In vitro biological screening of a critically endangered medicinal plant, Atropa acuminata Royle Ex Lindl of north western Himalaya.

Atropa acuminata Royle Ex Lindl (Atropa acuminata) under tremendous threat of extinction in its natural habitat. However, the antimicrobial, antileishmanial and anticancer effects of the plant's extracts have not been reported yet. In the current study, an attempt has been made to evaluate the pharmacological potential of this plant's extracts against microbes, Leishmania and cancer. The roots, stems and leaves of Atropa acuminata were ground; then, seven different solvents were used alone and in different ratios to prepare crude extracts, which were screened for pharmacological effects. The aqueous, methanolic and ethanolic extracts of all parts carried a broad spectrum of anti-bacterial activities, while no significant activity was observed with combined solvents. Three types of cytotoxicity assays were performed, i.e., haemolytic, brine shrimp and protein kinase assays. The aqueous extract of all the parts showed significant haemolytic activity while n-hexane extracts of roots showed significant activity against brine shrimp. The acetone extracts strongly inhibited protein kinase while the methanolic extracts exhibited significant cytotoxic activity of roots and stem. The anti-leishmanial assays revealed that the methanolic extract of leaves and roots showed significant activity. These findings suggest that this plant could be a potential source of natural product based drugs.

Microclimatic variation in UV perception and related disparity in tropane and quinolizidine alkaloid composition of Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger.

The aim of current research was to evaluate the physiological adjustment in three medicinal herbs viz., Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger to the winter period characterised by intense UV flux in Kashmir valley across the North Western Himalaya. Quinolizidine (QA) and tropane alkaloid (TA) concentrations were analysed in these herbs thriving at two different altitudes via GC-MS and correlated by PCA analysis. This study investigated the hypothesis that UV reflectance and absorbance at low temperatures are directly related to disparity in alkaloid accumulation. Among QAs in L. polyphyllus, ammodendrine and lupanine accumulated at higher concentration and exhibited significant variation of 186.36% and 95.91% in ammodendrine and lupanine respectively in both sites. Tetrahydrohombifoline displayed non-significant variation of about 9.60% irrespective of sites. Among tropane alkaloid (TA), hyoscyamine was recorded as the most abundant constituent irrespective of the plant and site while apotropine accumulated in lesser quantity in A. acuminata than H. niger. However, apotropine demonstrated significant variation of 175% among both sites. The final concentration of quinolizidine (QA) and tropane alkaloid (TA) reflects the interplay between reflectance and absorbance of UV radiation response field. These findings suggest that spectral response of UV light contributes directly to alkaloid biosynthesis.

Enhanced secretory activity of Atropa belladonna hairy root culture over-expressing ADP-ribosylation factor gene.

Agrobacterium-mediated transformation of leaf tissues of Atropa belladonna with an adenosine 5'-diphosphate (ADP)-ribosylation factor gene of carrot, arf-001, was performed employing pBCR82 as an expression vector. This vector co-expresses rol gene cluster together with arf-001, and thus, the transformed host cells were obtained as hairy roots. Two cell lines of the transformed belladonna were established as the liquid cultures of hairy root tissues, and expression of arf-001 and accumulation of its product in the cells were confirmed by RT-PCR and Western blot analyses, respectively. A marked increase in extracellular protein concentrations was observed in the transformed belladonna root cultures as compared with the controls transformed with an empty vector. However, the secretion of the proteins of the transformants was markedly reduced in the presence of a physiological concentration of monensin. These results suggest that over-expression of arf-001 in belladonna results in the enhancement of secretory activity in the transformed cells.