The Growth of Chlorella pyrenoidosa on Glycollate

The utilization of glycollate as a substrate for photoheterotrophicgrowth by a strain of Chlorella pyrenoidosa has been demonstrated.Glycollate stimulated growth of this alga in basal medium overthe pH range 4.0 to 8.0 in the light, but did not support growthin the dark. Stimulation of growth in the light occurred overa wide range of glycollate concentrations and was optimal at30 mM. Enzyme and inhibitor experiments suggested that the synthesisof cell constituents during growth on glycollate is achievedby the same pathway which operates during the metabolism ofexogenous glycollate by autotrophically-grown cells. For algaeto metabolize and grow on exogenous glycollate the cells mustbe readily permeable to this compound. When the cells readilytake up exogenous glycollate, the level of activity of enzymesin the cell, in particular glycollate:DCPIP oxidoreductase,may regulate the over-all rate of glycollate metabolism.     

The role of cotyledon metabolism in the establishment of quinoa (Chenopodium quinoa) seedlings growing under salinity

A growth chamber experiment was conducted to assess the effect of salinity on emergence, growth, water status, photosynthetic pigments, osmolyte accumulation, and ionic content of quinoa seedlings (Chenopodium quinoa). The aim was to test the hypothesis that quinoa seedlings are well adapted to grow under salinity due to their ability to adjust the metabolic functionality of their cotyledons. Seedlings were grown for 21 days at 250 mM NaCl from the start of the germination. Germination percentage and cotyledon area were not affected by salt whereas seedling height decreased 15%. FW increased in both control and salt-treated cotyledons, but the increase was higher under salinity. DW only increased in salt-treated cotyledons. The DW/FW ratio did not show significant differences between treatments. Relative water content, chlorophyll, carotenoids, lipids, and proteins were significantly lower under salinity. Total soluble sugars, sucrose and glucose concentrations were higher in salt-treated than in control cotyledons. Ion concentration showed a different distribution pattern. Na⁺ and Cl^? concentrations were higher under salinity, while an inverse result was observed for K⁺ concentration. Proline and glycinebetaine concentrations increased under salinity, but the increase was higher in the former than the latter. The osmoprotective role of proline, glycinebetaine, and soluble sugars is discussed.     

The Photometabolism of Acetate by Chlorella pyrenoidosa

Chlorella pyrenoidosa can utilize sodium acetate as a carbonsource for growth in the light. Growth proceeds under aerobicconditions both in the presence and in the absence of carbondioxide, but under anaerobic conditions only in its presence.The assimilation of acetate does not result from oxidation tocarbon dioxide followed by photosynthetic fixation because theproducts of ¹⁴C-acetate assimilation are different from theproducts of ¹⁴CO₂ fixation in the presence of unlabelled acetate. In aerobic conditions 10-⁶ M DCMU induces a pattern of acetateassimilation in the light similar to that in the dark. Thus,in the presence of DCMU in the light, less acetate carbon isincorporated into cells, particularly into lipids, polysaccharide,and protein, and more is released as carbon dioxide than inits absence. The effect of 4 x 10^-3 M MFA on acetate assimilationin the presence of 10^-6 M DCMU is the same in light and dark.Acetate assimilation is unaffected by desaspidine and sodiumbisulphite. The mean generation time of C. pyrenoidosa growing on acetatein the light under aerobic conditions is 20 hours. When 10^-5M DCMU is added the mean generation time is 60 hours, the sameas that for Chlorella growing on acetate in the dark. The activityof the enzymes of the glyoxylate cycle, isocitrate lyase (E.C.4.1.3.1.)and malate synthetase (E.C.4.1.3.2.) is repressed in the light,but activity of both enzymes increases markedly when DCMU isadded.     

