A solvent partitioning procedure for the separation of chlorophylls from their degradation products and carotenoid pigments

A simple liquid/liquid partitioning procedure was developed which employed aqueous acetonitrile and hexane, for the isolation of chlorophyll and pheophytin. This procedure separated these pigments from other interfering pigmented compounds. The efficacy of this solvent separation method was evaluated using commercially available chlorophylla, b, pheophytina, b, carotenoids, and algal pigment extracts. The recovery efficiencies of this solvent partitioning process for chlorophyll a and pheophytina have been shown to be 95–98% and 93–96%, respectively, furthermore, the chlorophylla fraction was practically free of any contaminating pigments. It appears that a more accurate assessment of chlorophylla and pheophytina can be accomplished employing liquid/liquid partitioning than with the present standard method.     

An improved method for extraction and separation of photosynthetic pigments

The method for extracting and separating hydrophobic photosynthetic pigments proposed by Katayama et al. (Japanese Journal of Phycology, 42, 71-77, 1994) has been improved to introduce it to student laboratories at the senior high school level. Silica gel powder was used for removing water from fresh materials prior to extracting pigments by a mixture of organic solvents that was also used for chromatographic separation of the pigments. A small silica gel thin-layer plate or a paper strip was used for separating the pigments. The improved method may be applicable to all kinds of plant materials including algae, is easier than most other methods, and can lead to more successful results in separating these pigments by both thin-layer chromatography and paper chromatography. The method has been carried out in student laboratories in some senior high schools and universities in Japan. The results indicate that this laboratory exercise is effective for students to recognise the unity and diversity of plants. Therefore, this laboratory seems to be useful for teaching plant systematics as well as for teaching photosynthesis.     

Algal and bacterial pigments in non-laminated lacustrine sediment: Studies of their sedimentation, degradation and stratigraphy

Abstract Natural phyto- and bacterioplankton populations of Lake Vechten (the Netherlands) were subjected to darkness under oxic and anoxic conditions at in situ temperatures in order to test the stability in time of their photosynthetic pigments. Furthermore, sedimentary fluxes and concentrations of pigments were estimated in (respectively) sediment trap catches and at the sediment-water interphase in order to measure the pigment breakdown upon burial into the sediment. The chlorophylls and most of the xanthophylls showed substantial losses of 20% to 60% in the incubation experiments as well as in the surficial sediment. β-Carotene, okenone and echinenone were most stable (2–10% losses); fucoxanthin and peridinin were degraded extensively; alloxanthin and zeaxanthin held an intermediate position as did the bacteriochlorophylls. Trends in sediment profiles of pigments were compared with limnological data obtained during the enhanced eutrophication of the lake. Evidence was provided that the β-carotene profile closely followed the increase of phytoplankton biomass. Although susceptible to substantial degradation, several profiles of pigments and of pigment ratios could be related in a qualitative way to biomass and to shifts in species composition which occurred as a result of the changing ecological conditions in the lake.     

A procedure for the separation and quantitation of tryptophan and amino sugars on the amino acid analyzer

Tryptophan, 5-methyl tryptophan, glucosamine, and galactosamine can be separated from each other and hydrolysis products including lysinoalanine by chromatography on a 6 × 260-mm column of W-3H resin. The column is developed at 70°C for 20 min with pH 3.95 (0.4 Na⁺) buffer, followed by pH 6.4 (1 Na⁺) buffer for 55 min using a Beckman 119 CL amino acid analyzer. The recovery of the internal standards, 5-methyl tryptophan and galactosamine, can then be used to correct for tryptophan and glucosamine losses, respectively. The procedure uses the column and buffers normally employed for protein hydrolysate analysis and does not require additional resin columns, special buffers, or flow rate changes.     

Spectrochromatography of photosynthetic pigments as a fingerprinting technique for microbial phototrophs

Abstract: The combination of chromatographic separation using reverse-phase high-performance liquid chromatography and continuous monitoring of the column eluate using a diode-array spectrophotometer allowed qualitative and quantitative pigment profiling of extracts of photosynthetic material in a single run. Carotenoids and the spectrally distinct types of chlorophyll and bacteriochlorophyll can be unambiguously identified even when imperfectly separated on the column. The resulting spectrochromatogram is a fingerprint useful for the rapid characterization of pure cultures or mixed populations. We have developed software to allow recording, manipulation, and presentation of the resulting spectrochromatograms and present results from photosynthetic microbes in pure and mixed cultures. We describe a number of approaches to the presentation of the resulting data in a readily comprehensible form.     

