Influence of
            <i>rol</i>
            Genes for Enhanced Biosynthesis of Potent Natural Products

Dilshad, Erum; Noor, Huma; Nosheen, Nabgha; Gilani, Syeda Rehab; Ali, Umar; Khan, Mubarak Ali

doi:10.1002/9781119640929.ch13

Cited by 5 publications

(5 citation statements)

References 71 publications

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“…rolA is found on all Ri plasmids and encodes a small protein with a basic isoelectric point whose expression showed a dramatic reduction in several classes of hormones (Ozyigit et al, 2013). rolB may play a critical role in the early stages of hairy-root induction and is the most powerful inducer of secondary metabolism (Dilshad et al, 2021). rolC is considered the most conserved of all rol genes and has a minor impact on root formation (Makhzoum et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

“…Most studies on the expression of rol and aux genes in hairy root cultures have focused on demonstrating their effect on specialized metabolism for increasing the production of phytochemicals, such as centelloside production in C. asiatica hairy root cultures (Dilshad et al, 2021), anthraquinones in rolAtransgenic tissues of Rubia cordifolia calli (Nemoto et al, 2009), or nicotine in tobacco hairy roots (Palazón et al, 1997). The rolB transformation was shown to induce resveratrol production in Vitis amurensis cells (Kiselev et al, 2007), and rolC gene expression was shown to be highly efficient for increasing the production of morphinan, tropane alkaloids (Cardillo et al, 2013;Hashemi and Naghavi, 2016), anthraquinones (Bulgakov et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

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Using machine learning to link the influence of transferred Agrobacterium rhizogenes genes to the hormone profile and morphological traits in Centella asiatica hairy roots

et al. 2022

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Hairy roots are made after the integration of a small set of genes from Agrobacterium rhizogenes in the plant genome. Little is known about how this small set is linked to their hormone profile, which determines development, morphology, and levels of secondary metabolite production. We used C. asiatica hairy root line cultures to determine the putative links between the rol and aux gene expressions with morphological traits, a hormone profile, and centelloside production. The results obtained after 14 and 28 days of culture were processed via multivariate analysis and machine-learning processes such as random forest, supported vector machines, linear discriminant analysis, and neural networks. This allowed us to obtain models capable of discriminating highly productive root lines from their levels of genetic expression (rol and aux genes) or from their hormone profile. In total, 12 hormones were evaluated, resulting in 10 being satisfactorily detected. Within this set of hormones, abscisic acid (ABA) and cytokinin isopentenyl adenosine (IPA) were found to be critical in defining the morphological traits and centelloside content. The results showed that IPA brings more benefits to the biotechnological platform. Additionally, we determined the degree of influence of each of the evaluated genes on the individual hormone profile, finding that aux1 has a significant influence on the IPA profile, while the rol genes are closely linked to the ABA profile. Finally, we effectively verified the gene influence on these two specific hormones through feeding experiments that aimed to reverse the effect on root morphology and centelloside content.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Using machine learning to link the influence of transferred Agrobacterium rhizogenes genes to the hormone profile and morphological traits in Centella asiatica hairy roots

et al. 2022

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“…By boosting beta-glucosidase activity, the rolC gene provides a gateway for the release of active cytokinins from their inactive counterparts [23]. Its versatile effects include roles in nicotine production in Nicotiana tabacum hairy roots [24], ginsenoside synthesis in Panax ginseng hairy roots [25], and resveratrol production in transgenic Vitis amurensis cell cultures [26].…”

Section: Discussionmentioning

confidence: 99%

Enhancing Centelloside Production in Centella asiatica Hairy Root Lines through Metabolic Engineering of Triterpene Biosynthetic Pathway Early Genes

