Molecular characterization of an adenylate cyclase gene of the cyanobacterium Spirulina platensis

A cyaA gene, encoding an adenylate cyclase, was isolated from a filamentous cyanobacterium, Spirulina platensis, by functional complementation of a cya mutant of Escherichia coli, defective in adenylate cyclase activity. The predicted gene product of cyaA contains a signal peptide-like domain, a putative sensor domain similar to the gene product of vsrA of Pseudomonas solanacearum, a putative membrane-spanning domain and an adenylate cyclase-like catalytic domain. Two other positive clones that complemented the E. coli mutant were isolated from the same cyanobacterium, suggesting that several cya genes are functioning in S. platensis.     

Involvement of Azopirillum brasilense plasmid DNA in the productio of indole acetic acid

Indole acetic acid (IAA) production in Azospirillum brasilense strain Sp245 is controlled by a 85 MDa plasmid naturally present in this bacterium. In the presence of L-tryptophan, anthranilic acid production and almost no IAA production occurs in a derivative strain harbouring a Tn5-Mob insertion in the 85 MDa plasmid. Agrobacterium tumefaciens strain GM19023, upon transfer of Tn5-Mob labelled 85 MDa plasmid of A. brasilense Sp245, gains the ability to produce anthranilic acid.     

Edible insects in China: Utilization and prospects

The use of edible insects has a long history in China, where they have been consumed for more than 2000 years. In general, the level of acceptance is high for the consumption of insects in China. Many studies on edible insects have been conducted in the last 20 years, and the scope of the research includes the culture of entomophagy and the identification, nutritional value, farming and breeding of edible insects, in addition to food production and safety. Currently, 324 species of insects from 11 orders are documented that are either edible or associated with entomophagy in China, which include the common edible species, some less commonly consumed species and some medicinal insects. However, only approximately 10 to 20 types of insects are regularly consumed. The nutritional values for 174 species are available in China, including edible, feed and medicinal species. Although the nutritional values vary among species, all the insects examined contain protein, fat, vitamins and minerals at levels that meet human nutritional requirements. Edible insects were, and continue to be, consumed by different ethnic groups in many parts of China. People directly consume insects or food products made from insects. The processing of products from insect protein powder, oil and chitin, and the development of healthcare foods has been studied in China. People also consume insects indirectly by eating livestock that were fed insects, which may be a more acceptable pathway to use insects in human diets. Although limited, the data on the food safety of insects indicate that insects are safe for food or feed. Incidences of allergic reactions after consuming silkworm pupae, cicadas and crickets have been reported in China. Insect farming is a unique breeding industry in rural China and is a source of income for local people. Insects are reared and bred for human food, medicine and animal feed using two approaches in China: the insects are either fully domesticated and reared completely in captivity or are partially raised in captivity, and the insect habitat is manipulated to increase production. Depending on the type of relationship the insect has with humans, plants and the environment, different farming strategies are used. The social and scientific communities must work together to promote the use of insects as food and feed.     

Thermal and chemical denaturation of Colocasia esculenta tuber agglutinin from α2β2 to unfolded state

The major tuber storage protein of Colocasia esculenta, is a monocot mannose-binding, widely used dietary lectin, containing two polypeptides of 12.0 and 12.4 kDa. By both gel filtration and dynamic light scattering at pH 7.2, the lectin has a α₂β₂ form of apparent molecular mass of 48.2 kDa and a hydrodynamic radius of 6.1 ± .2 nm; however, at pH 3, it migrates as αβ and has a reduced hydrodynamic radius of 4.6 ± .3 nm. Our circular dichroism spectroscopy studies show that the lectin retains approximately 100% of its secondary structure between pH 2–8, going down to ~90% for extreme acidic/alkaline conditions. The fluorescence emission maxima of 346 to 350 nm for pH 4 to 10 show that the tryptophan residues are relatively exposed. The unfolding is a simple two-state process, N₄ ? 4U, as seen in our denaturation scan profiles. These denaturation profiles, monitored separately by fluorescence, far-UV CD, and near-UV CD, are completely super imposable. Analyses of these profiles provide an estimate of several thermodynamic parameters at each guanidinium chloride concentration, including the melting temperature T_g, which is 346.9 K in 0 M, but lowers to 321.8 K in 3.6 M. Dimeric and tetrameric interfaces observed in the crystal structure for the same protein are used to rationalize solution data in some detail.     

