Respiration and nitrogenase activity in nodules ofCasuarina cunninghamiana and cultures ofFrankia sp. HFP020203: Effects of temperature and partial pressure of O2

The effects of time after exposure to acetylene and of nodule excision were examined using a flow-through system. After a transient depression in the rate of acetylene reduction that began about 1.5 min after exposure to acetylene, the rate recovered to 98% of the initial maximum value after 40 min. After nodule excision the rate stabilized to 90% of the initial maximum value observed in the intact plant.Excised nodules, measured at 6-min intervals in a closed system, with frequent changes of the gas mixture, were used for the remaining experiments. Acetylene reduction by the nodules increased rapidly as temperature was increased between 6 and 26°C. Between 26 and 36°C there was relatively little effect of temperature on acetylene reduction.Nodules and cultures ofFrankia were compared with respect to the effect of temperature and pO₂ (partial pressure of oxygen) on oxygen uptake. Cultures ofFrankia were grown on a nitrogen-free medium at either 0.3 kPa O₂ (vesicles absent) or 20 kPa O₂ (vesicles present). Oxygen uptake by nodules (vesicles absent) and by vesicle-containing cultures was strongly dependent on pO₂ at values below 20 kPa. This suggests the presence of a barrier to oxygen diffusion. Oxygen uptake was dependent on temperature as well as on pO₂, but the Q₁₀ was much larger for the cultures than for the nodules. This suggests that vesicles or related structures are not the source of the diffusion barrier in Casuarina nodules. Respiration by cultures ofFrankia lacking vesicles became O₂-saturated at low pO₂ values. Thus these cultures did not have a significant diffusion barrier. From these results it is concluded that nodules ofCasuarina cunninghamiana have a barrier to oxygen diffusion supplied by the host tissue and not byFrankia.     

Simple rapid identification of Candida albicans with emphasis on the differentiation between Candida albicans and Candida stellatoidea

Summary Subcultures ofC. albicans, made from Sabouraud agar, grown at room temperature for 48 hours, were inoculated into a 10 times saline dilution of Sabouraud liquid medium and left in the incubator for 45–60 minutes at 37° C, transferred to corn meal agar plates and incubated at 37° C for 18–24 hours.Small portions of the surface agar containing the yeasts from these plates were pressed under cover glasses and examined under the oil immersion lens.Under these conditions,C. albicans cultures were observed to produce only yeast-like cells, whereasC. stellatoidea cultures contained predominantly abundant, long, thin mycelia.     

The growth of Epidermophyton floccosum and E. stockdaleae at different temperatures

The ability of 17 strains of genus Epidermophyton (15 strains belonging to Epidermophyton floccosum, one to E. floccosum var. nigricans and one to E. stockdaleae) to grow at different temperatures (4 °C, 25 °C, 28 °C, 31 °C, 34 °C, 37 °C and 40 °C) was stated.The strains were inoculated on Sabouraud Dextrose Agar and regularly controled over a period of 14 days when the plates were incubated at 25 °C, 28 °C, 31 °C, 34 °C, 37 °C and 40 °C, and over a period of 70 days when the temperature was 4 °C. The optimal growth of E. floccosum was observed at 28 °C and 31 °C, and no signs of growth were recorded neither at 4 °C nor at 40 °C. The optimal development of E. stockdaleae was observed at 25 °C and 28 °C. This species grew from 4 °C to 31 °C.     

The influence of cultivar and temperature on carbohydrate and amino acid exudation from gram seeds and on pre-emergence damping-off byRhizoctonia bataticola

Summary Three gram varieties varying in susceptibility to pre-emergence damping-off caused byRhizoctonia bataticola were compared for amounts of seed exudation. The results showed that the greatest seed exudation occurred with the variety G-130 which is the most susceptible to pre-emergence damping-off compared with the resistant varieties BG-203 and C-214. Germinating gram seeds exuded greater quantities of soluble carbohydrate and amino acids when incubated at 35°C than at 15 or 25°C. Exudates from seeds germinated at 35°C stimulated more mycelial growth ofR. bataticola than did exudates from seeds incubated at 15 or 25°C. Evidence is presented suggesting that increased seed exudation is a major factor contributing to increased pre-emergence damping-off of gram seedlings byR. bataticola at high temperatures.     

