Analysis of photoprotection and apparent non-photochemical quenching of chlorophyll fluorescence in <Emphasis Type="Italic">Tradescantia</Emphasis> leaves based on the rate of irradiance-induced changes in optical transparence

The kinetics of irradiation-induced changes in leaf optical transparence (ΔT) and non-photochemical quenching (NPQ) of chlorophyll fluorescence in Tradescantia fluminensis and T. sillamontana leaves adapted to different irradiance in nature was analyzed. Characteristic times of a photoinduced increase and a dark decline of ΔT in these species were 12 and 20 min, respectively. The ΔT was not confirmed to be the main contributor to the observed middle phase of NPQ relaxation kinetics (τ = 10-28 min). Comparison of rate of photoinduced increase in ΔT and photosystem II quantum yield recovery showed that the former did not affect the tolerance of the photosynthetic apparatus (PSA) to irradiances up to 150 μmol PAR·m^–2·s^–1. Irradiance tolerance correlated with the rate of “apparent NPQ” induction. Considering that the induction of apparent NPQ involves processes significantly faster than ΔT, we suggest that the photoprotective mechanism induction rate is crucial for tolerance of the PSA to moderate irradiance during the initial stage of light acclimation (first several minutes upon the onset of illumination).     

Acclimation of shade-tolerant and light-resistant <Emphasis Type="Italic">Tradescantia</Emphasis> species to growth light: chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence,electron transport,and xanthophyll content

In this study, we have compared the photosynthetic characteristics of two contrasting species of Tradescantia plants, T. fluminensis (shade-tolerant species), and T. sillamontana (light-resistant species), grown under the low light (LL, 50–125 µmol photons m^?2 s^?1) or high light (HL, 875–1000 µmol photons m^?2 s^?1) conditions during their entire growth period. For monitoring the functional state of photosynthetic apparatus (PSA), we measured chlorophyll (Chl) a emission fluorescence spectra and kinetics of light-induced changes in the heights of fluorescence peaks at 685 and 740 nm (F ₆₈₅ and F ₇₄₀). We also compared the light-induced oxidation of P₇₀₀ and assayed the composition of carotenoids in Tradescantia leaves grown under the LL and HL conditions. The analyses of slow induction of Chl a fluorescence (SIF) uncovered different traits in the LL- and HL-grown plants of ecologically contrasting Tradescantia species, which may have potential ecophysiological significance with respect to their tolerance to HL stress. The fluorometry and EPR studies of induction events in chloroplasts in situ demonstrated that acclimation of both Tradescantia species to HL conditions promoted faster responses of their PSA as compared to LL-grown plants. Acclimation of both species to HL also caused marked changes in the leaf anatomy and carotenoid composition (an increase in Violaxanthin?+?Antheraxantin?+?Zeaxanthin and Lutein pools), suggesting enhanced photoprotective capacity of the carotenoids in the plants grown in nature under high irradiance. Collectively, the results of the present work suggest that the mechanisms of long-term PSA photoprotection in Tradescantia are based predominantly on the light-induced remodeling of pigment-protein complexes in chloroplasts.     

Integrated role of ROS and Ca<Superscript>+2</Superscript> in blue light-induced chloroplast avoidance movement in leaves of <Emphasis Type="Italic">Hydrilla verticillata</Emphasis> (L.f.) Royle

