Fungal DNA Barcoding
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Fungal DNA Barcoding
Fungal DNA barcoding is the process of identifying species of the biological kingdom Fungi through the amplification and sequencing of specific DNA sequences and their comparison with sequences deposited in a DNA barcode database such as the ISHAM reference database, or the Barcode of Life Data System (BOLD). In this attempt, DNA barcoding relies on universal genes that are ideally present in all fungi with the same degree of sequence variation. The interspecific variation, i.e., the variation between species, in the chosen DNA barcode gene should exceed the intraspecific (within-species) variation. A fundamental problem in fungal systematics is the existence of teleomorphic and anamorphic stages in their life cycles. These morphs usually differ drastically in their phenotypic appearance, preventing a straightforward association of the asexual anamorph with the sexual teleomorph. Moreover, fungal species can comprise multiple strains that can vary in their morphology or in tra ...
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Kingdom (biology)
In biology, a kingdom is the second highest taxonomic rank, just below domain. Kingdoms are divided into smaller groups called phyla (singular phylum). Traditionally, textbooks from Canada and the United States have used a system of six kingdoms (Animalia, Plantae, Fungi, Protista, Archaea/Archaebacteria, and Bacteria or Eubacteria), while textbooks in other parts of the world, such as Bangladesh, Brazil, Greece, India, Pakistan, Spain, and the United Kingdom have used five kingdoms (Animalia, Plantae, Fungi, Protista and Monera). Some recent classifications based on modern cladistics have explicitly abandoned the term ''kingdom'', noting that some traditional kingdoms are not monophyletic, meaning that they do not consist of all the descendants of a common ancestor. The terms ''flora'' (for plants), ''fauna'' (for animals), and, in the 21st century, '' funga'' (for fungi) are also used for life present in a particular region or time. Definition and associated terms Whe ...
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Ribosome
Ribosomes () are molecular machine, macromolecular machines, found within all cell (biology), cells, that perform Translation (biology), biological protein synthesis (messenger RNA translation). Ribosomes link amino acids together in the order specified by the codons of messenger RNA molecules to form polypeptide chains. Ribosomes consist of two major components: the small and large ribosomal subunits. Each subunit consists of one or more ribosomal RNA molecules and many ribosomal proteins (). The ribosomes and associated molecules are also known as the ''translational apparatus''. Overview The sequence of DNA that encodes the sequence of the amino acids in a protein is transcribed into a messenger RNA (mRNA) chain. Ribosomes bind to the messenger RNA molecules and use the RNA's sequence of nucleotides to determine the sequence of amino acids needed to generate a protein. Amino acids are selected and carried to the ribosome by transfer RNA (tRNA) molecules, which enter the riboso ...
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Basidiomycota
Basidiomycota () is one of two large divisions that, together with the Ascomycota, constitute the subkingdom Dikarya (often referred to as the "higher fungi") within the kingdom Fungi. Members are known as basidiomycetes. More specifically, Basidiomycota includes these groups: agarics, puffballs, stinkhorns, bracket fungi, other polypores, jelly fungi, boletes, chanterelles, earth stars, smuts, bunts, rusts, mirror yeasts, and '' Cryptococcus'', the human pathogenic yeast. Basidiomycota are filamentous fungi composed of hyphae (except for basidiomycota-yeast) and reproduce sexually via the formation of specialized club-shaped end cells called basidia that normally bear external meiospores (usually four). These specialized spores are called basidiospores. However, some Basidiomycota are obligate asexual reproducers. Basidiomycota that reproduce asexually (discussed below) can typically be recognized as members of this division by gross similarity to others, by ...
