Impact of Exogenous Elicitors on Artemisinin Production and Trichome Density in &lt;i&gt;Artemisia annua&lt;/i&gt; L. under Subtropical Conditions

Dangash, Alka; Pandya, Neeta; Bharillya, Ashish; Jhala, Ashween; JAIN, Dharamchand

doi:10.15835/nsb639109

Cited by 6 publications

(4 citation statements)

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“…Glandular trichomes are the sites of production of important phytochemicals, including artemisinin, as well as many others that have found numerous applications in the pesticide, pharmaceutical and flavor industries [ 50 , 51 ]. Artemisinin content has been directly related to the trichome index in previous studies [ 52 , 53 ].…”

Section: Discussionmentioning

confidence: 97%

Genetic Transformation of Artemisia carvifolia Buch with rol Genes Enhances Artemisinin Accumulation

et al. 2015

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The potent antimalarial drug artemisinin has a high cost, since its only viable source to date is Artemisia annua (0.01–0.8% DW). There is therefore an urgent need to design new strategies to increase its production or to find alternative sources. In the current study, Artemisia carvifolia Buch was selected with the aim of detecting artemisinin and then enhancing the production of the target compound and its derivatives. These metabolites were determined by LC-MS in the shoots of A. carvifolia wild type plants at the following concentrations: artemisinin (8μg/g), artesunate (2.24μg/g), dihydroartemisinin (13.6μg/g) and artemether (12.8μg/g). Genetic transformation of A. carvifolia was carried out with Agrobacterium tumefaciens GV3101 harboring the rol B and rol C genes. Artemisinin content increased 3-7-fold in transgenics bearing the rol B gene, and 2.3-6-fold in those with the rol C gene. A similar pattern was observed for artemisinin analogues. The dynamics of artemisinin content in transgenics and wild type A.carvifolia was also correlated with the expression of genes involved in its biosynthesis. Real time qPCR analysis revealed the differential expression of genes involved in artemisinin biosynthesis, i.e. those encoding amorpha-4, 11 diene synthase (ADS), cytochrome P450 (CYP71AV1), and aldehyde dehydrogenase 1 (ALDH1), with a relatively higher transcript level found in transgenics than in the wild type plant. Also, the gene related to trichome development and sesquiterpenoid biosynthesis (TFAR1) showed an altered expression in the transgenics compared to wild type A.carvifolia, which was in accordance with the trichome density of the respective plants. The trichome index was significantly higher in the rol B and rol C gene-expressing transgenics with an increased production of artemisinin, thereby demonstrating that the rol genes are effective inducers of plant secondary metabolism.

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Section: Discussionmentioning

confidence: 97%

Genetic Transformation of Artemisia carvifolia Buch with rol Genes Enhances Artemisinin Accumulation

et al. 2015

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“…As mentioned in a previous report that glandular trichomes are the sites of production of important phytochemicals including artemisinin and a mixture of chemicals that have been found to have an enormous array of uses in the pesticides, pharmaceutical and flavor industries [ 43 , 44 ]. Artemisinin content was found to be directly related to the trichome index in previous reports [ 45 , 46 ].…”

