Induction of Hairy Roots by Agrobacterium Rhizogenes and Growth of Hairy Roots In Vitro

2003 ◽  
pp. 279-288 ◽  
Author(s):  
Christopher S. Hunter ◽  
Steven J. Neill
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Roots In Vitro

scholarly journals Establishment of Highly Efficient Agrobacterium Rhizogenes-mediated Transformation for Stevia Rebaudiana Bertoni Explants

2016 ◽  
Vol 58 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Żaneta Michalec-Warzecha ◽  
Laura Pistelli ◽  
Francesca D’Angiolillo ◽  
Marta Libik-Konieczny
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Molecular Level ◽  
Stevia Rebaudiana ◽  
Culture Conditions ◽  
Light Conditions ◽  
Inoculum Concentration ◽  
Stevia Rebaudiana Bertoni ◽  
Pcr Method

Abstract Leaves and internodes from Stevia rebaudiana Bertoni plants growing in different conditions were used for transformation with two strains of Agrobacterium rhizogenes: ATCC 15384 and LBA 9402. Hairy roots formation was observed and the percentage of the transformed explants depended on the type of explant, time of inoculation and inoculum concentration. Inoculation of explants from ex vitro and in vitro plants with LBA 9402 strain led to higher efficiency of transformation than inoculation with ATCC 15384 strain. Growth rate of hairy roots in liquid culture was assessed under light and dark conditions. It was found that the growth of hairy roots decreased significantly under light conditions. Transformation of hairy roots growing in different culture conditions was confirmed at the molecular level using PCR method with primers constructed against rolB and rolC genes from A. rhizogenes.

The Use of Hairy Roots Transformed by Agrobacterium Rhizogenes for the Production of Plant Secondary Products in Vitro

1988 ◽  
pp. 425-429
Author(s):  
John D. Hamill ◽  
Adrian J. Parr ◽  
Cathie Martin ◽  
Nick J. Walton ◽  
Richard Robins ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Secondary Products

scholarly journals Induction of transgenic hairy roots in soybean genotypes by Agrobacterium rhizogenes‑mediated transformation

2011 ◽  
Vol 46 (9) ◽  
pp. 1070-1075 ◽  
Author(s):  
Ricardo Luís Mayer Weber ◽  
Maria Helena Bodanese‑Zanettini
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Fluorescent Protein ◽  
Root Culture ◽  
Transformation Frequency ◽  
Plant Production ◽  
Uniform Size ◽  
Soybean Genotypes

The objective of this work was to perform the screening of soybean genotypes as to their ability to respond to the induction of hairy roots by Agrobacterium rhizogenes‑mediated transformation. Four Brazilian soybean cultivars (BRSMG 68 Vencedora, BRS 137, Embrapa 48, and MG/BR 46 Conquista) and two North American ones adapted to Brazilian cropping conditions (Bragg and IAS‑5) were screened for their capacity to respond to A. rhizogenes in protocols for in vitro hairy root culture and ex vitro composite plant production. Four‑day‑old seedlings with uniform size were injected with A. rhizogenes harboring the plasmid p35S‑GFP. Seedlings expressing green fluorescent protein (GFP) in at least one hairy root were used to determine the transformation frequency. Using an axenic in vitro protocol, excised cotyledons from four‑day‑old seedlings were infected with A. rhizogenes harboring the pCAMBIA1301 plasmid, containing the gusA reporter gene. The transformation frequency and the number of days for hairy root emergence after bacterial infection (DAI) were evaluated. The transformation frequency and DAI varied according to the genotype. Cultivars MG/BR 46 Conquista and BRSMG 68 Vencedora are more susceptible to A. rhizogenes and can be recommended for transformation experiments.

scholarly journals PRODUÇÃO DE RAÍZES TRANSFORMADAS DE PIMENTA-LONGA POR INFECÇÃO COM AGROBACTERIUM RHIZOGENES

2017 ◽  
Vol 18 (4) ◽  
pp. 179
Author(s):  
José Nicomedes Júnior ◽  
Roberta Cristiane Ribeiro ◽  
Ricardo Luis Louro Berbara ◽  
Elvia Mariam Lis Martinez Stark ◽  
Wagner Campos Otoni ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots

