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agrobacterium rhizogenes

Tatiana V Matveeva
In most cases, the genetic engineering of plants uses Agrobacterium-mediated transformation to introduce novel genes. In nature, insertion of T-DNA into the plant genome and its subsequent transfer via sexual reproduction have been shown for several species in the genera Nicotiana, Ipomoea , and Linaria . A sequence homologous to T-DNA of the Ri plasmid of Agrobacterium rhizogenes was found in the genome of wild-type Nicotiana glauca (section Noctiflorae) more than 30 years ago and was named "cellular T-DNA" (cT-DNA)...
March 3, 2018: Current Topics in Microbiology and Immunology
Karthikeyan Alagarsamy, Lubobi Ferdinand Shamala, Shu Wei
Background: Camellia sinensis var. sinensis is widely grown for tea beverages that possess significant health promoting effects. Studies on tea plant genetics and breeding are hindered due to its recalcitrance to Agrobacterium -mediated genetic transformation. Among the possible reasons, oxidation of phenolics released from explant tissues and bactericidal effects of tea polyphenols during the process of transformation play a role in the plant recalcitrance. The aim of the present study was to alleviate the harmful effects of phenolic compounds using in - planta transformation...
2018: Plant Methods
Somaieh Zafari, Mohsen Sharifi, Najmeh Ahmadian Chashmi
The callus and hairy root cultures of Prosopis farcta were established to develop effective strategies to enhance its valuable and medicinally important flavonoid compounds. For callus induction, the hypocotyl, cotyledon and shoot explants were subjected to different plant hormones, naphthalene acetic acid (NAA), benzylaminopurine (BAP), kinetin and dichlorophenoxyacetic acid (2,4-D). Greater callus induction was obtained from hypocotyl explants on MS medium containing 3.0 mg L-1 NAA + 2.0 mg L-1 BAP. With the addition of 0...
February 19, 2018: Cytotechnology
Muthusamy Balasubramanian, Murugesan Anbumegala, Ramasamy Surendran, Muthukrishnan Arun, Girija Shanmugam
An efficient protocol for hairy root induction in radish was established by optimizing several parameters that affect the efficiency of Agrobacterium rhizogenes -mediated transformations. Explants wounded using sterile hypodermic needle, infected with Agrobacterium suspension (0.6 OD 600 ) for 10 min and co-cultivated in 1/2 MS medium containing acetosyringone (100 µM) for 2 days displayed maximum percentage of hairy root induction using MTCC 2364 (77.6%) and MTCC 532 (67.6%). On further experiments with MTCC 2364 initiated hairy roots, maximum biomass accumulation (fresh weight = 9...
February 2018: 3 Biotech
Zahra Shakeran, Mehrnaz Keyhanfar, Mustafa Ghanadian
The (-)-hyoscyamine, atropine and scopolamine (hyoscine) are three valuable tropane alkaloids while scopolamine is the most important member of this group for the pharmaceutical industry due to its higher demand compared to hyoscyamine and atropine. Scopolamine is an anticholinergic reagent with several therapeutic applications. In the current study, the hairy roots culture of Daturametel was used as an advantageous method for production of scopolamine. The hairy roots are formed by Agrobacterium rhizogenes and have genetic stability, high growth rate and lateral branching...
December 2017: Molecular Biology Research Communications
Victor P Bulgakov, Yulia V Vereshchagina, Dmitry V Bulgakov, Galina N Veremeichik, Yuri N Shkryl
The rolB plant oncogene of Agrobacterium rhizogenes perturbs many biochemical processes in transformed plant cells, thereby causing their neoplastic reprogramming. The oncogene renders the cells more tolerant to environmental stresses and herbicides and inhibits ROS elevation and programmed cell death. In the present work, we performed a proteomic analysis of Arabidopsis thaliana rolB-expressing callus line AtB-2, which represents a line with moderate expression of the oncogene. Our results show that under these conditions rolB greatly perturbs the expression of some chaperone-type proteins such as heat-shock proteins and cyclophilins...
February 2, 2018: Scientific Reports
Kalpana Nanjareddy, Manoj-Kumar Arthikala, Alma-Leticia Aguirre, Brenda-Mariana Gómez, Miguel Lara
The upstream sequences of gene coding sequences are termed as promoter sequences. Studying the expression patterns of promoters are very significant in understanding the gene regulation and spatiotemporal expression patterns of target genes. On the other hand, it is also critical to establish promoter evaluation tools and genetic transformation techniques that are fast, efficient, and reproducible. In this study, we investigated the spatiotemporal expression pattern of the rhizobial symbiosis-specific nodule inception (NIN) promoter of Phaseolus vulgaris in the transgenic hairy roots...
