Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Effects of Dietary Supplementation of Barodon on Growth Performance,Innate Immunity and Disease Resistance of Juvenile Olive Flounder,Paralichthys olivaceus,Against Streptococcus iniae

This study was conducted to evaluate the effects of dietary supplementation of Barodon, an anionic alkali mineral complex, on growth, feed utilization, humoral innate immunity and disease resistance of olive flounder. A basal experimental diet was used as a control and supplemented with 0.1, 0.2, 0.3, 0.4, or 0.5% Barodon. Triplicate groups of fish (26.4 ± 0.2 g) were fed one of the diets to apparent satiation twice daily for 10 wk. The growth performance was enhanced (P < 0.05) linearly and quadratically in fish fed diets containing Barodon compared with that in fish fed the control. Feed utilization was significantly improved by Barodon supplementation. Serum lysozyme and antiprotease activities were increased quadratically in Barodon fed groups. Also, significantly higher superoxide dismutase activity was found in Barodon‐fed fish. Dietary supplementation of 0.1–0.3% Barodon resulted in significant enhancement of fish disease resistance against Streptococcus iniae. The findings in this study indicate that dietary supplementation of Barodon can enhance growth, feed utilization, innate immunity, and disease resistance of olive flounder and that the optimum level seems to be 0.1% in diets.     

Does reverse growth dominance develop in old plantations of Eucalyptus saligna?

Patterns of stand growth and dominance among individual trees change through stand development. We tested a prediction that large, old plantations of Eucalyptus saligna (Sm.) in Hawaii and Brazil would show a pattern of “reverse” growth dominance, with the largest trees contributing more to stand biomass than to current stand increment. This pattern might develop if the largest trees accounted for a large portion of the total resource use of the stand, but were less efficient at utilizing resources to produce stemwood. The 70-year-old plantation in Hawaii had a stem mass of 610 Mg/ha, compared with 325 Mg/ha in the 66-year-old Brazil plantation. Cumulative frequency plots for both sites showed that dominant trees comprised a very large proportion of total stand mass; the largest 20% of the trees accounted for 50% and 30% of the total stand mass in Hawaii and Brazil, respectively. Contrary to expectations, the largest trees continued to produce a disproportionately large amount of total stand growth, with the largest 20% of trees accounting for 60% (Hawaii) and 40% (Brazil) of stand growth. The frequency distributions resulted in positive growth dominance coefficients (similar to Gini coefficients) of 0.14 for Hawaii and 0.09 for Brazil, rather than negative values that would indicate that reverse growth dominance. The failure of these monoculture Eucalyptus stands to develop reverse growth dominance contrasts starkly with a variety of studies with pine species, which tend to show neutral-to-reverse growth dominance. Indeed, continued dominance of large trees suggests the trees might respond well to silvicultural treatments such as thinning or fertilization. Future work that contrasts patterns of growth dominance among species will be important as a foundation for developing generalizations about trends in growth dominance with stand development.     

Quantification of DNA of citrus huanglongbing pathogen in diseased leaves using competitive PCR

A quantification system for huanglongbing pathogen using a competitive polymerase chain reaction method and image-analyzing software were developed to obtain precise results. Significant differences in the quantity of pathogen were thus determined in leaves of two citrus cultivars commonly cultivated in southern Vietnam. Less pathogen-related DNA was detected from the tissue of citrus cultivars that are believed to be more tolerant than susceptible cultivars. The quantification system will be used in studies on pathogen proliferation and movement inside citrus tissue.     

Effects of crushed conspecifics on growth and survival of Penaeus monodon Fabricius post larvae

In the present study, the hypothesis tested was that Penaeus monodon post‐larvae (PL) experience lower growth when exposed to crushed conspecifics, which was achieved by exposing individual P. monodon PL with abundant food for 4 weeks to a gradient from 0 to 100 crushed conspecific PL L^?1. Both dry weight (48.5±7.2 mg) and body size (28.0±1.3 mm) of animals exposed to 1 macerated PL L^?1 were significantly (P≤0.011) higher than those of animals in treatments with 0, 5 and 10 crushed PL L^?1 (average over treatments: 25.6±3.2 mg; 23.4±0.5 mm). All animals died within 1 week when exposed to 70 and 100 crushed PL L^?1, and within 3–4 weeks when exposed to 50 and 30 crushed PL L^?1. Exposure time affected mortality and it appeared that LC₅₀ values decreased from 60 to 13 crushed PL L^?1 from 1 to 4 weeks' exposure. Survival of P. monodon PL was negatively correlated to pH, biological oxygen demand, ammonia and nitrate. In conclusion, low dose of crushed conspecifics has a stimulatory effect on P. monodon PL, as larvae were heavier and larger, while high doses cause high mortality.     

