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Tytuł:
Odyssey of Agrobacterium T-DNA.
Autorzy:
Ziemienowicz, Alicja
Tematy:
Agrobacterium
DNA transfer
T-DNA
virulence
Pokaż więcej
Wydawca:
Polskie Towarzystwo Biochemiczne
Powiązania:
https://bibliotekanauki.pl/articles/1044091.pdf  Link otwiera się w nowym oknie
Opis:
Agrobacterium tumefaciens, a plant pathogen, is characterized by the unique feature of interkingdom DNA transfer. This soil bacterium is able to transfer a fragment of its DNA, called T-DNA (transferred DNA), to the plant cell where T-DNA is integrated into the plant genome leading to "genetic colonization" of the host. The fate of T-DNA, its processing, transfer and integration, resembles the journey of Odysseus, although our hero returns from its long trip in a slightly modified form.The soil bacterium, Agrobacterium tumefaciens, is a plant pathogen responsible for tumor induction on dicotyledonous plants due to its ability to transfer DNA to the plant cell (reviewed in: de la Cruz & Lanka, 1998; Gelvin, 2000; Hansen & Chilton, 1999; Lartey & Citovsky, 1997; Rossi et al., 1998; Zupan & Zambryski, 1997). In biotechnology this ability is widely used for plant transformation. During tumor induction Agrobacterium attaches to plant cells and then transfers part of its DNA to some of these cells. The transferred DNA (T-DNA) which resides on a large Ti (tumor inducing) plasmid, is processed within the bacterium and is exported to the plant where it becomes integrated into the plant genome (reviewed in: Sheng & Citovsky, 1996; Tinland & Hohn, 1995; Tinland, 1996). Proteins encoded by the virulence (vir) region of the Ti plasmid regulate T-DNA processing and transfer. Phenolic compounds derived from a wounded plant cell wall induce expression of the vir region genes. Virulence proteins recognize 25 bp imperfect direct repeats (border sequences) that define the T-DNA. In the presence of VirD1 protein, VirD2 cleaves the border sequence in a site- and strand-specific manner and subsequently becomes covalently attached to the 5' end of the nicked DNA. The nicked DNA is then displaced from the plasmid producing single-stranded T-DNA. The T-DNA-VirD2 complex and the VirE2 protein are believed to be transferred to the plant through a pilus-like structure containing VirB and VirD4 proteins. In the plant cell, T-DNA becomes coated with the single-stranded DNA-binding protein, VirE2. The T-DNA-protein complex is imported into the nucleus where the T-DNA is integrated into the nuclear genome. Expression of genes located on T-DNA leads to the formation of proteins involved in the production of auxins and cytokinins. These plant hormones cause the tumorous phenotype that is characterized by the ability of the plant cells to proliferate limitlessly and autonomously even in the absence of added phytohormones. Crown gall tumors are characterized by the production of opines (amino-acid derivatives). The biosynthesis of opines is catalyzed by opine synthases, which are encoded by the T-DNA. Opines formed in the tumors can be metabolized by the tumorigenic agrobacteria, but not by most of the other soil organisms. Thus, Agrobacterium creates for itself a favorable niche by genetic modification of plant cells, a process called "genetic colonization". All stages of this colonization, including chemotaxis, attachment, induction of virulence region, processing of T-DNA, T-DNA transfer, T-DNA integration, expression of T-DNA genes and changes in the plant phenotype, will be discussed in the following chapters. This will be an odyssey of T-DNA that leaves the Agrobacterium cell in the form of nucleic acid and returns from its journey in the form of opines, derivatives of amino acids (Fig. 1).
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Production of oleanolic acid glycosides by hairy root established cultures of Calendula officinalis L.