December 23, 2017: Journal of Visualized Experiments: JoVE
Galina Khafizova, Pavel Dobrynin, Dmitrii Polev, Tatiana Matveeva
OBJECTIVE: Nicotiana glauca (tree tobacco) is a naturally transgenic plant, containing sequences acquired from Agrobacterium rhizogenes by horizontal gene transfer. Besides, N. glauca contains a wide profile of alkaloids of medical interest. DATA DESCRIPTION: We report a high-depth sequencing and de novo assembly of N. glauca full genome and analysis of genome elements with bacterial origin. The draft genome assembly is 3.2 Gb, with N50 size of 31.1 kbp. Comparative analysis confirmed the presence of single, previously described gT insertion...
January 12, 2018: BMC Research Notes
Maheswor Gautam, Mohamed Elhiti, Inge S Fomsgaard
Hairy roots induced by Agrobacterium rhizogenes are well established models to study the metabolism of xenobiotics in plants for phytoremediation purposes. However, the model requires special skills and resources for growing and is a time-consuming process. The roots induction process alters the genetic construct of a plant and is known to express genes that are normally absent from the non-transgenic plants. In this study, we propose and establish a non-transgenic maize root model to study xenobiotic metabolism in plants for phytoremediation purpose using azoxystrobin as a xenobiotic compound...
December 26, 2017: Chemosphere
Shasmita, Manoj K Rai, Soumendra K Naik
Withania somnifera (L.) Dunal (family: Solanaceae), commonly known as "Indian Ginseng", is a medicinally and industrially important plant of the Indian subcontinent and other warmer parts of the world. The plant has multi-use medicinal potential and has been listed among 36 important cultivated medicinal plants of India that are in high demand for trade due to its pharmaceutical uses. The medicinal importance of this plant is mainly due to the presence of different types of steroidal lactones- withanolides in the roots and leaves...
December 26, 2017: Critical Reviews in Biotechnology
Przemysław Sitarek, Tomasz Kowalczyk, Patricia Rijo, Adam J Białas, Marzena Wielanek, Halina Wysokińska, Catarina Garcia, Monika Toma, Tomasz Śliwiński, Ewa Skała
This study examines the production of five phenolic acids (chlorogenic acid, neochlorogenic acid, ferulic acid, caffeic acid and p-coumaric acid) following over-expression of AtPAP1 transcription factor by four transgenic root clones of Leonurus sibiricus after Agrobacterium rhizogenes transformation. The AtPAP1 expression level was estimated by quantitative real-time PCR. High levels of phenolic acids were found in the transgenic roots of L. sibiricus and were determined by high-performance liquid chromatography-mass spectrometry analysis...
January 2018: Molecular Biotechnology
Ahmed M M Gabr, Hoda B Mabrok, Emam A Abdel-Rahim, Mohamed K El-Bahr, Iryna Smetanska
Hairy root culture is a promising alternative method for the production of secondary metabolites. In this study, transformed root of Linum usitatissimum was established using Agrobacterium rhizogenes A4 strain from root cultures for lignans, phenolic acids and antioxidant capacity determination. Total lignin content (secoisolariciresinol diglucoside, secoisolariciresinol and matairesinol) was 55.5% higher in transformed root cultures than in the non-transformed root culture. Secoisolariciresinol was detected in higher concentration (2...
November 20, 2017: Natural Product Research
Dimitri Neb, Arpita Das, Annette Hintelmann, Uwe Nehls
Composite poplars were used for ectomycorrhiza formation. Structurally normal mycorrhizas of transgenic roots revealed better fungal sugar support. Targeting fluorescent proteins to peroxisomes allowed easy in planta visualization of successful transformation. A bottle neck in ectomycorrhizal research is the time demand for generation of transgenic plants. An alternative strategy for such root-centered research might be the formation of the so-called composite plants, where transgenic roots are formed by non-transgenic shoots...
December 2017: Plant Cell Reports
S Ashok Prabhu, Buyani Ndlovu, Juanita Engelbrecht, Noëlani van den Berg
Avocado (Persea americana (Mill.)), an important commercial fruit, is severely affected by Phytophthora Root Rot in areas where the pathogen is prevalent. However, advances in molecular research are hindered by the lack of a high-throughput transient transformation system in this non-model plant. In this study, a proof-of-concept is demonstrated by the successful application of Agrobacterium rhizogenes-mediated plant transformation to produce composite avocado plants. Two ex vitro strategies were assessed on two avocado genotypes (Itzamna and A0...