Directional chloroplast photorelocation is a major physio-biochemical mechanism that allows these organelles to realign themselves intracellularly in response to the intensity of the incident light as an adaptive response. Signaling processes involved in blue light (BL)-dependent chloroplast movements were investigated in Hydrilla verticillata (L.f.) Royle leaves. Treatments with antagonists of actin filaments [2,3,5-triiodobenzoic acid (TIBA)] and microtubules (oryzalin) revealed that actin filaments, but not microtubules, play a pivotal role in chloroplast movement. Involvement of reactive oxygen species (ROS) in controlling chloroplast avoidance movement has been demonstrated, as exogenous H₂O₂ not only accelerated chloroplast avoidance but also could induce chloroplast avoidance even in weak blue light (WBL). Further support came from experiments with different ROS scavengers, i.e., dimethylthiourea (DMTU), KI, and CuCl₂, which inhibited chloroplast avoidance, and from ROS localization using specific stains. Such avoidance was also partially inhibited by ZnCl₂, an inhibitor of NADPH oxidase (NOX) as well as 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), a photosynthetic electron transport chain (ETC) inhibitor at PS II. However, methyl viologen (MV), a PS I ETC inhibitor, rather accelerated avoidance response. Exogenous calcium (Ca⁺²) induced avoidance even in WBL while inhibited chloroplast accumulation partially. On the other hand, chloroplast movements (both accumulation and avoidance) were blocked by Ca⁺² antagonists, La³⁺ (inhibitor of plasma membrane Ca⁺² channel) and ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA, Ca⁺² chelator) while LiCl that affects Ca⁺² release from endosomal compartments did not show any effect. A model on integrated role of ROS and Ca⁺² (influx from apolastic space) in actin-mediated chloroplast avoidance has been proposed.     

Antirestriction and antimodification activities of T7 Ocr: Effects of amino acid substitutions in the interface

The T7 antirestriction protein Ocr, encoded by 0.3 (ocr), specifically inhibits ATP-dependent type I restriction-modification systems. T7 0.3 (ocr) was cloned in pUC18. Ocr inhibited both restriction and modification activities of the type I restriction-modification system (EcoKI) in Escherichia coli K12. The Ocr F53D A57E mutant was obtained and proved to inhibit only restriction activity of EcoKI. The 0.3 (ocr) and Photorhabdus luminescens luxCDABE genes were cloned in pZ-series vectors with the P _ltetO-1 promoter, strongly controlled by the TetR repressor. The bioluminescence intensity and luciferase content varied up to 5000-fold in E. coli K12 MG1655Z1 tetR⁺ (pZE21-luxCDABE) cells, depending on the environmental concentration of the inductor anhydrotetracycline. The antirestriction activity of Ocr and Ocr F53D A57E was studied as a function of their concentration in the cell. The dissociation constant K _d, characterizing the binding with EcoKI, differed 1000-fold between Ocr and Ocr F53D A57E (10^?10 M versus 10^?7 M).     

Photosynthesis and primary production in Lake Kasumigaura (Japan) monitored monthly since 1981

This study reports the primary production of phytoplankton determined with a ¹³C tracer, and their related variables, in Lake Kasumigaura, a shallow, hyper-eutrophic lake, and the second largest lake in Japan. Measurements were conducted monthly from August 1981 to December 2013 at four stations within the lake. Monitoring was a component of the Lake Kasumigaura Long-term Environmental Monitoring program, conducted by the National Institute for Environmental Studies (NIES) since 1977. The program collects data on water quality, and plankton and benthic communities. Lake Kasumigaura is registered as a core site of the Japan Long-term Ecological Research Network (JaLTER), which is a member of the International Long-term Ecological Research Network (ILTER). This dataset includes daily primary production (Pzd gC m^?2 d^?1) and the six parameters required to calculate Pzd: maximum photosynthesis rate (P _max gC gC^?1 h^?1); light irradiance at the junction of the initial slope (α (gC gC^?1 h^?1) (μmol photon m^?2 s^?1)^?1) and P _max of the photosynthesis vs. irradiance (P vs. E) curve (E _k μmol photon m^?2 s^?1); attenuation coefficient of photosynthetically available radiation (PAR) (K _PAR m^?1); water depth at each sampling station (Z _b , m); dissolved inorganic carbon (DIC mgC L^?1) and particulate organic carbon concentrations (POC gC m^?3); and chlorophyll a amounts (Chl.a μg L^?1). Daily primary production was calculated by obtaining a P vs. E curve over a short-term incubation (approximately 1 h) in a water tank using in situ water temperature in the laboratory, based on the field conditions of the sampling date. The dataset has been used for ecological studies as well as for management studies on water quality and ecosystems. This dataset is unique among the available published papers concerning lakes or primary production in various ecosystems, collected over a long period of time and freely available.     

