Bio-interaction of Agrobacterium rhizogenes with Capsicum annuum L. (sweet variety) and Establishment Hairy Roots Cultures

2020 ◽  
Vol 1 (2) ◽  
pp. 30-33
Author(s):  
Qutaiba Al-Nema
Keyword(s):  
Capsicum Annuum ◽  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Future Research ◽  
Ms Medium ◽  
Transformed Roots ◽  
Transformed Hairy Roots ◽  
Petri Dishes ◽  
Suitable System ◽  
Modern Technologies

Genetic transformation using Agrobacterium is one of the techniques used to transfer desired genes to plants. This protocol is considered a short – cut to get transformed plants which could be an alternative method and suitable system compared with the classical methods. This study aimed to investigate the response of C. annuum seedlings (sweet variety) to the formation transformed hairy roots induced by A. rhizogenes strain R1601. Sterilized seeds of Capsicum annuum were inoculated with the Agrobacterium rhizogenes inoculum. The samples were then transferred to the surface of solidified MS medium. Hairy roots were developed at the inoculation sites and were enucleated 1.0-1.5 cm length and placed in 9.0 cm Petri-dishes containing 15 ml of agar solidified MS medium. Agropine test was performed according to the standard method. The inoculated seedlings showed a good response 90%. Transformed hairy roots were established at the injection sites within 10 days and these roots were easily grown on agar-solidified MS medium. The results are confirmed that these roots were transformed roots it in terms of positive agropine detection. The current study concluded that the biological interaction between Agrobacterium rhizogenes strain R1601 and Capsicum anuum L. seedlings, was successful. This study encourages future research to improve this plant by continuing and applying modern technologies to obtain genetically modified plants.

scholarly journals Bio-interaction of Agrobacterium rhizogenes with Capsicum annuum L. (sweet variety) and Establishment Hairy Roots Cultures

2020 ◽  
Vol 1 (2) ◽  
pp. 30-33
Author(s):  
Qutaiba AL-Nema
Keyword(s):  
Capsicum Annuum ◽  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Future Research ◽  
Ms Medium ◽  
Transformed Roots ◽  
Transformed Hairy Roots ◽  
Petri Dishes ◽  
Suitable System ◽  
Modern Technologies

Genetic transformation using Agrobacterium is one of the techniques used to transfer desired genes to plants. This protocol is considered a short – cut to get transformed plants which could be an alternative method and suitable system compared with the classical methods. This study aimed to investigate the response of C. annuum seedlings (sweet variety) to the formation transformed hairy roots induced by A. rhizogenes strain R1601. Sterilized seeds of Capsicum annuum were inoculated with the Agrobacterium rhizogenes inoculum. The samples were then transferred to the surface of solidified MS medium. Hairy roots were developed at the inoculation sites and were enucleated 1.0-1.5 cm length and placed in 9.0 cm Petri-dishes containing 15 ml of agar solidified MS medium. Agropine test was performed according to the standard method. The inoculated seedlings showed a good response 90%. Transformed hairy roots were established at the injection sites within 10 days and these roots were easily grown on agar-solidified MS medium. The results are confirmed that these roots were transformed roots it in terms of positive agropine detection. The current study concluded that the biological interaction between Agrobacterium rhizogenes strain R1601 and Capsicum anuum L. seedlings, was successful. This study encourages future research to improve this plant by continuing and applying modern technologies to obtain genetically modified plants.

scholarly journals Genetically transformed hairy roots producing agropine induced on Trigonella foenum-graecum L. plant by Agrobacterium rhizogenes 1601.

2013 ◽  
Vol 7 (1) ◽  
pp. 91-98
Author(s):  
M. M. Al-Mahdawe ◽  
M. K. Al-Mallah ◽  
A. O. Al-Attrakchii
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Root Formation ◽  
Paper Electrophoresis ◽  
Transformed Roots ◽  
Fresh Weight ◽  
Trigonella Foenum Graecum ◽  
Transformed Hairy Roots ◽  
Long Time ◽  
Trigonella Foenum Graecum L

Inoculation of leaves and stems explants excised from field-grown Trigonella foenum-graecum L. with engineered Agrobacterium rhizogenes 1601 inoculum of optical density 1.90, 2.06 and 1.96 led to the formation of hairy roots on these explants. The highest percent of infection with inoculum of O.D 1.90 was 20% in leaves, and was 53.3% in stems using inoculum of O.D 2.06. Infection percent was 50% in hypocotyl compared with non-inoculated explants. The results showed that inoculation of seedlings lacking roots were slow in hairy root formation and take long time compared with their formation in intact seedlings inoculated with the same inocula. Results of paper electrophoresis of these root proved the incidence of their genetic transformation. Moreover, these transformed roots grow happily in liquid WP medium with an average fresh weight 5.9g after four weeks of culture, whereas fresh weight of normal roots was 1.02g under the same conditions.

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.

