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 elevated CO2 concentration on bulbil germination and early seedling growth in Chinese yam under different air temperatures

The present study investigated the effects of elevated carbon dioxide concentration ([CO₂]) and air temperature on the germination of seed bulbils and the seedling vigour of two Chinese yam lines. Plants were grown under two [CO₂] levels, ambient and elevated (ambient + 200 μmol mol^?1), and two mean air temperature regimes, 22.2 °C (ambient + 1.4 °C) and 25.6 °C (ambient + 5.2 °C). Elevated [CO₂] did not affect bulbil germination under both air temperature regimes. During the early growth stage, the dry weight (DW) of leaves, vines, shoots, roots, belowground parts (roots + tubers) and whole plants were higher under elevated [CO₂] than ambient [CO₂] for both lines under the low- and high-temperature regimes. The values of vigour indexes (index I = germination % × seedling length and index II = germination % × seedling DW) were also higher under elevated [CO₂] than ambient [CO₂] for both lines. These results indicated that Chinese yam seedlings respond positively to elevated [CO₂] during the early growth stage. The above:belowground DW ratios were lower under elevated [CO₂] than ambient [CO₂] in seedlings with very small new tubers for both yam lines, indicating that elevated [CO₂] strongly affected the root growth in the early growth stage. The DWs of post-treatment seed bulbils were higher in the elevated [CO₂] under both air temperature regimes. The results showed that Chinese yam used a smaller amount of the reserves in seed bulbils under elevated [CO₂] than under ambient [CO₂].     

Effects of Fermentation Conditions Using Lactobacillus plantarum on the Charantin,Stigmasterol Glucoside and β-sitosterol Glucoside Contents of Bitter Gourd (Momordica charantia L.) Juice

Plant Foods for Human Nutrition - Bitter gourd fruits contain high amounts of charantin, stigmasterol glucoside and β-sitosterol glucoside, which have been shown to provide health benefits for...     

Differences in Ion Accumulation and Salt Tolerance among Glycine Accessions

Salinity reduces crop yield by limiting water uptake and causing ion‐specific stress. Soybean [Glycine max (L.) Merr.] is sensitive to soil salinity. However, there is variability among soybean genotypes and wild relatives for salt tolerance, suggesting that genetic improvement may be possible. The objective of this study was to identify differences in salt tolerance based on ion accumulation in leaves, stems and roots among accessions of four Glycine species. Four NaCl treatments, 0, 50, 75 and 100 mm , were imposed on G. max, G. soja, G. tomentella and G. argyrea accessions with different levels of salinity tolerance. Tolerant genotypes had less leaf scorch and a greater capacity to prevent Na⁺ and Cl^? transport from soil solution to stems and leaves than sensitive genotypes. Magnitude of leaf injury per unit increase in leaf Na⁺ or Cl^? concentrations was lower in tolerant than in susceptible accessions. Also, plant injury was associated more with Na⁺ rather than with Cl^? concentration in leaves. Salt‐tolerant accessions had greater leaf chlorophyll‐meter readings than sensitive genotypes at all NaCl concentrations. Glycine argyrea and G. tomentella accessions possessed higher salt tolerance than G. soja and G. max genotypes.     

Towards sustainable farming systems: Effectiveness and deficiency of the French procedure of sustainable agriculture

There is a widespread consensus about the importance of sustainable agriculture. In France, the Farming Orientation Law of July 1999 has set down a precise procedure for the implementation of sustainable agriculture: the contrat territorial d’exploitation – or territorial farm contract (TFC). This paper aims to analyse the TFC effectiveness. An analysis has been made of the TFCs signed in the Midi-Pyrenees Region in south-western France, using statistical analysis and qualitative surveys of a sample of farms. The results show that the most valuable effects have been mainly economic. Effects regarding social and environmental aspects were very limited. But from the point of view of an integrated approach, indirect effects of TFCs can be observed on social and environmental aspects. These results are explained, on the one hand, by farmers’ motivations guided by economic objectives, and on the other hand, by the dominance of professional farming organisations in the implementation of TFC’s procedure. Given these results, two ways of improving the elaboration procedure for TFCs are proposed. Firstly, technical improvements can be made to the diagnostic assessment of farms, and to the choice of actions addressed by the contract. Secondly, organisational improvements can be applied to TFCs elaboration and implementation process, in order to adopt a participatory approach which would involve all rural stakeholders.     