Production and characterization of fungal <Emphasis Type="Italic">β</Emphasis>-glucosidase and bacterial cellulases by tobacco chloroplast transformation

The high capacity of the chloroplast genome to integrate and express transgenes at high levels makes transplastomic technology a good option for overexpressing proteins of interest. This report presents the stable expression of β-glucosidase (bgl1 gene) from Aspergillus niger and two cellulases (celA and celB genes) from Thermotoga neapolitana into the chloroplast genome of tobacco. The pES6, pHM4, pHM5 and pHM6 vectors were derived from the pES4 plasmid containing bgl1, celA-celB, celA and celB synthetic genes, respectively. All of the genes were flanked by a synthetic rrn16 promoter and the 3′UTR from rbcL gene. The integration of the genes into intergenic regions rrn16 and 3′rps12 of the inverted repeats was confirmed by Southern blot analysis. Stable expression and processing of monocistronic mRNA were confirmed by Northern blot analysis, and protein functionality was analysed via enzymatic activity assay. The recombinant enzymes exhibited high enzymatic activity at pH 5 (β-glucosidase: 30.45 U mg^?1 of TSP, celA-celB 58 U mg^?1 of TSP, celA 49.10 U mg^?1 of TSP and celB 48.72 U mg^?1 of TSP). In addition, β-glucosidase exhibited high activity at 40 °C, whereas cellulases type A (celA) and type B (celB) showed high activity at 65 °C. NtpES6, NtpHM5 and NtpHM6 plants showed a similar phenotype compared with the wild type plants; however, NtpHM4 plants presented an abnormal phenotype with variegated leaves. This study, demonstrated that hydrolytic genes such as bgl1, celA and celB could be integrated and expressed correctly in the chloroplast genome. This work provides new information on methods and strategies for the expression of hydrolytic enzymes that are potentially useful for biotechnological applications using transplastomic plants.     

Assessment of photosynthetic potential of indoor plants under cold stress

Photosynthetic parameters including net photosynthetic rate (P_N), transpiration rate (E), water-use efficiency (WUE), and stomatal conductance (g_s) were studied in indoor C₃ plants Philodendron domesticum (Pd), Dracaena fragans (Df), Peperomia obtussifolia (Po), Chlorophytum comosum (Cc), and in a CAM plant, Sansevieria trifasciata (St), exposed to various low temperatures (0, 5, 10, 15, 20, and 25°C). All studied plants survived up to 0°C, but only St and Cc endured, while other plants wilted, when the temperature increased back to room temperature (25°C). The P_N declined rapidly with the decrease of temperature in all studied plants. St showed the maximum P_N of 11.9 μmol m^?2 s^?1 at 25°C followed by Cc, Po, Pd, and Df. E also followed a trend almost similar to that of P_N. St showed minimum E (0.1 mmol m^?2 s^?1) as compared to other studied C₃ plants at 25°C. The E decreased up to ≈4-fold at 5 and 0°C. Furthermore, a considerable decline in WUE was observed under cold stress in all C₃ plants, while St showed maximum WUE. Similarly, the g_s also declined gradually with the decrease in the temperature in all plants. Among C₃ plants, Pd and Po showed the maximum g_s of 0.07 mol m^?2 s^?1 at 25°C followed by Df and Cc. However, St showed the minimum g_s that further decreased up to ～4-fold at 0°C. In addition, the content of photosynthetic pigments [chlorophyll a, b, (a+b), and carotenoids] was varying in all studied plants at 0°C. Our findings clearly indicated the best photosynthetic potential of St compared to other studied plants. This species might be recommended for improving air quality in high-altitude closed environments.     

