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 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 Hairy Root Induction on Justicia gendarussa by Various Density of Agrobacterium rhizogenes strain LB 510

2017 ◽  
Vol 9 (1) ◽  
pp. 26
Author(s):  
Dwi Kusuma Wahyuni ◽  
Siti Nur Hafida ◽  
Tri Muji Ermayanti ◽  
Bambang Prajoga Eko Wardoyo ◽  
Hery Purnobasuki ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Solid Medium ◽  
Leaf Explants ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Root Induction ◽  
Hairy Root Induction ◽  
Industrial Needs

<p>Gandarusa (<em>Justicia gendarussa</em> Burm.f.) is an Indonesian medicinal plant that has many benefits as drug and male contracetive. For industrial needs, Gandarusa must be  available in large quantity. Hairy root culture is one of methode to produce phytochemistry compound. The objective of the study was to examine the effect of various density of <em>Agrobacterium rhizogenes</em> strain LB510 on hairy roots induction of gandarusa (<em>Justicia gendarussa </em>Burm.f.) leaf plant. Leaf explants were inoculated in MS liquid medium with various density of OD<sub>600</sub> = 0.1; 0.2; 0.3; 0.4; and 0.5. Explants were co-cultivated for 2 days on MS solid medium without any hormone then sub-cultured on MS solid medium containing antibiotic cefotaxim 300 ppm, in dark condition. The data were analyzed descriptively and statistically. The results showed that various density of <em>Agrobacterium rhizogenes</em> strain LB510 was affected the lenght of hairy roots induction of <em>J</em><em>.</em><em> gendarussa</em> Burm.f., but these was not effected toward lenght formation time and number of hairy root. The treatment of OD<sub>600</sub> 0.2 was the best treatment for hairy root induction on <em>Justicia gendarussa </em>Burm. f. This data could be used for optimized the quality of methode of hairy root induction. </p>

FACTORS AFFECTING THE REGENERATION OF PEPPER (CAPSICUM ANNUUM L.)

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1120G-1120
Author(s):  
J. L. Jacobs ◽  
C. T. Stephens
Keyword(s):  
Growth Hormone ◽  
Silver Nitrate ◽  
Callus Formation ◽  
Leaf Explants ◽  
Regeneration System ◽  
Factors Affecting ◽  
Nitrate Concentrations ◽  
Leaf Differentiation ◽  
Whole Plants

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.

A reliable and efficient protocol for induction of hairy roots in Agastache foeniculum

Biologia ◽  
2014 ◽  
Vol 69 (7) ◽  
Author(s):  
Elnaz Nourozi ◽  
Bahman Hosseini ◽  
Abbas Hassani
Keyword(s):  
Salicylic Acid ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Performance ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Biochemical Properties ◽  
Separate Experiment ◽  
Transformed Roots

AbstractHairy root culture system is a valuable tool to study the characteristics of gene expression, gene function, root biology, biochemical properties and biosynthesis pathways of secondary metabolites. In the present study, hairy roots were established in Anise hyssop (Agastache foeniculum) via Agrobacterium rhizogenes. Three strains of Agrobacterium rhizogenes (A4, A7 and 9435), were used for induction of hairy roots in four various explants (hypocotyl, cotyledon, one-month-old leaf and five-month-old leaf) of Anise hyssop. The highest frequency of transformation was achieved using A4 strain in one-month-old leaves (51.1%). The transgenic states of hairy root lines were confirmed by PCR (Polymerase chain reaction) method. High performance liquid chromatography analysis revealed that the production of rosmarinic acid (RA) in transformed roots of A. foeniculum was almost 4-fold higher than that of the non-transformed roots. In a separate experiment, hairy roots obtained from one-month-old leaves inoculated with A4 strain, were grown in liquid medium and the effects of different concentrations of salicylic acid (0.0, 0.01, 0.1 and 1 mM) and chitosan (0, 50, 100 and 150 mg L−1) (as elicitor) and sucrose (20, 30, 40 and 50 g L−1) on the growth of hairy roots were evaluated. The results showed that, 30 g L−1 sucrose and 100 mg L−1 chitosan increased the biomass of hairy root cultures and application of salicylic acid reduced the growth of hairy roots compared with control roots.

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.

