T-DNA Insertion Alters the Terpenoid Content Composition and Bioactivity of Transgenic Artemisia annua

Abstract

In this study, the interference of T-DNA insertion upon Agrobacterium-mediated transformation on the biochemical expression of the host genome is discussed. Plant extracts of transgenic Artemisia annua L. with or without an overexpressed farnesyl pyrophosphate synthase gene have been investigated for their bioactivity and metabolic profile in comparison with wild type A. annua. The highest antimicrobial activity against Staphylococcus aureus, Bacillus subtilis and Candida albicans was observed in the T253 transgenic lines. Moreover, the crude extract from T253 showed higher antimalarial activity against the Plasmodium faciparum K1 strain than those of the others. The terpenoid constituents and antimicrobial properties of the plant samples were grouped by hierarchical clustering analysis. The clustering showed that squalene is a putative compound that might be involved in increasing the bioactivity of the transgenic line. In addition, T253 had a triterpene content that was about twice as great as that of the T253-2 line, which had a higher content of sesquiterpenes. However, both lines were transformed by the same FPS gene. These results suggested that the different bioactive properties observed in each transgenic line may be caused by variations in their terpenoid composition, which is affected by T-DNA insertion at different positions in the host plant.

Variation in terpenoids in leaves of Artemisia annua grown under different LED spectra resulting in diverse antimalarial activities against Plasmodium falciparum

“…A similar result was observed in tomato leaf, with monoterpene α-pinene more abundant under blue spectrum [ 51 ]. In addition to light treatment, alterations of terpenoid composition and content in A. annua were reported by T-DNA insertions at different positions of the host genome [ 52 ]. A similar finding was demonstrated by overexpression of the gibberellin biosynthetic gene, as the proportion of monoterpenes in transgenic plants slightly increased compared with the proportion in wild type [ 37 ].…”

“…Several studies have focused on the synergistic effect of artemisinin with other terpenoid compounds against malarial parasites. A transgenic line with high squalene content but moderate level of artemisinin was reported to exhibit higher antimalarial activity than an artemisinin-rich line, suggesting that squalene was involved in increasing the bioactivity of crude compounds [ 52 ]. Thus, variations of terpenoid compounds in A. annua plants under different light conditions should be further considered for pharmaceutical evaluation.…”

“…GC-MS analysis was performed according to the previously reported protocol [ 52 ] using an Agilent 6890 N network gas chromatograph system (Agilent Technologies Inc., USA) equipped with HP-5 ms GC capillary columns (5% diphenyl 95% dimethylpolysiloxane, 30 m × 0.25 mm, film thickness 0.25 μm) and a 5973 N network mass selective detector (Agilent Technologies Inc., USA). Relative abundance of the compounds was quantified by comparing their peak areas with the internal standard (methyl heptadecanoate, C17) and represented as a relative content (%) [ 37 , 52 , 77 ]. Mass spectral information of the peaks was compared with the mass spectral library (Wiley 7 no.…”

Phytochemical Compounds in Arundo donax L. Rhizome and Antimicrobial Activities

“…The mass range of analyzed compounds detected at m/z 30-500 amu. 7 Screening against the WILEY07 library enabled characterization of the phytochemical compounds from A. donax L., with similarity of at least 90% warranting further analysis.…”