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.
Highlights
Plants have been tapped as ‘‘chemical factories’’ for their ability to produce phytochemicals of industrial and pharmaceutical importance from simple building blocks
Optimum transformation frequency of 61% was noted in explants of S. anacardium L. infected for 30 min in the bacterial suspension (Table 1)
Researchers have reported effect of infection time on transformation frequency using A. rhizogenes which depends on plant species
Summary
Plants have been tapped as ‘‘chemical factories’’ for their ability to produce phytochemicals of industrial and pharmaceutical importance from simple building blocks. The major drawback of extracting phytochemicals from normal plant tissue is that their growth and yield are significantly impacted due to environmental factors. The complex extraction procedure itself is detrimental to plant survival (Ono and Tian 2011). Various aspects and applications of hairy root cultures (HRCs), include phytochemicals, recombinant protein production, phytoremediation, molecular breeding and crop improvement, introduction of desirable foreign genes, rhizosphere physiology and biochemistry, metabolic engineering, bioreactor design and general overviews of the system (Ono and Tian 2011). HRCs receive more attention as biological matrices for producing valuable metabolites as they have several attractive features, including high genetic stability and relatively fast growth rates. Harvesting the roots for medicinal and chemical uses is destructive for the plants and there has been increasing interest in developing hairy root cultures from several medicinal plant species (Zhou et al 2011)
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