<i>Agrobacterium rhizogenes</i>-Mediated Transformation of<i>Artemisia annua</i>: Production of Transgenic Plants

Banerjee, Santanu; Zehra, M.; Gupta, Madhuri; Kumar, Susheel

doi:10.1055/s-2006-957737

Cited by 23 publications

(13 citation statements)

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“…However, ALDH1 was almost not detectable in hairy roots, which may lead to the formation of artemisinic acid and arteannuin B instead of artemisinin. These compounds have been isolated from hairy root cultures of A. annua [45]. …”

Section: Resultsmentioning

confidence: 99%

Relative expression of genes of terpene metabolism in different tissues of Artemisia annuaL

et al. 2011

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BackgroundRecently, Artemisia annua L. (annual or sweet wormwood) has received increasing attention due to the fact that the plant produces the sesquiterpenoid endoperoxide artemisinin, which today is widely used for treatment of malaria. The plant produces relatively small amounts of artemisinin and a worldwide shortage of the drug has led to intense research in order to increase the yield of artemisinin. In order to improve our understanding of terpene metabolism in the plant and to evaluate the competition for precursors, which may influence the yield of artemisinin, we have used qPCR to estimate the expression of 14 genes of terpene metabolism in different tissues.ResultsThe four genes of the artemisinin biosynthetic pathway (amorpha-4,11-diene synthase, amorphadiene-12-hydroxylase, artemisinic aldehyde ∆11(13) reductase and aldehyde dehydrogenase 1) showed remarkably higher expression (between ~40- to ~500-fold) in flower buds and young leaves compared to other tissues (old leaves, stems, roots, hairy root cultures). Further, dihydroartemisinic aldehyde reductase showed a very high expression only in hairy root cultures. Germacrene A and caryophyllene synthase were mostly expressed in young leaves and flower buds while epi-cedrol synthase was highly expressed in old leaves. 3-Hydroxy-3-methyl-glutaryl coenzyme A reductase exhibited lower expression in old leaves compared to other tissues. Farnesyldiphosphate synthase, squalene synthase, and 1-deoxy-D-xylulose-5-phosphate reductoisomerase showed only modest variation in expression in the different tissues, while expression of 1-deoxy-D-xylulose-5-phosphate synthase was 7-8-fold higher in flower buds and young leaves compared to old leaves.ConclusionsFour genes of artemisinin biosynthesis were highly expressed in flower buds and young leaves (tissues showing a high density of glandular trichomes). The expression of dihydroartemisinic aldehyde reductase has been suggested to have a negative effect on artemisinin production through reduction of dihydroartemisinic aldehyde to dihydroartemisinic alcohol. However, our results show that this enzyme is expressed only at low levels in tissues producing artemisinin and consequently its effect on artemisinin production may be limited. Finally, squalene synthase but not other sesquiterpene synthases appears to be a significant competitor for farnesyl diphosphate in artemisinin-producing tissues.

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Section: Resultsmentioning

confidence: 99%

Relative expression of genes of terpene metabolism in different tissues of Artemisia annuaL

et al. 2011

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“…1B and C; Table 1). Spontaneous regeneration of plants from hairy root cultures has already been reported in Pelargonium (Pellegrineschi et al 1994;Pellegrineschi and Mariani 1996;Banerjee et al 1997; Perez-Morphe-Balch and Ochoa-Alejo 1987).…”

Section: Resultsmentioning

confidence: 77%

Rose-scented geranium (Pelargonium sp.) generated by Agrobacterium rhizogenes mediated Ri-insertion for improved essential oil quality