蛇足石杉中产石杉碱甲内生真菌的分离和鉴定

【目的】从野生蛇足石杉(Huperzia serrata)中分离筛选产石杉碱甲的内生真菌。【方法】采用薄层层析及高效液相色谱法对内生真菌代谢产物进行测定和分析以期分离获得产石杉碱甲菌株,运用形态及ITS序列分析方法对产石杉碱甲菌株进行鉴定,并利用连续传代方法考察菌株遗传稳定性。【结果】经筛选获得一株产石杉碱甲内生真菌NSH-5,经形态学鉴定及ITS序列分析鉴定为轮枝镰孢菌(Fusarium verticillioides),其石杉碱甲产量为11.76 mg/100 m L,菌株经20次连续传代后遗传稳定。【结论】NSH-5菌株为一株具有产石杉碱甲能力的轮枝镰孢菌,该菌株的发现为生物合成石杉碱甲提供了新的菌种资源。     

In situ photosynthetic differentiation of the green algal and the cyanobacterial photobiont in the crustose lichen Placopsis contortuplicata

In situ photosynthetic activity in the green algal and the cyanobacterial photobionts of Placopsis contortuplicata was monitored within the same thallus using chlorophyll a fluorescence methods. It proved possible to show that the response to hydration of the green algal and the cyanobacterial photobionts is different within the same thallus. Measurements of the photochemical efficiency of PS II, Fv/Fm, reveal that in the dry lichen thallus photosynthetic activity could be induced in the green algal photobiont by water vapour uptake, in the cyanobacterial photobiont only if it was hydrated with liquid water. However, rates of apparent electron flow through PS II as well as rates of CO₂ gas exchange were suboptimal after hydration with water vapour alone and maximum rates could only be observed when the thallus was saturated with liquid water. The differences in the waterrelated photosynthetic performance and different light response curves of apparent electron transport rate through PS II indicate that the two photobionts act highly independently of each other. It was shown that the cyanobacteria from the cephalodia in P. contortuplicata act as photobiont. The rate of electron flow through PS II was found to be saturated at 1500 mol photon m^–2 s^–1, despite a considerable increase of non-photochemical quenching in the green algal photobiont which is lacking in the cyanobacterial photobiont. No evidence of photoinhibition could be found in either photobiont. Pronounced competition between the green algal and the cyanobacterial thallus can be observed in the natural habitat, indicating that the symbiosis in P. contortuplicata should be regarded as a very variable adaptation to the extreme environmental conditions in the maritime Antarctic.Abbreviations DR dark respiration - ETR apparent rate of electron flow of PS II (=F/Fm×PFD) - F difference in yield of fluorescence and maximal Fm and steady state Fs under ambient light - Fo minimum level of fluorescence yield in dark-adapted state - Fo minimum level of fluorescence yield after transient darkening and far-red illumination - Fm maximum level of dark-adapted fluorescence yield - Fm maximum yield of fluorescence under ambient light - Fs yield of fluorescence at steady state - Fv difference in minimum fluorescence and maximum fluorescence in dark-adapted state - NP net photosynthesis - NPQ coefficient for non-photochemical quenching - PAR photosynthetically active radiation (400–700 nm) - PFD photon flux density in PAR - PS II photosystem II - qN coefficient for non-photochemical quenching - qP coefficient for photochemical quenching     

A new procedure for the purification of spinach leaf photosynthetic fructose-1,6-bisphosphatase by affinity chromatography on mercaptoethylamine-Sepharose

A new purification procedure for spinach leaf fructose-1,6-bisphosphatase is proposed, which includes the use of affinity chromatography on mercaptoethylamine-Sepharose. A homogeneous preparation of the enzyme can be obtained in 48 hr, with a specific activity of 67 U/mg and a yield of 23%. The method may also be useful for the purification of other thioredoxin-activated chloroplast enzymes.     

Quinone and pheophytin in the photosynthetic reaction center II from spinach chloroplasts

Prenylquinones and pheophytin a in a preparation of photosynthetic reaction center II from spinach chloroplasts were chemically determined. Each reaction center II had two molecules, each of plastoquinone-9 and pheophytin a, but practically no phylloquinone, α-tocopherylquinone or α-tocopherol.     

A new isolate of the rhodospirillum fulvum group and its photosynthetic pigments

Summary The present paper reports the isolation of an obligate phototrophic bacterium which belongs to the Rhodospirillum fulvum-group on the basis of its colour, morphology, nutritional requirements and strictly anaerobic nature. p-Aminobenzoic acid was shown to be the only growth factor required. Rhsp. fulvum synthesizes bacteriochlorophyll a as its only chlorophyllous pigments. The carotenoid composition comprises lycopene (I) and rhodopin (II) as the major components; traces of 1,2,1,2-tetrahydro-1,1-dihydroxy-lycopene (III) were also present.

