Correlation of chlorophyll a and total carotenoid concentrations with coral bleaching from locations on the Pacific coast of Mexico

Diving campaigns were undertaken to study the bleaching phenomenon in the Mexican Central Pacific and to catalogue the degree to which bleaching affected three coralline reefs (la Boquita, Carrizales, and Tenacatita). Bleaching evaluations of Pocillopora capitata were conducted by assessing the concentrations of chlorophyll a and total carotenoids from samples along submarine transect lines. We identified that the Reffs Boquita and Tenacatita locations were predominantly composed of bleached and pale coral populations. The concentrations of chlorophyll a within the areas of the different populations were greater at sites of greater depth compared with those at the surface. In contrast, the total carotenoid concentrations in normal pigmented coral at greater depth were significantly higher (P?<?0.05) compared with both levels of discolouration at both depths. The chlorophyll a/total carotenoid ratios in these populations are discussed as a potential mechanism responsible for the poor condition of these corals as a result of environmental stress.     

Accumulation of di-n-butylphthalate in plants and its effect on pigment and protein content

Seedlings of radish ( Raphanus salivas L. cv. Cherry Belle) and wheat ( Triticum aestivum L. cv. Starke II, Weibull) placed in air containing vapour of di- n -butyl-phthalate accumulate phthalate to a concentration that is 10⁶ times that of the surrounding air within 3 days. Particularly high accumulation is found in the cuticular and wax layers. Plants earlier found not to respond to di- n -butylphthalate (wheat) show toxic symptoms (carotenoid and chlorophyll deficiency in strong light, conco-mittant with chloroplast destruction and swollen mitochondria) when the phthalate concentration is further increased by addition to the nutrient medium. Phthalate-affected, white leaves lack the protein moieties normally belonging to the light harvesting complex.     

Differences in the capacity for radiationless energy dissipation in the photochemical apparatus of green and blue-green algal lichens associated with differences in carotenoid composition

Green algal lichens, which were able to form zeaxanthin rapidly via the de-epoxidation of violaxanthin, exhibited a high capacity to dissipate excess excitation energy nonradiatively in the antenna chlorophyll as indicated by the development of strong nonphotochemical quenching of chlorophyll fluorescence (F_M, the maximum yield of fluorescence induced by pulses of saturating light) and, to a lesser extent, F_O (the yield of instantaneous fluorescence). Blue-green algal lichens which did not contain any zeaxanthin were incapable of such radiationless energy dissipation and were unable to maintain the acceptor of photosystem II in a low reduction state upon exposure to excessive photon flux densities (PFD). Furthermore, following treatment of the thalli with an inhibitor of the violaxanthin de-epoxidase, dithiothreitol, the response of green algal lichens to light became very similar to that of the blue-green algal lichens. Conversely, blue-green algal lichens which had accumulated some zeaxanthin following long-term exposure to higher PFDs exhibited a response to light which was intermediate between that of zeaxanthin-free blue-green algal lichens and zeaxanthin-containing green algal lichens. Zeaxanthin can apparently be formed in blue-green algal lichens (which lack the xanthophyll epoxides, i.e. violaxanthin and antheraxanthin) as part of the normal biosynthetic pathway which leads to a variety of oxygenated derivatives of β-carotene during exposure to high light over several days. We conclude that the pronounced difference in the capacity for photoprotective energy dissipation in the antenna chlorophyll between (zeaxanthin-containing0 green algal lichens and (zeaxanthin-free) blue-green algal lichens is related to the presence or absence of zeaxanthin, and that this difference can explain the greater susceptibility to high-light stress in lichens with blue-green phycobionts.     

Effect of chlorocholine chloride and gibberellic acid on the anthocyanin synthesis in radish seedlings

Anthocyanin synthesis in radish ( Raphanus sativus L. cv. Scarlet Globe) seedlings after treatment with chlorocholine chloride (CCC) and gibberellic acid (GA) has been investigated. CCC promotes and GA₃ inhibits the synthesis. When both substances are given together, CCC reverses the inhibition caused by GA₃. Simultaneous external feeding of anthocyanin precursors (sucrose and phenylalanine) reverses the GA₃ inhibition. A higher amount of total free amino acids, in particular phenylalanine, was present in CCC-treated seedlings compared to controls grown on distilled water. The amount of phenylalanine was lower in seedlings treated with both CCC and GA₃ as compared to seedlings treated with CCC alone, and total free sugars (reducing plus non-reducing) was lower in CCC treated seedlings than in controls grown on distilled water. We conclude that CCC and GA3 control the anthocyanin synthesis at the level of precursors.     