Ciguatoxin in moray eels raising the risk for seafood safety in Viet Nam

Fisheries Science - Recently, suspected ciguatera fish poisoning (CFP) cases caused by the consumption of moray eels, popular seafood for locals and tourists, have been reported in Viet Nam....     

Reverse-phase liquid chromatographic determination of benzoic and sorbic acids in foods

An isocratic liquid chromatographic (LC) technique is described for the determination of benzoic acid and sorbic acid in foods such as beverages, fruits, seafood, vegetables, sauces, and dairy, bakery, and confectionery products. A C18 column is used with methanol-phosphate buffer (5 + 95) as mobile phase and 4-hydroxyacetanilide or 3,5-dinitrobenzoic acid as internal standard. Sample preparation is simple, rapid, and produces a sample extract that has a minimum effect on the column performance and life. Specificity of the method was checked against common food additives such as L-ascorbic acid, caffeine, artificial sweeteners (saccharin, cyclamate, aspartame), antioxidants (BHT, BHA) and artificial colors. Also described are 2 procedures for confirmation of the preservatives, using either redox reaction of sorbic acid with potassium permanganate or gas chromatography/mass spectrometry. Mean recoveries of 90-105% were obtained with a precision of 1-6% and a detection limit of 20 mg/kg for the 2 preservatives.     

A domain in TNF receptors that mediates ligand-independent receptor assembly and signaling

A conserved domain in the extracellular region of the 60- and 80-kilodalton tumor necrosis factor receptors (TNFRs) was identified that mediates specific ligand-independent assembly of receptor trimers. This pre-ligand-binding assembly domain (PLAD) is physically distinct from the domain that forms the major contacts with ligand, but is necessary and sufficient for the assembly of TNFR complexes that bind TNF-alpha and mediate signaling. Other members of the TNFR superfamily, including TRAIL receptor 1 and CD40, show similar homotypic association. Thus, TNFRs and related receptors appear to function as preformed complexes rather than as individual receptor subunits that oligomerize after ligand binding.     

Anti-Inflammatory Components of the Starfish Astropecten polyacanthus

Inflammation is important in biomedical research, because it plays a key role in inflammatory diseases including rheumatoid arthritis and other forms of arthritis, diabetes, heart disease, irritable bowel syndrome, Alzheimer’s disease, Parkinson’s disease, allergies, asthma, and even cancer. In the present study, we describe the inhibitory effect of crude extracts and steroids isolated from the starfish Astropecten polyacanthus on pro-inflammatory cytokine (Interleukin-12 (IL-12) p40, interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α)) production in lipopolysaccharide (LPS)-stimulated bone marrow-derived dendritic cells (BMDCs). Among those tested, compounds 5 and 7 showed potent inhibitory effects on the production of all three pro-inflammatory cytokines with IC₅₀ values ranging from 1.82 ± 0.11 to 7.00 ± 0.16 μM. Potent inhibitory activities were also observed for compound 1 on the production of IL-12 p40 and IL-6 with values of 3.96 ± 0.12 and 4.07 ± 0.13 μM, respectively, and for compounds 3 and 4 on the production of IL-12 p40 with values of 6.55 ± 0.18 and 5.06 ± 0.16 μM, respectively. Moreover, compounds 2 (IC₅₀ = 34.86 ± 0.31 μM) and 6 (IC₅₀ = 79.05 ± 2.05 μM) exhibited moderate inhibitory effects on the production of IL-12 p40, whereas compounds 3 (IC₅₀ = 22.80 ± 0.21 μM) and 4 (IC₅₀ = 16.73 ± 0.25 μM) moderately inhibited the production of TNF-α and IL-6, respectively.     

Productivity of Sargassum linearifolium in potassium fortified inland saline water under laboratory conditions

Growing aquatic species in inland saline water (ISW) is one way to reduce the adverse impact of ISW to agriculture farms. This 84‐day laboratory‐trial was conducted to study the growth of Sargassum linearifolium cultured in ocean water (OW), ISW, ISW fortified with potassium equivalent to 100% (ISW100), 66% (ISW66) and 33% (ISW33) of potassium in OW at 35 g/L. The biomass and cumulative specific growth rate (SGR) of S. linearifolium increased significantly (p < .05) with increased potassium in ISW until 56 days and then declined. The ISW100 and OW resulted in similar growth patterns and yielded peak biomass at day 42, proving static biomass for the next 28 days before declining. The biomass of S. linearifolium cultured in ISW and ISW33 significantly (p < .05) decreased and was lower than in ISW100 and died after day 56. The SGR of S. linearifolium in OW, ISW100 and ISW66 levelled off and showed no difference during the first 56 days. The S. linearifolium biomass and SGR negatively and significantly (p < .05) correlated with the concentrations of nitrate, phosphate in all waters. The increased potassium concentration in ISW similar to its concentration in SW brought the growth of S. linearifolium cultured to a level that was similar in OW.