Expression of the yeast mevalonate kinase gene in trangenic tobacco

The coding region of the yeast mevalonate kinase gene (ERG12), under the control of the cauliflower mosaic virus (CaMV) 35S promoter, has been inserted in tobacco (Nicotiana tabacum cv. Paraguay Bell) using an Agrobacterium tumefaciens binary vector system. Integration and expression of the ERG12 chimaeric gene was demonstrated in several independent transformants in which specific mevalonate kinase (MK) activity in young plantlets was increased by about 60% on average. The expression of this MK gene was accompanied by phenotypical modifications, such as acceleration of regenerating processes, lateral bud growth, and peculiar flowering behaviour. A higher chlorophyll content all along the plant development, paralleled by an unusual starch accumulation in the leaves of young plantlets and, later, in roots of full-grown plants, was also detected. Overexpression of the MK gene led also to a stronger inhibition of cytokinin-induced plant growth by methyl jasmonate in transgenic plants. All these events may be interpreted as a possible modification of the hormonal balance in transgenic tobaccos.     

Fluorescence from low-energy chlorophylls in Photosystem I of Synechocystis sp. PCC 6803 at physiological temperatures

Fluorescence spectra from Photosystem I (PS I) are measured from 25 to –5 °C on a PS II-less mutant of the cyanobacterium Synechocystis sp. PCC 6803. Emission from antenna chlorophylls (Chls) with energy levels below that of the reaction center, or low-energy Chls (LE Chls), is resolved verifying their presence at physiological temperatures. The 25°C spectrum is characterized by peaks at 688 and 715 nm. As temperature decreases, fluorescence at 688 nm decreases while at 715 nm it increases. The total fluorescence yield does not change. The temperature dependent spectra are fit to a sum of two basis spectra. At 25°C, the first basis spectrum has a major peak at 686 nm and a minor peak at 740 nm. This is attributed to fluorescence from the majority or bulk antenna Chls. The second basis spectrum has a major peak at 712 nm, with shoulders at 722 and 770 nm. It characterizes fluorescence from a small number of LE Chls. A progressive shift to the red in the fluorescence spectra occurs as the temperature is decreased. The temperature dependence in the relative amount of fluorescence from the bulk and LE Chls is fit using a two-component energy transfer model at thermal equilibrium.     

Quantification of photosystem I and II in different parts of the thylakoid membrane from spinach

Electron paramagnetic resonance (EPR) was used to quantify Photosystem I (PSI) and PSII in vesicles originating from a series of well-defined but different domains of the thylakoid membrane in spinach prepared by non-detergent techniques. Thylakoids from spinach were fragmented by sonication and separated by aqueous polymer two-phase partitioning into vesicles originating from grana and stroma lamellae. The grana vesicles were further sonicated and separated into two vesicle preparations originating from the grana margins and the appressed domains of grana (the grana core), respectively. PSI and PSII were determined in the same samples from the maximal size of the EPR signal from P700⁺ and Y_D, respectively. The following PSI/PSII ratios were found: thylakoids, 1.13; grana vesicles, 0.43; grana core, 0.25; grana margins, 1.28; stroma lamellae 3.10. In a sub-fraction of the stroma lamellae, denoted Y-100, PSI was highly enriched and the PSI/PSII ratio was 13. The antenna size of the respective photosystems was calculated from the experimental data and the assumption that a PSII center in the stroma lamellae (PSIIβ) has an antenna size of 100 Chl. This gave the following results: PSI in grana margins (PSIα) 300, PSI (PSIβ) in stroma lamellae 214, PSII in grana core (PSIIα) 280. The results suggest that PSI in grana margins have two additional light-harvesting complex II (LHCII) trimers per reaction center compared to PSI in stroma lamellae, and that PSII in grana has four LHCII trimers per monomer compared to PSII in stroma lamellae. Calculation of the total chlorophyll associated with PSI and PSII, respectively, suggests that more chlorophyll (about 10%) is associated with PSI than with PSII.     

Study of phytoplankton characteristics in Tolo Harbour,Hong Kong,by HPLC analysis of chemotaxonomic pigments

Spatial and seasonal characteristics of phytoplankton in Tolo Harbour, Hong Kong, were studied by microscopic observation of phytoplankton samples and HPLC analysis of chemotaxonomic pigments. Diatoms dominated the phytoplankton. Common diatoms included Skeletonema costatum and species of Cerataulina, Leptocylindrus, Pseudo-nitzschia and Thalassiosira. Dinoflagellates occurred sporadically and mainly in the inner part of the harbour. The dinoflagellate Scrippsiella trochoidea was the causative organism for the red tide occurrences in March, April and September 2001. Significant positive correlations between fucoxanthin and diatoms and between peridinin and dinoflagellates suggested that fucoxanthin and peridinin were valuable chemotaxonomic markers for diatoms and dinoflagellates, respectively. Analysis of pigment ratios revealed that red tide events caused by dinoflagellates were marked by increase in the value of PERI:chl a and decrease in the value of FUCO:chl a. Increase in the value of FUCO:chl a also revealed the presence of a dense population of Pseudo-nitzschia that was not indicated by increase in chlorophyll a and fucoxanthin concentrations. Pigment analysis also revealed the presence of cyanobacteria, silicoflagellates, cryptophytes and green algae in the surface waters of Tolo Harbour.     