Section: Resultsmentioning

confidence: 73%

Enhanced artemisinin yield by expression of rol genes in Artemisia annua

Dilshad

Cusidó

Palazón

et al. 2015

Malar J

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BackgroundDespite of many advances in the treatment of malaria, it is still the fifth most prevalent disease worldwide and is one of the major causes of death in the developing countries which accounted for 584,000 deaths in 2013, as estimated by World Health Organization. Artemisinin from Artemisia annua is still one of the most effective treatments for malaria. Increasing the artemisinin content of A. annua plants by genetic engineering would improve the availability of this much-needed drug.MethodsIn this regard, a high artemisinin-yielding hybrid of A. annua produced by the centre for novel agricultural products of the University of York, UK, was selected (artemisinin maximally 1.4 %). As rol genes are potential candidates of biochemical engineering, genetic transformation of A. annua with Agrobacterium tumefaciens GV3101 harbouring vectors with rol B and rol C genes was carried out with the objective of enhancement of artemisinin content. Transgenic lines produced were analysed by the LC–MS for quantitative analysis of artemisinin and analogues. These high artemisinin yielding transgenics were also analysed by real time quantitative PCR to find the molecular dynamics of artemisinin enhancement. Genes of artemisinin biosynthetic pathway were studied including amorphadiene synthase (ADS), cytochrome P450, (CYP71AV1) and aldehyde dehydrogenase 1 (ALDH1). Trichome-specific fatty acyl-CoA reductase 1(TAFR1) is an enzyme involved in both trichome development and sesquiterpenoid biosynthesis and both processes are important for artemisinin biosynthesis. Thus, real time qPCR analysis of the TAFR1 gene was carried out, and trichome density was determined.ResultsTransgenics of rolB gene showed two- to ninefold (the decimal adds nothing in the abstract, please simplify to two- to ninefold) increase in artemisinin, 4–12-fold increase in artesunate and 1.2–3-fold increase in dihydroartemisinin. Whereas in the case of rol C gene transformants, a fourfold increase in artemisinin, four to ninefold increase in artesunate and one- to twofold increase in dihydroartemisinin concentration was observed. Transformants with the rol B gene had higher expression of these genes than rol C transformants. TAFR1 was also found to be more expressed in rol gene transgenics than wild type A. annua, which was also in accordance with the trichome density of the respective plant.ConclusionThus it was proved that rol B and rol C genes are effective in the enhancement of artemisinin content of A. annua, rol B gene being more active to play part in this enhancement than rol C gene.

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“…Larger leaf size and glandular trichome size in polyploid plants indicate the possibility of polyploid plants producing higher artemisinin. The study by Dangash et al [37] concluded that artemisinin secreted in glandular trichomes, the trichome density, and trichome size are directly proportional to the percent artemisinin content. So, the higher the trichome, the amount and size, the higher will be artemisinin content.…”

Section: The Effect Of Polyploidy Induction On Artemisinin Content In...mentioning

confidence: 99%

Changing of Morphological, Anatomical, Cytological Characteristic and Artemisinin Content in Artemisia cina by Colchicine Treatment

Herawati¹,

Pudjihartati²,

Setiawan³

2022

Proceedings of the 4th International Conference on Life Sciences and Biotechnology (ICOLIB 2021)

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The induction of artificial polyploidy has been considered a valuable tool that can improve many plants' genetics and change their morphological, anatomical, and physiological characteristics. Artemisia cina plants' polyploidy is induced through shoot culture to increase artemisinin content. This research consisted of stages of explant propagation, polyploid induction, and acclimatization. The colchicine concentrations of 100 mg/L, 150 mg/L, 200 mg/L and application times of 24 h, 48 h, 72 h, and 96 h were the treatments given. Descriptive analysis was used to analyze the chromosome determination. The number of alive explants after induction was analyzed at a 5% level using analysis of variant (ANOVA). The paired T-test was also conducted at a 5% level to determine the significance of the plant height, leaf area, stomata, chlorophyll content, glandular trichomes density, and glandular trichomes size and content of artemisinin between the two treatments on the observed data. In Colchicines, treatment with concentrations of 100, 150, and 200 mg/L and time application of 24, 48, 72, and 96 h can induce polyploidy on A. cina. Colchicines' improved concentration and application time are of lower percentage in living explants. Eight plants are the highest number of polyploidy crops on colchicines treatment of 100 mg/L for 72 h. The resulting ploidy level varies on 2n = 2x = 16, 2n = 3x = 24, 2n = 4x = 36, 2n = 5x = 40, and 2n = 6x = 48. The highest level of polyploidy 2n = 4x = 32 is 20% in the treatment Colchicines 100 mg/L for 72 h. The modal number of chromosome x obtained in this study was = 8. The ploidy level's highest plant height, leaf area, chlorophyll content, and stomata sizes were 2n = 4x = 32. The number of stomata and glandular trichomes on polyploidy plants was smaller than the diploid plants. Polyploidy plants generally have larger leaves, stomata, and glandular trichome sizes than diploid plants. However, the number of stomata and glandular trichomes is less than in diploid plants. Artemisinin content on polyploidy plants is higher than on diploid plants.

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Impact of Exogenous Elicitors on Artemisinin Production and Trichome Density in <i>Artemisia annua</i> L. under Subtropical Conditions

Cited by 6 publications

References 20 publications

Genetic Transformation of Artemisia carvifolia Buch with rol Genes Enhances Artemisinin Accumulation

Genetic Transformation of Artemisia carvifolia Buch with rol Genes Enhances Artemisinin Accumulation

Enhanced artemisinin yield by expression of rol genes in Artemisia annua

Changing of Morphological, Anatomical, Cytological Characteristic and Artemisinin Content in Artemisia cina by Colchicine Treatment

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