KINETICS OF GROWTH AND DEATH IN ANABAENA FLOS-AQUAE (CYANOBACTERIA) UNDER LIGHT LIMITATION AND SUPERSATURATION

The kinetics of population growth and death were investigated in Anabaena flos-aquae (Lyngb.) Bréb grown at light intensities ranging from limitation to photoinhibition (5 W·m⁻² to 160 W·m⁻²) in a nutrient-replete turbidostat. Steady-state growth rate (μ, or dilution rate, D) increased with light intensity from 0.44·day⁻¹ at a light intensity of 5 W·m⁻² to 0.99·day⁻¹ at 20 W·m⁻² and started to decrease above about 22 W·m⁻², reaching 0.56·day⁻¹ at 160 W·m⁻². The Haldane function of enzyme inhibition fit the growth data poorly, largely because of the unusually narrow range of saturation intensity. However, it produced a good fit (P < 0.001) for growth under photoinhibition. Anabaena flos-aquae died at different specific death rates (γ) below and above the saturation intensity. When calculated as the slope of a v_x⁻¹ and D⁻¹ plot, where v_x and D are cell viability (or live cell fraction) and dilution rate, respectively; γ was 0.047·day⁻¹ in the range of light limitation and 0.103·day⁻¹ under photoinhibition. Live vegetative cells and heterocysts, either in numbers or as a percentage of the total cells, showed a peak at the saturation intensity and decreased at lower and higher intensities. The ratio of live heterocysts to live vegetative cells increased with intensity when light was limiting but decreased when light was supersaturating. In cells growing at the same growth rate, the ratio was significantly lower under light inhibition than under subsaturation and the cell N:C ratio was also lower under inhibition. The steady-state rate of dissolved organic carbon (DOC) production increased with light intensity. However, its production as a percentage of the total C fixation was lowest at the optimum intensity and increased as the irradiance decreased or increased. The rate and percentage was significantly higher under photoinhibition than limitation in cells growing at the same growth rate. About 22% of the total fixed carbon was released as DOC at the highest light intensity. No correlation was found between the number of dead cells and DOC.     

EFFECTS OF POTASSIUM ON THE PHOTOSYNTHETIC RECOVERY OF THE TERRESTRIAL CYANOBACTERIUM,NOSTOC FLAGELLIFORME (CYANOPHYCEAE) DURING REHYDRATION1

Effects of potassium on the photosynthetic recovery of Nostoc flagelliforme (Berk. & Curtis) Bornet & Flahault were investigated to determine its exact role during rehydration. Potassium enhanced recovery of the ability to reduce the primary quinone‐type acceptor (Q_A) and plastoquinone (PQ) pool and the area over the fluorescence rise curve was increased by 127%. The proportions of closed PSII reaction centers at phases J and I and the net rate of closure of PSII reaction centers were decreased by, respectively, 19%, 8%, and 23% with the addition of potassium, due to changes in the ability of PSII for multiple turnovers needed to reduce the PQ pool. Potassium significantly enhanced the probability of electron transfer beyond Q_A and the recovery of electron transport flux per PSII reaction center. Electron transport from water to methyl viologen for samples rehydrated in K⁺‐free BG₁₁ medium was 54% of those with the addition of potassium. However, electron flow from water to p‐benzoquinone and from reduced 2,6‐dichlorophenol‐indophenol to methyl viologen showed little change with the addition of potassium. The fast phase and slow phase of millisecond delayed light emission and the ATP content for samples rehydrated in K⁺‐free BG₁₁ medium were, respectively, 71.6%, 50.7%, and 77.1% of those with the addition of potassium. These suggested that potassium affected electron transfer from PQ to plastocyanin through the cytochrome b₆f complex and the proton motive force across the thylakoid membranes, probably reflecting its role in charge balance during H⁺ transport by the cytochrome b₆f complex.     