CHARACTERIZATION AND PHYLOGENETIC POSITION OF THE ENIGMATIC GOLDEN ALGA PHAEOTHAMNION CONFERVICOLA: ULTRASTRUCTURE, PIGMENT COMPOSITION AND PARTIAL SSU rDNA SEQUENCE

The morphology, ultrastructure, photosynthetic pigments, and nuclear-encoded small subunit ribosomal DNA (SSU rDNA) were examined for Phaeothamnion confervicola Lagerheim strain SAG119.79. The morphology of the vegetative filaments, as viewed under light microscopy, was indistinguishable from the isotype. Light microscopy, including epifluorescence microscopy, also revealed the presence of one to three chloroplasts in both vegetative cells and zoospores. Vegetative filaments occasionally transformed to a palmelloid stage in old cultures. An eyespot was not visible in zoospores when examined with light microscopy, but small droplets, similar to eyespot droplets, were apparent beneath the shorter flagellum when cells were viewed with electron microscopy. Zoospores had two flagella that were laterally inserted in the cell approximately one-third of the cell length from the apex. The longer flagellum was directed anteriorly and the shorter flagellum was directed posteriorly. Electron microscopy revealed the presence of tubular tripartite flagellar hairs on the longer flagellum, but no lateral filaments were found on the tripartite hairs. The general organization of the flagellar root system was similar to that of zoospores belonging to the Xanthophyceae and Phaeophyceae. However, the transitional region of the flagella contained a transitional helix with four to six gyres. Microtubular root R₁ consisted of six microtubules at its proximal end and one microtubule at its distal end. Roots R₂ and R₄ consisted of one microtubule each and root R₃ consisted of two microtubules. No rhizoplast was found. Thin-layer chromatography revealed the presence of fucoxanthin, diadinoxanthin, neoxanthin, and heteroxanthin as well as chlorophylls a, c₁ and c₂. High-performance liquid chromatography revealed the presence of fucoxanthin, diadinoxanthin, diatoxanthin, heteroxanthin, and β,β-carotene as well as chlorophylls a and c. The complete sequence of the SSU rDNA could not be obtained, but a partial sequence (1201 bases) was determined. Parsimony and neighbor-joining distance analyses of SSU rDNA from Phaeothamnion and 36 other chromophyte algae (with two Öomycete fungi as the outgroup) indicated that Phaeothamnion was a weakly supported (bootstrap = <50%, 52%) sister taxon to the Xanthophyceae representatives and that this combined clade was in turn a weakly supported (bootstrap = <50%, 67%) sister to the Phaeophyceae. Based upon ultrastructural observations, pigment analysis, and SSU rDNA phylogenetic analysis, Phaeothamnion is not a member of the Chrysophyceae and should be classified as incertae sedis with affinities to the Xanthophyceae and Phaeophyceae.     

UV-A Irradiation Guards the Photosynthetic Apparatus Against UV-B-Induced Damage

In clusterbean leaves UV-B radiation caused a reduction in contents of chlorophylls and carotenoids and in the efficiency of photosystem 2 photochemistry. The degree of damage was reduced when UV-A accompanied the UV-B radiation. This indicates the counteracting effect of UV-A radiation against UV-B-induced impairment.     

Effect of high irradiance and high temperature on chloroplast composition and structure of Dioscorea zingiberensis

High irradiance (HI) and high temperature (HT) increased in chloroplasts the content of monogalactosyldiacylglycerol (MGDG) and decreased the contents of digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol (SQDG), and phosphatidylinositol (PI). HI and HT accelerated the transformation of DGDG to MGDG. The contents of unsaturated fatty acids in chloroplasts increased, while those of saturated fatty acids decreased. The contents of total carotenoids, neoxanthin, violaxanthin, lutein, and -carotene increased first, then decreased. The content of chlorophyll decreased. HI caused the unfolding of thylakoids that was not resumed after a 72-h recovery.This revised version was published online in March 2005 with corrections to the page numbers.     

Effects of CO<Subscript>2</Subscript> concentration on acclimatization and physiological responses of two cultivars of carob tree

This study reports survival and physiological responses of micropropagated Ceratonia siliqua L. cvs. Galhosa and Mulata plants during ex vitro acclimatization under ambient (AC; 330 mol mol^–1) or elevated (EC; 810 mol mol^–1) CO₂ concentration and a photosynthetic photon flux density of 125 mol m^–2 s^–1. CO₂ enrichment during acclimatization did not improve survival rate that was around 80 % for both treatments. Eight weeks after ex vitro transplantation, photosynthetic capacity and apparent quantum yield in acclimatized leaves were higher in comparison with those in in vitro-grown leaves, without any significant difference between CO₂ treatments. Chlorophyll content increased after acclimatization. However, EC led to a decrease in the total amount of chlorophyll in new leaves of both cultivars, compared to those grown at AC. Soluble sugars and starch contents were not markedly affected by growth EC, although starch had significantly increased after transfer to ex vitro conditions. EC induced an increase in the stem elongation and in the effective life of leaves, and a decrease in the number of new leaves.