Genetic risk factors for myocardial infarction more clearly manifest for early age of first onset

Epidemiological genetics established that heritability in determining the risk of myocardial infarction (MI) is substantially greater when MI occurs early in life. However, the genetic architecture of early-onset and late-onset MI was not compared. We analyzed genotype frequencies of SNPs in/near 20 genes whose protein products are involved in the pathogenesis of atherosclerosis in two groups of Russian patients with MI: the first group included patients with age of first MI onset <60 years (N?=?230) and the second group with onset ≥60 years (N?=?174). The control group of corresponding ethnicity consisted of 193 unrelated volunteers without cardiovascular diseases (93 individuals were over 60 years). We found that in the group of patients with age of onset <60 years, SNPs FGB rs1800788*T, TGFB1 rs1982073*T/T, ENOS rs2070744*C and CRP rs1130864*T/T were associated with risk of MI, whereas in patients with age of onset ≥60 years, only TGFB1 rs1982073*T/T was associated with risk of MI. Using APSampler software, we found composite markers associated with MI only in patients with early onset: FGB rs1800788*T?+?TGFB1 rs1982073*T; FGB rs1800788*T?+?LPL rs328*C?+?IL4 rs2243250*C; FGB rs1800788*T?+?ENOS rs2070744*C (Fisher p values of 1.4?×?10^?6 to 2.2?×?10^?5; the permutation p values of 1.1?×?10^?5 to 3.0?×?10^?4; ORs?=?2.67–2.54). Alleles included in the combinations were associated with MI less significantly and with lower ORs than the combinations themselves. The result showed a substantially greater contribution of the genetic component in the development of MI if it occurs early in life, and demonstrated the usefulness of genetic testing for young people.     

Effect of low irradiance on the photosynthetic performance and spiking of Phalaenopsis

Lowering irradiance can delay the flower stalk, i.e., spike development, in order to schedule flowering time of Phalaenopsis; however, the effect on photosynthetic performance and spiking inhibition remains poorly understood. We compared light and shade treatments of Phalaenopsis aphrodite subsp. formosana in order to determine how limiting light affects day-night changes in the photosynthetic capacity of leaves and the carbon pool of leaves and stems resulting in delayed spiking. The low irradiance treatment [20 μmol(photon) m^?2 s^?1] for six weeks did not affect potential functions of photosynthetic apparatus estimated by chlorophyll a fluorescence analysis, but it significantly reduced the net CO₂ uptake and O₂ evolution rates, carbohydrate and organic acid concentrations, and amplitudes of CAM activity in new and fully expanded leaves of Phalaenopsis and delayed the spiking compared with the control kept at 150 μmol(photon) m^?2 s^?1. The shortened stem contained a remarkably high sucrose concentration, accounting for more than 80% of total soluble sugars for both treatments throughout the day. Moreover, the sucrose concentration was unaffected by the lowering of irradiance. The relationship between the sucrose content and spiking seemed to be loose; the major factor(s) for spiking in Phalaenopsis remained to be ascertained as the flower stalk bud is attached to the shortened stem.     

The aromatic cytokinin <Emphasis Type="Italic">meta-</Emphasis>topolin promotes in vitro propagation,shoot quality and micrografting in <Emphasis Type="Italic">Corylus colurna</Emphasis> L.

The role of 4.1 or 8.2 μM meta-topolin (mT) on shoot multiplication, rooting and ex vitro acclimatization of micropropagated Corylus colurna L., a promising non-suckering rootstock for hazelnut (Corylus avellana L.), was examined in comparison to N⁶-benzyladenine (BA), the most used cytokinin in tissue culture of Corylus spp. The influence of 8.2 μM mT and BA on photosynthetic pigments content and antioxidant enzymes activity, catalase (CAT) and guaiacol peroxidase (POD), in regenerated shoots, and on the preparation of the rootstock for micrografting was also evaluated. The highest shoot multiplication was recorded on medium containing 8.2 μM mT and an overall positive effect of mT on growth and quality of micropropagated shoots was found. The highest chlorophyll a content (1.236 mg g^?1 fresh weight, FW) and chlorophyll a/b ratio (2.48), and the lowest total carotenoids content (0.292 mg g^?1 FW) and CAT activity (25.8 μmol min^?1 mg^?1 protein) were detected after 8.2 μM mT application, while no significant differences were found in chlorophyll b content and POD activity between the two cytokinins. The best rhizogenesis response (98% for 4.1 μM and 100% for 8.2 μM mT) and ex vitro acclimatization competence (higher than 78%) were exhibited from shoots multiplied on mT. Furthermore, the multiplication of rootstock on mT allowed obtaining the highest (70%) response of successful micrografting. The present findings provide the first evidence of the successful applicability of mT in C. colurna tissue culture and development of micrografted plantlets.     

