scholarly journals Influences of various factors on hairy root induction in Agastache foeniculum (Pursh) Kuntze

2016 ◽  
Vol 107 (1) ◽  
pp. 45 ◽  
Author(s):  
Elnaz NOUROZI ◽  
Bahman HOSSEINI ◽  
Abbas HASSANI

<em>Agrobacterium rhizogenes</em> is known as a natural tool of genetic engineering in many plant species. For the first time, hairy root induction in <em>Agastache foeniculum</em> using <em>A. rhizogenes</em>,<em> </em>rosmarinic acid content<em> </em>and the effect of different culture media and inoculation methods on hairy root growth rate were investigated. Hairy root culture of <em>A. foeniculum</em> was established by inoculation of the 1-month-old leaf explant with A4 strain of <em>A. rhizogenes</em> and the effectiveness of light – dark conditions and two inoculation methods (immersion and injection) were tested. Furthermore, in immersion method, the effects of inoculation time (3, 5 and 7 min) on root induction were investigated. In the second part of the study, the hairy root culture of <em>A. foeniculum</em> was studied using different<em> </em>basal culture media (MS, 1/2 MS and B5). Rosmarinic acid content in hairy roots and non- transformed roots was analyzed using high-performance liquid chromatography (HPLC). There was no significant difference between various inoculation methods in the ability of hairy roots induction. Observations showed that percentage of hairy root induction was higher when the explants were immersed for 5 min in bacterial suspension. Light conditions displayed the highest hairy root induction rates compared with dark condition. Various culture media are different in terms of types and amounts of nutrients and have influence on growth rate. The maximum growth rate (1.61 g fr wt/50 ml) of hairy roots were obtained in 1/2 MS medium. Rosmarinic acid content in transformed roots (213.42 µg/g dry wt) was significantly higher than non-transformed roots (52.28 µg/ g dry wt).

2012 ◽  
Vol 40 (1) ◽  
pp. 125 ◽  
Author(s):  
Afsaneh SAMADI ◽  
Jirair CARAPETIAN ◽  
Reza HEIDARI ◽  
Morad JAFARI ◽  
Abdollah HASSANZADEH GORTTAPEH

Transgenic hairy root system is a promising source of secondary metabolites in medicinal plants with high pharmaceutical value.For the first time, hairy roots were established in different explants of Linum mucronatum, an anti-cancer agent producing plant, via amikimopine type strain of Agrobacterium rhizogenes, ‘A13’. The percentage of hairy root induction varied from 0 to 60% depended onthe explants and hypocotyl (including cotyledonary node) explants were found to be highly susceptible to A. rhizogenes infection withthe highest (60%) rate of hairy root induction. four different Murashige and Skoog (MS)-based liquid culture media were used for wellestablishment of hairy roots. Hairy root growth medium D (HRGM-D) containing hormone-free MS basal medium with an extra oneday pre-incubation period at 35°C was found to be more efficient for profuse growth (fresh weight; 8500 mg per 25 ml culture medium)of hairy roots. Hairy root system presented in this study may offer a suitable platform for optimization and production of satisfactorylevel of aryltetralin lignans like podophyllotoxin and its derivatives from L. mucronatum.


2021 ◽  
Vol 07 ◽  
Author(s):  
Chang-Qi Hao ◽  
Shuai-Run Wang ◽  
Yi Wang ◽  
Xin-Yi Hou ◽  
Ya-Xuan Jiang ◽  
...  

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.


2014 ◽  
Vol 4 (2) ◽  
pp. 107-112 ◽  
Author(s):  
Vinay Kumar ◽  
Dnyanada Desai ◽  
Varsha Shriram

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1080C-1080
Author(s):  
Fredy R. Romero ◽  
David J. Hannapel ◽  
Kathleen Delate