Alcalde,

Palazon,

Bonfill

et al. 2023

Plants

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Centella asiatica is a medicinal plant with a rich tradition of use for its therapeutic properties. Among its bioactive compounds are centellosides, a group of triterpenoid secondary metabolites whose potent pharmacological activities have attracted significant attention. Metabolic engineering has emerged as a powerful biotechnological tool to enhance the production of target compounds. In this study, we explored the effects of overexpressing the squalene synthase (SQS) gene and transcription factor TSAR2 on various aspects of C. asiatica hairy root lines: the expression level of centelloside biosynthetic genes, morphological traits, as well as squalene, phytosterol, and centelloside content. Three distinct categories of transformed lines were obtained: LS, harboring At-SQS; LT, overexpressing TSAR2; and LST, simultaneously carrying both transgenes. These lines displayed noticeable alterations in morphological traits, including changes in branching rate and biomass production. Furthermore, we observed that the expression of T-DNA genes, particularly aux2 and rolC genes, significantly modulated the expression of pivotal genes involved in centelloside biosynthesis. Notably, the LS lines boasted an elevated centelloside content but concurrently displayed reduced phytosterol content, a finding that underscores the intriguing antagonistic relationship between phytosterol and triterpene pathways. Additionally, the inverse correlation between the centelloside content and morphological growth values observed in LS lines was countered by the action of TSAR2 in the LST and LT lines. This difference could be attributed to the simultaneous increase in the phytosterol content in the TSAR2-expressing lines, as these compounds are closely linked to root development. Overall, these discoveries offer valuable information for the biotechnological application of C. asiatica hairy roots and their potential to increase centelloside production.

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“…In addition, it affects auxin metabolism and auxin perception (Otten, 2018). Thus, it plays an essential role in the early stages of hairy root induction (Dilshad et al, 2021). Adventitious roots induced by the rolB gene produce large lateral roots in tissue culture.…”

Section: Effect Of Rol Genesmentioning

confidence: 99%

“…However, such hypotheses were withdrawn (Mauriel et al, 1991;Nilsson et al, 1993). rolB gene is also considered the gene that mostly influences plant secondary metabolism and activates plant defense reactions out of all rol genes (Bulgakov et al, 2002;Kiselev et al, 2007;Dilshad et al, 2021). Its expression increases tolerance to biotic and abiotic stresses (Veremeichik et al, 2012;Bulgakov et al, 2013;Arshad et al, 2014) and enhances resistance against fungi (Arshad et al, 2014).…”

Section: Effect Of Rol Genesmentioning

confidence: 99%

Bacteria – Plant Interaction: Transgenic Roots for Human Health

Matvieieva,

Bohdanovych,

Duplij

2023

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plasmid -Tumor induced plasmid vir genesvirulence genes rol genesroot loci or root locus genes T-DNAregion (area) of transport DNA (T-DNA), which is transferred to plant cells by the action of virulence genes (vir) from the same plasmid IAAindoleacetic acid orfopen reading frame mismikimopine synthase sequences iaaHindole-3-acetamide hydrolase Acsagrocinopine synthase CFUcoliny forming unit DMBQ -2,6-dimethoxy-1,4-benzoquinone UV light -Ultraviolet light LBleft border RBright border CRISPR -Clustered Regularly Interspaced Short Palindromic Repeats Cas9 -CRISPR-associated nuclease MSCmultiple cloning sites Tra/trbtranscriptional activators B markerbacterial selectable marker Propromoter Repreporter gene FDSfarnesyl diphosphate synthase gene GFPgreen fluorescent protein ifn-α2bhuman interferon-α2b SMssecondary metabolites SH -Schenk & Hildebrandt medium MS -Murashige & Skoog medium B5 -Gamborg medium SODsuperoxide dismutase GPXglutathione peroxidase PALphenylalanine ammonia-lyase gene CHIchalcone isomerase gene FLSflavonol synthase gene DPPH -2,2-diphenyl-1-picrylhydrazyl ABTS -2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) ZFNszinc finger nucleases NLSnuclear localization signal p35S -Cauliflower Mosaic Virus promoter GUS FW β-glucuronidase, a product of the uidA gene, which converts a soluble colorless substrate 5-bromo-4-chloro-3-indolyl-β-D-glucuronic acid into an insoluble colored product chloro-bromoindigo FWfresh weight RSAradical scavenging activity CTABcetyltrimethylammonium bromide AgNPssilver nanoparticles

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Influence of rol Genes for Enhanced Biosynthesis of Potent Natural Products

Cited by 5 publications

References 71 publications

Using machine learning to link the influence of transferred Agrobacterium rhizogenes genes to the hormone profile and morphological traits in Centella asiatica hairy roots

Using machine learning to link the influence of transferred Agrobacterium rhizogenes genes to the hormone profile and morphological traits in Centella asiatica hairy roots

Enhancing Centelloside Production in Centella asiatica Hairy Root Lines through Metabolic Engineering of Triterpene Biosynthetic Pathway Early Genes

Bacteria – Plant Interaction: Transgenic Roots for Human Health

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