Este trabalho teve por objetivo a obtenção de raízes transgênicas (hairy roots) de Piper hispidinervium (pimenta-longa), espécie reconhecida pela produção de óleo essencial rico em Safrol, metabólito secundário utilizado com fins terapêuticos e industrial. As raízes foram produzidas a partir da infecção de explantes in vitro de pimenta-longa com Agrobacterium rhizogenes (Estirpe R1601). Os resultados mostraram que a estirpe R1601 de A. rhizogenes foi eficiente para inoculação baseado na imersão de explantes em meio de cultura liquido, com a bactéria em crescimento, o que representou maior praticidade, menor índice de contaminação, maior taxa de infecção e menores danos aos explantes. A resposta morfológica foi a mesma entre os clones, não permitindo a distinção dos mesmos. Explantes de folha de pimenta-longa, inoculados com a estirpe R1601, formaram raízes que apresentaram crescimento nos subcultivos subseqüentes. Foi possível realizar a extração de óleo essencial produzido pela cultura de raízes de forma semelhante a plantas sob cultivo, sendo o metanol, o melhor solvente extrator do óleo essencial produzido.

scholarly journals TÉCNICAS DE TRANSFORMAÇÃO E LINHAGENS De Agrobacterium rhizogenes NA PRODUÇÃO DE HAIRY ROOTS DE Ocimum basilicum L.

Nativa ◽  
2018 ◽  
Vol 6 (3) ◽  
pp. 266
Author(s):  
José Nicomedes Júnior ◽  
Roberta Cristiane Ribeiro ◽  
Luis Louro Berbara ◽  
Elvia Mariam Lis Martinez Stark ◽  
Wagner Campos Otoni ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Leaf Explants ◽  
Laboratory Strain ◽  
Root Culture ◽  
Ocimum Basilicum ◽  
Best Response ◽  
Agrobacterium Rhizogenes Strains ◽  
Whole Plants

O manjericão é uma planta medicinal e seus metabólitos especiais são utilizados com fins terapêuticos e industriais. Este trabalho teve por objetivo avaliar técnicas e linhagens de Agrobacterium rhizogenes na produção de raízes transformadas de duas variedades de Ocimum basilicum L., Dark Opal (roxo) e Minete Anão (verde). Verificou-se o efeito da inoculação com linhagens de A. rhizogenes selvagens (A4, 8196, 9402, 2659, 2659 G, 17242, LBA, 15834) ou transformadas (R1601) em plantas inteiras ou explantes (folhas e segmentos de caule) de manjericão, cultivados em casa de vegetação ou in vitro. A inoculação com as linhagens LBA e 8196 possibilitou redução no número de explantes de folha de manjericão-verde oxidados, quando comparado ao controle. Dentre as linhagens selvagens, as que proporcionaram melhor indução na produção de raízes transformadas foram A4, LBA e 8196, respectivamente. Entretanto, a cepa R1601 apresentou a melhor resposta. Raízes foram eficientemente induzidas mediante a inoculação de explantes de manjericão-roxo e manjericão-verde e expressaram o fenótipo típico de raízes em cabeleira (hairy roots). As culturas clonais de manjericão-verde apresentaram rápido crescimento em meio de cultura livre de reguladores de crescimento.Palavras-chave: agrobactéria, cultura de raízes, manjericão, plantas medicinais. TRANSFORMATION TECHNIQUES AND Agrobacterium rhizogenes Strains In The Production Of Hairy Roots Of Ocimum basilicum L. ABSTRACT:Basil is a medicinal plant and its special metabolites are used for therapeutic or industrial purposes. The aim of this work was to evaluate Agrobacterium rhizogenes techniques and strains in the production of hairy roots of two varieties of Ocimum basilicum L., Dark Opal (purple) and Dwarf Racer (green). The effects of inoculation of A. rhizogene (A4, 8196, 9402, 2659, 2659, 2659 G, 17242, LBA, 15834) or (R1601) lines on whole plants or explants (leaves and stem segments) of basil, grown in a greenhouse or in vitro, in the production of hairy roots and calluses made by subcultures. Inoculation with the LBA and 8196 lines allowed a reduction in the number of oxidized basil-green leaf explants when compared to the control. Among the wild strains, those that had best ability to induce hairy root of hairy roots were A4, LBA and 8196, respectively. However, the R1601 laboratory strain had the best response. Roots were efficiently induced by the inoculation of explants basil-purple and basil-green and expressed the typical phenotype of hairy roots. Green-basal clonal cultures showed rapid growth in culture medium free of growth regulators.Keywords: agrobacterim, basil, medicinal plants, root culture. DOI:

scholarly journals Soybean hairy roots produced in vitro by Agrobacterium rhizogenes-mediated transformation

