Partial purification,characterization, and thermal and high-pressure inactivation of pectin methylesterase from carrots (Daucus carrota L.)

Pectin methylesterase (PME) from carrots (Daucus carrota L.) was extracted and purified by affinity chromatography on a CNBr-Sepharose 4B-PME inhibitor column. A single protein and PME activity peak was obtained. A biochemical characterization in terms of molar mass (MM), isoelectric points (pI), and kinetic parameters of carrot PME was performed. In a second step, the thermal and high-pressure stability of the enzyme was studied. Isothermal and combined isothermal-isobaric inactivation of purified carrot PME could be described by a fractional-conversion model.     

RAPD variability in rice (Oryza sativa L.) plants derived from desiccation-tolerant calli

Genetic modification from selfed progenies of 18 rice (Oryza sativa L.) plants regenerated from callus tissues which survived desiccation, were investigated at the DNA level using the random amplified polymorphic DNA (RAPD) method. Twelve 10-mer random primers were used to amplify DNA of progenies from the regenerated plants, and a total of 228 PCR products and 1780 DNA fragments were obtained by primers, generating between four to thirteen major bands. The size of the amplified fragments ranged from 0.2 to 2.55 kb. The results showed that 10 out of 12 primers produced polymorphic bands, two primers (RA31 and RA185) showed no polymorphism among plants tested. A dendrogram of the genetic distance was constructed based on their polymorphism, demonstrating that somaclonal variation exists in rice plants regenerated from callus which survived the desiccation treatment. Part of this variation can be useful in rice breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Distribution of the mutant allele of the DMRT3 gene associated with ambling gaits in Japanese native horse populations

There are currently eight native horse populations in Japan, namely, Hokkaido, Kiso, Noma, Taishu, Misaki, Tokara, Miyako, and Yonaguni horses. Since locomotion traits, including gaitedness, are important for riding and packing horses, the genetic properties associated with these traits could be informative for understanding the characteristics and history of these horses. In this study, we investigated the distribution of the mutant allele of DMRT3 gene (DMRT3:p.Ser301Ter) associated with ambling gaits in the Japanese native horse. We also examined haplotypes of SNPs in the 83‐kb region including DMRT3 gene by genotyping four SNPs in this region. The results revealed the presence of DMRT3:p.Ser301Ter in the Hokkaido and Yonaguni populations at allele frequencies of 0.18 and 0.02, respectively, and the observed haplotype associated with DMRT3:p.Ser301Ter was estimated as the most common haplotype in the horses in the world. Since DMRT3:p.Ser301Ter has been hypothesized to spread across Eurasian continent from Medieval England after 850 to 900 CE, our findings of the presence of DMRT3:p.Ser301Ter with the common haplotype in the Japanese native horses will provide a new insight into the history of the Japanese native horse, such as considerable level of gene flow from Eurasian continent after 850 to 900 CE.     

Controlled reproduction of an important indigenous fish species, Spinibarbus denticulatus (Oshima, 1926), in Southeast Asia

Aquaculture of Spinibarbus denticulatus (Oshima, 1926), a fish species indigenous to North Vietnam and Eastern China, is constrained by lack of fingerlings for stocking ponds and cages. As these fish do not naturally breed in captivity, carp pituitary extract (CPE), luteinizing hormone-releasing hormone analogue (LHRHa) with domperidone (DOM) and human chorionic gonadotropin (HCG) were administered at various doses to induce ovulation. A first set of experiments evaluated the response to LHRHa (30, 40 and 50 μg kg⁻¹) with or without DOM (10 mg kg⁻¹), CPE (20, 30 and 40 mg kg⁻¹) and HCG (3000, 4000 and 5000 IU kg⁻¹). A second set of experiments evaluated the dose response to LHRHa (30, 35, 40 and 50 μg kg⁻¹) primed with 6 mg kg⁻¹ of CPE, and HCG (3000, 3500, 4000, 5000 IU kg⁻¹) primed with 6 mg kg⁻¹ of CPE. The treatment groups were compared with each other and the control (injected with 0.9% saline solution). Only 25% and 50% ovulation resulted when treated with LHRHa at 40 and 50 μg kg⁻¹, whereas 100% ovulation was achieved with an addition of DOM to LHRHa. Both 30 and 40 mg kg⁻¹ CPE induced 100% ovulation. However, HCG (4000 and 5000 IU kg⁻¹) induced ovulation in only 33% of females. When primed with CPE, the minimum dose of LHRHa required was 35 μg kg⁻¹ to achieve 70% ovulation. Priming HCG with CPE also resulted in 100% ovulation, and the minimum effective dose of HCG to induce ovulation was 3500 IU kg⁻¹ with 60% ovulation. Fertilization and hatch rates observed in this study with different hormonal stimulation were high (80–93%). The results indicate that while the use of combined hormone strategy has no apparent advantage over a single hormone strategy, LHRHa+DOM (40 μg kg⁻¹+10.0 mg kg⁻¹) and CPE (30 mg kg⁻¹) are most effective in consistently inducing ovulation and thus can be used for commercial hatchery production of S. denticulatus larvae.     