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Zusammenfassung Es wird die Isolierung eines obligat phototrophen Bakteriums beschrieben, welches auf Grund seiner Farbe, Morphologie, Nährstoff-Bedürfnisse und streng anaeroben Lebensweise in die Rhodospirillum fulvum-Gruppe eingeordnet wird. Als einzigen Wachstumsfaktor benötigt der Stamm p-Aminobenzoesäure. Rhsp. fulvum bildet Bacteriochlorophyll a als einzigen chlorophyllartigen Farbstoff. Die Hauptmenge der Carotinoide besteht aus Lycopin (I) und Rhodopin (II); es sind außerdem Spuren von 1,2,12-Tetrahydro-1,1-dihydroxy-Lycopin (III) nachgewiesen.

     

Spatial covariance in resources affects photosynthetic rate and water potential,but not the growth of Glechoma longituba fragments

Glechom longituba is a widespread clonal plant and usually experiences diverse patchiness of its growing sites. The hypothesis was tested that spatial covariance in resources differentially affects the growth and physiology of G. longituba. Plants were exposed to two patchy habitats where above- and below-ground resources were either positively or negatively associated, and to four control habitats. When equal amounts of resources were given, G. longituba fragments from two patchy habitats had similar biomass, root weight ratio, leaf weight ratio, fluorescence yield, specific petiole length, and specific root length; fragments under reciprocal patchiness significantly increased photosynthetic rate and decreased leaf water potential. Biomass of clones grown in patchy habitats was equal or less than that of counterparts from the control habitats, not supporting the notion that clonal growth is more advantageous in patchy habitats than in uniform habitats. In addition, no evidence was detected for spatial division of labour because biomass allocation to roots and leaves was similar in patchy habitats compared with the control habitats.     

薄层层析指导下的嗜碱放线菌菌株YIMGQ-14次生代谢产物研究

在微生物资源的开发研究过程中,利用TLC方法,配合TLC数据库的检索,对嗜碱放线菌菌株YIMGQ14的次生代谢产物进行了研究。并从中分离纯化出两个具有生物活性的专利化合物“Umycin C”和“Umycin B”。结果证明：相对于常规的生物活性筛选,TLC法在微生物次生代谢产物研究中具有快速、简便、经济的特点,适合于小量常规筛选的目的。通过进一步对相关的TLC数据库进行构建和补充,将能极大地改善该方法的使用效率。     

Kinos of Eucalyptus species and their acid degradation products

The kinos of Eucalyptus astringens, E. lehmannii and E. platypus contain mainly polymerized leucocyanidin, appreciable amounts of dihydrokaempferol 3-rhamnoside and trace amounts of other components, The acid degradation products of different kinos contained components not present in the products from wood and bark. These include phloroglucinol and two other components whose properties are described.     

干旱胁迫下内生真菌感染对黑麦草光合色素和光合产物的影响

以含有内生真菌的黑麦草 (L olium perenne L.)种子为材料 ,采用加热处理方式构建内生真菌非感染的黑麦草种群 ,通过比较内生真菌感染 (EI)和非感染 (EF)植株在正常条件下和干旱胁迫条件下叶片相对水分含量、叶绿素、可溶性糖和淀粉含量等指标的差异 ,探讨黑麦草 EI和 EF种群对干旱胁迫的适应性差异。结果表明 :在中度胁迫后期 ,EI植株叶片的 RWC显著高于 EF植株 ,即 EI植株的保水能力更强。轻度水分胁迫下 ,内生真菌感染可使其宿主植物的可溶性糖含量增加 ,以增强宿主的渗透调节能力 ,随着干旱胁迫强度的加大 ,内生真菌的这一增益效应不再起作用 ,此时 ,宿主植物将更多的光合产物——淀粉积累于体内 ,以度过不良环境。第 2年春天 EI和 EF种群的恢复生长情况进一步表明 ,经过中度干旱胁迫后 ,EI种群的恢复更为迅速。生物量的大小是植物种群净光合作用能力的直接体现 ,研究中在中度干旱胁迫条件下 ,黑麦草 EI种群的生物量显著高于EF种群 ,但从光合色素的变化来看 ,相同水分状况下 EI和 EF植株的 Chla、Chlb以及 Car的变化趋势比较接近 ,这说明内生真菌感染并未缓解干旱胁迫对光合色素的破坏 ,内生真菌可能通过其它途径来改善宿主植物的光合能力     