The Effect of Hydroxymethanesulphonate on Photosynthesis in Chlorella pyrenoidosa

Photosynthesis under conditions known to favour glycollate excretionby algae did not result in glycollate excretion in a strainof Chlorella pyrenoidosa unless an inhibitor of glycollate oxidase,-hydroxypyridin-2yl-methane sulphonate (-HPMS), was present.This inhibitor increased the total amount of glycollate presentin the supernatant from the cells during photosynthetic carbondioxide fixation and gave accumulation of ¹⁴C in glycollateduring ¹⁴CO₂ fixation under conditions favouring glycollatesynthesis. At pH 8.3 -HPMS did not stimulate photosynthetic¹⁴CO₂ fixation in C. pyrenoidosa as occurs with some algae.Photoassimilation of acetate was inhibited by -HPMS, and thiswas shown to result from acetyl-CoA synthetase inhibition by-HPMS.     

增强UV-B对蛋白核小球藻中NO释放的影响

研究了蛋白核小球藻在增强UV—B辐射下NO信号的产生。结果表明：NO释放量与UV—B的强度相关。在藻类细胞培养液中加入NOS的竞争性抑制剂硝基精氨酸(LNNA),不能抑制NO的释放。加入NO清除剂乙酰半胱氨酸(NAC),能明显减少NO的释放。在无氮和有氮培养基中NO同样的释放。这些结果说明NO生成途径不经过依赖L-精氨酸的NOS和依赖NO3^-和硝酸还原酶,NO的底物不是L-精氨酸和NO3^-在无氮和有氮培养基中同样有NO3^-的形成,NAC减少NO2^-的释放,说明生成了NO2^-有可能是NO释放后生成的。     

Construction of a quinoa (Chenopodium quinoa Willd.) BAC library and its use in identifying genes encoding seed storage proteins

Quinoa (Chenopodium quinoa Willd.) is adapted to the harsh environments of the Andean Altiplano region. Its seeds have a well-balanced amino acid composition and exceptionally high protein content with respect to human nutrition. Quinoa grain is a staple in the diet of some of the most impoverished people in the world. The plant is an allotetraploid displaying disomic inheritance (2n=4x=36) with a di-haploid genome of 967 Mbp (megabase pair), or 2C=2.01 pg. We constructed two quinoa BAC libraries using BamHI (26,880 clones) and EcoRI (48,000 clones) restriction endonucleases. Cloned inserts in the BamHI library average 113 kb (kilobase) with approximately 2% of the clones lacking inserts, whereas cloned inserts in the EcoRI library average 130 kb and approximately 1% lack inserts. Three plastid genes used as probes of high-density arrayed blots of 73,728 BACs identified approximately 2.8% of the clones as containing plastid DNA inserts. We estimate that the combined quinoa libraries represent at least 9.0 di-haploid nuclear genome equivalents. An average of 12.2 positive clones per probe were identified with 13 quinoa single-copy ESTs as probes of the high-density arrayed blots, suggesting that the estimate of 9.0× coverage of the genome is conservative. Utility of the BAC libraries for gene identification was demonstrated by probing the library with a partial sequence of the 11S globulin seed storage protein gene and identifying multiple positive clones. The presence of the 11S globulin gene in four of the clones was verified by direct comparison with quinoa genomic DNA on a Southern blot. Besides serving as a useful tool for gene identification, the quinoa BAC libraries will be an important resource for physical mapping of the quinoa genome.     

Effects of Phenylethyl Alcohol on Chlorella pyrenoidosa

Asynchronous as well as synchronous cultures were used to study the effect of phenylethyl alcohol on Chlorella pyrenoidosa. The following observations were made:

• 1 Autotrophic growth and the synthesis of DNA and chlorophyll were inhibited more than 80% by 8.4 mM phenylethyl alcohol when added to synchronous suspensions containing autospores.
• 2 Autospore formation did not occur when fully grown cells capable of division were transferred to 4.2 mM phenylethyl alcohol.
• 3 Photosynthetic oxygen evolution of asynchronous cultures was inhibited more than 70 % by 21 mM phenylethyl alcohol.
• 4 Endogenous respiration in the dark was stimulated by 8.4 mM, while higher concentrations were inhibitory.
• 5 Glucose respiration in the dark was inhibited by all concentrations in the range from 8.4 mM to 21 mM. Assimilation of glucose in darkness was retarded by phenylethyl alcohol at concentrations which gave maximal stimulation of endogenous respiration.

It is concluded that phenylethyl alcohol is not a specific inhibitor of DNA synthesis. The prime effect appears to be on respiration and photosynthesis.     