Between ‘state’ and ‘society’: Commune authorities and the environment in Vietnam's craft villages

Maintaining a liveable environment in Vietnam's polluted craft villages is a daily challenge for state authorities and residents. Neighbouring urban populations demand that the state effectively curtails and manages pollution, while local residents prioritise their livelihoods and routinely flout regulations. The commune official, tasked with the seemingly impossible task of environmental regulation, occupies a fraught position, torn between the imperatives and constraints of craft producers and state regulatory demands. This study of water pollution in northern Vietnam's craft villages finds that commune officials' conflicted role in environmental governance is a central factor in the failure of the current environmental governance regime, and reflects the internally conflicted nature of the Vietnamese state.     

Assessment of genetic diversity and population structure of Vietnamese indigenous cattle populations by microsatellites

Cattle play a very important role in agriculture and food security in Vietnam. A high level of cattle diversity exists and serves different needs of Vietnamese cattle keepers but has not yet been molecularly characterized. This study evaluates the genetic diversity and structure of Vietnamese indigenous cattle populations, using microsatellite markers. A total of 410 individuals from six indigenous cattle populations and an exotic breed was characterized using 27 microsatellite markers A total of 362 alleles was detected and the number of alleles per locus ranged from 8 (INRA005 and ILSTS005) to 17 (ETH185). The level of gene diversity was high indicated by a mean expected heterozygosity (He) across populations and loci of 0.73. Level of inbreeding (mean F_IS=0.05) and genetic differentiation (mean F_ST=0.04) was moderate. The phylogenetic tree based on Reynolds genetic distance reflected geographic distances. Structure analysis indicated five homogeneous clusters. The Brahman, Lang Son, Ha Giang and U Dau Riu cattle were assigned to independent clusters while Nghe An, Thanh Hoa and Phu Yen cattle were grouped in a single cluster. We conclude that Vietnamese indigenous cattle have high levels of genetic diversity and distinct genetic structures. Based on these results, we recommend that for conservation homogenous populations (Nghe An, Thanh Hoa and Phu Yen) can be grouped to reduce costs and U Dau Riu, Lang Son and Ha Giang populations should be conserved separately to avoid loss of genetic diversity.     

Weed flora of water rice in the Red River Delta,Vietnam

The occurrence of weeds in water rice was surveyed in the Red River Delta, Vietnam during spring and summer rice-growing seasons in 1995 and 1996. Sixty different weeds from 19 plant families were recorded. The most important plant families as weeds of rice were Poaceae and Cyperaceae. The most important weed however was Rotala indica (Willd.) Koehne (Lythraceae) followed by Echinochloa crus-galli (L.) Beauv. and Cyperus difformis L. A brief comparison of this rice weed flora and those of Australia and California was made.     