2017: PloS One
Yuri N Shkryl, Galina N Veremeichik, Dmitriy G Kamenev, Tatiana Y Gorpenchenko, Yulia A Yugay, Dmitriy V Mashtalyar, Aleksander V Nepomnyaschiy, Tatiana V Avramenko, Aleksandr A Karabtsov, Vladimir V Ivanov, Victor P Bulgakov, Sergey V Gnedenkov, Yury N Kulchin, Yury N Zhuravlev
In the present investigation, transgenic tobacco callus cultures and plants overexpressing the silicatein gene LoSilA1 from marine sponge Latrunculia oparinae were obtained and their bioreduction behaviour for the synthesis of silver nanoparticles (AgNPs) was studied. Synthesized nanoparticles were characterized using UV-visible spectroscopy, Fourier transformed infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), atomic flame electron microscopy (AFM) and nanoparticle tracking analysis (NTA)...
October 12, 2017: Artificial Cells, Nanomedicine, and Biotechnology
Ke Chen, Léon Otten
Agrobacterium rhizogenes causes hairy root growth on a large number of plant species. It does so by transferring specific DNA fragments (T-DNA) from its root-inducing plasmid (pRi) into plant cells. Expression of T-DNA genes leads to abnormal root growth and production of specific metabolites (opines) which are taken up by the bacterium and used for its growth. Recent work has shown that several Nicotiana, Linaria, and Ipomoea species contain T-DNA genes from A. rhizogenes in their genomes. Plants carrying such T-DNAs (called cellular T-DNA or cT-DNA) can be considered as natural transformants...
2017: Frontiers in Plant Science
Heping Shi, Yuanfeng Zhu, Po Keung Eric Tsang, Cheuk Fai Stephen Chow, Zhen'ao Yu, Shengqin Huang
To use hairy roots for producing medicinal ingredients of Phytolacca americana L. we studied the factors influencing the induction and in vitro culture. Hairy roots could be incited from the veins of cut surface (morphological lower) of P. americana L. leaf explants around 18 days after infection with the strain of Agrobacterium rhizogenes ATCC15834. The highest rooting rate, 70%, was obtained when leaf explants were pre-cultured for 1 day, infected for 20 min, and co-cultured for 4 days. The transformation was confirmed by PCR amplification of rolC of Ri plasmid and silica gel thin-layer chromatography of opines from P...
February 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
Mohamed A El-Esawi, Amr Elkelish, Hosam O Elansary, Hayssam M Ali, Mohamed Elshikh, Jacques Witczak, Margaret Ahmad
Lactuca serriola L. is a herbaceous species, used for human nutrition and medicinal purposes. The high antioxidant capacity of L. serriola indicates the possibility of enhancing its edible and health potential by increasing the flavonoid and phenolic contents. The present study aimed at enhancing the production of phenolics and flavonoids by hairy root cultures in Lactuca serriola transformed with Agrobacterium rhizogenes strain AR15834 harbouring the rolB gene. The genetic transformation of rolB in transformed roots was validated, and rolB expression level was evaluated using real-time qPCR analysis...
2017: Oxidative Medicine and Cellular Longevity
Poornima Devi Gurusamy, Holger Schäfer, Siva Ramamoorthy, Michael Wink
Hairy root culture is a potential alternative to conventional mammalian cell culture to produce recombinant proteins due to its ease in protein recovery, low costs and absence of potentially human pathogenic contaminants. The current study focussed to develop a new platform of a hairy root culture system from Nicotiana tabacum for the production of recombinant human EPO (rhEPO), which is regularly produced in mammalian cells. The human EPO construct was amplified with C-terminal hexahistidine tag from a cDNA of Caco-2 cells...
2017: PloS One
Jing An, Zhenmin Hu, Benning Che, Haiying Chen, Bingjun Yu, Weiming Cai
The Panax ginseng TIP gene PgTIP1 was previously demonstrated to have high water channel activity by its heterologous expression in Xenopus laevis oocytes and in yeast; it also plays a significant role in growth of PgTIP1-transgenic Arabidopsis plants under favorable conditions and has enhanced tolerance toward salt and drought treatment. In this work, we first investigated the physiological effects of heterologous PgTIP1 expression in soybean cotyledon hairy roots or composite plants mediated by Agrobacterium rhizogenes toward enhanced salt tolerance...
2017: Frontiers in Plant Science
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