Sennosides A and B Production by Hairy Roots of Senna alata (L.) Roxb.

2006 ◽  
Vol 61 (5-6) ◽  
pp. 367-371 ◽  
Author(s):  
Waraporn Putalun ◽  
Suwat Pimmeuangkao ◽  
Wanchai De-Eknamkul ◽  
Hiroyuki Tanaka ◽  
Yukihiro Shoyama
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Ms Medium ◽  
Strain Atcc ◽  
Dark Condition ◽  
Medium Effects ◽  
Half Strength ◽  
Senna Alata

Hairy roots of Senna alata transformed with Agrobacterium rhizogenes, strain ATCC 15834 were induced and grown in half-strength Murashige and Skoog (MS) medium. Effects of sucrose contents and hormones on the growth and sennosides A, B production were investigated. Hairy roots cultured on hormone-free half-strength MS medium containing 5% sucrose under dark condition mostly stimulated the growth of hairy roots and increased the content of sennosides A and B yielding (169 ± 4) and (34 ± 3) μg g-1 dry wt, respectively.

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.

scholarly journals Specific activity of thymidine nucleotide biosynthetic enzymes in hairy roots extracts of Sesamum indicum L. transformed by two strains of Agrobacteruim rhizogenes

2014 ◽  
Vol 8 (2) ◽  
pp. 64-72
Author(s):  
Nihal E. Al- Taee ◽  
Sajida A. Abood ◽  
Mozahim K. Al-Mallah
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Dihydrofolate Reductase ◽  
Specific Activity ◽  
Sesamum Indicum ◽  
Dna And Rna ◽  
Transformed Hairy Roots ◽  
Natural Vector ◽  
Activity Of Enzymes ◽  
Control Samples

The study concluded induction of transformed hairy roots from leaves and decapitated seedlings of Sesamum indicum L. using two strains of Agrobacterium rhizogenes considered as natural vector of transformation. The strain R1601 stimulated roots on leaves and seedlings during 20 days of inoculation placed on solidified Arnon and Hoagland medium. Whereas they involved 12 days when inoculated with the strain R15834. Generally strain R15834 was efficient in inducing these roots and their numbers than strain R1601 which approached 54.4% and 41.6% respectively. The results indicated an increase in the specific activity of enzymes Thymidlate synthase (TS), Dihydrofolate reductase (DHFR), Serine hydroxy methyl transfrase (SHMT) in extract of transformed hairy roots producing agropine by stain R15834 and approach 4.610, 1.057, 0.480 µmolmin mg of protein respectively compared with the activity of 1.256, 0.097, 0.125µ molmin mg in the control samples. This was coupled with an increase in amount of DNA and RNA that approached 105, 1020 µg per gram respectively compared to 44, 462 µg per gram in control samples. The transformation of these hairy roots was pointed out through the separation of agropine spots from their extracts when electrophoreted in the presence of standared agropine.

scholarly journals A Stable Agrobacterium rhizogenes-Mediated Transformation of Cotton (Gossypium hirsutum L.) and Plant Regeneration From Transformed Hairy Root via Embryogenesis

2020 ◽  
Vol 11 ◽  
Author(s):  
Min-Long Cui ◽  
Chen Liu ◽  
Chun-Lan Piao ◽  
Chuan-Liang Liu
Keyword(s):  
Gossypium Hirsutum ◽  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Gene Function ◽  
Fluorescent Protein ◽  
Function Analysis ◽  
Green Fluorescent Protein Gene ◽  
Pcr Analysis ◽  
Cotton Transformation ◽  
Transformed Hairy Roots

Genetic transformation is a powerful tool to study gene function, secondary metabolism pathways, and molecular breeding in crops. Cotton (Gossypium hirsutum L.) is one of the most important economic crops in the world. Current cotton transformation methods take at least seven to culture and are labor-intensive and limited to some cultivars. In this study, we first time achieved plantlet regeneration of cotton via embryogenesis from transformed hairy roots. We inoculated the cotyledon explants of a commercial cultivar Zhongmian-24 with Agrobacterium rhizogenes strain AR1193, harboring a binary vector pBI-35S::GFP that contained the NPT II (neomycin phosphotransferase) gene and the GFP (green fluorescent protein) gene as a fluorescent marker in the T-DNA region. 82.6% explants produced adventitious roots, of which 53% showed GFP expression after transformation. 82% of transformed hairy roots produced embryonic calli, 12% of which regenerated into stable transformed cotton plants after 7 months of culture. The integration of GFP in the transformed cotton genomes were confirmed by PCR (Polymerase chain reaction) and Southern blot analysis as well as the stable expression of GFP were also detected by semi-quantitative RT-PCR analysis. The resultant transformed plantlets were phenotypically, thus avoiding Ri syndrome. Here we report a stable and reproducible method for A. rhizogenes-mediated transformation of cotton using cotyledon as explants, which provides a useful and reliable platform for gene function analysis of cotton.

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