Echinacea is one of the best-selling medicinal plants in the United States. It was historically harvested from wild populations, but its demand has increased so significantly that commercial production has become a necessity to supply the increasing demand and to protect wild populations. The medicinal properties of echinacea are associated with secondary metabolites that are produced mainly in the roots. Hairy roots, induced by the Ri plasmid of Agrobacterium rhizogenes, have been produced in other crops as alternative sources of secondary metabolites that commonly are produced and synthesized in the roots of mature plants. This method of production offers some advantages over traditional agricultural systems, such as the possibility of producing novel compounds year-round. The overall goal of this project is to explore the utility of hairy root cultures (mediated by A. rhizogenes) as an efficient, alternative, and enriched source of secondary compounds with medicinal properties, such as alkamides, flavonoids, and caffeic acid derivatives. We have been successful in transforming roots from E. angustifolia, E. pallida, and E. purpurea plants, and confirming the presence of rol ABC genes in hairy roots using molecular techniques. Roots from control plants show no active growth under dark conditions, whereas transformed roots from E. pallida and E. purpurea show a low degree of branching with a slow growth rate on solid media under darkness. However, transformed E. angustifolia show a faster growth rate and higher degree of branching under the same conditions. Currently, we are working on the optimization of the growing conditions of the transformed roots and will proceed to the biochemical analysis phase of the project.


Agronomy ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1170
Author(s):  
Chrysanthi Foti ◽  
Ourania Pavli

Previous efforts to transform lentil have been considerably hampered by the crop’s recalcitrant nature, giving rise to particularly low transformation and regeneration frequencies. This study aimed at optimizing an Agrobacterium rhizogenes-mediated transformation protocol for the generation of composite lentil plantlets, comprised of transgenic hairy roots and wild-type shoots. Transformation was performed by inoculating the cut hypocotyl of young lentil seedlings, while optimization involved the use of different bacterial strains, namely R1000, K599 and Arqua, and protocols differing in media composition with respect to the presence of acetosyringone and MES. Composite plantlets had a transgenic hairy root system characterized by an increased number of hairy roots at the hypocotyl proximal region, occasionally showing plagiotropic growth. Overall findings underline that transformation frequencies are subject to the bacterial strain, media composition as well as their combined effect. Among strains tested, R1000 proved to be the most capable of hairy root formation, while the presence of both acetosyringone and MES in inoculation and culture media yielded considerably higher transformation rates. The transgenic nature of hairy roots was demonstrated by the Ri T-DNA-mediated transfer of the rolB2 gene and the simultaneous absence of the virCD sequence of A. rhizogenes. Our findings provide strong evidence that A. rhizogenes-mediated transformation may be employed as a suitable approach for generating composite seedlings in lentil, a species whose recalcitrance severely hampers all efforts addressed to transformation and whole plant regeneration procedures. To the best of our knowledge, this is the first report on the development of a non-laborious and time-efficient protocol for the generation of transgenic hairy roots in lentil, thus providing an amenable platform for root biology and gene expression studies in the context of improving traits related to biotic and abiotic stress tolerance.


2017 ◽  
Vol 17 (3) ◽  
pp. 136-142
Author(s):  
Woo Tae Park ◽  
Thanislas Bastin Baskar ◽  
Sun Kyung Yeo ◽  
Nam Il Park ◽  
Jong Seok Park ◽  
...  

Author(s):  
Dwi Kusuma Wahyuni ◽  
Siti Nur Hafida ◽  
Tri Muji Ermayanti ◽  
Bambang Prajoga Eko Wardoyo ◽  
Hery Purnobasuki ◽  
...  

<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>


2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Mohamed A. El-Esawi ◽  
Amr Elkelish ◽  
Hosam O. Elansary ◽  
Hayssam M. Ali ◽  
Mohamed Elshikh ◽  
...  

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. Expression levels of flavonoid biosynthesis genes (CHI, PAL, FLS, and CHS) were assessed in the hairy and nontransformed roots. Results showed higher expression levels in the transgenic roots than in the nontransformed ones (p<0.01). Transgenic hairy roots exhibited a 54.8–96.7% increase in the total phenolic content, 38.1–76.2% increase in the total flavonoid content, and 56.7–96.7% increase in the total reducing power when compared with the nontransgenic roots (p<0.01). DPPH results also revealed that the transgenic hairy roots exhibited a 31.6–50% increase in antioxidant potential, when compared to normal roots. This study addressed the enhancement of secondary metabolite biosynthesis by hairy root induction in L. serriola.


Biologia ◽  
2014 ◽  
Vol 69 (7) ◽  
Author(s):  
Elnaz Nourozi ◽  
Bahman Hosseini ◽  
Abbas Hassani

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.


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