Optimization of photosynthesis,growth, and biochemical composition of the microalga <Emphasis Type="Italic">Rhodomonas salina</Emphasis>—an established diet for live feed copepods in aquaculture

The cryptophyte Rhodomonas salina is widely used as feed for copepod cultures. However, culturing conditions to obtain high-quality algae have not yet been efficiently optimized. Therefore, we aimed to develop a cultivation protocol for R. salina to optimize its nutritional value and provide technical recommendations for later large-scale production in algal photobioreactors. We studied photosynthesis, growth, pigments, fatty acid (FA) and free amino acid (FAA) composition of R. salina cultured at different irradiances (10–300 μmol photons m^?2 s^?1) and nutrient availability (deficiency and excess). The optimal range of irradiance for photosynthesis and growth was 60–100 μmol photons m^?2 s^?1. The content of chlorophylls a and c decreased with increasing irradiance while phycoerythrin peaked at irradiances of 40–100 μmol photons m^?2 s^?1. The total FA content was maximal at optimal irradiances for growth, especially under nutrient deficiency. However, highly unsaturated fatty acids, desired components for copepods, were higher under nutrient excess. The total FAA content was highest at limiting irradiances (10–40 μmol photons m^?2 s^?1) but a better composition with a higher fraction of essential amino acids was obtained at saturated irradiances (60–140 μmol photons m^?2 s^?1). These results demonstrate that quality and quantity of FA and FAA of R. salina can be optimized by manipulating the irradiance and nutrient conditions. We suggest that R. salina should be cultivated in a range of irradiance 60–100 μmol photons m^?2 s^?1 and nutrient excess to obtain algae with high production and a balanced biochemical composition as feed for copepods.     

Investigating species boundaries using DNA and morphology in the mite <Emphasis Type="Italic">Tyrophagus curvipenis</Emphasis> (Acari: Acaridae), an emerging invasive pest,with a molecular phylogeny of the genus <Emphasis Type="Italic">Tyrophagus</Emphasis>

Mites of the genus Tyrophagus (Acari: Acaridae) are among the most widespread and common mites, inhabiting diverse natural and anthropogenic habitats. Some species are pests of agricultural products and stored food and/or live in house dust, causing allergies to humans. We sequenced 1.2 kb of the mitochondrial COI gene for 38 individuals belonging to seven species of Tyrophagus, including T. curvipenis, T. putrescentiae, T. fanetzhangorum, T. longior, T. perniciosus, and T. cf. similis. Molecular phylogenetic analyses (1) recovered two major clades corresponding to the presence or absence of eyespots, and (2) separated all included morphological species. Tyrophagus curvipenis and T. putrescentiae had the lowest between-species genetic distances (range, mean?±?SD): 14.20–16.30, 15.17?±?0.40 (K2P). The highest within-species variation was found in T. putrescentiae 0.00–4.33, 1.78?±?1.44 (K2P). In this species, we recovered two distinct groups; however, no geographical or ecological dissimilarities were observed between them. Based on our analyses, we document important morphological differences between T. curvipenis and T. putrescentiae. For the first time, we record the occurrence of T. curvipenis in the New World and suggest that it may be an emerging pest as it is currently spreading in agricultural produce.     

