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EcoSal Plus Dec 2022In the late 1950s, a number of laboratories took up the study of plasmids once the discovery was made that extrachromosomal antibiotic resistance (R) factors are the... (Review)
Review
In the late 1950s, a number of laboratories took up the study of plasmids once the discovery was made that extrachromosomal antibiotic resistance (R) factors are the responsible agents for the transmissibility of multiple antibiotic resistance among the enterobacteria. The use of incompatibility for the classification of plasmids is now widespread. It seems clear now on the basis of the limited studies to date that the number of incompatibility groups of plasmids will likely be extremely large when one includes plasmids obtained from bacteria that are normal inhabitants of poorly studied natural environments. The presence of both linear chromosomes and linear plasmids is now established for several species. One of the more fascinating developments in plasmid biology was the discovery of linear plasmids in the 1980s. A remarkable feature of the Ti plasmids of Agrobacterium tumefaciens is the presence of two DNA transfer systems. A definitive demonstration that plasmids consisted of duplex DNA came from interspecies conjugal transfer of plasmids followed by separation of plasmid DNA from chromosomal DNA by equilibrium buoyant density centrifugation. The formation of channels for DNA movement and the actual steps involved in DNA transport offer many opportunities for the discovery of proteins with novel activities and for establishing fundamentally new concepts of macromolecular interactions between DNA and specific proteins, membranes, and the peptidoglycan matrix.
Topics: Plasmids; Agrobacterium tumefaciens; Plant Tumor-Inducing Plasmids; Bacteria; DNA, Bacterial
PubMed: 35373578
DOI: 10.1128/ecosalplus.esp-0028-2021 -
Biotechnology Advances Nov 2015Agrobacterium tumefaciens is widely used as a versatile tool for development of stably transformed model plants and crops. However, the development of Agrobacterium... (Review)
Review
Agrobacterium tumefaciens is widely used as a versatile tool for development of stably transformed model plants and crops. However, the development of Agrobacterium based transient plant transformation methods attracted substantial attention in recent years. Transient transformation methods offer several applications advancing stable transformations such as rapid and scalable recombinant protein production and in planta functional genomics studies. Herein, we highlight Agrobacterium and plant genetics factors affecting transfer of T-DNA from Agrobacterium into the plant cell nucleus and subsequent transient transgene expression. We also review recent methods concerning Agrobacterium mediated transient transformation of model plants and crops and outline key physical, physiological and genetic factors leading to their successful establishment. Of interest are especially Agrobacterium based reverse genetics studies in economically important crops relying on use of RNA interference (RNAi) or virus-induced gene silencing (VIGS) technology. The applications of Agrobacterium based transient plant transformation technology in biotech industry are presented in thorough detail. These involve production of recombinant proteins (plantibodies, vaccines and therapeutics) and effectoromics-assisted breeding of late blight resistance in potato. In addition, we also discuss biotechnological potential of recombinant GFP technology and present own examples of successful Agrobacterium mediated transient plant transformations.
Topics: Agrobacterium tumefaciens; Biotechnology; Gene Transfer Techniques; Plant Proteins; Plants; Transformation, Genetic
PubMed: 25819757
DOI: 10.1016/j.biotechadv.2015.03.012 -
ACS Synthetic Biology Nov 2014Plant Synthetic Biology requires robust and efficient methods for assembling multigene constructs. Golden Gate cloning provides a precision module-based cloning...
Plant Synthetic Biology requires robust and efficient methods for assembling multigene constructs. Golden Gate cloning provides a precision module-based cloning technique for facile assembly of multiple genes in one construct. We present here a versatile resource for plant biologists comprising a set of cloning vectors and 96 standardized parts to enable Golden Gate construction of multigene constructs for plant transformation. Parts include promoters, untranslated sequences, reporters, antigenic tags, localization signals, selectable markers, and terminators. The comparative performance of parts in the model plant Nicotiana benthamiana is discussed.
