Optimizing culture conditions for establishment of hairy root culture of Semecarpus anacardium L.

Abstract

Semecarpus anacardium L. is a tree species which produces secondary metabolites of medicinal importance. Roots of the plant have been traditionally used in folk medicines. Different strains of Agrobacterium rhizogenes (A4, ATCC15834 and LBA 9402) were used for induction of hairy roots in in vitro grown tissues of the plant. Hairy root initiation was observed after 25–30 days of infection. Optimum transformation frequency of 61% was achieved on leaf explants with ATCC15834 strain. Infection time of 30 min resulted in greater transformation frequency compared to 10 and 20 min, respectively. The hairy roots cultured in growth regulator-free semi-solid woody plant medium differentiated into callus. Whole shoots infected with ATCC 15834 were found to produce more transformants upon co-cultivation for 4 (65%) and 5 (67%) days. Induction of hairy roots in stem explants infected with ATCC 15834 was lower (52%) compared to leaves (62%) after 4 days of co-cultivation. In A4 and LBA9402 strains transformation efficiency was 49 ± 2.8% and 36 ± 5.7% in shoots after 4 days of co-cultivation. Transformation frequency was higher in ATCC15834 strain, irrespective of explants. The hairy roots of S. anacardium elongated slowly upon transfer to half-strength liquid medium. After 3–4 passages in liquid medium slender hairy roots started differentiating which were separated from the original explants. Visible growth of the roots was observed in hormone-free liquid medium after 2–3 months of culturing. Polymerase chain reaction with gene-specific primers from rol A, B and C genes confirms the positive transformation events.

Influence of different Agrobacterium rhizogenes strains on hairy root induction and analysis of phenolic and flavonoid compounds in marshmallow (Althaea officinalis L.)

“…Total flavonoid content was determined following a method by Park et al (2008). In a test tube, 4 ml of methanolic extracts, 0.15 ml of NaNO2 (0.5 M) and 0.15 ml of AlCl3.6H2O (0.3 M) were mixed.…”

“…Several studies have focused on the production, purification and enhancement of these compounds in plants (Pistelli et al 2010;Guerriero et al 2018). Moreover, the extraction of phytochemicals from plant tissues is subject to limitations due to the impact of ecological conditions on their growth and yield (Panda et al 2017).…”

Developing a Sufficient Protocol for the Enhancement of α-Glucosidase Inhibitory Activity by Urena lobata L. Aeroponic Hairy Roots Using Exogenous Factors, a Precursor, and an Elicitor

“…Hairy root induction and root growth is affected by different factors such as plant age, explants, Agrobacterium strains, infection time, media composition, and co-inoculation time [15]. In our previous study, we established a protocol for AHRs cultures of Urena lobata [13].…”

Optimizing hairy root production from explants of Phyllanthus hainanensis, a shrub used for traditional herbal medicine

“…Whereas, prolonged infection time may increase the risk of explant tissues being contaminated by the bacteria, which causes local necrotic lesions or death of the explant. The optimal infection time varies between 10 min for B. monnieri [32] , 20 min for V. vinifera [26] and 30 min for Semecarpus anacardium [33] . Our results showed that hairy root induction had a unimodal pattern with infection time with maximum induction rate (38.9%) recorded at 20 min ( Fig.…”

“…After inoculation, an incubation period is necessary for the Ri plasmid of the bacterium to fully integrate into the genome of the host. The optimal incubation period varies from 2 d for V. vinifera [26] to 4 d for Semecarpus anacardium [33] . In P. hainanensis, incubating the inoculated explants for 0 (control) to 5 d resulted in up to a 14fold variation in hairy root induction rate between treatments (Fig.…”