Abstract
Plant cell and organ cultures of Helianthella quinquenervis, a medicinal plant whose roots are used by the Tarahumara Indians of Chihuahua, Mexico, to relieve several ailments, were established to identify and quantify some chromenes with biological activity, such as encecalin, and to evaluate their potential for biotechnological production. Gas chromatography–mass spectrometry (GC-MS) analysis corroborated the presence of quantifiable amounts of encecalin in H. quinquenervis cell cultures (callus and cell suspensions). In addition, hairy roots were obtained through three transformation protocols (prick, 45-s sonication and co-culture), using wild type Agrobacterium rhizogenes A4. After three months, cocultivation achieved the highest percentage of transformation (66%), and a comparable production (FW) of encecalin (110 μg/g) than the sonication assay (120 μg/g), both giving far higher yields than the prick assay (19 μg/g). Stable integration of rolC and aux1 genes in the transformed roots was confirmed by polymerase chain reaction (PCR). Hairy roots from cocultivation (six months-old) accumulated as much as 1086 μg/g (FW) of encecalin, over three times higher than the cell suspension cultures. The production of encecalin varied with growth kinetics, being higher at the stationary phase. This is the first report of encecalin production in hairy roots of H. quinquenervis, demonstrating the potential for a future biotechnological production of chromenes.
Highlights
IntroductionGray (Asteraceae) is a perennial herb that grows in montane and subalpine coniferous forest of the Rocky Mountains of the western United States of America and Canada where it is known as the aspen sunflower, and in pine-oak forests of the Sierra Madre of Chihuahua and
Wild plants of H. quinquenervis proved to be susceptible to infection by A. rhizogenes strain A4 using three protocols: prick, sonication and co-culture
Several important chromenes were identified in diverse in vitro cultures: encecalin in both hairy roots and callus cultures, and euparin only in the former. This is the first report of hairy root cultures in H. quinquenervis
Summary
Gray (Asteraceae) is a perennial herb that grows in montane and subalpine coniferous forest of the Rocky Mountains of the western United States of America and Canada where it is known as the aspen sunflower, and in pine-oak forests of the Sierra Madre of Chihuahua and. The Tarahumara Indians of Chihuahua know it as rarésoa [2] or corsonero (probably derived from escorzonera) and employ it medicinally. Dry and powdered roots are used to alleviate coughs and muscle aches and to treat gastrointestinal diseases and ulcers [3]. Crude extracts of H. quinquenervis roots have shown antimicrobial activity against Trichophyton mentagrophytes, one of the fungi that cause ringworm infections in humans [4], and antiprotozoal activity [5].
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