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Dikarya
Dikarya is a subkingdom of Fungi that includes the divisions Ascomycota and Basidiomycota, both of which in general produce dikaryons, may be hypha, filamentous or unicellular, but are always without flagella. The Dikarya are most of the so-called "higher fungi", but also include many anamorphic species that would have been classified as Mold (fungus), molds in historical literature. Phylogenetically the two divisions regularly group together. In a 1998 publication, Thomas Cavalier-Smith referred to this group as the Neomycota. Phylogeny The 2007 classification of Kingdom Fungi is the result of a large-scale collaborative research effort involving dozens of mycologists and other scientists working on fungal Taxonomy (biology), taxonomy. It recognizes seven phylum, divisions within the Fungi, two of which—the Ascomycota and the Basidiomycota—are contained within a branch representing subkingdom Dikarya. The cladogram depicts the major fungal Taxon, taxa and their relationship ...
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Polymerase Chain Reaction
The polymerase chain reaction (PCR) is a method widely used to make millions to billions of copies of a specific DNA sample rapidly, allowing scientists to amplify a very small sample of DNA (or a part of it) sufficiently to enable detailed study. PCR was invented in 1983 by American biochemist Kary Mullis at Cetus Corporation. Mullis and biochemist Michael Smith (chemist), Michael Smith, who had developed other essential ways of manipulating DNA, were jointly awarded the Nobel Prize in Chemistry in 1993. PCR is fundamental to many of the procedures used in genetic testing and research, including analysis of Ancient DNA, ancient samples of DNA and identification of infectious agents. Using PCR, copies of very small amounts of DNA sequences are exponentially amplified in a series of cycles of temperature changes. PCR is now a common and often indispensable technique used in medical laboratory research for a broad variety of applications including biomedical research and forensic ...
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Primer (molecular Biology)
A primer is a short, single-stranded nucleic acid used by all living organisms in the initiation of DNA synthesis. A synthetic primer is a type of oligo, short for oligonucleotide. DNA polymerases (responsible for DNA replication) are only capable of adding nucleotides to the 3’-end of an existing nucleic acid, requiring a primer be bound to the template before DNA polymerase can begin a complementary strand. DNA polymerase adds nucleotides after binding to the RNA primer and synthesizes the whole strand. Later, the RNA strands must be removed accurately and replaced with DNA nucleotides. This forms a gap region known as a nick that is filled in using a ligase. The removal process of the RNA primer requires several enzymes, such as Fen1, Lig1, and others that work in coordination with DNA polymerase, to ensure the removal of the RNA nucleotides and the addition of DNA nucleotides. Living organisms use solely RNA primers, while laboratory techniques in biochemistry and mole ...
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DNA Barcode
DNA barcoding is a method of species identification using a short section of DNA from a specific gene or genes. The premise of DNA barcoding is that by comparison with a reference library of such DNA sections (also called "sequences"), an individual sequence can be used to uniquely identify an organism to species, just as a supermarket scanner uses the familiar black stripes of the UPC barcode to identify an item in its stock against its reference database. These "barcodes" are sometimes used in an effort to identify unknown species or parts of an organism, simply to catalog as many taxa as possible, or to compare with traditional taxonomy in an effort to determine species boundaries. Different gene regions are used to identify the different organismal groups using barcoding. The most commonly used barcode region for animals and some protists is a portion of the cytochrome ''c'' oxidase I (COI or COX1) gene, found in mitochondrial DNA. Other genes suitable for DNA barcoding are ...
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Translation (biology)
In biology, translation is the process in living Cell (biology), cells in which proteins are produced using RNA molecules as templates. The generated protein is a sequence of amino acids. This sequence is determined by the sequence of nucleotides in the RNA. The nucleotides are considered three at a time. Each such triple results in the addition of one specific amino acid to the protein being generated. The matching from nucleotide triple to amino acid is called the genetic code. The translation is performed by a large complex of functional RNA and proteins called ribosomes. The entire process is called gene expression. In translation, messenger RNA (mRNA) is decoded in a ribosome, outside the nucleus, to produce a specific amino acid chain, or polypeptide. The polypeptide later protein folding, folds into an Activation energy, active protein and performs its functions in the cell. The polypeptide can also start folding during protein synthesis. The ribosome facilitates decoding ...