Temperature-sensitive plant mutants isolated in vitro

Summary Conditional-lethal, temperature-sensitive plant mutants have been isolated using a simple protoplast cloning method. The leaf protoplasts used were obtained from sterile, haploid shoot cultures of Nicotiana plumbaginifolia. Recessive mutations are described at three loci: ts1, ts2 and ts3. The mutations are lethal when either tissue cultures or plants are incubated at 33°C but not at 26°C.     

Microbial cell responses to a non-ionic surfactant

Summary The effects of a non-ionic surfactant, Pluronic F-68, on growth of microbial cell cultures have been studied. Growth ofSaccharomyces cerevisiae at 30°C or 37°C as measured by viable cell counts was unaffected by culture with pluronic. However, corresponding absorbance measurements forS. cerevisiae incubated with 5–10% pluronic were lower than controls at both temperatures. Absorbance ofE. coli cultures was also significantly reduced by incubation with 5.0–10.0% pluronic at both temperatures although viable counts again revealed no significant inhibition of growth.     

Studies on Aspergilli XVI. Effect of pH,temperature, and carbon and nitrogen interaction

Summary The effect of pH, temperature, and carbon and nitrogen interaction on the growth and sporulation ofAspergillus nidulans (Eidam)Wint.,A. rugulosus Thom &Raper,A. variecolor (Berk. &Br.)Thom &Raper andA. quadrilineatus was studied. All the moulds could grow on a wide range of pH (2.0 to 12.0) but the growth was poor on too acid and too alkaline media. Best growth ofA. rugulosus, A. quadrilineatus, andA. violaceus was seen at pH 6.5 and that ofA. nidulans andA. variecolor at pH 7.0. In general maximum production of perithecia was recorded between pH 6.0 and 8.0.All the above species ofAspergillus under study could grow between a temperature range of 10° C–48° C, but the growth was poor at 10° C and 48° C. The present moulds showed good growth at 20° C, 25°C, and 30° C. At 40° CA. nidulans andA. rugulosus showed moderate growth while the rest of the Aspergilli attained good growth. Temperatures between 20° C–30° C favoured excellent perithecial production.In general, little improvement in growth was noted on media containing good carbon and nitrogen sources. Malic acid was found to be useless when supplied singly. But, poor growth was recorded when supplied in combination with amino acids, amide, and peptone. This was due to the fact that these N sources also supplied carbon for their metabolism.     

Influence of temperature upon growth and sporulation in two species of Phytophthora

The paper deals with the influence of temperature on the growth and sporulation of two species ofPhytophthora, viz.,P. palmivora Butl. andP. parasitica Dast. var.macrospora Ashby, the causal agents of fruit rots ofAchras sapota L. andAnona squamosa L. respectively. Germination of sporangia at different temperatures were also undertaken. There was marked variation in growth and sporulation of these two organisms. Isolate C (Phytophthora palmivora) showed no growth at 5° and 35°C, scanty growth at 10° and 32.5° with an optimum temperature between 26–28°C. On the other hand, Isolate S (Phytophthora parasitica var.macroscora) showed no growth at 10°C, but slight growth even at 37°C. Eight days exposure at 37°C completely stopped the growth of this Isolate. It showed best growth at 30°C and hence this was its optimum temperature. In general, Isolate C sporulated abundantly at all temperatures tested but reached its maximum at 25°C. On the other hand Isolate S showed best growth but failed to sporulate at any of the temperatures in 98 hours growth, although it sporulated freely when the incubation period extended up to two weeks. On the basis of temperature toleration the twoPhytophthora isolates are distinguished from each other as two different species. This confirms the earlier observations and nomenclature criterion as emphasized and formulated byTucker (1931). In the germination studies, it was observed that the indirect germination with the formation of abundant zoospores started from 5° and continued even up to 35°C, reaching maximum at 20°C. High temperature was not favourable for indirect germination. As the temperature proceeded increasing, the percentage of direct germination by formation of germ tubes also increased. Direct germination was observed from 10° which continued up to 37°C, with a maximum reach at 30°C. This confirms the epidemic of fruit rots in nature during monsoon season which is prevalent with the persistence of high humidity and rainfall.Taken from a thesis submitted by the author for the degree of Master of Science in the Faculty of Agriculture, Poona University, India.     