Autorzy:
Długosz, Marek
Wiktorowska, Ewa
Wiśniewska, Anita
Pączkowski, Cezary
Tematy:
Marigold
Agrobacterium rhizogenes
hairy roots
triterpenic saponins
oleanolic acid
transformation
Pokaż więcej
Wydawca:
Polskie Towarzystwo Biochemiczne
Powiązania:
https://bibliotekanauki.pl/articles/1039552.pdf  Link otwiera się w nowym oknie
Opis:
In order to initiate hairy root culture initiation cotyledons and hypocotyls of Calendula officinalis L. were infected with Agrobacterium rhizogenes strain ATCC 15834 or the same strain containing pCAMBIA 1381Z vector with β-glucuronidase reporter gene under control of promoter of NIK (Nematode Induced Kinase) gene. The efficiency of induction of hairy roots reached 33.8% for cotyledons and 66.6% for hypocotyls together for both transformation experiments. Finally, eight control and nine modified lines were established as a long-term culture. The hairy root cultures showed the ability to synthesize oleanolic acid mainly (97%) as glycosides; control lines contained it at the average 8.42 mg · g-1 dry weight in tissue and 0.23 mg · dm-3 in medium; modified lines: 4.59 mg · g-1 for the tissue, and 0.48 mg · dm-3 for the medium. Additionally lines showed high positive correlation between dry/fresh weight and oleanolic acid concentration in tissue. Using the Killiani mixture in acidic hydrolysis of oleanolic acid glycosides released free aglycones that were partially acetylated in such conditions.
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Optimization of transient Agrobacterium-mediated gene expression system in leaves of Nicotiana benthamiana
Autorzy:
Wydro, Mateusz
Kozubek, Edward
Lehmann, Przemysław
Tematy:
RNA silencing
Nicotiana benthamiana
viral suppressor
Agrobacterium-mediated transient expression
Pokaż więcej
Wydawca:
Polskie Towarzystwo Biochemiczne
Powiązania:
https://bibliotekanauki.pl/articles/1041239.pdf  Link otwiera się w nowym oknie
Opis:
Here we report on a simple and reproducible system of Agrobacterium-mediated transient gene expression assay that utilizes infiltration of young Nicotiana benthamiana leaves. Although some of the phenomena described in this paper have been already reported by other researchers, here we have further developed them. The highest level of transient gfp gene expression was detected in the youngest leaves of N. benthamiana infiltrated with A. tumefaciens strains AGL0 and EHA105 precultured in the presence of 450-600 µM acetosyringone. Although the maximum level of transient gfp gene expression was restricted presumably by RNA silencing, it was completely suppressed in the presence of the viral protein HC-Pro. The transient expression system described here can be used to identify new viral suppressors of RNA silencing, for detailed analysis of unidentified genes and for industrial production of proteins in plants as well.
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Transformation of wild Solanum species resistant to late blight by using reporter gene gfp and msh2 genes
Autorzy:
Rakosy-Tican, Lenuta
Aurori, Adriana
Aurori, Cristian M.
Ispas, Gabriela
Famelaer, Ivan
Tematy:
Agrobacterium tumefaciens mediated transformation
DNA mismatch repair
gfp
nptII marker gene
Pokaż więcej
Wydawca:
Instytut Hodowli i Aklimatyzacji Roślin
Powiązania:
https://bibliotekanauki.pl/articles/55928632.pdf  Link otwiera się w nowym oknie
Opis:
Green fluorescent protein (gfp) reporter gene and nptII marker gene were used to optimize Agrobacterium tumefaciens (agro) mediated transformation of wild Solanum genotypes resistant to late blight. Different genotypes of Solanum bulbocastanum, S. chacoense, S. microdontum and S. verrucosum were assessed for their regeneration ability on MS based media and for agro-mediated transformation. As the first step reporter genes were used to optimize transformation protocol for each species and then the transfer of genes involved in mismatch repair of DNA were attempted in Solanum chacoense. For transformation, either leaf or stem fragments were used. It was shown that gfp is a valuable and elegant tool for monitoring the efficiency of transformation or the occurrence of chimera in all genotypes. Transformation efficiency was dependent on a plant genotype. A number of genotypes have been successfully transformed and they expressed constitutively the bright green fluorescence of gfp without any side effects. The most recalcitrant species proved to be S. microdontum, which did not regenerate plants although different media and phytohormones had been used. The best protocol for S. chacoense transformation was also found to work in the transfer of msh2 genes. Msh2 isolated from Arabidopsis was used and transferred either as mutated (Apa) or antisense (As) gene. The integration of msh2-mutated gene into S. chacoense genome was demonstrated by PCR amplification and confirmed by RT-PCR for some of the putative transgenic clones. The implications of mismatch repair in homologous recombination and its importance for potato improvement are discussed. 
