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     

Neuroprotective Metabolites from Vietnamese Marine Derived Fungi of Aspergillus and Penicillium Genera

Low molecular weight secondary metabolites of marine fungi Aspergillus flocculosus, Aspergillus terreus and Penicillium sp. from Van Phong and Nha Trang Bays (Vietnam) were studied and a number of polyketides, bis-indole quinones and terpenoids were isolated. The structures of the isolated compounds were determined by 1D and 2D NMR and HR-ESI-MS techniques. Stereochemistry of some compounds was established based on ECD data. A chemical structure of asterriquinone F (6) was thoroughly described for the first time. Anthraquinone (13) was firstly obtained from a natural source. Neuroprotective influences of the isolated compounds against 6-OHDA, paraquat and rotenone toxicity were investigated. 4-Hydroxyscytalone (1), 4-hydroxy-6-dehydroxyscytalone (2) and demethylcitreoviranol (3) have shown significant increasing of paraquat- and rotenone-treated Neuro-2a cell viability and anti-ROS activity.     

Milk vetch dwarf virus infection in the Solanaceae and Caricaceae families in Southeast Asia

Milk vetch dwarf virus (MDV) is an important member of the genus Nanovirus and is transmitted by the aphid Aphis craccivora. MDV has multiple single-stranded DNA genome components, each approximately 1 kb, and two or three alpha-satellite molecules. It mainly infects plants of the legume family Fabaceae. Recently, papaya (Carica papaya) collected in Yesan, South Korea, displaying symptoms of leaf yellowing and dwarfism, was identified as a new host for MDV. To examine the geographical distribution of MDV, papaya samples with symptoms were harvested in South Korea, Vietnam, and Taiwan in August 2018, along with tomato and pepper samples grown in adjacent fields in Vietnam. The results revealed the presence of MDV not only in papaya but also in pepper and tomato. This MDV infection in members of the Solanaceae family was confirmed by Southern blot hybridization performed using a PCR product of segment S as a probe. Based on sequence analysis of three MDV components (M, S, and C3), we verified the presence of three different isolates of MDV in these three countries and homology between sequences of isolates from papaya and from members of the Solanaceae from Vietnam. Taken together, our results clearly demonstrate MDV infection in Vietnam and Taiwan for the first time and confirm that MDV can infect economically important pepper and tomato.     

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.     

Evaluation of smallholder pig production systems in North Vietnam: Pig production management and pig performances

This study investigated the productive adaptability of pig breeds under different smallholder production conditions in Vietnam, comparing an indigenous with a Vietnamese improved breed. Fieldwork was conducted in four villages with different remoteness in North Vietnam from 2001 to 2002, in 64 households keeping the improved Mong Cai or indigenous Ban as sow breeds and progeny for fattening. Four visits per farm yielded 234 structured interviews. Reproductive performances were derived from 135 litters. Individual weights of pigs (n = 755) were obtained. Data were analysed by regression, linear and generalised linear, especially loglinear models.

Two distinct pig production systems were identified, that differed in remoteness, market access, resource availability, distribution of pig breeds and pig production intensity. Higher performances of 1.4 and 1.5 litters year^− 1, 8.4 and 8.4 piglets weaned litter^− 1, 66.6 and 93.0 kg piglets weaned sow^− 1 year^− 1, and 136 and 177 g day^− 1 ADG were found in two villages near town with semi-intensive production conditions and a high rate of improved Mong Cai sows and MC and LW × MC offspring in the observed population. Lower performances of 1.1 and 1.1 litters year^− 1, 4.6 and 5.5 piglets weaned litter^− 1, 20.5 and 30.9 kg piglets weaned sow^− 1 year^− 1, and 66 and 85 g day^− 1 ADG were observed in parallel to higher incidences of indigenous Ban pigs away from town under extensive conditions. Total live weight offtake per household per year was higher near town and in one village distant to town. MC and LW × MC remained even under demand-driven conditions below the performance potential reported for improved genotypes. Under resource-driven conditions with a saving-oriented production pattern, the higher-yielding but more demanding Mong Cai might not be an efficient production alternative as a further performance reduction can be assumed. A considerable live weight output was observed under resource-driven conditions and with a higher percentage of crossbred LW × Ban offspring in the population, possibly representing a promising production alternative. Factors influencing the performance expression of pigs, and the suitability of different methods for the assessment of performance parameters on-farm are discussed.     

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.     

A new ent-kaurane diterpenoid from Croton tonkinensis leaves

A new ent-kaurane diterpenoid (1) was isolated from the leaves of Croton tonkinensis. The structure of 1 was determined as ent-7beta-hydroxy-15-oxokaur-16-en-18-ol from spectroscopic methods.