2018 ◽  
Vol 6 (2) ◽  
pp. 162-171 ◽  
Author(s):  
Li Chen ◽  
Yupeng Cai ◽  
Xiujie Liu ◽  
Chen Guo ◽  
Shi Sun ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots

Agrobacterium rhizogenes-Transformed Roots of Medicago truncatula for the Study of Nitrogen-Fixing and Endomycorrhizal Symbiotic Associations

2001 ◽  
Vol 14 (6) ◽  
pp. 695-700 ◽  
Author(s):  
Aurélien Boisson-Dernier ◽  
Mireille Chabaud ◽  
Fernand Garcia ◽  
Guillaume Bécard ◽  
Charles Rosenberg ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Medicago Truncatula ◽  
Hairy Root ◽  
Hairy Roots ◽  
Glomus Intraradices ◽  
Subsequent Development ◽  
Nitrogen Fixing ◽  
Root Explants ◽  
Symbiotic Associations

Medicago truncatula, a diploid autogamous legume, is currently being developed as a model plant for the study of root endosymbiotic associations, including nodulation and mycorrhizal colonization. An important requirement for such a plant is the possibility of rapidly introducing and analyzing chimeric gene constructs in root tissues. For this reason, we developed and optimized a convenient protocol for Agrobacterium rhizogenes-mediated transformation of M. truncatula. This unusual protocol, which involves the inoculation of sectioned seedling radicles, results in rapid and efficient hairy root organogenesis and the subsequent development of vigorous “composite plants.” In addition, we found that kanamycin can be used to select for the co-transformation of hairy roots directly with gene constructs of interest. M. truncatula composite plant hairy roots have a similar morphology to normal roots and can be nodulated successfully by their nitrogen-fixing symbiotic partner, Sinorhizobium meliloti. Furthermore, spatiotemporal expression of the Nod factor-responsive reporter p MtENOD11-gusA in hairy root epidermal tissues is indistinguishable from that observed in Agrobacterium tumefaciens-transformed lines. M. truncatula hairy root explants can be propagated in vitro, and we demonstrate that these clonal lines can be colonized by endomycorrhizal fungi such as Glomus intraradices with the formation of arbus-cules within cortical cells. Our results suggest that M. truncatula hairy roots represent a particularly attractive system with which to study endosymbiotic associations in transgenically modified roots.

scholarly journals Desarrollo de un sistema de cultivo in vitro de raíces pilosas de Phyllantus acumminatus, para la producción de compuestos con actividad biológica

2018 ◽  
Vol 31 (3) ◽  
Author(s):  
Giovanni Garro-Monge ◽  
Karol Jiménez-Quesada ◽  
Raquel Pérez-Méndez
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots

Las plantas del género Phyllanthus son actualmente investigadas por su habilidad para producir compuestos fenólicos con alta capacidad antioxidante. Además, diversos estudios han revelado que estas plantas poseen metabolitos (filantósidos y lignanos) que pueden inducir la apoptosis celular, lo que les proporciona un gran potencial para posibles tratamientos de prevención contra el cáncer. En este trabajo, se llevó a cabo la introducción in vitro de semillas de P. acumminatus; se logró el cultivo in vitro del material vegetal, y las vitro-plantas fueron utilizadas en ensayos de inducción de raíces pilosas, mediante transformación genética con Agrobacterium rhizogenes cepa Ar15834. El tratamiento más favorable para la inducción de estos cultivos organogénicos consistió en el uso de estacas de aproximadamente 1-2 cm, a las cuales se les hicieron cortes transversales con un bisturí, para luego ser inoculadas en el cultivo bacteriano (OD600: 0.6-1).  Las estacas fueron colocadas en medio M & S semisólido al 50% de sales y vitaminas, sin reguladores de crecimiento, por un período de 48 h. Transcurrido este tiempo, las estacas fueron micropropagadas en medio M & S al 100% de sales y vitaminas, complementado con 500 µg/l del antibiótico Ceftriaxona® para erradicar la bacteria. Se logró el  cultivo de las raíces pilosas en medio M & S al 100% de sales y vitaminas, sin reguladores de crecimiento. Por medio del uso de microscopía de luz, se logró la caracterización morfológica de las raíces establecidas, las cuales coinciden plenamente con las típicas raíces pilosas o hairy roots obtenidas por medio del sistema de agro-infección, según la literatura consultada.

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