Modelling photosynthesis in shallow algal production ponds

Shallow ponds with rapidly photosynthesising cyanobacteria or eukaryotic algae are used for growing biotechnology feedstock and have been proposed for biofuel production but a credible model to predict the productivity of a column of phytoplankton in such ponds is lacking. Oxygen electrodes and Pulse Amplitude Modulation (PAM) fluorometer technology were used to measure gross photosynthesis (P _G) vs. irradiance (E) curves (P _G vs. E curves) in Chlorella (chlorophyta), Dunaliella salina (chlorophyta) and Phaeodactylum (bacillariophyta). P _G vs. E curves were fitted to the waiting-in-line function [P _G = (P _Gmax × E/E_opt) × exp(1 — E/E_opt)]. Attenuation of incident light with depth could then be used to model P _G vs. E curves to describe P _G vs. depth in pond cultures of uniformly distributed planktonic algae. Respiratory data (by O2-electrode) allowed net photosynthesis (P _N) of algal ponds to be modelled with depth. Photoinhibition of photosynthesis at the pond surface reduced P _N of the water column. Calculated optimum depths for the algal ponds were: Phaeodactylum, 63 mm; Dunaliella, 71 mm and Chlorella, 87 mm. Irradiance at this depth is ≈ 5 to 10 μmol m^?2 s^?1 photosynthetic photon flux density (PPFD). This knowledge can then be used to optimise the pond depth. The total net P _N [μmol(O2) m^?2 s^?1] were: Chlorella, ≈ 12.6 ± 0.76; Dunaliella, ≈ 6.5 ± 0.41; Phaeodactylum ≈ 6.1 ± 0.35. Snell’s and Fresnel’s laws were used to correct irradiance for reflection and refraction and thus estimate the time course of P _N over the course of a day taking into account respiration during the day and at night. The optimum P _N of a pond adjusted to be of optimal depth (0.1–0.5 m) should be approximately constant because increasing the cell density will proportionally reduce the optimum depth of the pond and vice versa. Net photosynthesis for an optimised pond located at the tropic of Cancer would be [in t(C) ha^?1 y^?1]: Chlorella, ≈ 14.1 ± 0.66; Dunaliella, ≈ 5.48 ± 0.39; Phaeodactylum, ≈ 6.58 ± 0.42 but such calculations do not take weather, such as cloud cover, and temperature, into account.     

Survey of Philippine coffee beans for the presence of ochratoxigenic fungi

In 2012 to 2014, Philippine green coffee beans from Coffea arabica in Benguet and Ifugao; Coffea canephora var. Robusta in Abra, Cavite, and Ifugao; and Coffea liberica and Coffea excelsea from Cavite were collected and assessed for the distribution of fungi with the potential to produce ochratoxin A (OTA). The presence of fungal species was evaluated both before and after surface sterilization. There were remarkable ecological and varietal differences in the population of OTA-producing species from the five provinces. Aspergillus ochraceus, A. westerdijkiae, and Penicillium verruculosum were detected from Arabica in Benguet and Ifugao while Aspergillus carbonarius, Aspergillus niger, and Aspergillus japonicus were isolated in Excelsa, Liberica, and Robusta varieties from Abra, Cavite, and Davao. Contamination by Aspergillus and Penicillium species was found on 59 and 19 %, respectively, of the 57 samples from five provinces. After disinfection with 1 % sodium hypochlorite, the levels of infection by Aspergillus and Penicillium fell to 40 and 17 %, respectively. A total of 1184 fungal isolates were identified to species level comprising Aspergillus sections Circumdati (four species), Clavati (one), Flavi (one), Fumigati (one), Nigri (three), and Terrie (one). Within section Circumdati, 70 % of A. ochraceus produced OTA as high as 16238 ng g^?1 while 40 % of A. westerdijkiae produced maximum OTA of 36561 ng g^?1 in solid agar. Within section Nigri, 16.76 % of A. niger produced OTA at the highest 18439 ng g^?1, 10 % of A. japonicus at maximum level of 174 ng g^?1, and 21.21 % of A. carbonarius yielded maximum OTA of 1900 ng g^?1. Of the 12 species of Penicillium isolated, P. verruculosum was ochratoxigenic, with a maximum OTA production of 12 ng g^?1.     

