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Abstract

We explored the use of the medicinally important plant Centella asiatica for expression of hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) strain AF2240. HN protein is the principal target for subunit vaccine development against NDV. The full-length HN gene was cloned into a plant expression construct driven by the CaMV 35S promoter and C-terminal fusion of green fluorescence protein (GFP) as reporter system. The recombinant expression construct was transformed via particle bombardment into C. asiatica callus. Transformants were screened using GFP and selected on MS medium supplemented with 15 mg/l hygromycin. The ~1.8 kb HN mRNA transcript was detected on the putative transformants using RT-PCR. The presence of HN protein expression was further confirmed through dot blot analysis using anti-NDV chicken serum. Here we report, for the first time, the use of a novel medicinal plant as a new platform for HN protein expression.
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Abstract

We determined the level of flavonoids, citric acid and ascorbic acid in hips of rose species from the Caninae section occurring in Poland. We performed phytochemical analyses of 75 samples representing 11 species: Rosaagrestis Savi, R. canina L., R. dumalis Bechst., R. glauca Pourret, R. inodora Fries, R. jundzillii Besser, R. rubiginosa L., R. sherardii Davies, R. tomentosa Sm., R. villosa L. and R. zalana Wiesb. Flavonoid content was determined spectrophotometrically, and organic acid concentrations by HPLC. The content of the studied compounds varied greatly. Interspecific differences in the amount of flavonoids and ascorbic acid were highly significant. The most common species, Rosa canina, showed low average content of vitamin C (0.51 g/100 g of dry matter) and flavonoids (41 mg/100 g DM) and high content of citric acid (3.48 g/100 g DM). Ascorbic acid was highest in R. villosa hips (avg. 2.25 g/100 g DM), flavonoids were highest in R. rubiginosa (72 mg/100 g DM), and citric acid was highest in R. tomentosa (4.34 g/100 g DM). Flavonoid level correlated negatively with the amount of citric acid (r=-0.47, p<0.001). Cluster analysis of rose species based on the content of the investigated compounds confirmed the validity of the division of sect. Caninae into three subsections: Rubiginosae, Vestitae and Rubrifoliae. The phytochemical variation of these roses reflects their probable phylogenetic relationships as determined from morphology.
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