Life history,resting egg formation,and hatching may explain the temporal-geographical distribution ofArtemia strains in the Mediterranean basin

Life history traits are presented for the sexual species (Artemia tunisiana) and for parthenogenetic diploid and tetraploid strainsA. parthenogenetica reared at 15 °, 24 ° and 29.5 °C.In laboratory cultures, we present evidence that the seasonal appearance of the sexualArtemia tunisiana (the dominant winter-spring population), and of two parthenogenetic populations ofArtemia (the dominant spring-summer populations) in certain Spanish saltworks is controlled by temperature through its effect on reproductive and survival traits. Minimum and maximum reproductive output and survival for the sexual and parthenogenetic populations, respectively, occurred at a typical temperature (24 °C) of the late-spring season when the sexual population is replaced by parthenogenetic forms. Furthermore, the high production and hatchability of cysts from the sexual population at low temperatures (15 °C), and of the parthenogenetic populations at middle temperature (24 °C), indicate the role of dormancy as an adaptation regulating seasonal occurrence.     

A rapid pigmentation test for identification ofCryptococcus neoformans

A rapid pigmentation test for identification ofCryptococcus neoformans is described. The method is based on the formation of a characteristic, mousegrey to violaceous-black pigment when shake cultures ofC. neoformans in a phosphate-buffered,l-DOPA — ferric citrate medium are incubated at 37 C for one hour.This work was carried out in part with financial support from the Council of Scientific and Industrial Research, New Delhi.     

Observations on the effects of incubation at inhibitory temperature on subsequent hatching of anostracan cysts

Hatching progressively for cysts ofBranchinecta lindahli Packard when they were incubated for 6 and 12 hours at a temperature known to inhibit hatching (30 °C) prior to transfer to a favorable hatching temperature of 15 °C. Cyst ofThamnocephalus platyurus Packard, by contrast, hatched normally at 28 °C after transfer from 48, 72, and 96 hour incubation periods at the inhibitory low temperature of 10 °C.     

Tapesia yallundae collected from wheat stubble in South Africa

Tapesia yallundae var.yallundae is newly recorded from wheat stubble collected near Moorresburg in the western Cape Province of South Africa. Apothecia were observed on wheat stubble incubated for 8 months at 10°C under near-ultraviolet light. Single ascospore isolates produced colonies typical of the Wheat (W)-type in culture. Apothecia were also induced after single-conidial isolates were mated on wheat stubble and incubated at 10–15°C for 6 months. All single-conidial isolates were of the W-pathotype (Ramulispora herpotrichoides var.herpotrichoides).     

The effect of low pressure-homogenized lactic cultures on the development of proteolysis in cheese slurry

