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.     

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.     

Use of urea-molasses-multinutrient block and urea-treated rice straw for improving dairy cattle productivity in Vietnam

After conducting a preliminary survey, a feeding trial was carried out to determine the effect of urea-molasses-multinutrient block (UMMB) and urea-treated rice straw (UTRS) as a feed supplement on the productivity of dairy cows. Sixty Holstein-Friesian crossbred cows on 11 smallholder farms were divided equally into control, UMMB and UTRS supplementation groups. Milk yield and feed intake were recorded daily. Milk fat content, body weight and body condition score (BSC) of each cow were determined at two week intervals. Milk samples for progesterone analysis were collected once a week commencing one month after parturition. Data were recorded for date of onset of ovarian activity, estrus, insemination, and conception rate. Milk production increased by 10.3-11.9% and milk fat content increased by 3-5%, therefore, profit for farmers increased by US $0.55-0.73 per cow per day (exchange rate US $1 = VN $11,000). The intervals from calving to onset of ovarian activity (91-94 days), to estrus (110-114 days), to conception (121-122 days) and the calving interval (13.4-13.6 months) in the trial groups were significantly shorter than those in the control group (112, 135, 152 days and 14.4 months, respectively.     

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.     

Structural characteristics of cell walls of kenaf (<Emphasis Type="Italic">Hibiscus cannabinus </Emphasis>L.) and fixation of carbon dioxide

 Kenaf (Hibiscus cannabinus) plants are widely known for their contribution to the global and regional environment because of their ability to fix CO₂. On the other hand, some scientists have doubts about CO₂ fixation by kenaf and have misgivings about the effect of kenaf on the ecosystem. We have characterized the structural characteristics of cell walls of bast fibers, cores, roots, and leaves of kenaf during the maturation of plants and investigated the rate of photosynthesis. During maturation of the kenaf plant the cellulose (bast fiber 52–59%, core 44–46%) and lignin (bast fiber 9.3–13.2%, core 18.3–23.2%) contents increased significantly. The aromatic composition of the lignin of bast fiber was significantly different from that of the core lignin and of other plants. The lignin of bast fiber appears similar to pure syringyl lignin. Fixation of CO₂ by kenaf plants and their contribution to the global environment are discussed. A significatly high rate of photosynthesis of kenaf plants was observed compared to that of woody plants in Japan, but the amount of CO₂ fixation depends on the characteristics of the plantation. If the kenaf was planted in high density, about twice as much CO₂ was fixed as was fixed by trees in a tropical rain forest. Received: April 22, 2002 / Accepted: July 24, 2002 Acknowledgments This project was supported by the Science and Technology Agency (STA) fellowship of the Japan International Science and Technology Exchange Center (JISTEC), which has been successfully applied by Dr. Shuji Hosoya, Forestry and Forest Products Research Institute. We thank Dr. Toshio Sumizono and Mr. Masao Sakurai, Forestry and Forest Products Research Institute, for their kind help in determining the rate of photosynthesis and cultivating the kenaf plants, respectively. We also express our appreciation to Dr. Quang Hung Le, College of Agriculture and Forestry, Ho Chi Minh City, Vietnam for offering information about the cultivation of kenaf at Thu Duc District, Ho Chi Minh City.     

Crinum L. (Amaryllidaceae)

Alkaloids isolated from Crinum species have been reviewed for the period 1985-2000. Non-nitrogenous compounds have been surveyed for the first time. Botanical classification and biological activity are discussed. Spectral data literature sources are listed.