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

Thi Thanh Nhan Pham  Thi Ngoc Lan Nguyen  Thi Ha Bui  Huu Quan Nguyen  Thi Tam Nguyen  Van Son Le 《The Journal of Horticultural Science and Biotechnology》2019,94(5):591-598

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  相似文献   

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

Elena V. Girich  Anton N. Yurchenko  Olga F. Smetanina  Phan Thi Hoai Trinh  Ngo Thi Duy Ngoc  Mikhail V. Pivkin  Roman S. Popov  Evgeny A. Pislyagin  Ekaterina S. Menchinskaya  Ekaterina A. Chingizova  Shamil S. Afiyatullov  Ekaterina A. Yurchenko 《Marine drugs》2020,18(12)

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.  相似文献   

Ouabain-sensitive Na/K-ATPase activity increases during embryogenesis in the giant freshwater prawn Macrobrachium rosenbergii     

Marcy N Wilder  Do Thi Thanh Huong  Atsuro Okuno  Muharijadi Atmomarsono  Wei-Jun Yang 《Fisheries Science》2001,67(1):182-184

  相似文献   

Na/K-ATPase activity during larval development in the giant freshwater prawn Macrobrachium rosenbergii and the effects of salinity on survival rates     

Do Thi  Thanh HUONG  Vidya  JAYASANKAR  Safiah  JASMANI  Hisako  SAIDO-SAKANAKA  Andrew J.  WIGGINTON  Marcy N.  WILDER 《Fisheries Science》2004,70(3):518-520

  相似文献   

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

Chang‐Hoon Shin  Hien Thi Dieu Bui  Samad Rahimnejad  Ji‐Hoon Cha  Byung‐Woo Yoo  Bo‐Kyeun Lee  Hyung‐Jin Ahn  Soo‐Il Choi  Yun‐Jeong Choi  Yong‐Ho Park  Jeong‐Dae Kim  Kyeong‐Jun Lee 《Journal of the World Aquaculture Society》2014,45(3):258-268

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.  相似文献   

Is N‐feedback involved in the regulation of nitrogenase activity in Medicago truncatula?     

Saad Sulieman  Joachim Schulze  Karl H. Mühling  Lam-Son Phan Tran 《植物养料与土壤学杂志》2020,183(1):42-45

To determine whether senescing leaves provoke an active nitrogen (N) remobilization that results in the reduction of nitrogenase activity, 60% of Medicago truncatula lower leaves were either darkened or individually excised for two weeks. Although a considerable amount of N was remobilized, N₂ fixation activity was found to be increased to maintain the N source/sink balance, indicating an absence of the negative N‐feedback regulation of nitrogenase activity in the senescing M. truncatula.  相似文献   

Prevalence of cpb2, encoding beta2 toxin,in Clostridium perfringens field isolates: correlation of genotype with phenotype   总被引：4，自引：0，他引：4  

Bueschel DM  Jost BH  Billington SJ  Trinh HT  Songer JG 《Veterinary microbiology》2003,94(2):121-129

Beta2 toxin, encoded by the cpb2 gene, has been implicated in the pathogenesis of porcine, equine and bovine enteritis by type A Clostridium perfringens. By incorporating primers to cpb2 into a multiplex genotyping PCR, we screened 3270 field isolates of C. perfringens. Of these, 37.2% were PCR positive for the cpb2 gene. The majority of isolates from cases of porcine enteritis were positive for cpb2 (>85%), and this was even more true for C. perfringens isolated from cases of porcine neonatal enteritis (91.8%). In contrast, isolates from normal pigs only contained cpb2 in 11.1% of cases. The correlation between enteritis in other animal species and the presence of cpb2 was not so strong. cpb2 was found in 21.4% of C. perfringens isolates from cattle with enteritis, and in 47.3% of isolates from calves with enteritis or abomastitis. The prevalence of cpb2 varied with genotype, with type A isolates being positive for this gene in 35.1% of cases. Furthermore, enterotoxigenic type D or type E strains almost always carried cpb2. We cloned a 6xHIS-tagged beta2 (HIS-beta2) and used this protein to raise antiserum against beta2. Culture supernatants from 68 cpb2-positive and 13 cpb2-negative strains were tested for the presence of beta2 by Western blotting. In cpb2-positive isolates of porcine origin, beta2 was almost always detected (96.9%). However, in cpb2-positive isolates from other animal species, only 50.0% expressed beta2 protein. The high rate of cpb2-positivity among strains from neonatal pigs with enteritis and the high correlation of genotype with phenotype, supports the contention that beta2 toxin plays a role in the pathogenesis of these infections. However, it may be important to consider the use of an additional method for the detection of beta2 toxin in non-porcine cpb2-positive isolates when making claims about the role of beta2 in enteritis in non-porcine species.  相似文献   

PCR detection and analysis of Pasteurella multocida from the tonsils of slaughtered pigs in Vietnam     

Townsend KM  Hanh TX  O'Boyle D  Wilkie I  Phan TT  Wijewardana TG  Trung NT  Frost AJ 《Veterinary microbiology》2000,72(1-2):69-78

A total of 36 tonsil swab samples were collected from healthy swine prior to slaughter at the abattoirs in Can tho and Tien giang provinces of Southern Vietnam. The presence of Pasteurella multocida in these samples was detected by the combination of direct cultivation and isolation, mouse inoculation and the polymerase chain reaction (PM-PCR). P. multocida was detected in 16 samples by PCR, with 17 strains ultimately isolated. All samples were negative for serogroup B by HSB-PCR and conventional serotyping, with isolates identified as A:3, D:1 or D:3. In addition, all samples were determined to be negative for the P. multocida toxin (PMT). Characterisation of isolated P. multocida by REP-PCR and biotyping revealed nine distinct REP profiles and seven biotypes among the 17 isolates. Some correlation was seen with P. multocida isolated from a previous Australian outbreak of acute swine pasteurellosis, and those isolated from fowl cholera outbreaks in Vietnamese poultry.  相似文献   

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

Kunimasa Kawabe  Nguyen Thi Ngoc Truc  Bui Thi Ngoc Lan  Le Thi Thu Hong  Masatoshi Onuki 《Journal of General Plant Pathology》2006,72(6):355-359

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.  相似文献   

A new ent-kaurane diterpenoid from Croton tonkinensis leaves     

Minh PT  Ngoc PH  Taylor WC  Cuong NM 《Fitoterapia》2004,75(6):552-556

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.  相似文献