores of crystal-forming strains could be activated, and this phenotype was independent of whether crystals were present with the spores in the activation solution. The spores of a B. thuringiensis var. kurstaki strain that is temperature sensitive for protoxin accumulation could be activated by the alkaline solution when produced at the permissive temperature, whereas spores produced at the nonpermissive temperature were not activated. The results indicate that protoxin in the spore coat is responsible for the alkaline-activation phenotype and may serve an ecological function for the organism.     

Induction of mutation in the cyanobacteriumAnabaena doliolum: A strain-specific property

Mutations with respect to the sporulating character in the cyanobacteriumAnabaena doliolum (AdS and AdB strains) were induced after treatment with acriflavin, acridine organge 9-aminoacridine, N-methyl-N′-nitro-N-nitrosoguanidine, ethyl methanesulfonate, hydroxylamine, nitrous acid, low pH (pH 4.2) and elevated temperature (65±1 °C). Exposure to higher temperature was most effective in inducing nonsporulating mutants in both strains. Uptake of acridine dyes, inactivation and mutability with respect to sporulation of two strains of cyanobacteriumA. doliolum revealed that the mutagen uptake could be directly correlated with the frequency of induced mutations but that survival and mutability are independent strain-specific properties.     

Induction of Oligosporogeneous and Asporogeneous Mutants of Blue-green Alga, Anabaena doliolum by Acriflavine and Acridine Orange

Acriflavine and acridine orange were highly effective in producingmutants defective in spore differentiation in the blue-greenalga, Anabaena doliolum. Acridine orange produced both oligosporogeneous(Osp) and asporogeneous (Sp^–) mutants in high yield withthe frequency ranging from 2.4 to 21 per 10⁴ survivors, whereaswith acriflavine no oligosporogeneous mutants were detected,although it produced non-sporulating (Sp^–) mutants withthe frequency of 1.4 per 10⁴ survivors. Mutants isolated throughthese dyes were stable and showed no tendency towards spontaneousor induced reversion. Behaviour of various mutants indicatedtheir being blocked at different stages of sporulation withdifferent mutagenic sites. Induction of high numbers of Ospand Sp^– mutants after acridine dye treatment indicatesthe involvement of an extrachromosomal determinant in sporulationin Anabaena doliolum. Ohgosporogeneous mutants, asporogeneous mutants, Anabaena doliolum, blue-green algae, mutagenesis, acriflavine, acridine orange     

A Mutational Study of the Nitrogen-fixing Blue-Green Alga Anabaena doliolum

A heterocystous, non-nitrogen-fixing mutant of the nitrogen-fixingblue-green alga Anabaena doliolum has been isolated followingtreatment with nitrosoguanidine and UV radiation. Some reversiblevariations in the habit and morphology of the alga were inducedfollowing its treatment with nitrosoguanidine.     

Growth,cell division and sporulation in mycobacteria

Bacteria have the ability to adapt to different growth conditions and to survive in various environments. They have also the capacity to enter into dormant states and some bacteria form spores when exposed to stresses such as starvation and oxygen deprivation. Sporulation has been demonstrated in a number of different bacteria but Mycobacterium spp. have been considered to be non-sporulating bacteria. We recently provided evidence that Mycobacterium marinum and likely also Mycobacterium bovis bacillus Calmette–Guérin can form spores. Mycobacterial spores were detected in old cultures and our findings suggest that sporulation might be an adaptation of lifestyle for mycobacteria under stress. Here we will discuss our current understanding of growth, cell division, and sporulation in mycobacteria.     

A Biochemical Study of Spore Germination in the Blue-green Alga Anabaena doliolum

The spores of Anabaena doliolum formed in light (light spores)and after transfer to darkness (dark spores) are biochemicallydifferent in that the light spores contain chlorophyll a andphycocyanin, while dark spores seem to lack them. The apparentbiosyntheses accompanying dark-spore germination seem to proceedin the following order: RNA, chlorophyll a, phycocyanin andDNA. Results of chloramphenicol treatment indicate that proteinsynthesis precedes RNA synthesis. The biosynthetic events followingRNA synthesis show a requirement for light.     

Re-Exposure of Tapes at High Temperature and Pressure in the Lycopodium Clavatum Spore Exine

Lamellations are visualizable through the staining commonly used in transmission electron microscopy during exine formation on Lycopodium and other spores, and the nexine of pollen grains. The lamellations so exposed consist-of dark tapes at either side of an unstained (white) line. Neither tapes nor white lines are visualizable in the exine of mature spores of Lycopodium. The continued presence of lamellations having tape-white line spacing has been demonstrated with inorganic tracers in the nexine of pollen in which lamellations otherwise appeared to be absent. Through high contrast staining methods for TEM we have observed lamellations in the residual exine following heat treatment (350[ddot]C) of mature spores of Lycopodium clavatum. The surface of these residual exines was etched by treatment with hot 2-aminoethanol and filaments were observed to protrude from the etched surfaces. The residual exine stained darkly. Lycopodium spores heated to 300[ddot]C at 1 kb pressure had long filaments exposed at the surface of the residual exine (sporopollenin). Sections of the pellet remaining after heat and pressure treatment also included bundles of closely parallel filaments and masses of isolated filaments. The filaments were stained while the exine residue, assumed to include sporopollenin, was not. Isolated filaments produce a stable metachromasia with toluidine blue indicating the presence of many closely spaced sites of negative charge. The staining of intact exines with basic dyes may result from anionic sites on filaments embedded within the exine rather than being due to sporopollenin. The results of our experiments indicate that the filaments are more resistant to heat and pressure and 2-aminoethanol than is sporopollenin. We propose that the trilamellar elements commonly called tapes and white lines might be composed of two filaments bridged by polybasic molecules.     