Topics: Agrobacterium tumefaciens; Cloning, Molecular; Genetic Engineering; Genetic Vectors; Models, Genetic; Synthetic Biology; Nicotiana
PubMed: 24933124
DOI: 10.1021/sb4001504 -
Plant Communications Apr 2024Plant genetic transformation strategies serve as essential tools for the genetic engineering and advanced molecular breeding of plants. However, the complicated...
Plant genetic transformation strategies serve as essential tools for the genetic engineering and advanced molecular breeding of plants. However, the complicated operational protocols and low efficiency of current transformation strategies restrict the genetic modification of most plant species. This paper describes the development of the regenerative activity-dependent in planta injection delivery (RAPID) method based on the active regeneration capacity of plants. In this method, Agrobacterium tumefaciens is delivered to plant meristems via injection to induce transfected nascent tissues. Stable transgenic plants can be obtained by subsequent vegetative propagation of the positive nascent tissues. The method was successfully used for transformation of plants with strong regeneration capacity, including different genotypes of sweet potato (Ipomoea batatas), potato (Solanum tuberosum), and bayhops (Ipomoea pes-caprae). Compared with traditional transformation methods, RAPID has a much higher transformation efficiency and shorter duration, and it does not require tissue culture procedures. The RAPID method therefore overcomes the limitations of traditional methods to enable rapid in planta transformation and can be potentially applied to a wide range of plant species that are capable of active regeneration.
Topics: Plants, Genetically Modified; Agrobacterium tumefaciens; Ipomoea batatas
PubMed: 38243598
DOI: 10.1016/j.xplc.2024.100822 -
Biochemistry and Cell Biology =... Jun 2019is a well studied phytopathogen given its various applications for deciphering host-pathogen interactions, bacterial communication, and capacity to transfer DNA... (Review)
Review
is a well studied phytopathogen given its various applications for deciphering host-pathogen interactions, bacterial communication, and capacity to transfer DNA fragments into host cells via a membrane protein system, the type IV secretion system (T4SS). T4SS mechanism is similar to the one responsible for antibiotic resistance gene transmission, and new knowledge gained could be applied to other organisms using such a mechanism. As well, is of economic importance in biotechnology due to its capacity to generate genetically modified plants. harbours a plasmid known as Ti plasmid encoding T4SS function genes used for transferring genetic information and plant colonization. In this review, the authors describe the molecular basis of infection, from detection of host signals, to the description of different regions of Ti plasmid key to infection, ending with substrate transfer through bacterial wall. [Journal translation].
Topics: Agrobacterium tumefaciens; Host-Parasite Interactions; Plant Diseases; Type IV Secretion Systems
PubMed: 30142282
DOI: 10.1139/bcb-2018-0160 -
Current Topics in Microbiology and... 2018Agrobacterium exopolysaccharides play a major role in the life of the cell. Exopolysaccharides are required for bacterial growth as a biofilm and they protect the... (Review)
Review
Agrobacterium exopolysaccharides play a major role in the life of the cell. Exopolysaccharides are required for bacterial growth as a biofilm and they protect the bacteria against environmental stresses. Five of the exopolysaccharides made by A. tumefaciens have been characterized extensively with respect to their structure, synthesis, regulation, and role in the life of the bacteria. These are cyclic-β-(1, 2)-glucan, cellulose, curdlan, succinoglycan, and the unipolar polysaccharide (UPP). This chapter describes the structure, synthesis, regulation, and function of these five exopolysaccharides.