The pathway of glycollate utilization in Chlorella pyrenoidosa

1. Exogenous glycollate was rapidly metabolized in both the light and the dark by photoautotrophically grown Chlorella pyrenoidosa. 2. The incorporation of (14)C from [1-(14)C]glycollate by these cells was inhibited by the tricarboxylic acid-cycle inhibitors monofluoroacetate, diethylmalonate and arsenite, and also by alpha-hydroxypyrid-2-ylmethanesulphonate and isonicotinylhydrazine. 3. Short-term kinetic experiments showed over 80% of the total (14)C present in the soluble fraction from the cells to be in glycine and serine after 10s. This percentage decreased with time whereas the percentage radioactivity in glycerate increased for up to 30s then remained steady. The percentage of the total radioactivity present in citrate increased over the experimental period. Malate was the only other tricarboxylic acid-cycle intermediate to become labelled. 4. The kinetic and inhibitor experiments supported the following pathway of glycollate incorporation: glycollate --> glyoxylate --> glycine --> serine --> hydroxypyruvate --> glycerate --> 3-phosphoglycerate --> 2-phosphoglycerate --> phosphoenolpyruvate --> pyruvate --> acetyl-CoA. 5. The specific activities of the enzymes catalysing this metabolic sequence in cell-free extracts were great enough to account for the observed rate of glycollate metabolism of 0.25mumol/h per mg dry wt. of cells in the light.     

Algicidal effect of bromine and chlorine on Chlorella pyrenoidosa

Chlorella pyrenoidosa was found to grow rapidly in tap water. Peak growth was reached after 2 to 3 days. Chlorine and bromine, added to such water, were shown to be effective inhibitors of algal growth. Bromine and bromamine were primarily algicidal, whereas chlorine and chloramines were mainly algistatic. It is assumed that the mechanisms of action of these halogens on Chlorella are not the same.     

Phenological growth stages of quinoa (Chenopodium quinoa) based on the BBCH scale

Quinoa (Chenopodium quinoa) is a pseudocereal native from the Andean region of South America that has increased in importance worldwide. Quinoa is now considered an alternative to traditional crops in a climate change scenario, considering its ability to adapt to marginal soils, droughts and frosts. Despite the interesting agronomic and nutritional features of this crop, research into quinoa is characterised by individual attempts to define its phenological stages without an international consensus. A unique criterion to quantify the phenology of quinoa could become a useful tool for researchers and plant breeders in future work by standardising this information for international cooperation. In this article, a proposed scale of the phenological growth stages of quinoa based on the BBCH coding system (Biologische Bundesanstalt Bundessortenamt und CHemische Industrie) was developed. Growth stages were described utilising the decimal code of the BBCH system, and figures were included for the most representative stages.     

藜麦及其资源开发利用

藜麦Chenopodium quinoa Willd.英文名:quinoa,原产于南美洲安第斯山区,是印加土著居民的主要传统食物,至今已有5 000~7 000多年的利用和种植历史。古代印加人将它称之为"粮食之母"。藜麦在20世纪80年代,被美国宇航局用于宇航员的太空食品。联合国粮农组织认为藜麦是唯一的单一植物即可满足人体基本营养需求的食物,正式推荐藜麦为最适宜人类的完美的全营养食品。本文对藜麦的植物形态、生态特性、营养价值以及在我国种植展望作了综合报道。     

Ricinosomes provide an early indicator of suspensor and endosperm cells destined to die during late seed development in quinoa (Chenopodium quinoa)

Background and Aims

In mature quinoa (Chenopodium quinoa) seeds, the lasting endosperm forms a micropylar cone covering the radicle. The suspensor cells lie within the centre of the cone. During the final stage of seed development, the cells of the lasting endosperm accumulate protein and lipids while the rest are crushed and disintegrated. Both the suspensor and endosperm die progressively from the innermost layers surrounding the embryo and extending towards the nucellar tissue. Ricinosomes are endoplasmic reticulum-derived organelles that accumulate both the pro-form and the mature form of cysteine endopeptidase (Cys-EP), first identified in castor bean (Ricinus communis) endosperm during germination. This study sought to identify associations between the presence of ricinosomes and programmed cell death (PCD) hallmarks in suspensor and endosperm cells predestined to die during quinoa seed development.