Technology and Application of Hairy Root Culture in Monocotyledons

2021 ◽  
Vol 07 ◽  
Author(s):  
Chang-Qi Hao ◽  
Shuai-Run Wang ◽  
Yi Wang ◽  
Xin-Yi Hou ◽  
Ya-Xuan Jiang ◽  
...  
Keyword(s):  
Medicinal Plants ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Food Crops ◽  
Root Induction ◽  
Hairy Root Induction ◽  
Selection Of

Background: Hairy root culture has been widely used in the production of metabolites in dicotyledons, and a large number of food crops and medicinal plants in monocotyledons need to be developed, but there are many difficulties in the induction of hairy roots in monocotyledons. The purpose of this paper is to introduce the inducing methods, influencing factors and application of hairy roots in monocotyledons, and to promote the development of hairy root system in monocotyledons. Methods: The mechanism of action of Agrobacterium rhizogenes and the current situation of hairy root induction, induction methods and influencing factors of monocotyledons were summarized so as to provide convenience for efficient acquisition of hairy root of monocotyledons. Results: Monocotyledons are not easy to produce phenols, cells are prone to lignification, adverse differentiation and selective response to Agrobacterium rhizogenes strains. It is proposed that before induction, plant varieties and explants should be selected, and different infection strains should be screened. In the process of hairy root induction, exogenous inducers such as acetosyringone can be added. Although these factors can provide some help for the induction of hairy roots in monocotyledons, we still need to pay attention to the disadvantages of monocotyledons from dicotyledons at the cellular level. Conclusion: A large number of food crops and medicinal plants are monocotyledons. Hairy root culture can be used to help the breeding and production of medicinal substances. Therefore, it is necessary to pay attention to the selection of varieties and explants, the selection of Agrobacterium rhizogenes and the addition of acetosyringone in the process of hairy root induction so as to improve the production efficiency and facilitate the development and utilization of monocotyledons.

scholarly journals In vitro Propagation of Vanda testacea (Lindl.) Reichb.f. – A Rare Orchid of High Medicinal Value

2010 ◽  
Vol 19 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Saranjeet Kaur ◽  
K.K. Bhutani
Keyword(s):  
Leaf Explants ◽  
Subsequent Development ◽  
Individual Treatment ◽  
Chemical Stimulus ◽  
Medicinal Value ◽  
Best Response ◽  
High Regeneration ◽  
Regeneration Response ◽  
High Regeneration Frequency

Foliar explants of Vanda testacea (Lindl.) Reichb. f. were cultured on Mitra (M) medium with 1.0 mg/l BAP, Kn  each and 1.0 mg/l NAA individually and in combination for initiation of regeneration response, proliferation of regenerants and subsequent development of plantlets. Juvenility of the tissues and chemical stimulus were important factors in initiating the regeneration response in the explants. The relatively older leaf explants (>1cm in length) remained recalcitrant to regeneration the representing younger ones (<1cm in length) responded to certain chemical regimes. BAP, Kn individually in the medium should direct PLB regeneration whereas when used with NAA, the explants showed callus proliferation and further differentiated into PLBs. An individual treatment with NAA (1.0 mg/l) impaired the response frequency and delayed further morphogenetic processes leading to plantlet development. The best response in the explants (in terms of high regeneration frequency, early initiation, PLB proliferations, and plantlet development) was observed in 1.0 mg/l BAP alone/with 1.0 mg/l NAA + activated charcoal. Plantlets were transferred to pots containing epiphytic compost (1 charcoal : 1 brick pices : 1 bats). Nearly 75% of plantlets survival was recorded.  Key words: In vitro, Orchid, Vanda testacea, Micropropagation, Protocorm-like bodies, callus D.O.I. 10.3329/ptcb.v19i1.4077 Plant Tissue Cult. & Biotech. 19(1): 1-7, 2009 (June) 

Effects of Agrobacterium rhizogenes strains and other parameters on production of isoflavonoids in hairy roots of Pueraria candollei Grah. ex Benth. var. candollei

2012 ◽  
Vol 111 (3) ◽  
pp. 315-322 ◽  
Author(s):  
Pimsiri Danphitsanuparn ◽  
Panitch Boonsnongcheep ◽  
Thanaphol Boriboonkaset ◽  
Yupyn Chintapakorn ◽  
Sompop Prathanturarug
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Pueraria Candollei ◽  
Agrobacterium Rhizogenes Strains
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