Preliminary screening of antioxidant and cytotoxic potential of green seaweed,Halimeda opuntia (Linnaeus) Lamouroux

Marine natural products have displayed numerous advantageous effects on biological activities, including antioxidants and cytotoxicity. The total lipids, carotenoids, chlorophyll a and b content, total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity of methanolic crude extract of the green seaweed Halimeda opuntia were all measured in this study. The TPC of the extracts was determined according to the Folin-Ciocalteu method, yielding a result of 55.04 ± 0.98 mg GAE/g of extract. As determined by the aluminium chloride colorimetric method, the TFC of the extract was 40.02 ± 0.02 mg QE/g of extract. Antioxidant activity was determined by using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay with different concentrations that ranged between 200 and 1000 µg/mL, noted H. opuntia as the highest in DPPH reduction (63.61 %) at 1000 µg/mL concentration. Total antioxidant capacity (TAC) of the extract was 57.36 ± 0.004 mg AAE/g extract at concentration of 1.0 mg/mL. The cytotoxic activity of this seaweed was pre-screened against a panel of cell lines including estrogen receptor-positive human breast adenocarcinoma (MCF-7), estrogen negative human breast adenocarcinoma (MDA-MB-231), human colorectal adenocarcinoma (HT-29), human hepatocellular carcinoma (HepG2), and mouse embryonic fibroblast (3T3) using the MTT assay. The content of total lipids in H. opuntia was 1.60 ± 0.002 %. Total carotenoids were 115.57 ± 0.98 µg/g, while chlorophyll a and b were 148.73 ± 2.60 µg/g and 290.83 ± 9.46 µg/g, respectively. In terms of cytotoxicity activity, methanolic extract of H. opuntia was found to be highly cytotoxic to MCF-7 cells, with an IC₅₀ of 25.14 ± 1.02 g/mL, and slightly less so to 3T3 cells (IC₅₀ 65.23 ± 0.25 µg/mL). This study's findings suggest that natural pigments (carotenoids and chlorophyll), phytochemicals like phenolic and flavonoid compounds found in this species may play an important role and could be used as a natural cancer treatment.     

An update on carotenoid biosynthesis in algae: phylogenetic evidence for the existence of two classes of phytoene synthase

Carotenoids play crucial roles in structure and function of the photosynthetic apparatus of bacteria, algae, and higher plants. The entry-step reaction to carotenoid biosynthesis is catalyzed by the phytoene synthase (PSY), which is structurally and functionally related in all organisms. A comparative genomic analysis regarding the PSY revealed that the green algae Ostreococcus and Micromonas possess two orthologous copies of the PSY genes, indicating an ancient gene duplication event that produced two classes of PSY in algae. However, some other green algae (Chlamydomonas reinhardtii, Chlorella vulgaris, and Volvox carteri), red algae (Cyanidioschyzon merolae), diatoms (Thalassiosira pseudonana and Phaeodactylum tricornutum), and higher plants retained only one class of the PSY gene whereas the other gene copy was lost in these species. Further, similar to the situation in higher plants recent gene duplications of PSY have occurred for example in the green alga Dunaliella salina/bardawil. As members of the PSY gene families in some higher plants are differentially regulated during development or stress, the discovery of two classes of PSY gene families in some algae suggests that carotenoid biosynthesis in these algae is differentially regulated in response to development and environmental stress as well.     

Contribution of hydroxymethylbutenyl diphosphate synthase to carotenoid biosynthesis in bacteria and plants

The methylerythritol 4-phosphate (MEP) pathway synthesizes the precursors of carotenoids and other isoprenoids in bacteria and plant plastids. Despite recent progress in the identification of rate-determining steps, the relative contribution of most pathway enzymes to flux control remains to be established. In this work we investigated whether upregulated levels of hydroxymethylbutenyl diphosphate synthase (HDS) could increase the metabolic flux through this pathway, as judged by endpoint (carotenoid) measurements. Unlike other MEP pathway enzymes, however, increasing the levels of an active HDS protein in carotenoid-producing Escherichia coli cells and transgenic Arabidopsis thaliana plants did not result in an enhanced accumulation of MEP-derived isoprenoids. Our data suggest that enhanced flux through the MEP pathway for peak demand periods in bacteria and plastids does not require increased HDS activity.     