RELATIONSHIP OF GROWTH AND PHOTOSYNTHESIS WITH COLONY SIZE IN AN EDIBLE CYANOBACTERIUM,GE‐XIAN‐MI NOSTOC (CYANOPHYCEAE)1

Growth and photosynthesis of an edible cyanobacterium, Ge‐Xian‐Mi (Nostoc), were investigated with differently sized colonies. Both photosynthesis and growth were dependent on the colony size. Compared with larger ones, smaller colonies grew faster regardless of the levels of light and temperature for culture and showed higher values of maximal net photosynthetic rate, apparent quantum yield, light‐saturating and compensating points, and dark respiration. The ratios of chl a content and mass to surface area of a colony increased and that of chl a to mass or mass to volume of a colony decreased with increased colonial sizes. A Ge‐Xian‐Mi colony appeared to increase its chl a content per surface area, enhancing the light‐shading effect; however, at the same time it decreased its mass density on a volume basis, minimizing the enhanced effects of shading and diffusion barrier caused by the thickening outer layer with increasing colony size during growth.     

Diversity of macrofungi and exploitation of edible mushroom resources in the National Park “Appennino Lucano,Val D'Agri,Lagonegrese” (Italy)

An investigation on the macrofungal diversity of the National Park “Appennino Lucano, Val D'Agri, Lagonegrese” (Basilicata, southern Italy) was carried out, together with an evaluation of wild edible mushrooms in agro-forest ecosystems and their possible exploitation as a new source of food and revenue in rural and under-developed areas of this region. An unpublished list of 249 mushroom taxa (229 Basidiomycota and 20 Ascomycota), mostly belonging to the genera Tricholoma, Tuber, Russula, Amanita, Boletus, Lactarius, Mycena, Agaricus, Clitocybe and Hygrophorus, growing in this important natural area is provided together with ecological characterization of the recorded specimens. Inonotus obliquus is a new record for Basilicata, while Mycena bresadolana is reported for the first time from southern Italy. The presence of Amaurodon mustialaënsis, an extremely rare species of the northern hemisphere reported for the first time on white fir, is noteworthy. Among the previous findings, 116 species of edible mushrooms are presented and discussed in view of their relative abundance and edibility.     

Role of external calcium in homeostasis of intracellular pH in the cyanobacterium Anabaena sp. strain PCC7120 exposed to low pH

The role of external Ca²⁺ in the homeostasis of intracellular pH (pHi) of Anabaena sp. strain PCC7120 in response to a decrease in the external pH (pHex) has been studied in cell suspensions. Increase in cytoplasmic pH after acid shock is dependent on the presence of Ca²⁺ in the medium. The observed Ca²⁺-mediated alkalization of the cytoplasm depends on the extent of the shift in external pH. Acid pH shifts resulted in an increased permeability of the cytoplasmic membrane to protons, which could be reversed by increasing the concentration of Ca²⁺ in the medium. Thus, the ability of Ca²⁺ to increase cytoplasmic pH might be correlated with an inhibition of net proton uptake by increasing concentrations of external Ca²⁺ under these conditions. This combined response resulted in the generation and maintenance of a larger pH gradient (ΔpH) at acid external pH values. All Ca²⁺ channel blockers tested, such as verapamil and LaCl₃, inhibited the observed Ca²⁺-mediated response. On the other hand, the Ca ionophore calcimycin (compound A23187) was agonistic, and stimulated both cytoplasmic alkalization and inhibition of net proton uptake. The protonophorous uncoupler carbonylcyanide m -chlorophenyl hydrazone, inhibited this Ca²⁺-mediated response, whereas monensin, an inhibitor of the Na⁺/H⁺ antiporter, had no significant effect. The results of the present study suggest that an influx of Ca²⁺ from the extracellular space is required for the regulation of cytoplasmic pH in Anabaena sp. strain PCC7120 exposed to low external pH values.     

生物信息学在食用菌研究中的应用

食用菌因其富含多种氨基酸及微量元素等物质,具有较高的营养价值和药用价值,越来越受到人们的关注和喜爱。我国作为最重要的食用菌生产国,食用菌生产规模不断扩大,产量也在逐年提高。为了更好地发展食用菌产业,迫切需要在传统的食用菌产业链,如优良品种选育及栽培生产中融入新技术。生物信息学作为一门研究分析生物生命结构的技术门类,通过运用数学、计算机科学等工具揭示了数据所蕴含的生物学意义,极大地促进了生命科学研究的发展,也为食用菌更深入的研究与应用提供了技术保障。本文从食用菌育种及种质资源调查、病虫害防治、基因组学、食用菌安全等几方面阐述了生物信息学在食用菌领域的具体应用,对生物信息学在食用菌及农业领域的发展进行了展望,以期为促进食用菌研究和生产发展提供参考。