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28S Ribosomal RNA
28S ribosomal RNA is the structural ribosomal RNA (rRNA) for the large subunit (LSU) of eukaryotic cytoplasmic ribosomes, and thus one of the basic components of all eukaryotic cells. It has a size of 25S in plants and 28S in mammals, hence the alias of 25S–28S rRNA. Combined with 5.8S rRNA to the 5' side, it is the eukaryotic nuclear homologue of the prokaryotic 23S and mitochondrial 16S ribosomal RNAs. Use in phylogeny The genes coding for 28S rRNA are referred to as 28S rDNA. The comparison of the sequences from these genes are sometimes used in molecular analysis to construct phylogenetic trees, for example in protists, fungi, insects, arachnids, tardigrades, and vertebrates. Structure The 28S rRNA is typically 4000–5000 nt long. Some eukaryotes cleave 28S rRNA into two parts before assembling both into the ribosome, a phenomenon termed the "hidden break". Databases Several databases provide alignments and annotations of LSU rRNA sequences for compar ...
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LSU RRNA
Large subunit ribosomal ribonucleic acid (LSU rRNA) is the largest of the two major RNA components of the ribosome Ribosomes () are molecular machine, macromolecular machines, found within all cell (biology), cells, that perform Translation (biology), biological protein synthesis (messenger RNA translation). Ribosomes link amino acids together in the order s .... Associated with a number of ribosomal proteins, the LSU rRNA forms the large subunit of the ribosome. The LSU rRNA acts as a ribozyme, catalyzing peptide bond formation. Characteristics Use in phylogenetics LSU rRNA sequences are widely used for working out evolutionary relationships among organisms, since they are of ancient origin and are found in all known forms of life. See also * SSU rRNA: the small subunit ribosomal ribonucleic acid. References {{ribosome subunits Ribosomal RNA Protein biosynthesis ...
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18S Ribosomal RNA
18S ribosomal RNA (abbreviated 18S rRNA) is a part of the ribosomal RNA in eukaryotes. It is a component of the Eukaryotic small ribosomal subunit (40S) and the cytosolic homologue of both the 12S ribosomal RNA, 12S rRNA in mitochondria and the 16S ribosomal RNA, 16S rRNA in plastids and prokaryotes. Similar to the prokaryotic 16S rRNA, the genes of the 18S ribosomal RNA have been widely used for Phylogenetics, phylogenetic studies and biodiversity screening of eukaryotes. Research history Along with the 28S ribosomal RNA, 28S and 5.8S ribosomal RNA, 5.8S rRNA in eukaryotes, the 18S rRNA was early identified as integral structural element of ribosomes which were first characterized by their sedimentation properties and named according to measured Svedberg, Svedberg units. Given its ubiquitous presence in eukaryotic life, the evolution of the 18S rRNA was soon proposed as marker for phylogenetics, phylogenetic studies to resolve the evolution of eukaryotes. Structure and function ...
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SSU RRNA
Small subunit ribosomal ribonucleic acid (SSU rRNA) is the smaller of the two major RNA components of the ribosome. Associated with a number of ribosomal proteins, the SSU rRNA forms the small subunit of the ribosome. It is encoded by SSU- rDNA. Characteristics Use in phylogenetics SSU rRNA sequences are widely used for determining evolutionary relationships among organisms, since they are of ancient origin and are found in all known forms of life. See also *LSU rRNA Large subunit ribosomal ribonucleic acid (LSU rRNA) is the largest of the two major RNA components of the ribosome Ribosomes () are molecular machine, macromolecular machines, found within all cell (biology), cells, that perform Translation (b ...: the large subunit ribosomal ribonucleic acid. References {{Ribosome subunits Ribosomal RNA Protein biosynthesis ...
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