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Transformation of wild Solanum species resistant to late blight by using reporter gene gfp and msh2 genes
Autorzy:
Rakosy-Tican, Lenuta
Aurori, Adriana
Aurori, Cristian M.
Ispas, Gabriela
Famelaer, Ivan
Tematy:
Agrobacterium tumefaciens mediated transformation
DNA mismatch repair
gfp
nptII marker gene
Pokaż więcej
Wydawca:
Instytut Hodowli i Aklimatyzacji Roślin
Powiązania:
https://bibliotekanauki.pl/articles/2198986.pdf  Link otwiera się w nowym oknie
Opis:
Green fluorescent protein (gfp) reporter gene and nptII marker gene were used to optimize Agrobacterium tumefaciens (agro) mediated transformation of wild Solanum genotypes resistant to late blight. Different genotypes of Solanum bulbocastanum, S. chacoense, S. microdontum and S. verrucosum were assessed for their regeneration ability on MS based media and for agro-mediated transformation. As the first step reporter genes were used to optimize transformation protocol for each species and then the transfer of genes involved in mismatch repair of DNA were attempted in Solanum chacoense. For transformation, either leaf or stem fragments were used. It was shown that gfp is a valuable and elegant tool for monitoring the efficiency of transformation or the occurrence of chimera in all genotypes. Transformation efficiency was dependent on a plant genotype. A number of genotypes have been successfully transformed and they expressed constitutively the bright green fluorescence of gfp without any side effects. The most recalcitrant species proved to be S. microdontum, which did not regenerate plants although different media and phytohormones had been used. The best protocol for S. chacoense transformation was also found to work in the transfer of msh2 genes. Msh2 isolated from Arabidopsis was used and transferred either as mutated (Apa) or antisense (As) gene. The integration of msh2-mutated gene into S. chacoense genome was demonstrated by PCR amplification and confirmed by RT-PCR for some of the putative transgenic clones. The implications of mismatch repair in homologous recombination and its importance for potato improvement are discussed. 
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Improved transformation of Agrobacterium assisted by silver nanoparticle
Autorzy:
Benny, Amala
Alex, Swapny
Soni, K.B.
Anith, K.N.
Kiran, A.G.
Viji, M.M.
Tematy:
silver nanoparticles
Agrobacterium transformation
calcium chloride
competent cells
freeze thaw
transformation efficiency
Pokaż więcej
Wydawca:
Polska Akademia Nauk. Czasopisma i Monografie PAN
Powiązania:
https://bibliotekanauki.pl/articles/16648111.pdf  Link otwiera się w nowym oknie
Opis:
In transgenic plant development, the low transformation efficiency of Agrobacterium with exogenous DNA is the major constraint, and hence, methods to improve its transformation efficiency are needed. Recently, nanoparticlemediated gene transfer has evolved as a key transformational tool in genetic transformation. Since silver nanoparticles (AgNPs) can induce pores on the cell membrane, their efficacy in the improvement of conventional calcium chloride freeze-thaw technique of transformation of Agrobacterium was explored in this study. Agrobacterium cells in the exponential growth phase were exposed to different concentrations of AgNPs (0.01, 1, 5, 10, and 20 mg/l), and the half-maximal effective concentration (EC50) was determined via Probit analysis using the SPSS software. Transformation efficiency of AgNPs alone and in combination with calcium chloride was compared with that of the conventional calcium chloride freeze-thaw technique. AgNPs at a concentration of 0.01 mg/l in combination with calcium chloride (20 mM) showed a ten fold increase in the transformation efficiency (3.33 log CFU (colony-forming unit/microgram of DNA) of Agrobacterium tumefaciens strain EHA 105 with plasmid vector pART27 compared with the conventional technique (2.31 log CFU/μg of DNA). This study indicates that AgNPs of size 100 nm can eliminate the freeze-thaw stage in the conventional Agrobacterium transformation technique, with a 44% improvement in efficiency. The use of AgNPs (0.01 mg/l) along with 20 mM calcium chloride was found to be an economically viable method to improve the transformation of Agrobacterium with exogenous plasmid DNA.
Dostawca treści:
Biblioteka Nauki
Artykuł
Wyświetlanie 1-10 z 16

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