Saxena

Banerjee

Laiq-ur-Rahman³

et al. 2007

Plant Cell Tiss Organ Cult

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Transgenic plants of rose-scented geranium (Pelargonium graveolens cv. Hemanti) have been produced from Agrobacterium rhizogenes (strains A 4 and LBA9402) mediated hairy root cultures. Amongst the explants tested, leaves were most responsive followed by the petioles and internodal segments, respectively. The A 4 strain performed better for all the three explants both in terms of frequency of response and time requirement for hairy root induction. Transgenic shoots could be obtained by spontaneous regeneration without intervening callus phase amongst 16% and 12% root lines of A 4 and LBA 9402 origin, respectively, or they were induced in 29% and 22% hairy root lines of A 4 and LBA9402 origin, respectively, with different hormonal supplementation. These transgenic plants showed 30% survival as against 90% of their control under the confined environment of glasshouse. The transgenic plants were of similar morphotype having increased branching, higher number of leaves with increased dentations, short and round stature, highly branched root system and absence of leaf wrinkling. These transgenic plants showed opine positive results even after 5 months of their transfer to the glasshouse. The essential oil compositions of 81% of these transgenics were qualitatively similar to that of the wild type parent. However, two transgenic plants showed increase in concentrations of geraniol and geranyl esters signifying improved oil quality with respect to the citronellol:geraniol ratio. These two oils having better olfactory value represent an improvement over that of the wild type parent from the commercial point of view.

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“…However, only a few studies so far have been carried out to study the regulation of this pathway by TFs-encoding genes. Since the first reports of successful Agrobacterium -mediated transformation of A. annua in the 1990s ( Vergauwe et al, 1996 ; Banerjee et al, 1997 ), these transformation protocols have been optimized ( Han et al, 2005 ). Generally, TFs regulate the expression of a certain number of genes from specific and/or related pathways ( Borevitz et al, 2000 ); therefore loss and gain of the function of TFs has arisen as a promising biofarming approach for more efficiently regulation of secondary metabolite production ( Verpoorte and Memelink, 2002 ; Petersen, 2007 ) ( Figure 3 ).…”

Section: Artemisia Annua Biofarming Approaches Using Metabolmentioning

confidence: 99%

Molecular Farming in Artemisia annua, a Promising Approach to Improve Anti-malarial Drug Production

Pulice¹,

Pelaz

Matías‐Hernández

2016

Front. Plant Sci.

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Malaria is a parasite infection affecting millions of people worldwide. Even though progress has been made in prevention and treatment of the disease; an estimated 214 million cases of malaria occurred in 2015, resulting in 438,000 estimated deaths; most of them occurring in Africa among children under the age of five. This article aims to review the epidemiology, future risk factors and current treatments of malaria, with particular focus on the promising potential of molecular farming that uses metabolic engineering in plants as an effective anti-malarial solution. Malaria represents an example of how a health problem may, on one hand, influence the proper development of a country, due to its burden of the disease. On the other hand, it constitutes an opportunity for lucrative business of diverse stakeholders. In contrast, plant biofarming is proposed here as a sustainable, promising, alternative for the production, not only of natural herbal repellents for malaria prevention but also for the production of sustainable anti-malarial drugs, like artemisinin (AN), used for primary parasite infection treatments. AN, a sesquiterpene lactone, is a natural anti-malarial compound that can be found in Artemisia annua. However, the low concentration of AN in the plant makes this molecule relatively expensive and difficult to produce in order to meet the current worldwide demand of Artemisinin Combination Therapies (ACTs), especially for economically disadvantaged people in developing countries. The biosynthetic pathway of AN, a process that takes place only in glandular secretory trichomes of A. annua, is relatively well elucidated. Significant efforts have been made using plant genetic engineering to increase production of this compound. These include diverse genetic manipulation approaches, such as studies on diverse transcription factors which have been shown to regulate the AN genetic pathway and other biological processes. Results look promising; however, further efforts should be addressed toward optimization of the most cost-effective biofarming approaches for synthesis and production of medicines against the malaria parasite.

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Agrobacterium rhizogenes-Mediated Transformation ofArtemisia annua: Production of Transgenic Plants

Cited by 23 publications

References 9 publications

Relative expression of genes of terpene metabolism in different tissues of Artemisia annuaL

Relative expression of genes of terpene metabolism in different tissues of Artemisia annuaL

Rose-scented geranium (Pelargonium sp.) generated by Agrobacterium rhizogenes mediated Ri-insertion for improved essential oil quality

Molecular Farming in Artemisia annua, a Promising Approach to Improve Anti-malarial Drug Production

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