Agrobacterium rhizogenes-mediated transformation of a multi-medicinal herb,Boerhaavia diffusaL.: optimization of the process and anti-microbial activity against bacterial pathogens causing urinary tract infections

Abstract

Hairy root cultures (HRCs) of Boerhaavia diffusa L., an ethno-medicinally important herb, were established via explant co-cultivation with Agrobacterium rhizogenes. Several key factors influencing transformation efficiency were optimized. Rhizoclones showing prolific growth were maintained through successive subcultures in solidified Murashige & Skoog's medium without growth regulators (MS0) under diffused illumination. PCR analysis of the rhizoclones demonstrated positive amplification of rolB and rolC genes of pRi T L -DNA. Reverse transcription-PCR (RT-PCR) and opine assay revealed the presence and expression of the pRi T R -DNA borne ags gene. The methanol and dichloromethane (M:DM) extracts of hairy roots (HRs) and natural roots (NRs) were screened against a wide range of bacterial pathogens causing human urinary tract infections (UTIs). The MIC and MBC values of HRs were markedly lower when compared to NRs against microorganisms causing UTIs. Extracts of HRs were more effective than those of NRs not only against wild type strains but also, and more importantly, against multi-drug resistant strains. Establishment of fast-proliferating renewable HRCs of B. diffusa would provide a direct source of anti-microbial phytocompounds against several pathogenic bacteria, including resistant strains, causing UTIs.

Advances in Boerhaavia diffusa hairy root technology: a valuable pursuit for identifying strain sensitivity and up-scaling factors to refine metabolite yield and bioactivity potentials

“…The presently observed exclusive susceptibility of leaf explants and total failure of stem segments towards A. rhizogenes-mediated hairy root induction contradict the earlier findings of Sahu et al (2013Sahu et al ( , 2014, who have demonstrated notable susceptibility of intermodal segments of in vivo-grown B. diffusa plants towards the peak limit of hairy root induction. The disparity in the maturity levels of the utilized explants could be one of the causes for the observed difference in susceptibility of the nodal segments, as the maturation status of explants predominantly impacts the availability of functionally competent cells (as well as their internal hormonal levels), which essentially serve as ideal targets for hairy root induction (Potrykus 1990).…”

“…Different solvents have been used to screen the antibacterial property of both root and aerial parts of B. diffusa with variable results involving both Gram-negative and Gram-positive bacteria (Murti et al 2010;Ramachandra et al 2012;Malhotra et al 2013;Sahu et al 2013). Enhanced antimicrobial activity of RBH hairy root clone than that of its control roots against S. aureus has corroborated a previous report demonstrating better performance of Maytenus senegalensis hairy roots than the control roots against the same bacterium (Jain et al 2008).…”

“…It is however disheartening to note that the hairy root culture system is relatively underutilized in B. diffusa and phytochemically the same remained uncharted so far. Only a handful of reports are presently available concerning the establishment of both adventitious root culture as well as A. rhizogenes-mediated hairy root cultures in this plant (Shrivastava and Padhya 1995;Jenifer et al 2012;Sahu et al 2013), which assigns ample scope for further studies.…”

Biological investigation of novel metal complexes of 2-amino-4-substituted phenylthiazole Schiff bases

“…The faecal collected from Institute of Medical Sciences & SUM Hospital, Bhubaneswar, Odisha. Cryptococcus neoformans was obtained from the University Department of Pharmaceutical Sciences, Utkal University 22 . Amoxicillin (Reference Antibiotic: RA) and ampicillin (RA 1 ) were used as reference antibacterial antibiotics, and fluconazole (RA) was used as a reference antifungal antibiotic.…”

Determination of in Vivo Biological Activities of Dodonaea Viscosa Flowers Against CCL4 Toxicity in Albino Mice With Bioactive Compound Detection

“…One of the targets of mission 2035 by WHO is to make the world TB free by the end of 2035 leaving no patient with persistent TB [19]. Many plant species have always shown potential in ancient history to treat the medical disorder with promising therapeutic agents [20]. The knowledge about single chemical component encompassed by medicinal plant plays a critical role from method of extraction to understanding pharmacological assay and possible toxicity.…”