Induction of Pyridine Alkaloid Formation in Transformed Root Cultures of Nicotiana tabacum

Hairy root cultures of Nicotiana tabacum were set up by excised root tips with Agrobacterium rhizogenes. The successful transformation was confirmed by analyzing rolC and virC genes using polymerase chain reaction (PCR). Hairy root cultures were employed to study the formation of pyridine alkaloids, mainly nicotine. The transformed cultures were incubated with potential elicitors, such as methyljasmonate, quercetin and salicylic acid, in order to stimulate the biosynthesis of pyridine alkaloids. Profile and amounts of pyridine alkaloids were analyzed using capillary GLC-MS. Treatment of the cultures with methyljasmonate (50 μM) increased the alkaloid accumulation ca. 7-fold up to a level of 0.58 mg/g fresh weight as compared to untreated controls. Quercetin (200 μM) enhanced the alkaloid production ca. 4-fold (0.34 mg/g fresh weight) within 24 h. In contrast salicylic acid in all tested concentrations decreased the alkaloid level to 1 μg/g fresh weight. Also the inhibitory effect of salicylic acid on the elicitation effect of methyljasmonate and quercetin was investigated.

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Induction of Tropane Alkaloid Formation in Transformed Root Cultures of Brugmansia suaveolens (Solanaceae)

Zeitschrift für Naturforschung C ◽

10.1515/znc-2004-11-1216 ◽

2004 ◽

Vol 59 (11-12) ◽

pp. 863-867 ◽

Cited By ~ 25

Author(s):

Rawia Zayed ◽

Michael Wink

Keyword(s):

Salicylic Acid ◽

Hairy Root ◽

Tropane Alkaloids ◽

Tropane Alkaloid ◽

Hairy Root Cultures ◽

Root Cultures ◽

Fresh Weight ◽

Root Tips ◽

Brugmansia Suaveolens ◽

Set Up

Hairy root cultures of Brugmansia suaveolens were set up by infection of root tips with Agrobacterium rhizogenes. The successful transformation was confirmed by analysing rolC and virC genes using polymerase chain reaction (PCR). Hairy root cultures were employed to study the formation of tropane alkaloids, such as hyoscyamine. The transformed cultures were incubated with potential elicitors, such as methyljasmonate, quercetin and salicylic acid in order to stimulate the biosynthesis of tropane alkaloids. Profile and amounts of tropane alkaloids were analysed using capillary GLC-MS. At least 18 different tropane alkaloids could be identified. Treatment of the cultures with 200 μᴍ methyljasmonate increased the alkaloid accumulation 25-fold up to a level of 1 mg/g fresh weight as compared to untreated controls. Quercetin enhanced the alkaloid production 10 fold (0.4 mg/g fresh weight) within 24 h. In contrast 100 μᴍ salicylic acid decreased alkaloids to a level of 1 μg/g fresh weight.

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Hairy Roots

Advances in Environmental Engineering and Green Technologies - Molecular Plant Breeding and Genome Editing Tools for Crop Improvement ◽

10.4018/978-1-7998-4312-2.ch008 ◽

2021 ◽

pp. 219-252

Keyword(s):

Secondary Metabolites ◽

Hairy Root ◽

Hairy Roots ◽

High Growth ◽

Root Culture ◽

Hairy Root Culture ◽

High Growth Rate ◽

Hairy Root Cultures ◽

Root Cultures ◽

Transformed Root Cultures

Agrobacterium rhizogenes induces hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection, when cultured in hormone free medium, show high growth rate and genetic stability. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Several elicitation methods can be used to further enhance the production and accumulation of secondary metabolites. Thus, hairy root culture offer promise for high production and productivity of valuable secondary metabolites in many plants. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for pharmaceutical industry. Hairy root cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites, and for phytoremediation due to their abundant neoplastic root proliferation property. Various applications of hairy root cultures and potential problems associated with them are discussed in this chapter.

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Targeted profiling reveals metabolic perturbations in cryptogein-cotransformed hairy root cultures of Nicotiana tabacum

Acta Physiologiae Plantarum ◽

10.1007/s11738-020-03155-4 ◽

2020 ◽

Vol 42 (11) ◽

Author(s):

Mahesh Kumar ◽

Sumita Jha ◽

Adinpunya Mitra

Keyword(s):

Nicotiana Tabacum ◽

Hairy Root ◽

Hairy Root Cultures ◽

Root Cultures ◽

Targeted Profiling

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Enhanced production of valtrate in hairy root cultures of Valeriana jatamansi Jones by methyl jasmonate, jasmonic acid and salicylic acid elicitors

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha48211891 ◽

2020 ◽

Vol 48 (2) ◽

pp. 839-848

Author(s):

Shuang ZHAO ◽

Hong TANG

Keyword(s):

Salicylic Acid ◽

Jasmonic Acid ◽

Methyl Jasmonate ◽

Hairy Root ◽

Hairy Roots ◽

Hairy Root Cultures ◽

Root Cultures ◽

Potent Inducer ◽

Valeriana Jatamansi ◽

Pharmacologically Active

Valtrate is a pharmacologically active epoxyiridoid ester found in the roots and rhizomes of Valeriana jatamansi Jones. The plant produces only small amounts of this metabolite naturally, and so induction of hairy roots as well as elicitation can be useful to increase its commercial production. In this study, strain R1601 of Agrobacterium rhizogenes was used to induce hairy roots in V. jatamansi, and stable hairy root cultures of V. jatamansi were established successfully. The influence of three exogenous elicitors including methyl jasmonate (MJ), jasmonic acid (JA) and salicylic acid (SA) on valtrate production in the hairy root cultures of V. jatamansi was also investigated, and the 25-day-old hairy root cultures were treated with different concentrations of the elicitors at exposure time of 7 days. This present study showed that MJ (100 mg/L) highly promoted valtrate production at 7 days after elicitation, to a level of 3.63 times higher than that of non-elicited control. SA did not significantly increase the production of valtrate. This is the first-time study to assess the elicitation of hairy root cultures to promote valtrate biosynthesis in V. jatamansi and the resulting experiments demonstrated that MJ was indeed a potent inducer of valtrate biosynthesis.

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