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Agroinfiltration
Agroinfiltration is a method used in plant biology and especially lately in plant biotechnology to induce transient expression of genes in a plant, or isolated leaves from a plant, or even in cultures of plant cells, in order to produce a desired protein. In the method, a suspension of '' Agrobacterium tumefaciens'' is introduced into a plant leaf by direct injection or by vacuum infiltration, or brought into association with plant cells immobilised on a porous support (plant cell packs), whereafter the bacteria transfer the desired gene into the plant cells via transfer of T-DNA. The main benefit of agroinfiltration when compared to the more traditional plant transformation is speed and convenience, although yields of the recombinant protein are generally also higher and more consistent. The first step is to introduce a gene of interest to a strain of ''Agrobacterium tumefaciens''. Subsequently, the strain is grown in a liquid culture and the resulting bacteria are washed and s ...
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Agroinfiltration
Agroinfiltration is a method used in plant biology and especially lately in plant biotechnology to induce transient expression of genes in a plant, or isolated leaves from a plant, or even in cultures of plant cells, in order to produce a desired protein. In the method, a suspension of '' Agrobacterium tumefaciens'' is introduced into a plant leaf by direct injection or by vacuum infiltration, or brought into association with plant cells immobilised on a porous support (plant cell packs), whereafter the bacteria transfer the desired gene into the plant cells via transfer of T-DNA. The main benefit of agroinfiltration when compared to the more traditional plant transformation is speed and convenience, although yields of the recombinant protein are generally also higher and more consistent. The first step is to introduce a gene of interest to a strain of ''Agrobacterium tumefaciens''. Subsequently, the strain is grown in a liquid culture and the resulting bacteria are washed and s ...
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Transient Expression
Transient expression, more frequently referred to "transient gene expression", is the temporary expression of genes that are expressed for a short time after nucleic acid, most frequently plasmid DNA encoding an expression cassette, has been introduced into eukaryotic cells with a chemical delivery agent like calcium phosphate (CaPi) or polyethyleneimine (PEI). However, unlike "stable expression," the foreign DNA does not fuse with the host cell DNA, resulting in the inevitable loss of the vector after several cell replication cycles. The majority of transient gene expressions are done with cultivated animal cells. The technique is also used in plant cells; however, the transfer of nucleic acids into these cells requires different methods than those with animal cells. In both plants and animals, transient expression should result in a time-limited use of transferred nucleic acids, since any long-term expression would be called "stable expression." Methodology varies depending on t ...
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Agroinfiltration Uing A Promoter--GUS Construct With And Without P19
Agroinfiltration is a method used in plant biology and especially lately in plant biotechnology to induce transient expression of genes in a plant, or isolated leaves from a plant, or even in cultures of plant cells, in order to produce a desired protein. In the method, a suspension of ''Agrobacterium tumefaciens'' is introduced into a plant leaf by direct injection or by vacuum infiltration, or brought into association with plant cells immobilised on a porous support (plant cell packs), whereafter the bacteria transfer the desired gene into the plant cells via transfer of T-DNA. The main benefit of agroinfiltration when compared to the more traditional plant transformation is speed and convenience, although yields of the recombinant protein are generally also higher and more consistent. The first step is to introduce a gene of interest to a strain of ''Agrobacterium tumefaciens''. Subsequently, the strain is grown in a liquid culture and the resulting bacteria are washed and sus ...
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Nicotiana Benthamiana
''Nicotiana benthamiana'', colloquially known as benth or benthi, is a species of ''Nicotiana'' indigenous to Australia. It is a close relative of tobacco. A synonym for this species is ''Nicotiana suaveolens'' var. ''cordifolia'', a description given by George Bentham in ''Flora Australiensis'' in 1868. This was transferred to ''Nicotiana benthamiana'' by Karel Domin in ''Bibliotheca Botanica'' (1929), honoring the original author in the specific epithet. History The plant was used by people of Australia as a stimulant, containing nicotine and other alkaloids, before the introduction of commercial tobacco ('' N. tabacum'' and '' N. rustica''). Indigenous names for it include ''tjuntiwari'' and ''muntju''. It was first collected on the north coast of Australia by Benjamin Bynoe on a voyage of in 1837. Description The herbaceous plant is found amongst rocks on hills and cliffs throughout the northern regions of Australia. Variable in height and habit, the species may be er ...
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Stomata
In botany, a stoma (from Greek ''στόμα'', "mouth", plural "stomata"), also called a stomate (plural "stomates"), is a pore found in the epidermis of leaves, stems, and other organs, that controls the rate of gas exchange. The pore is bordered by a pair of specialized parenchyma cells known as guard cells that are responsible for regulating the size of the stomatal opening. The term is usually used collectively to refer to the entire stomatal complex, consisting of the paired guard cells and the pore itself, which is referred to as the stomatal aperture. Air, containing oxygen, which is used in respiration, and carbon dioxide, which is used in photosynthesis, passes through stomata by gaseous diffusion. Water vapour diffuses through the stomata into the atmosphere in a process called transpiration. Stomata are present in the sporophyte generation of all land plant groups except liverworts. In vascular plants the number, size and distribution of stomata varies widely. ...
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Small Interfering RNA
Small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, is a class of double-stranded RNA at first non-coding RNA molecules, typically 20-24 (normally 21) base pairs in length, similar to miRNA, and operating within the RNA interference (RNAi) pathway. It interferes with the expression of specific genes with complementary nucleotide sequences by degrading mRNA after transcription, preventing translation. Text was copied from this source, which is available under Creative Commons Attribution 4.0 International License Structure Naturally occurring siRNAs have a well-defined structure that is a short (usually 20 to 24- bp) double-stranded RNA (dsRNA) with phosphorylated 5' ends and hydroxylated 3' ends with two overhanging nucleotides. The Dicer enzyme catalyzes production of siRNAs from long dsRNAs and small hairpin RNAs. siRNAs can also be introduced into cells by transfection. Since in principle any gene can be knocked down by a syntheti ...