铁皮石斛叶色突变体的叶绿体超微结构、光合色素和叶绿素荧光特性的研究

以铁皮石斛(Dendrobium officinale Kimura et Migo)(‘TP35’)经自然突变的白绿杂色突变体(‘TP-MA’)和太空诱变的绿黄杂色突变体(‘TP-MG’)为材料,研究不同光照强度(0、50、100μmol·m^-2·s^-1)处理后,植株叶片的叶绿体超微结构、光合色素含量以及叶绿素荧光动力学参数的变化规律,并阐明叶色突变体与正常植株光合特性的差异。结果显示,‘TP-MA’和‘TP-MG’的叶绿体形态均发生了一定程度的缺失,且叶绿体分布不均匀、无规则,基粒片层结构不完整且排列疏松,基本与其表型性状相一致。‘TP-MA’光合色素的含量和叶绿素荧光参数F v/F m、ΦPSⅡ及F v′/F m′等显著低于‘TP35’,但非光化学淬灭系数(NPQ和q P)则相对较高;‘TP-MG’的光合色素含量及叶绿素荧光参数均低于‘TP35’,但差异不显著,且对较强的光照(100μmol·m^-2·s^-1)条件具有一定的适应性。研究结果表明不同光强处理后,铁皮石斛叶色突变体的叶绿体结构和光合生理指标均发生了不同程度的变化,且对植株的叶绿素含量及荧光参数产生一定影响,过低和过高的光照强度均不利于植株的正常生长。     

Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L.under drought strees conditions

Endophyte-infected fED seeds of Lolium perenne L.were used to attain endophyte-free fED population by heating the seeds at 43℃ for 15 min and then 57℃ for 25 min.Relative water content(RWC),chlorophyll,soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant.Under severe stress.RWC of EI leaf was significantly higher than that Of EF leaf,i.e.EI plants took more advantages over EF plants in water-holding ability.Under mild stress,endophytc could enhance soluble sugars in host plants to improve their osmotic ability.With stress intensification,the improvement of endophyte no longer existed,and more photosynthetic products(such as starch)accumulated in EI plants to survive through the undesirable condidons.In the next spring,EI populations will recover more rapidly than EF populations.The biomass of a population is closely related to its photosynthesis.Under severe stress,EI population significantly accumulated more biomass than EF population.As far as photosynthetic pigments were concerned,contents of Chla,Chlb and Car of EI plants were close to those of EF plants,which suggested that endophyte infection didn't alleviate photosynthetic pigments from being destroyed by drought stress,and endophyte might improve photosynthesis ability of its host plant in other ways.     

A two-step extraction procedure for concentrating acidic organic volatiles in aqueos solution prior to gas chromatographic head-space analysis,as exemplified by short-chain fatty acids produced by Bacillus cereus

A concentration procedure for short-chain fatty acids in aqueos solution is described, utilizing extraction by diethyl ether followed by re-extraction of fatty acid salts from the ether phase into a small amount of aqueous NaOH. The samples were acidified and analyzed by automatic head-space gas chromatography using a fused silica capillary column. The method's usefulness for trace analysis of short-chain fatty acids was exemplified in a study on broth cultures of Bacillus cereus where acetic, isobutyric and isovaleric acids could be readily detected after only 12 h of incubation.     

Sampling of solid spent agar media for gas chromatography of diffused acidic fermentation products

Abstract Sampling and processing of solid agar media for gas chromatographic analysis is described. Small pieces of agar are cut from a 'sterile' area of the plate leaving the actual bacterial growth undisturbed for further tests. It is shown that diffused fermentation acids are adequately represented in these samples to permit their use for phenotypic and taxonomic characterization of isolates. Because of the established qualitative and quantitative correspondence of fermentation patterns from agar-grown and from broth-grown bacteria, respectively, the method is offered as a rapid alternative for the sampling of spent culture broths. In addition, the sampling of spent agar plugs seems to hold promise for the study of defined mixed cultures and of mixed growth from primary plates.     

A new approach for quantitation of receptors and their ligands

A novel and sensitive method for quantitating receptors and their ligands is described, using the calf uterine estrogen receptor and estradiol as a model system. When ligand conjugated to malate dehydrogenase is incubated with the estrogen receptor, the enzyme is inhibited proportionately to the concentration of receptor. However, receptor saturated with free ligand has almost no effect on the ligand-conjugated enzyme. The assay can detect as little as three femtomoles of receptor and one femtomole of estradiol and can, in principle, be applied to any receptor-ligand interaction.