Topics: Agrobacterium tumefaciens; Biofilms; Polysaccharides, Bacterial
PubMed: 29992358
DOI: 10.1007/82_2018_100 -
Methods in Molecular Biology (Clifton,... 2015
Topics: Agrobacterium tumefaciens; Genetic Engineering; Transformation, Genetic
PubMed: 25568905
DOI: 10.1007/978-1-4939-1658-0 -
Transgenic Research Apr 2023Agrobacterium tumefaciens-mediated plant transformation has become routine work across the world to study gene function and the production of genetically modified... (Review)
Review
Agrobacterium tumefaciens-mediated plant transformation has become routine work across the world to study gene function and the production of genetically modified plants. However, several issues hamper the transformation process in a profound way, both directly and indirectly. One of the major concerns is the overgrowth of Agrobacterium, which occasionally appears after the co-cultivation phase of the explant. This phenomenon is reported in several species and seems to spoil the whole transformation process. There are multiple approaches being employed to counter this unwanted growth of bacteria in a few plant species. In reality, once the overgrowth appears, it becomes nearly impossible to cure it. Hence, for the prevention of this phenomenon, numerous factors are regulated. These factors are: explant nature, A. tumefaciens strain, T-DNA vector, co-cultivation (time and condition), acetosyringone, washing medium, antibiotics (type, concentration, combination, incubation period), etc. In this article, we discuss these factors based on available reports. It can be of immense help in formulating viable strategies to control A. tumefaciens overgrowth.
Topics: Agrobacterium tumefaciens; Plants; Transformation, Genetic; Plants, Genetically Modified
PubMed: 36806963
DOI: 10.1007/s11248-023-00338-w -
Current Topics in Microbiology and... 2018During the last decade, small noncoding RNAs (ncRNAs) have emerged as essential post-transcriptional regulators in bacteria. Nearly all important physiological and... (Review)
Review
During the last decade, small noncoding RNAs (ncRNAs) have emerged as essential post-transcriptional regulators in bacteria. Nearly all important physiological and stress responses are modulated by ncRNA regulators, such as riboswitches, trans-acting small RNAs (sRNAs), and cis-antisense RNAs. Recently, three RNA-seq studies identified a total of 1534 candidate ncRNAs from Agrobacterium tumefaciens, a pathogen and biotechnology tool for plants. Only a few ncRNAs have been functionally characterized in A. tumefaciens, and some of them appear to be involved in virulence. AbcR1 regulates multiple ABC transporters and modulates uptake of a quorum-sensing inhibitor produced by plants. RNA1111, a Ti plasmid-encoded sRNA, might regulate the dispersal of the Ti plasmid and virulence. In addition, a chromosomally encoded sRNA Atr35C is induced by the vir gene regulator VirG and its expression is affected by iron, manganese, and hydrogen peroxide, suggesting a possible role in oxidative stress responses and Agrobacterium-plant interactions. Progress in ncRNA functional analysis is slow, likely resulting from innate challenges, such as poor sequence conservation and imperfect base-pairing between sRNAs and mRNAs, which make computational target predictions inefficient. Advances in single-cell-based RNA-seq and proteomics approaches would provide valuable tools to reveal regulatory networks involving ncRNA regulators.
Topics: Agrobacterium tumefaciens; Gene Expression Regulation, Bacterial; RNA, Bacterial; RNA, Messenger; RNA, Small Untranslated
PubMed: 29556823
DOI: 10.1007/82_2018_84 -
Methods in Molecular Biology (Clifton,... 2015Agrobacterium tumefaciens-mediated transformation of sour chgahtvy (Prunus cerasus L.) "Montmorency" and sweet cherry rootstocks "Gisela 6" and "Gisela 7" (P. cerasus ×...
Agrobacterium tumefaciens-mediated transformation of sour chgahtvy (Prunus cerasus L.) "Montmorency" and sweet cherry rootstocks "Gisela 6" and "Gisela 7" (P. cerasus × P. canescens) is described. Briefly, leaf explants from in vitro shoots are cocultivated with A. tumefaciens either directly (for "Gisela 6" and "Gisela 7") or after pretreatment (for "Montmorency") on cocultivation medium; selection and regeneration of transformed shoots are carried out on selection medium containing 50 mg/L kanamycin (Km) and 250 mg/L timentin (or cefotaxime) for 3-5 months. In this protocol, the optimal media for shoot proliferation and shoot regeneration from leaf explants are genotype dependent.
Topics: Agrobacterium tumefaciens; Coculture Techniques; Environment, Controlled; Genetic Engineering; Plant Leaves; Plant Roots; Prunus; Regeneration; Transformation, Genetic
PubMed: 25416255
DOI: 10.1007/978-1-4939-1658-0_12