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RNA Interference
RNA interference (RNAi) is a biological process in which RNA molecules are involved in sequence-specific suppression of gene expression by double-stranded RNA, through translational or transcriptional repression. Historically, RNAi was known by other names, including ''co-suppression'', ''post-transcriptional gene silencing'' (PTGS), and ''quelling''. The detailed study of each of these seemingly different processes elucidated that the identity of these phenomena were all actually RNAi. Andrew Fire and Craig C. Mello shared the 2006 Nobel Prize in Physiology or Medicine for their work on RNAi in the nematode worm '' Caenorhabditis elegans'', which they published in 1998. Since the discovery of RNAi and its regulatory potentials, it has become evident that RNAi has immense potential in suppression of desired genes. RNAi is now known as precise, efficient, stable and better than antisense therapy for gene suppression. Antisense RNA produced intracellularly by an expression vector m ...
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Bunyaviridae
''Bunyavirales'' is an order of segmented negative-strand RNA viruses with mainly tripartite genomes. Member viruses infect arthropods, plants, protozoans, and vertebrates. It is the only order in the class ''Ellioviricetes''. The name ''Bunyavirales'' derives from Bunyamwera, where the original type species ''Bunyamwera orthobunyavirus'' was first discovered. ''Ellioviricetes'' is named in honor of late virologist Richard M. Elliott (virologist), Richard M. Elliott for his early work on bunyaviruses. Bunyaviruses belong to the fifth group of the Baltimore classification, Baltimore classification system, which includes viruses with a Sense (molecular biology), negative-sense, single-stranded RNA genome. They have an Viral envelope, enveloped, spherical virion. Though generally found in arthropods or rodents, certain viruses in this order occasionally infect humans. Some of them also infect plants. In addition, there is a group of bunyaviruses whose replication is restricted to a ...
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Tospovirus
''Orthotospovirus'' is a genus of negative-strand RNA viruses, in the family ''Tospoviridae'' of the order ''Bunyavirales,'' which infects plants. Tospoviruses take their name from the species '' Tomato spotted wilt orthotospovirus'' (TSWV) which was discovered in Australia in 1919. TSWV remained the only known member of the family until the early 1990s when genetic characterisation of plant viruses became more common. There are now at least twenty species in the genus with more being discovered on a regular basis. Member viruses infect over eight hundred plant species from 82 different families. Genome Tospoviruses have a negative-sense, single-strand RNA genome. The genome resembles that of the genus ''Phlebovirus''. It is linear and is 17.2 kb in size. It is divided into three segments termed S (2.9kb), M (5.4kb), and L (8.9kb). The M and S RNA segments encode for proteins in an ambisense direction. Transmission Tospoviruses are arboviruses usually vectored by thrips. At ...
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Tomato
The tomato is the edible berry of the plant ''Solanum lycopersicum'', commonly known as the tomato plant. The species originated in western South America, Mexico, and Central America. The Mexican Nahuatl word gave rise to the Spanish word , from which the English word ''tomato'' derived. Its domestication and use as a cultivated food may have originated with the indigenous peoples of Mexico. The Aztecs used tomatoes in their cooking at the time of the Spanish conquest of the Aztec Empire, and after the Spanish encountered the tomato for the first time after their contact with the Aztecs, they brought the plant to Europe, in a widespread transfer of plants known as the Columbian exchange. From there, the tomato was introduced to other parts of the European-colonized world during the 16th century. Tomatoes are a significant source of umami flavor. They are consumed in diverse ways: raw or cooked, and in many dishes, sauces, salads, and drinks. While tomatoes are fruits†...
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Tomato Spotted Wilt Virus
Transmission and lifespan TSWV, which is transmitted by thrips, causes serious losses in economically important crops and it is one of the most economically devastating plant viruses in the world. The circulative propagative transmission of TSWV is carried out by at least ten different species of thrips. The most common species is ''Frankliniella occidentalis'' (western flower thrips) as it is the vector that predominantly transmits TSWV globally and in greenhouses. The rapid developmental and reproductive rate of the thrips contributes to the spread of TSWV. The amount of time it takes for insects to acquire the virus (acquisition period) and the amount of time it takes for the virus to move from the insect to the plant (inoculation) for TSWV varies depends on the vector species. For ''Frankliniella occidentalis'', the acquisition and inoculation of TSWV can be as short as 5 minutes. However, the acquisition and inoculation periods for optimal transmission is 21.3 hours and 42 ...
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Tomato Bushy Stunt Virus
Tomato bushy stunt virus (TBSV) is a virus of the tombusvirus family. It was first reported in tomatoes in 1935 and primarily affects vegetable crops, though it is not generally considered an economically significant plant pathogen. Depending upon the host, TBSV causes stunting of growth, leaf mottling, and deformed or absent fruit. The virus is likely to be soil-borne in the natural setting, but can also transmitted mechanically, for example through contaminated cutting tools. TBSV has been used as a model system in virology research on the life cycle of plant viruses, particularly in experimental infections of the model host plant ''Nicotiana benthamiana''. Host range TBSV has a broad host range under experimental conditions and has been reported to infect over 120 plant species spanning 20 families. However, under natural conditions its range is much narrower and generally comprises crop vegetables and ornamental plants. It was first identified in tomato plants and also has ...
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