Summary The aim of this study was to determine the effect of low pressure-homogenization of lactic acid bacteria (LAB) on the development of proteolysis in the slurry medium. For the slurry, the milk was pasteurized at 65 °C for 30 min, cooled to 32 °C and coagulated. The curd obtained was blended; the dry matter was adjusted to 30% by adding distilled water, placed into the flasks and autoclaved. The LAB Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and Lactobacillus helveticus were used in cheese slurry. Homogenization was performed at 30 MPa and 40 °C. The cheese slurries were incubated with and without homogenized cultures at 9 and 30 °C for up to 72 h. During incubation, the changes in trichloroacetic acid-soluble nitrogen (TCA-SN) and phosphotungstic acid-soluble nitrogen (PTA-SN) as well as pH were monitored. The results showed that pH development was slower in the slurries to which homogenized culture was added. Higher TCA-SN and PTA-SN values were obtained from the slurries incubated at 30 °C. Moreover, higher TCA-SN and PTA-SN values were found in the slurries incubated with homogenized mesophilic culture and Lb. helveticus (P<0.05). The results suggested that homogenization of the cultures was a promising method for the acceleration of cheese ripening.     

Leishmania donovani: In vitro culture and [1H] NMR characterization of amastigote-like forms

WhenLeishmania donovani promastigote forms, were cultured in TC-199 medium at 28°C and subsequently incubated at 38°C, they turned into aflagellate (amastogote-like) forms. A return of the incubation-culture temperature to 28°C these amastigote-like forms to revert to promastigotes. The amastigotes obtained by heat-shock, were viable and retained antigenic capacity being recognized by the sera of naturally infected patients. These forms, remained also capable of multiplying inside the J-774A. 1 macrophages. When the amastigote-like forms are kept in culture at 38°C retained their rounded appearance and their biological characteristics for more than 3 months subculturing every 6 days. These amastigote-like forms, when used for subcultures at 28°C, transformed into promastigotes capable of multiplying, as flagellate forms. The amastigotelike forms obtainedin vitro can be used in biochemical studies related to chemotherapy and immunology studies, as part of an effort to combat this parasite. The end-products of of glycolysis were studied in both the amastigote-like and promastigote forms ofL. donovani, by proton magnetic resonance analysis of the culture media. Alanine, succinate, and acetate, were predominant, and to a lesser extent pyruvate, glycine and D-lactate. Our results suggest that both forms ofLeishmania use different biochemical strategies to obtain their energy.     

An ultrastructural study of septal dissolution inSchizophyllum commune

A strain ofSchizophyllum commune carrying a mutation in theB-mating factor (B-mut) shows septal dissolution when grown at 30° C for 2 to 3 days. The septa are intact if the organism is grown at 25° C for the same time, but begin to break down within 1 h after transfer to 30° C. At the ultrastructural level the dolipore swelling is the first part of the septal apparatus to be degraded, closely followed by the disorganization of the parenthesomes. A progressive thinning of the septal cross-wall produces an enlargement of the septal aperture sufficient to allow the passage of nuclei. It appears that degradative enzymes are probably carried to the site of septal dissolution in vesicles derived from endoplasmic reticulum in the area of the septal apparatus.     

A new variety ofTalaromyces wortmannii and some observation onTalaromyces assiutensis

A new variety ofTalaromyces, T. wortmannii var.sublevisporus, is described and illustrated. It is characterized by producing nearly smooth-walled ascospores. The other characters are almost identical with those of the type variety. Based on observations of several strains including the authentic ones,T. assiutensis is considered a finely rough-spored variety ofT. trachyspermus.     

The effect of different temperatures on fatty-acid synthesis and polyunsaturation in cell suspension cultures

Cell suspension cultures of Catharanthus roseus G. Don, Glycine max (L.) Merr. and Nicotiana tabacum L. were incubated with [¹⁴C]acetate, [¹⁴C]oleic acid and [¹⁴C]linoleic acid at five different temperatures ranging from 15 to 35° C. When the incubation temperature was increased, [¹⁴C]acetate was incorporated preferentially into [¹⁴C]palmitate, with a concomitant drop in [¹⁴C]oleate formation. Between 15 and 20° C, [¹⁴C]oleic acid accumulated in C. roseus cells. In all cultures, optimum desaturation of [¹⁴C]oleic acid to [¹⁴C]linoleic acid occurred between 20 and 25° C, and in G. max this was also the optimal range for desaturation of [¹⁴C]linoleic acid to [¹⁴C]linolenic acid. Elongation of [¹⁴C]palmitic acid was inhibited when cultures grown at 15° C for 25 h were subsequently incubated with [¹⁴C]acetate at 25° C. [¹⁴C]oleic acid accumulated in G. max and C. roseus cultures grown at 35° C for 25 h and subsequently incubated at 25° C. Desaturation of [¹⁴C]oleic acid increased up to 25° C, but then decreased or leveled off depending on the cell line and on the temperature prior to incubation.     