Responses in residual feed intake in lines of Large White pigs selected for growth rate on restricted feeding (measured on ad libitum individual feeding)

Residual feed intake (RFI) represents the deviation of the actual ad libitum food consumption of each animal from that predicted from combination of growth rate, backfat and metabolic body weight measurements. After 4 years of divergent selection of pigs in lines for either high or low postweaning growth rate on restricted feeding, the high line exhibited a significant reduction in RFI relative to the low line. This indicated, to some extents, a lower energy requirement for maintenance in the high than in the low line, possibly because of reduced physical activity of the animals. Estimates of genetical parameters showed that RFI was moderately heritable and those genetic correlations of RFI with carcass backfat and food conversion ratio were moderate to highly positive.     

Seasonal effects on the aerobic bacterial and fungal conjunctival flora of normal thoroughbred brood mares in Florida

Objective To evaluate seasonal effects on the presence or absence of fungal and aerobic bacterial flora of the conjunctival fornix of normal Florida Thoroughbred horses. Sample population Both eyes of 100 horses. Procedure Horses with normal anterior segment ophthalmic examinations from three farms in north central Florida were included. Each animal had the ventral conjunctival fornix of each eye swabbed with sterile culturettes. Samples were taken in October, January, April, and July (1999–2000). Aerobic and fungal cultures were plated. Bacterial cultures were reviewed at 24 and 48 h. Fungal cultures were reviewed weekly for 4 weeks. Logistic regression analysis with season as a factor and age of the horse as a covariate was performed. Statistical significance was set at P < 0.01. Results Horses ranged from 3 to 24 years of age, with a median age of 9 years. Twenty‐four genera of bacteria and 35 genera of fungi were recovered. Corynebacterium sp., Staphylococcus sp., Bacillus sp. and Moraxella sp. were the bacteria most frequently isolated. Mold species, dematiaceous mold species, Chrysosporium sp., Cladosporium sp., and Aspergillus sp. were the most frequently recovered fungi. Season did not have a significant effect on the presence of microorganisms isolated for individual horses adjusted for age. Younger horses had an increased incidence of gram‐negative rods and fungal isolates. The number of bacteria and fungi isolated are not uniform across seasons. Conclusion There were no significant differences between the number or type of organisms cultured during the sampling seasons in normal Florida horses. A large range of normal bacterial and fungal flora were isolated from these horses. The number of bacteria and fungi isolated are not uniform across seasons. The likelihood of detecting an organism depends on the horses’ age.     

The epidemiology of nematode and fluke infections in cattle in the Red River Delta in Vietnam

Over a period of 13 months, faecal samples were collected monthly from approximately 45 cattle over 3 months of age. Additionally, 74 calves of 1-2 months were sampled to determine the presence of Toxocara vitulorum eggs. Individual egg counts and infective strongyle larvae from pooled faecal samples were examined. Post-mortem worm counts were carried out on six groups of tracer calves (n=12) that had been kept for 4 weeks on pasture in and around the village studied. The following helminths were identified: T. vitulorum, Cooperia punctata, C. pectinata, C. oncophora, Oesophagostomum radiatum, Trichostrongylus axei, T. colubriformis, Haemonchus spp., Fasciola spp. and Paramphistomum spp. In 8% of the samples collected from young calves, individual egg counts for T. vitulorum were found indicative for pathogenic worm burdens. Strongyle egg counts and worm counts indicated that transmission is low without a distinct seasonality. In animals of 3-9 months old, a strongyle egg count peak can be demonstrated which at a higher age steadily and significantly decreased. In faecal cultures Cooperia spp. were most prominent in all age groups throughout the year with the exception of the period September-November when Haemonchus spp. and Oesophagostomum spp. were most prevalent. Fasciola spp. eggs were found in 22% of the collected faecal samples and the egg counts were low indicating that the intensity of Fasciola spp. infection is mild. Based on the present data, regular anthelmintic treatments seem not to be justified, except for a single treatment at the age of 2 weeks against toxocariosis.