城市污泥和化肥对水稻土种植的通菜中邻苯二甲酸酯（PAEs）的影响

对水稻土施用城市污泥和化肥盆栽通菜,应用GC／MS联机检测技术对通菜中6种邻苯二甲酸酯化合物(PAEs)进行分析,探讨施肥对通菜中PAEs含量的影响,结果表明,各处理通菜中6种PAEs化合物的总含量(∑PAEs)在2．129～7.111mg·kg^-1之间,依次为广州污泥+化肥(7．111mg·kg^-1)>广州污泥(4．767mg·kg^-1)>佛山污泥(3;569mg·kg^-1)>佛山污泥+化肥(3．305mg·kg^-1)>化肥(2．638mg·kg^-1)>空白对照(2．129mg·kg^-1)。显示了施肥造成通菜中∑PAEs不同程度的提高,各处理通菜中的PAEs均以个别化合物为主,其中空白对照、化肥、佛山污泥和广州污泥处理的通菜中邻苯二甲酸丁基苄基酯(BBP)占∑PAEs的40％～81％。佛山污泥+化肥处理的通菜中邻苯二甲酸正二丁酯(DnBP)占∑PAEs的56％,而广州污泥+化肥处理的通菜中邻苯二甲酸丁基苄基酯(BBP)、邻苯二甲酸正二辛酯(DnOP)和邻苯二甲酸(2-乙基己基)酯(DEHP)各占∑PAEs的30％左右,通菜中∑PAEs和多数化合物的含量不同程度地大于其在根系中的含量。     

Detection of unusual carotenoid esters in fresh mango (Mangifera indica L. cv. 'Kent')

The carotenoid pattern of mango cv. 'Kent' was investigated by LC-(APcI)MS analyses. In solvent extracts from the mesocarp an unusual carotenoid ester was identified as violaxanthin dibutyrate. For unequivocal identification of butyric acid by an independent method, total lipids were isolated by solvent extraction from the fruit flesh and analyzed by GC after saponification and subsequent methylation. Thus, evidence of butyric acid (1.6 area%) was provided. To the best of our knowledge, this is the first report on a xanthophyll dibutyrate in plants. Additionally, further carotenoid peaks were tentatively assigned to 9-cis-violaxanthin and neochrom or luteoxanthin, respectively, by their UV/vis and MS data of the saponified extracts.     

城市污泥与稻草堆肥中邻苯二甲酸酯（PAEs）的研究

将广州城市污泥与稻草进行翻堆、接菌-翻堆、连续通气和间歇通气4种方式的堆肥,应用GC／MS技术对堆肥中6种属于USEPA优控污染物的邻苯二甲酸醇化合物(PAEs)进行分析,探讨堆肥产物中PAEs的含量分布以及不同方式堆肥对PAEs的降解效果,结果表明,4种方式堆肥中PAEs总含量(∑PAEs)在9．815～17．832mg·kg^-1之间,依次为翻堆(17．832mg·kg^-1)>接菌-翻堆(13．927mg·kg^-1)>间隙通气(10．765mg·kg^-1)>连续通气(9．815mg·kg^-1),堆肥中PAEs以邻苯二甲酸正二辛酯(DhOP)为主,占∑PAEs的82．2％～89．696,不同方式堆肥中∑PAEs的降解率为连续通气(45．71％)>间隙通气(40．4696)>接菌-翻堆(22．97％)>翻堆(1．3796)(平均降解率为27．63％),其中邻苯二甲酸二乙醇(DEP)、邻苯二甲酸正二丁酯(DnBP)和邻苯二甲酸丁基苄基酯(BBP)的降解率分别为95．7696～98．6896、79．5696～99．46％和87．42％～98．42％;但邻苯二甲酸二甲酯(DMP)和邻苯二甲酸正二辛酯的含量反而增加,邻苯二甲酸(2-乙基己基)酯(DEHP)在所有堆肥中均未检出。     

Parameters affecting productivity in the lipase-catalysed synthesis of sucrose palmitate

