Establishment of Persicaria minor hairy roots and analysis of secreted β-caryophyllene in medium broth

Abstract

A method for hairy root induction of Persicaria minor, was developed and investigated, and a qualitative evaluation of the sesquiterpenes in the culture medium was performed. The transgenic status was confirmed by PCR analysis using rolB and virD specific primers. The efficiency with which four different strains of Agrobacterium rhizogenes (A4, ATCC43056, ATCC15834 and ATCC13333) and different concentrations of acetosyringone induced hairy roots from in vitro leaf segments of P. minor was evaluated. Strains A4 and ATCC13333 were able to induce hairy root in an average of 6 days of cocultivation and exhibited significantly higher transformation efficiencies, 20 and 21.3 %, respectively, compared to the other strains in the presence of 100 lM acetosyringone. Half-strength Murashige-Skoog basal medium supplemented with Gamborg vitamins supported the greatest increase (35-fold) of growth index of hairy roots relative to the initial weight after 8 weeks of culture. GC-MS results showed that b-caryophyllene was the main sesquiterpenes detected and was quantified with standards, with total peak areas of 7.934. Results suggested that hairy roots secreted sesquiterpenes into the medium while preserving viable hairy roots as a source of specialized metabolites.

Influence of cotyledon, hypocotyl extracts and authentic andrographolide on selective Agrobacterium rhizogenes strains growth: A deterrent to hairy root induction in Andrographispaniculata (Burm.f.) Wall. ex Nees

“…In the present work, cotyledon explant was found best for transformation; in contrast previous studies showed that hypocotyl explant were successful for transformation experiments (Wahby et al, 2013;Sudha et al, 2013). Leaf explants were used for the induction of hairy roots in Persicaria minor (Ashraf et al, 2015) and Salvia castanea (Li et al, 2016).…”

Potential of the genetically transformed root cultures of Plumbago europaea for biomass and plumbagin production

“…MS medium may be too nutritious for hairy root growth and secondary metabolite accumulation due to the lower concentration of thiamine. Hence, the vitamins of MS medium can be replaced with B5 vitamins . As shown in Figure , hairy roots cultured in the ½ MS‐B5 medium yielded the highest plumbagin content of 3.3 mg g −1 DW (Figure ) in comparison to hairy roots in other media.…”

“…Fresh weight (0.5 ± 0.05 g) of root was moved to 100 mL of liquid 1/2 MS‐B5 medium supplemented with 30 g L −1 sucrose . The cultures were incubated on a rotary shaker in the dark for 6 weeks.…”

“…Generation of transgenic roots might be a promising approach in future owing to their specific characteristics including higher genetic and biochemical constancy, easy maintenance, fast growth, ability to synthesize a range of secondary metabolites, growth in PGRs‐free media, high lateral branching and plagiotropism . As previously reported, numerous elements affect the transformation frequency in various plants such as bacterial strain, type of explant, co‐cultivation media, co‐cultivation period, inoculation media, inoculation temperature, type and concentration of carbohydrate source, seedling age, genotype variation and different nutrient components . In hairy root system, some factors including medium composition, light and temperature have significant impacts on growth and production of natural products, which are frequently species dependent .…”

“…In hairy root system, some factors including medium composition, light and temperature have significant impacts on growth and production of natural products, which are frequently species dependent . Previous reports focused mainly on a number of factors such as composition of nutrients medium (eg, salts concentration), temperature and light to achieve maximum growth rates of roots and efficient biosynthesis of natural products . A few studies have been described the stimulation of transgenic hairy root growth and plumbagin synthesis including Plumbago indica and Plumbago zeylanica .…”