Associations of <Emphasis Type="Italic">CTLA4</Emphasis> +49 A/G Dimorphism and HLA-DRB1*/DQB1* Alleles With Type 1 Diabetes from South India

The aim of present study was to elucidate the association of CTLA4 +49 A/G and HLA-DRB1*/DQB1* gene polymorphism in south Indian T1DM patients. The patients and controls (n?=?196 each) were enrolled for CTLA4 and HLA-DRB1*/DQB1* genotyping by RFLP/PCR-SSP methods. The increased frequencies of CTLA4 ‘AG’ (OR?=?1.99; p?=?0.001), ‘GG’ (OR?=?3.94; p?=?0.001) genotypes, and ‘G’ allele (OR?=?2.42; p?=?9.26?×?10^?8) were observed in patients. Reduced frequencies of ‘AA’ (OR?=?0.35; p?=?7.19?×?10^?7) and ‘A’ (OR?=?0.41; p?=?9.26?×?10^?8) in patients revealed protective association. Among HLA-DRB1*/DQB1* alleles, DRB1*04 (OR?=?3.29; p?=?1.0?×?10^?5), DRB1*03 (OR?=?2.81; p?=?1.9?×?10^?6), DQB1*02:01 (OR?=?2.93; p?=?1.65?×?10^?5), DQB1*02:02 (OR?=?3.38; p?=?0.0003), and DQB1*03:02 (OR?=?7.72; p?=?0.0003) were in susceptible association. Decreased frequencies of alleles, DRB1*15 (OR?=?0.32; p?=?2.55?×?10^?7), DRB1*10 (OR?=?0.45; p?=?0.002), DQB1*06:01 (OR?=?0.43; p?=?0.0001), and DQB1*05:02 (OR?=?0.28; p?=?2.1?×?10^?4) in patients were suggested protective association. The combination of DRB1*03+AG (OR?=?5.21; p?=?1.4?×?10^?6), DRB1*04+AG (OR?=?2.14; p?=?0.053), DRB1*04+GG (OR?=?5.21; p?=?0.036), DQB1*02:01+AG (OR?=?4.44; p?=?3.6?×?10^?5), DQB1*02:02+AG (OR?=?20.9; p?=?9.5?×?10^?4), and DQB1*02:02+GG (OR?=?4.06; p?=?0.036) revealed susceptible association. However, the combination of DRB1*10+AA (OR?=?0.35; p?=?0.003), DRB1*15+AA (OR?=?0.22; p?=?5.3?×?10^?7), DQB1*05:01+AA (OR?=?0.45; p?=?0.007), DQB1*05:02+AA (OR?=?0.17; p?=?1.7?×?10^?4), DQB1*06:01+AA (OR?=?0.40; p?=?0.002), and DQB1*06:02+AG (OR?=?0.34; p?=?0.001) showed decreased frequency in patients, suggesting protective association. In conclusion, CTLA4/HLA-DR/DQ genotypic combinations revealed strong susceptible/protective association toward T1DM in south India. A female preponderance in disease associations was also documented.     

Adaptive photosynthetic strategies of the Mediterranean maquis species according to their origin