The industrial application of lipases for the synthesis of sucrose esters is usually limited by its low productivity, as we need a medium where a polar reagent (the sugar) and a non-polar fatty acid donor are soluble and able to react in the presence of the biocatalyst. In this work, we have studied the problems encountered when trying to increase the volumetric productivity of sucrose esters. The synthesis of sucrose palmitate was performed in 2-methyl-2-butanol:dimethylsulfoxide mixtures by transesterification of different palmitic acid donors with sucrose, catalysed by the immobilized lipase from Candida antarctica B (Novozym 435). A protocol for substrate preparation different from that previously reported was found to improve the reaction rate. Several parameters, such as sucrose and acyl donor loadings, the percentage of DMSO in the mixture and the nature of acyl donor, were investigated. Under the best experimental conditions (15% DMSO, 0.1 mol l^?1 sucrose, 0.3 mol l^?1 vinyl palmitate), a maximum of 45 g l^?1 sucrose palmitate was obtained in 120 h. Using methyl or ethyl palmitate, the highest productivity was 7.3 g l^?1 in 120 h using 20% DMSO with 0.2 mol l^?1 sucrose and 0.6 mol l^?1 acyl donor. The formation of free fatty acid, and the effect of the percentage of DMSO on the monoester/diester selectivity were also studied. To our knowledge, this is the first report on enzymatic synthesis of sucrose esters of long fatty acids using alkyl esters as acyl donors.     

类胡萝卜素产生菌深红酵母(Rhodotorula rubra)的培养条件的优化研究

本文研究深红酵母及其产生的类胡萝卜素的培养优化条件,并对其进行了小型发酵试验,结果表明pH值对深红酵母和类胡萝卜素有影响,初始pH值越低,色素的积累量越高,发酵过程中控制pH值能有效地增加色素积累的速度和初始色素积累量,但随着发酵时间的延长,色素积累量逐渐下降。     

Production of phytoene, a carotenoid, and induction of connexin 26 in transgenic mice carrying the phytoene synthase gene crtB

Carotenoids have been recognized as chemopreventive agents against human diseases, such as cancer and cardiovascular disease. Mammalians utilize carotenoids supplied from their food since they are unable to perform the de novo synthesis of carotenoids. We previously created mammalian cultured cells producing phytoene, a type of carotenoid, and showed that these cells acquired resistance against oxidative stress and oncogenic transformation. In the present study, we established a transgenic mouse line, carrying the crtB gene encoding phytoene synthase, which could produce phytoene endogenously. It was found that connexin 26 was induced in these phytoene-producing mice. Since it is known that carotenoids enhance gap junctional communication by inducing the expression of connexin genes, the present data suggest that the induction of connexin 26 in phytoene-producing mice may play a role in controlling cell-to-cell communication. Phytoene-producing mice provide a useful system in which to investigate the in vivo function of the carotenoid phytoene.     

The role of vanadium in green plants

In a series of experiments, it is demonstrated that the trace element vanadium (4·10^-7 g-at/l as NH₄VO₃) has a considerable positive influence on the synthesis of -aminolevulinic acid (-ALA) in the autotrophically growing green algaChlorella pyrenoidosa, the effect being visible by an enhanced output of the amino acid into the culture medium in presence of levulinic acid (LA). The level of intracellularly accumulated -ALA, however, is not changed in presence of the metal. The V-effect on exogenous found -ALA is suppressed, when LA is added to the nutrient medium at low pH (pH 5), although V-uptake into the algal cells is not disturbed by LA. As demonstrated in culture media with various nitrogen sources (urea, partially hydrolized urea, ammonium salts), the development of the pH during the cultivation time is important for the presentation of the V-effect on -ALA. It is suggested that vanadium acts as a catalyst in the conversion of 4,5-dioxovaleric acid to -ALA by transamination.Abbreviations -ALA -aminolevulinic acid - LA levulinic acid - DOVA 4,5-dioxovaleric acid     

Biodegradation of phthalic acid esters (PAEs) in soil bioaugmented with acclimated activated sludge

The degradation characteristics of four phthalic acid esters (PAEs), i.e. di-methyl phthalate (DMP), di-ethyl phthalate (DEP), di-n-butyl phthalate (DBP) and di-n-octyl phthalate (DOP) in the soil augmented with acclimated sludge was investigated in order to assess the efficacy of bioaugmentation as a strategy for remediating PAEs-contaminated soil and correlate the degradation rate of PAEs with their alkyl chain length. The results demonstrated that PAEs with shorter alkyl chain, that is, DMP and DEP could be degraded more quickly than DBP and DOP. The degradation of four PAEs in the soil conformed to a first-order reaction kinetic equation. The half-lives of PAEs degradation decreased significantly with increasing carbon number of the alcohol moiety. Half-lives decreased from 2.29 days for DMP to 28.4 days for DOP when the carbon number of alkyl chain increased from one for DMP to eight for DOP. The degradation rate of PAEs and the corresponding half-lives could correlate with the alkyl chain length and their octanol–water partition coefficients (K_ow) quite well for the four PAEs tested in this study.     

