Agrobacterium tumefaciens and Agrobacterium rhizogenes transformed roots of the parasitic plant Triphysaria versicolor retain parasitic competence

Planta ◽  
2006 ◽  
Vol 225 (5) ◽  
pp. 1059-1071 ◽  
Author(s):  
Alexey Tomilov ◽  
Natalya Tomilova ◽  
John I. Yoder
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Agrobacterium Rhizogenes ◽  
Parasitic Plant ◽  
Transformed Roots

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 Transgenic root cultures of Gentiana punctata L.

2014 ◽  
Vol 68 (4) ◽  
pp. 275-280 ◽  
Author(s):  
Branka Vinterhalter ◽  
Vladimir Orbović ◽  
Dragan Vinterhalter
Keyword(s):  
Growth Rate ◽  
Agrobacterium Rhizogenes ◽  
Basal Medium ◽  
Shoot Cultures ◽  
Root Cultures ◽  
Transformed Roots ◽  
Flower Buds ◽  
Transgenic Root ◽  
Short Internodes

Shoot cultures of <em>Gentiana punctata</em> L. were inoculated with suspension of <em>Agrobacterium rhizogenes</em> strain A4 M70GUS. Hairy roots which appeared 2-3 weeks later were cultured on hormone-free, liquid, WPM (Lloyd and McCown 1980) basal medium for more than 5 years (60 subcultures). Growth rate of transformed roots was higher than the growth rate of nontransformed roots. Spontaneous shoot regeneration occured only in three culture vessels in subcultures No. 40 and 42. Plants had phenotype characteristics typical for <em>A. rhizogenes</em> transformed plants including: wrincled leaves, short internodes, plagiotropic roots and in general their growth rate was reduced. These plants also manifested precocious formation of flower buds without vernalization and flowering under in vitro conditions. Flowers were pale yellow, the same as in the standard phenotype.

scholarly journals Agrobacterium rhizogenes-Mediated Transformation of the Parasitic Plant Phtheirospermum japonicum

PLoS ONE ◽  
2011 ◽  
Vol 6 (10) ◽  
pp. e25802 ◽  
Author(s):  
Juliane K. Ishida ◽  
Satoko Yoshida ◽  
Masaki Ito ◽  
Shigetou Namba ◽  
Ken Shirasu
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Parasitic Plant

scholarly journals Successful genetic transformation in date palm (Phoenix dactylifera)

2014 ◽  
Vol 11 (2) ◽  
pp. 171-176 ◽  
Author(s):  
L Hassan
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Agrobacterium Rhizogenes ◽  
Reporter Gene ◽  
Hairy Roots ◽  
Date Palm ◽  
Binary Vector ◽  
Nptii Gene ◽  
Neomycin Phosphotransferase ◽  
Gus Gene ◽  
Nos Terminator

The introduction of foreign genes into most of the Phoenix spp using recombinant DNA technology is not a straight forward task. In Phoenix spp application of this technology towards successful transformation proved to be a more difficult one – so far no report on the successful regeneration of transgenic date palm plants has been published. We developed an efficient and reproducible variety-independent method for producing transgenic date palm (Phoenix spp) via Agrobacterium-mediated transformation. Agrobacterium rhizogenes strains LBA 9402 were used and for cotransformation experiments the strain LBA 9402 with the binary vector pBIN19 with the p35S GUS INT gene was used. Off-shoot segments from different Phoenix spp cultivars were infected with Agrobacterium rhizogenes. The development of ‘hairy roots’ at a high frequency only on infected tissue pieces showed that transformation is possible. Various parameters like, effect of different genotypes on root initiation, root number and root length have been studied. Regeneration of transformed root cultures to plantlets was also attempted. Histochemical GUS assay and polymerase chain reaction analysis of hairy roots confirmed the presence of GUS gene. Agrobacterium tumifaciensmediated transformation was also performed using the leaves of off-shoot explants. Agrobacterium tumefaciens strains: I) GV3101 with the vir plasmid pMP90 the strain C58C1 ATHV with the vir-plasmid pTiBo542 (=pEHA101; Hood et al. 1986) was used. The nptII gene (neomycin phosphotransferase) was used as a selectable marker gene. The ?-Glucuronidase-gene (GUS-Gene: Jefferson et al. 1987) under control of the Ubi- and 35S-Promotors, with an Intron (Vancanneyt et al. 1990), was used as the reporter gene. We also used the genetically engineered Agrobacterium tumefaciens strain LBA4404 as a vector for infection in the transformation experiment, which contains plasmid pBI121 of 14 KDa (binary vector). This binary vector contains following genes within the right border (RB) and left border (LB) region of the construct: The udiA gene (Jefferson, 1986) predetermining GUS (?-glucuronidase), driven by CaMV promoter and NOS terminator. This reporter gene can be used to assess the efficiency of transformation. The nptII gene (Herrera-Estrella et al., 1983) encoding neomycin phosphotransferase II (nptII) conferring kanamycin resistance, driven by NOS promoter and NOS terminator. The bacterium also contains plasmid pAL4404 which is a disarmed Ti plasmid (132 KDa) containing the virulence genes. For the confirmation of transgenes, calli were taken from the growing callus mass for DNA isolation. PCR- and Southern analysis was performed to determine the integration and the copy number of the transgene. The GUS-test was performed to demonstrate ß-glucuronidase expression. The transgenic plantlets were kept in a hardening room for four weeks and they will be transferred to a growth chamber with controlled environment for further establishment. DOI: http://dx.doi.org/10.3329/jbau.v11i2.19841 J. Bangladesh Agril. Univ. 11(2): 171-176, 2013

scholarly journals Complementation of Agrobacterium tumefaciens tumor-inducing aux mutants by genes from the TR-region of the Ri plasmid of Agrobacterium rhizogenes

1986 ◽  
Vol 83 (18) ◽  
pp. 6935-6939 ◽  
Author(s):  
I. A. Offringa ◽  
L. S. Melchers ◽  
A. J. G. Regensburg-Tuink ◽  
P. Costantino ◽  
R. A. Schilperoort ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Agrobacterium Rhizogenes ◽  
Ri Plasmid

Induction of hairy roots in Arnica montana L. by Agrobacterium rhizogenes

2013 ◽  
Vol 8 (5) ◽  
pp. 470-479 ◽  
Author(s):  
Mariya Petrova ◽  
Ely Zayova ◽  
Mariana Vlahova
Keyword(s):  
Genetic Transformation ◽  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Culture Media ◽  
Pcr Analysis ◽  
Transformed Roots ◽  
Fresh Weight ◽  
Transformation Techniques ◽  
Nutrient Media ◽  
Arnica Montana

AbstractThe induction of hairy roots in Arnica montana L. by Agrobacterium rhizogenes mediated system was established. The frequency of genetic transformation varied from 4.8 to 12% depended on method of infection. The cefotaxime at concentration of 200 mg/l proved to suppress effectively the growth of A. rhizogenes after co-cultivation. Among the three tested nutrient media: Murashige and Skoog (MS), Gamborg’s (B5) and Schenk and Hildebrandt (SH), MS medium was superior for growth and high biomass production of transformed roots compared to other culture media. After culturing for 40 days the fresh weight of clone T4 increased 7.6 fold over the non-transformed roots. The transfer of rol A, rol B and rol C genes into Arnica genome was confirmed by PCR analysis. Established genetic transformation techniques in A. montana efficiently provided and generated a large number of transformed roots — an excellent system for studying gene function and could be used for the production of secondary metabolites synthesized in roots.

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