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Agrobacterium and Plant Genetic Engineering
Published in Yoshikatsu Murooka, Tadayuki Imanaka, Recombinant Microbes for Industrial and Agricultural Applications, 2020
Masami Sekine, Atsuhiko Shinmyo
Agrobacterium rhizogenes is a closely related species of A. tumefaciens, and causes hairy root disease on dicotyledonous plants. This bacterium also contains a large plasmid, called the Ri (root-inducing) plasmid. The T-DNA region of the Ri plasmid is transferred to the plant genome, presumably much like the Ti plasmid T-DNA transformation [73,74].
Establishment of hairy root system of transgenic IRT1 brassica campestris L. and preliminary study of its effect on cadmium enrichment
Published in International Journal of Phytoremediation, 2023
Xiaoyu Liu, Wenxuan Li, Menghua Wang, Yushen Cao, Teng Zhang, Honggang Hu, Xiyu Cheng, Qiong Yan
The results of DNA gel electrophoresis (Figure 2) showed that a specific DNA fragment of 423 bp, namely the rolB gene, was successfully amplified using the transgenic hairy root genome (lanes 4-6) . The rolB gene is related to the formation of hairy roots and is the most critical gene in the process of Ri plasmid transformation. The presence or absence of rolB can be used as a marker for the successful construction of hairy roots. Using the transgenic hairy root genome as template, it is amplified another specific DNA fragment of about 471 bp (lanes 10-12), namely the IRT1 gene, which was the same as the amplification result using the recombinant plasmid pRI101-IRT1 as a template. Using B. campestris L. genome as the negative control, no fragments (rol B and IRT1) were amplified (lanes 3 and 9). Therefore, the above experiments proved that the rolB and IRT1 genes were integrated into the genome of B. campestris L. hairy roots.
Establishment and elicitation of transgenic root culture of Plantago lanceolata and evaluation of its anti-bacterial and cytotoxicity activity
Published in Preparative Biochemistry & Biotechnology, 2021
Samaneh Rahamouz-Haghighi, Khadijeh Bagheri, Ali Sharafi, Hossein Danafar
One of the plant diseases is hairy root (HR) which is caused by Agrobacterium rhizogenes.[1] HR occurs when root loci (rol) genes harbored by the root inducing plasmid (Ri) of A. rhizogenes integrate into the nuclear genome of the infected plant cells. It is believed that numerous factors impact the frequency of A. rhizogenes mediated transformation in host plants like bacterial strains, Acetosyringone as well as salt concentration in co-cultivation media.[2] The high growth rate of transformed root cultures is very attractive for the industrial production of secondary metabolites.[3] In a study, Sudha et al. mentioned that the effects of endogenous hormones and bacterial strains reacted to the rol gene products might lead to tumors.[4] The T-DNA regions, chromosomal virulence (chv) and virulence (vir) genes, are essential for DNA transference from bacteria to the host plant cells. The rol A, B, C and D genes are supported by TL-DNA that separates the Ri-plasmid of A. rhizogenes from the Ti-plasmid in A. tumefaciens.[5,6] The rolB induced HR and plays a key role in the secretion of active auxin performed by indoxyl-β-oxidase activity.[7] On the other hand, rolA, rolC, rolD, and other open reading frames act together in order to promote root induction.[8]
Comparative analysis of glucosinolate production in hairy roots of green and red kale (Brassica oleracea var. acephala)
Published in Preparative Biochemistry and Biotechnology, 2019
Do Manh Cuong, Sang Un Park, Chang Ha Park, Nam Su Kim, Sun Ju Bong, Sook Young Lee
Hairy root cultures of kale were initiated using A. rhizogenes strain R1000 for inducing extensive root development at wounding sites on kale leaves. Green and red kale hairy roots were established at wounding sites on leaves at 5 to 7 days of inoculation with A. rhizogenes strain R1000. These hairy roots began to grow more rapidly over the next 14 to 21 days and showed extensive branching, whereas, the non-transformed roots failed to grow. One fast-growing clone of green or red kale was selected and grown on 1/2× MS agar supplemented with 250 mg/L of cefotaxime (Figure 1A,C). After consecutive transfers to fresh medium over 30 days, hairy roots were transferred to the liquid culture (Figure 1B,D). Formation of the hairy roots, used in this study, typically depends on the rol gene of the Ri-plasmid in A. rhizogenesis, which is responsible for inducing hairy roots in plants. To confirm the insertion of rol genes, PCR analysis was performed using primers for amplifying rolA, rolB, rolC, and rolD gene fragments (Table 2). The hairy root samples generated fragments of the expected size for rolA, rolB, rolC, and rolD, whereas, the wild-type (control) root sample did not generate any fragment. These results confirmed that the hairy roots contained the rol genes of the Ri-plasmid (Figure 2).