The role of vanadium in green plants

Cells of Chlorella pyrenoidosa, derived from vanadium free agar slants, respond with great sensitivity to microamounts of vanadium, added as NH₄VO₃ to autotrophic liquid cultures. Between 0.01 and 1 g V per litre nutrient medium (2·10^-10-2·10^-8g-at/l), the algae respond with a continuous increase in dry weight. At higher V-concentrations, further enhancement in biomass is accompanied by a additional increase in chlorophyll content. Maximum V-effect on both parameters was found to be at 500g V/l (10^-5 g-at/l). Dry weight as well as chlorophyll content of Chlorella are decreased by concentrations above 25 mg V/l; 100 mg V/l (2·10^-3 g-at/l) stop growth and cause death of the cells. The toxic threshold for the V-content in the algae was determined to be at 150–200 g V/g (3–4·10^-6 g-at/g) dry weight.Two different pH-optima for a positive vanadium action on dry weight and chlorophyll biosynthesis were established, the first at pH 7, the other in the range pH 7.5–8. Two sites of vanadium action in green algae are discussed.Part I: Arch. Microbiol. 105, 77–82 (1975)     

沉默lycB基因对雨生红球藻类胡萝卜素合成代谢的影响

雨生红球藻是一种淡水浮游单细胞绿藻, 逆境条件下可积累大量的类胡萝卜素。番茄红素是类胡萝卜素中的一种, 是类胡萝卜素合成代谢中的一个重要中间产物。番茄红素β-环化酶(LycB)是催化番茄红素形成β-胡萝卜素的关键酶。文章以杜氏盐藻lycB基因为干扰序列, 构建了含卡那霉素与阿特拉津双抗性的RNAi载体p1301-BS-RNAi。将其电转化进雨生红球藻细胞, 经抗性筛选、基因组PCR及RT-PCR筛选, 获得了16个独立的干扰株系。选取生长良好的7个进行高光诱导, 发现其番茄红素含量增加了99.4%, β-胡萝卜素含量减少了48.4%, 即通过异源的lycB-RNAi基因沉默可抑制番茄红素向β-胡萝卜素的转化。对比分析发现, 番茄红素增加量仅是β-胡萝卜素减少量的5%, 表明因lycB-RNAi抑制而产生的番茄红素的95%又被其他通路转换成了其他代谢产物, 因此要实现雨生红球藻番茄红素含量的大幅增长, 需协同调控其他代谢通路。     

A two-photon excitation study on the role of carotenoid dark states in the regulation of plant photosynthesis

Plants are exposed to sun light intensities that vary rapidly over several orders of magnitude during a typical day. It is known that the regulation of photosynthetic activity under these circumstances is essential for the survival and fitness of natural and gene modified plants. A quick balancing between utilization and dissipation of absorbed light energy ensures optimized levels of CO₂ fixation and protection from photo damage by excessive light-irradiation. Despite intensive investigations the biophysical mechanisms of these regulation processes are still poorly understood. Potentially involved singlet states of carotenoids are optically “dark” and so far it was impossible to investigate their role directly in living plants by conventional absorption or fluorescence spectroscopy. Here, we show by selective two-photon excitation of the carotenoid dark states in planta that a dominant part of the regulation is correlated with a substantial change in the energy transfer between these states and the chlorophylls (Chl). The results support a considerable role of the molecular gear shift model in which a reversible and step-wise enzymatic modification of the electronic structure of xanthophyll carotenoids enables a switching between carotenoid-to-Chl light-harvesting and Chl-to-carotenoid quenching. The shifting can be observed in real time in any plant. Treatment with the xanthophyll cycle inhibitor dithiothreitol slowed down both the light adaptation and the carotenoid–Chl energy flow changes to the same extent. Based on these results, we propose a biophysical quenching model in which both carotenoid dark states and radical cations contribute to the dissipation of excessive excitation energy. An erratum to this article can be found at