In consideration of their origin the adaptive strategies of the evergreen species of the Mediterranean maquis were analysed. Rosmarinus officinalis L., Erica arborea L., and Erica multiflora L. had the lowest net photosynthetic rate (P_N) in the favourable period [7.8±0.6 μmol(CO₂) m^?2s^?1, mean value], the highest P_N decrease (on an average 86 % of the maximum) but the highest recovery capacity (>70 % of the maximum) at the first rainfall in September. Cistus incanus L. and Arbutus unedo L. had the highest P_N during the favourable period [15.5±5.2 μmol(CO₂) m^?2s^?1, mean value], 79 % decrease during drought, and a lower recovery capacity (on an average 54 %). Quercus ilex L., Phillyrea latifolia L., and Pistacia lentiscus L. had an intermediate P_N in the favourable period [9.2±1.3 μmol(CO2) m^?2s^?1, mean value], a lower reduction during drought (on an average 63 %), and a range from 62 % (Q. ilex and P. latifolia) to 39 % (P. lentiscus) of recovery capacity. The Mediterranean species had higher decrease in P_N and stomatal conductance during drought and a higher recovery capacity than the pre-Mediterranean species. Among the pre-Mediterranean species, P. latifoliahad the best adaptation to long drought periods also by its higher leaf mass per area (LMA) which lowered leaf temperature thus decreasing transpiration rate during drought. Moreover, its leaf longevity determined a more stable leaf biomass during the year. Among the Mediteranean species, R. officinalis was the best adapted species to short drought periods by its ability to rapidly recover. Nevertheless, R. officinalis had the lowest tolerance to high temperatures by its P_N dropping below half its maximum value when leaf temperature was over 33.6°C. R. officinalismay be used as a bioindicator species of global change.     

<Emphasis Type="Italic">Bruguiera hainesii</Emphasis>, a critically endangered mangrove species,is a hybrid between <Emphasis Type="Italic">B. cylindrica</Emphasis> and <Emphasis Type="Italic">B. gymnorhiza</Emphasis> (Rhizophoraceae)

Bruguiera hainesii (Rhizophoraceae) is one of the two Critically Endangered mangrove species listed in the IUCN Red List of Threatened Species. Although the species is vulnerable to extinction, its genetic diversity and the evolutionary relationships with other Bruguiera species are not well understood. Also, intermediate morphological characters imply that the species might be of hybrid origin. To clarify the genetic relationship between B. hainesii and other Bruguiera species, we conducted molecular analyses including all six Bruguiera species using DNA sequences of two nuclear genes (CesA and UNK) and three chloroplast regions (intergenic spacer regions of trnL-trnF, trnS-trnG and atpB-rbcL). For nuclear DNA markers, all nine B. hainesii samples from five populations were heterozygous at both loci, with one allele was shared with B. cylindrica, and the other with B. gymnorhiza. For chloroplast DNA markers, the two haplotypes found in B. hainesii were shared only by B. cylindrica. These results suggested that B. hainesii is a hybrid between B. cylindrica as the maternal parent and B. gymnorhiza as the paternal one. Furthermore, chloroplast DNA haplotypes found in B. hainesii suggest that hybridization has occurred independently in regions where the distribution ranges of the parental species meet. As the IUCN Red List of Threatened Species currently excludes hybrids (except for apomictic plant hybrids), the conservation status of B. hainesii should be reconsidered.     

Efficiency of spinosad, <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> and <Emphasis Type="Italic">Trichogramma brassicae</Emphasis> against the tomato leafminer in greenhouse

The objective of this study was to evaluate the efficacy of three eco-friendly control agents, either singly or in a pairwise combination, for the control of the tomato leafminer, Tuta absoluta (Meyrick) (Lep: Gelechiidae). They include the naturally derived pesticide spinosad, a commercially available formulation of Bacillus thuringiensis var. Kurstaki (Bt), and a native population of Trichogramma brassicae Bezdenko (Hym: Trichogrammatidae). Tomato plants containing the T. absoluta were treated with one of the seven following treatments in a greenhouse: (1) a single release of T. brassicae against the eggs; (2) two applications of Bt (2 kg ha^?1); (3) and (4) one application of spinosad at two rates (60 and 120 g a.i. ha^?1); (5) T. brassicae release?+?Bt spray; (6) T. brassicae release?+?spinosad spray; and (7) spinosad spray?+?Bt spray. The highest mortality rate was recorded for the spinosad?+?Bt (88.33?±?1.43%) and T. brassicae?+?spinosad (78.33?±?3.74%) combinations, respectively; while the lowest mortality rate was obtained through the single application of T. brassicae (31.67?±?4.84%). Based on our results, the Bt and spinosad seem to be suitable candidates for combination with other biological and cultural techniques towards an integrated management of the tomato leafminer.