The interactive effects of biochar and cow manure on rice growth and selected properties of salt-affected soil

Salt-affected soil induces detrimental influences on paddy rice (Oryza sativa L.) growth and ameliorating the influences could be done with organic amendments, such as animal manure and biochar. The aims of the current study are: (1) to examine the interactive effects of biochar and cow manure on rice growth and on selected properties of salt-affected soil, and (2) to identify potential mechanisms related to the amendments. Saline-sodic soil was used for a net house experiment with two experimental factors: biochar (no-biochar, rice-husk, and -straw biochar) and cow manure (with and without cow manure). Without the manure, addition of both rice-hush and – straw biochar significantly increased rice growth, whereas a combination of individual biochar with manure did not show a positive synergistic effect. The interactive effect of two factors was not significant on available P and exchangeable K concentrations, but the main effects of the two factors were significant. Biochar addition resulted in higher soil cation exchange capacity (CEC) (28.8 to 29.0 cmol_c kg^?1) than the control (25.6 cmol_c kg^?1), but manure addition did not. Improved nutrient availabilities such as P and K, as well as CEC are among the potential mechanisms accounting for the enhanced rice growth with biochar.     

Marker-Assisted Selection of Xa21 Conferring Resistance to Bacterial Leaf Blight in indica Rice Cultivar LT2

Bacterial leaf blight of rice (BLB), caused by Xanthomonas oryzae pv. oryzae, is one of the most destructive diseases in Asian rice fields. A high-quality rice variety, LT2, was used as the recipient parent. IRBB21, which carries the Xa21 gene, was used as the donor parent. The resistance gene Xa21 was introduced into LT2 by marker-assisted backcrossing. Three Xoo races were used to inoculate the improved lines following the clipping method. Eleven BC₃F₃ lines carrying Xa21 were obtained based on molecular markers and agronomic performance. The 11 lines were then inoculated with the three Xoo races. All the 11 improved lines showed better resistance to BLB than the recipient parent LT2. Based on the level of resistance to BLB and their agronomic performance, five lines (BC₃F₃ 5.1.5.1, BC₃F₃ 5.1.5.12, BC₃F₃ 8.5.6.44, BC₃F₃ 9.5.4.1 and BC₃F₃ 9.5.4.23) were selected as the most promising for commercial release. These improved lines could contribute to rice production in terms of food security.     

Downregulation of Kinesin Spindle Protein Inhibits Proliferation,Induces Apoptosis and Increases Chemosensitivity in Hepatocellular Carcinoma Cells

Background: Kinesin spindle protein (KSP) plays a critical role in mitosis. Inhibition of KSP function leads to cell cycle arrest at mitosis and ultimately to cell death. The aim of this study was to suppress KSP expression by specific small-interfering RNA (siRNA) in Hep3B cells and evaluate its anti-tumor activity. Methods: Three siRNA targeting KSP (KSP-siRNA #1-3) and one mismatched-siRNA (Cont-siRNA) were transfected into cells. Subsequently, KSP mRNA and protein levels, cell proliferation, and apoptosis were examined in both Hep3B cells and THLE-3 cells. In addition, the chemosensitivity of KSP-siRNA-treated Hep3B cells with doxorubicin was also investigated using cell proliferation and clonogenic survival assays. Results: The expression of endogenous KSP at both mRNA and protein levels in Hep3B cells was higher than in THLE-3 cells. In Hep3B cells, KSP-siRNA #2 showed a further downregulation of KSP as compared to KSP-siRNA #1 or KSP-siRNA #3. It also exhibited greater suppression of cell proliferation and induction of apoptosis than KSP-siRNA #1 or KSP-siRNA #3; this could be explained by the significant downregulation of cyclin D1, Bcl-2, and survivin. In contrast, KSP-siRNAs had no or lower effects on KSP expression, cell proliferation and apoptosis in THLE-3 cells. We also noticed that KSP-siRNA transfection could increase chemosensitivity to doxorubicin in Hep3B cells, even at low doses compared to control. Conclusion: Reducing the expression level of KSP, combined with drug treatment, yields promising results for eradicating hepatocellular carcinoma (HCC) cells in vitro. This study opens a new direction for liver cancer treatment. Key Words: Apoptosis, Chemosensitivity, Doxorubicin, Hepatocellular carcinoma (HCC) cells, Kinesin spindle protein (KSP)