Methods

A structural study using light microscopy and transmission electron microscopy was performed. To detect the presence of Cys-EP, both western blot and in situ immunolocalization assays were carried out using anti-R. communis Cys-EP antibody. A TUNEL assay was used to determine DNA fragmentation.

Results and Conclusions

Except for the one or two cell layers that constitute the lasting endosperm in the mature seed, ricinosomes were found in suspensor and endosperm cells. These cells were also the site of morphological abnormalities, including misshapen and fragmented nuclei, vesiculation of the cytosol, vacuole collapse and cell wall disorganization. It is proposed that, in suspensor and endosperm cells, the early detection of Cys-EP in ricinosomes predicts the occurrence of PCD during late seed development.     

The Kinetics of Extracellular Glycollate Production by Chlorella pyrenoidosa

Extracellular glycollate is liberated by Chlorella pyrenoidosaduring growth in medium bubbled with air or 3 per cent carbondioxide in air. With air the rate of release of glycollate percell decreases, with 3 per cent carbon dioxide it increases,with increase in cell number. Glycollate is released duringshort-term experiments when C. pyrenoidasa, grown under lowlight and high carbon dioxide, is transferred suddenly to highlight and low carbon dioxide. No other combination of thesefactors produces a comparable release of glycollate. The quantityof glycollate released in short-term experiments increases exponentiallywith the relative growth-rate of the culture from which thecells are derived. A crucial condition for maximum glycollaterelease is that growth of the culture prior to the experimentshould not be limited by carbon-dixoide concentration. The effectof pH is related to its effect on growth-rate; i.e. C. pyrenoidosahas a lower relative growth-rate at pH 8.3 and produces correspondinglyless glycollate than faster growing cultures at pH 6.4. Duringshort-term experiments under high light and low carbon dioxidethe rate of glycollate release drops after 50–100 minutessuggesting exhaustion of the glycollate precursor.     

金藜麦耐盐性分析及营养评价

对我国沿海地区新收集种质资源金藜麦(Chenopodium quinoa Willd.)进行了耐盐性及营养品质评价。结果表明:金藜麦在对盐胁迫相对敏感的芽期和苗期表现出相对较高的耐盐性;子粒蛋白质含量为14.2%,蛋白营养价值优于牛奶以及小麦、水稻、玉米、大豆等作物;子粒中富含维生素B、E等以及钙、锰、铁、铜、锌等矿质元素,特别是钙含量高达190.16 mg/100g,是小米钙含量的35倍;且金藜麦子粒含有丰富的必需脂肪酸,如亚油酸(3.58 g/100g)和亚麻酸(0.44 g/100g),天然抗氧化剂维生素E含量为7.66 mg/100g。这些研究结果表明,新收集的金藜麦种质资源具有较高的营养价值和耐盐性,将为我国藜麦研究和种植提供重要的种质资源。     

Effect of phenolic acids on growth of Chlorella pyrenoidosa

Static bioassays were conducted on Chlorella pyrenoidosa using vanillic acid, syringic acid, and 4-hydroxybenzoic acid. At 0.1 mM, vanillic and 4-hydroxybenzoic acid stimulated cultural growth compared to the control. For both compounds, concentrations of 0.3 mM and 0.4 mM were initially inhibitory, then after 3–5 days became stimulatory compared to control. Bioassays with syringic acid at 0.1 mM, 0.3 mM, and 0.4 mM were all stimulatory. At 0.5 mM–2.5 mM, syringic acid resulted in 100% mortality in C. pyrenoidosa. Bacteria-free cultures of C. pyrenoidosa were stimulated by vanillic acid at 0.1 mM and inhibited at 0.4 mM with no shift in response observed. It was suggested that degradation of test material is responsible for the shift from inhibition to stimulation. All three compounds are reported to change from enzyme synergists to antagonists as concentrations are reduced. Because these compounds are major components of humic material, it is suggested that they have the potential to confound studies involving interaction of toxins and humics.