Über die weite Verbreitung lichenisierter Sippen vonDictyochloropsis und die systematische Stellung vonMyrmecia reticulata (Chlorophyta)

Catinaria grossa is lichenized withDictyochloropsis splendida var.gelatinosa, var. nova. When cultured isolated from the fungus the cells of this variety are covered individually by thick gelatinous envelopes. The phycobionts ofMegalospora gompholoma subsp.gompholoma andM. atrorubicans subsp.australis belong to a new variety ofD. symbiontica, i.e. var.pauciautosporica, which preferentially reproduces by zoo- and aplanospores. The phycobiont ofPseudocyphellaria aurata does not belong toMyrmecia reticulata as formerly thought, but toDictyochloropsis symbiontica. Specimens from one locality in Tenerife and from New Zealand are lichenized with a strain ofD. symbiontica var.symbiontica, those from another locality in Tenerife with a strain ofD. symbiontica var.pauciautosporica. These strains differ in certain characters from other lichenized strains of these varieties.

     

A taxonomic study of KoreanSmilacina

Several taxonomic problems of KoreanSmilacina were examined by statistical methods, based on morphological data. Consequently,S. davurica described in Korean flora differs greatly from originalS. davurica, and closely related toS. bicolor morphologically. Therefore, the present authors treated this new taxon as a variety ofS. bicolor, S. bicolor var.flavovirens N. S. Lee et J. Y. Kim var. nov (“Yondusomdae” in Korean). The distribution in South Korea ofS. bicolor var.bicolor is known as only Mt. Chiri, but this species is collected in Mt. Dukyou, also. The bounds of morphological variations ofS. japonica var.japonica, the short plant size and no hairs on the stem, were revealed. The morphological characteristics ofS. japonica var.japonica andS. japonica var.mandshurica is obscure to distinguish. According to the present study,S. japonica var.mandshurica differs fromS. japonica var.japonica in plant size, size of adaxial leaf epidermal cell, ratio of style/ovary length and shape of stigma. The unrecorded taxon in Korean flora,S. robusta, was given a new Korean name as “Kunsomdae.” AlthoughS. japonica var.mandshurica is the variety ofS. japonica var.japonica, it is more closely related toS. robusta than toS. japonica var.japonica.     

Production of 3-nitropropionic acid byArthrinium species

3-Nitropropionic acid (NPA) is a known toxic metabolite produced by many species ofAstragalus of the Leguminosae family. In 1992, Liu et al. reported that some species ofArthrinium were the etiological fungi of moldy sugarcane poisoning (MSP) occurring in China. The toxic metabolite NPA produced byArthrinium was the main causative agent. Ten percent (88/884) of the MSP patients died, and many were left with lifelong disabilities. In this study, an accurate, sensitive, and rapid method for quantifying NPA produced byArthrinium species in sugarcane juice was developed. The method includes extraction, thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC). Fourteen strains ofArthrinium obtained from the American Type Culture Collection (ATCC) were examined for NPA-producing ability. Eight strains (57.1%), including three ofA. sacchari, two ofA. phaeospermum, and one each ofA. terminalis, A. aureum, andA. sereaensis, were positive for NPA production. The highest level of NPA produced byA. sacchari ATCC 76981 was 1.7 mg/ml when cultured at 24°C for 2 weeks. The optimum temperature for the production of NPA toxin was 24°C. The optimum pH was around pH 5.0. NPA production was higher in stationary cultures than in those shaken at 120 rpm.