The Genetic Control of Cell Growth and Development in Yeast Saccharomyces cerevisiae: Disturbed Sporulation in Diploids with a Decreased Activity of the Ras/cAMP Signal Transduction Pathway

Seven haploid strains (four with the MAT mating type and three with the MATa mating type) were selected from the Peterhof genetic collection of yeast. Previous phenotypic analysis assigned six of these strains to a physiological group of strains with changed activity of the Ras/cAMP signal transduction pathway. The haploids were crossed, and the resulting 12 diploids showed higher glycogen accumulation, tolerance to heat shock and nitrogen starvation, and sporulation in complete media. Ten of the diploids expressed the hypersporulation phenotype (higher sporulation efficiency). The phenotypic characters of these ten diploids suggested a reduced activity of the Ras/cAMP pathway. All 12 diploids were tested for sporulation and production of two groups of asci (those with one or two spores and those with three or four spores) as dependent on culture conditions (21, 30, or 34°C; standard sporulation medium or a complete medium containing potassium acetate or glycerol in place of glucose). Sporulation proved to depend on temperature and medium composition. The results are collated with the data on yeast phenotypes associated with a lower activity of the Ras/cAMP signal transduction pathway.     

Ultrastructure of Minchinia Sp. Spores from Shipworms (Teredo Spp.) in the Western North Atlantic, with a Discussion of Taxonomy of the Haplosporidiidae

Electron microscopy of haplosporidan spores from Teredo navalis and T. furcifera revealed 4 distinct membrane-bound extensions, 1 apical extension opposite the opercular hinge, 1 terminal and 2 opposing lateral extensions. These extensions were not continuous with the spore wall, but contained microtubule-like structures and degrading epispore cytoplasm. No other known species in the family Haplosporidiidae is characterized by spores possessing four epispore extensions. There are currently two genera in this family, Minchinia and Haplosporidium. The genus Minchinia includes spores such as those of M. chitonis which bear two epispore cytoplasm extensions. Spores of the genus Haplosporidium have been characterized by spore wall derived filaments. A 3rd group of haplosporidan species with spores ornamented by wrappings have traditionally also been assigned to the genus Haplosporidium. Based on the presence of epispore cytoplasm extensions rather than spore wall filaments, the haplosporidan of Teredo spp. can be placed in the genus Minchinia.     

Germination and physiological properties of Frankia spores

Spores from four Frankia strains were isolated and purified to homogeneity. The purified spores were biochemically and physiologically characterized and compared to vegetative cells. Frankia spores exhibited low levels of endogenous respiration that were at least ten-fold lower than the endogenous respiration rate of vegetative cells. The macromolecular content of purified spores and vegetative cells differed. One striking difference among the Frankia spores was their total DNA content. From DAPI staining experiments, only 9% of strain ACN1^AG spore population contained DNA. With strains DC12 and EuI1c, 92% and 67% of their spore population contained DNA. The efficiency of spore germination was correlated to the percentage of the spore population containing DNA. These results suggest that the majority of strain ACN1^AG spores were immature or nonviable. The presence of a solidifying agent inhibited the initial stages of spore germination, but had no effect once the process had been initiated. The optimal incubation temperature for spore germination was 25°C and 30°C for strains DC12 and EuI1c, respectively. A mild heat shock increased the efficiency of spore germination, while root extracts also stimulated spore germination. These results suggest that strains DC12 and EuI1c may be suitable strains for further germination and genetic studies.     

Occurrence of the gametophyte of Agarum clathratum (Laminariales, Phaeophyceae) as an endophyte in Orculifilum denticulatum (Gigartinales, Rhodophyceae)

An endophytic filamentous brown alga, growing in the red alga Orculifilum denticulatum Lindstrom, was collected from the north‐east Pacific, near Juneau, Alaska. Within the host tissue, its branched filaments formed a network in the space between the filaments of the host tissues embedded in the host intercellular substances. Cells of the filaments contained many discoid chloroplasts without pyrenoids. Neither microscopic morphological observation nor culturing was sufficient to reveal the specific identity or even the familial affinity of the alga; in contrast, molecular phylogenetic analysis of its rbcL gene and rDNA ITS sequences showed that it was the gametophyte of Agarum clathratum Dumortier (Laminariales). There are few reports of laminarialean gametophytes in nature; this is the first report actually identifying the species of laminarialean gametophyte in a red alga.     

酒鬼酒酿造环境异常球菌属菌株的生物学特征及系统发育分析

研究对分离自馥郁香型白酒酒鬼酒制曲和发酵车间空气的8株异常球菌属(Deinococcus)菌株的特征及系统发育分析进行研究。采用添加体积分数10%酒鬼酒混合浸汁的营养琼脂培养基分离,挑选并纯化与异常球菌属菌株表型相似的菌株,考察其相关生理生化特征,并进行基于16S rRNA基因序列的系统发育分析。通过表型特点筛选,得到23株异常球菌属疑似菌株。系统发育分析表明,23株菌株中有8株属于异常球菌属,归属于该属的4个种(D.radiotolerans、D.daejeonensis、D.ficus和D.yunweiensis)。形态观察和生理生化实验表明,这些异常球菌属菌株在内生孢子、革兰染色、过氧化氢酶实验、生长温度范围、生长pH范围、多聚物水解等特征方面,与它们系统发育关系最为密切的典型菌株之间存在不同程度的差异。