Efficient Production of Gossypol from Hairy Root Cultures of Cotton (Gossypium hirsutum L.)

Abstract

A protocol for induction and establishment of Agrobacterium rhizogenes mediated hairy root culture of Gossypium hirsutum was developed through infection with the A4 strain and co-cultivation on hormone-free semi-solid MS medium with B5 vitamins. It resulted in the emergence of hairy roots from the leaf explants, 21 days after infection. The transformation of hairy roots was established by PCR amplification of rol B and rol C genes of the Ri plasmid. All root lines expressed gossypol, although distinct inter-clonal quantitative variations were noticed. Five independent hairy root lines were studied for their growth kinetics as well as gossypol production. The yield potentials of one of them superseded others, as well as the non-transformed, in-vitro grown control roots. The content of gossypol in hairy roots reached a level of 2.43 mg/g DW. It was 4.5 times higher than in vitro and 1.47 times higher than in vivo grown roots of G. hirsutum. Selection of high gossypol producing hairy root lines of G. hirsutum can provide an alternative source for large-scale production of gossypol.

Evaluation of Total Phenolic Compounds and Insecticidal and Antioxidant Activities of Tomato Hairy Root Extract

“…The clonal nature of the individually selected hairy root lines resulting from site-specific insertion of Ri T-DNA, and the difference in the incorporated copy numbers might finally be responsible for the interclonal variation in terms of growth . Profuse adventitious root growth was attained after 21 days of growth in various medicinal plants and reached its maximum up to 35 days of culture . Similar observations of decline in growth were observed in Plumbago zeylanica, Plumbago kurroa, and Gossypium hirsutum. ,, The decline in growth is attributed to the secretion of secondary metabolites and other phenolics in the medium, which hamper the adventitious growth of root hairs.…”

“…Profuse adventitious root growth was attained after 21 days of growth in various medicinal plants and reached its maximum up to 35 days of culture . Similar observations of decline in growth were observed in Plumbago zeylanica, Plumbago kurroa, and Gossypium hirsutum. ,, The decline in growth is attributed to the secretion of secondary metabolites and other phenolics in the medium, which hamper the adventitious growth of root hairs.…”

“…30 Similar observations of decline in growth were observed in Plumbago zeylanica, Plumbago kurroa, and Gossypium hirsutum. 17,30,31 The decline in growth is attributed to the secretion of secondary metabolites and other phenolics in the medium, which hamper the adventitious growth of root hairs.…”

Distinct defensive activity of phenolics and phenylpropanoid pathway genes in different cotton varieties toward chewing pests

“…For histochemical analysis, the fresh young and mature leaves were stained using vanillin-HCl for phenolics and 4-dimethylaminocinnamaldehyde (DMACA) for condensed tannins. 26 Bio-chemical analysis of gossypol, carbohydrate and protein profiling All biochemical assays were executed using Lambda 35 UV/ Vis spectrometer. Extraction and estimation of gossypol were done according to Verma et al (2009).…”

Enhanced Secondary Metabolite Production in Hairy Root Cultures Through Biotic and Abiotic Elicitors

“…Hairy root culture (HRC), however, plays important functions in the enhancement of secondary metabolite production in plants, as it provides controlled conditions for the plant propagation and a very good substitute for the whole plant material extraction [6][7][8][9][10][11], since tissue-specific compounds can easily be extracted by the plant root culture [8,12]. Besides, the higher proliferation rates, controlled conditions, shorter life cycles, etc.…”