Optimization of micropropagation protocol and elicitation of luteolin and rutin biomolecules using phenylalanine and chitosan in root callus of Rumex hastatus D. Don

Abstract

Rumex hastatus D. Don, a traditional medicinal plant known for its, antitumorous, antiangiogenic and antibacterial potential is continuously under threat of extinction due to over exploitation and climatic changes. Phytochemical analysis of different plant parts showed that roots had the maximum quantity of two bioactive molecules, luteolin and rutin and thus was selected for establishment of aseptic culture.In vitro technology has been employed for developing an e cient micropropagation protocols for its conservation, large scale production as well elicitation of the valuable bioactive compounds. Murashige and Skoog's medium supplemented with 5µM TDZ proved best for developing greenish-brown and compact calluses in 98% root cultures within 30 days. Such callus pieces on subculture to same but fresh medium differentiated an average of 10.50 ± 1.29 shoots which were elongated on MS basal medium. Such excised shoots, rooted in MS + 0.1 µM IBA medium with an average of 12.03 ± 2.49 roots per shoot.These plantlets were successfully acclimatized under greenhouse conditions and elds and their genetic delity was evaluated employing SRAP and SCoT markers which revealed their high similarity with mother plant proving their true-to-type nature. Interestingly, the in vitro regenerants showed elevated levels of phenols, avonoids, luteolin and rutin content and higher antioxidant activities over the mother plant as done by phosphomolybdenum and DPPH assays. For elicitation of bioactive compounds, 100 mg/L phenylalanine was best for enhancing optimum luteolin content (407.18 ± 10.58 µg/g d.w.) over control (22.10 ± 0.57 µg/g d.w.) almost, 17.42-fold, whereas for rutin, 25 mg/L chitosan proved most effective enhancing its synthesis up to 1169.07 ± 46.49 µg/g d.w. over control (312.41 ± 12.42 µg/g d.w.), i.e., 2.74-fold. To the best of our knowledge, this is our rst report on micropropagation and elicitation of luteolin and rutin in Rumex hastatus through root culture.

Critical review on Rumex dentatus L. a strong pharmacophore and the future medicine: Pharmacology, phytochemical analysis and traditional uses

“… Roots Used for treating astringent, cutaneous and purgative disorders Chopra et al, 1986. [ 53 ], Manandhar et al, 2002 [ 54 ] Root Paste Used to relieve headache Manandhar, 2002 [ 54 ] Roots and Leaves Used to treat asthma, jaundice, foot and mouth infections, fever, cough, weakness and scabies Pakistan Sing et al, 2023 [ 61 ] Whole plant Anti-inflammatory, astringent, antitumor, and diuretic, antidermatitis, cholagogue, tonic and laxative agents Suleyman et al, 1999 [ 62 ]., Litvinenko and Muzych Kina, 2003 [ 63 ], Demirezer, 1993 [ 64 ] Roots Used as a folk medicine for treating ascariasis, eczema, diarrhea and constipation. Olowokudejo et al, 2008 [ 65 ] Rumex dentatus Roots and leaves bactericidal, anti-inflammatory, antitumor, anthelmintic, astringent, and anti-dermatitis, its roots are also used in folk medicine for treating acariasis, eczema, diarrhea, and constipation Egypt Abou et al, 2013 [ 66 ] Roots and leaves Bactericidal, astringent, anti-dermatitis, diuretic, cholagogue, tonic and laxative agents India Humeera et al, 2013 [ 50 ] …”