Neurospora crassa

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Neurospora crassa
Neurospora crassahyphae.jpg
Scientific classification OOjs UI icon edit-ltr.svg
Domain: Eukaryota
Kingdom: Fungi
Division: Ascomycota
Class: Sordariomycetes
Order: Sordariales
Family: Sordariaceae
Genus: Neurospora
Species:
N. crassa
Binomial name
Neurospora crassa
Shear & B.O. Dodge

Neurospora crassa is a type of red bread mold of the phylum Ascomycota. The genus name, meaning 'nerve spore' in Greek, refers to the characteristic striations on the spores. The first published account of this fungus was from an infestation of French bakeries in 1843. [1]

Neurospora crassa is used as a model organism because it is easy to grow and has a haploid life cycle that makes genetic analysis simple since recessive traits will show up in the offspring. Analysis of genetic recombination is facilitated by the ordered arrangement of the products of meiosis in Neurospora ascospores. Its entire genome of seven chromosomes has been sequenced. [2]

Neurospora was used by Edward Tatum and George Wells Beadle in their experiments for which they won the Nobel Prize in Physiology or Medicine in 1958. Beadle and Tatum exposed N. crassa to x-rays, causing mutations. They then observed failures in metabolic pathways caused by errors in specific enzymes. This led them to propose the "one gene, one enzyme" hypothesis that specific genes code for specific proteins. Their hypothesis was later elaborated to enzyme pathways by Norman Horowitz, also working on Neurospora. As Norman Horowitz reminisced in 2004, [3] "These experiments founded the science of what Beadle and Tatum called 'biochemical genetics'. In actuality, they proved to be the opening gun in what became molecular genetics and all developments that have followed from that."

In the 24 April 2003 issue of Nature , the genome of N. crassa was reported as completely sequenced. [4] The genome is about 43 megabases long and includes approximately 10,000 genes. There is a project underway to produce strains containing knockout mutants of every N. crassa gene. [5]

In its natural environment, N. crassa lives mainly in tropical and sub-tropical regions. [6] It can be found growing on dead plant matter after fires.

Neurospora is actively used in research around the world. It is important in the elucidation of molecular events involved in circadian rhythms, epigenetics and gene silencing, cell polarity, cell fusion, development, as well as many aspects of cell biology and biochemistry.


The sexual cycle

Neurospora crassa life cycle. The haploid mycelium reproduces asexually by two processes: (1) simple proliferation of existing mycelium, and (2) formation of conidia (macro- and micro-) which can be dispersed and then germinate to produce new mycelium. In the sexual cycle, mating can only occur between individual strains of different mating type, A and a. Fertilization occurs by the passage of nuclei of conidia or mycelium of one mating type into the protoperithecia of the opposite mating type through the trichogyne. Fusion of the nuclei of opposite mating types occurs within the protoperithecium to form a zygote (2N) nucleus. Neurospora crassa life cycle.jpg
Neurospora crassa life cycle. The haploid mycelium reproduces asexually by two processes: (1) simple proliferation of existing mycelium, and (2) formation of conidia (macro- and micro-) which can be dispersed and then germinate to produce new mycelium. In the sexual cycle, mating can only occur between individual strains of different mating type, A and a. Fertilization occurs by the passage of nuclei of conidia or mycelium of one mating type into the protoperithecia of the opposite mating type through the trichogyne. Fusion of the nuclei of opposite mating types occurs within the protoperithecium to form a zygote (2N) nucleus.

Sexual fruiting bodies (perithecia) can only be formed when two mycelia of different mating type come together (see Figure). Like other Ascomycetes, N. crassa has two mating types that, in this case, are symbolized by A and a. There is no evident morphological difference between the A and a mating type strains. Both can form abundant protoperithecia, the female reproductive structure (see Figure). Protoperithecia are formed most readily in the laboratory when growth occurs on solid (agar) synthetic medium with a relatively low source of nitrogen. [7] Nitrogen starvation appears to be necessary for expression of genes involved in sexual development. [8] The protoperithecium consists of an ascogonium, a coiled multicellular hypha that is enclosed in a knot-like aggregation of hyphae. A branched system of slender hyphae, called the trichogyne, extends from the tip of the ascogonium projecting beyond the sheathing hyphae into the air. The sexual cycle is initiated (i.e. fertilization occurs) when a cell (usually a conidium) of opposite mating type contacts a part of the trichogyne (see Figure). Such contact can be followed by cell fusion leading to one or more nuclei from the fertilizing cell migrating down the trichogyne into the ascogonium. Since both A and a strains have the same sexual structures, neither strain can be regarded as exclusively male or female. However, as a recipient, the protoperithecium of both the A and a strains can be thought of as the female structure, and the fertilizing conidium can be thought of as the male participant.

The subsequent steps following fusion of A and a haploid cells, have been outlined by Fincham and Day [9] and Wagner and Mitchell. [10] After fusion of the cells, the further fusion of their nuclei is delayed. Instead, a nucleus from the fertilizing cell and a nucleus from the ascogonium become associated and begin to divide synchronously. The products of these nuclear divisions (still in pairs of unlike mating type, i.e. A/a) migrate into numerous ascogenous hyphae, which then begin to grow out of the ascogonium. Each of these ascogenous hypha bends to form a hook (or crozier) at its tip and the A and a pair of haploid nuclei within the crozier divide synchronously. Next, septa form to divide the crozier into three cells. The central cell in the curve of the hook contains one A and one a nucleus (see Figure). This binuclear cell initiates ascus formation and is called an "ascus-initial" cell. Next the two uninucleate cells on either side of the first ascus-forming cell fuse with each other to form a binucleate cell that can grow to form a further crozier that can then form its own ascus-initial cell. This process can then be repeated multiple times.

After formation of the ascus-initial cell, the A and a nucleus fuse with each other to form a diploid nucleus (see Figure). This nucleus is the only diploid nucleus in the entire life cycle of N. crassa. The diploid nucleus has 14 chromosomes formed from the two fused haploid nuclei that had 7 chromosomes each. Formation of the diploid nucleus is immediately followed by meiosis. The two sequential divisions of meiosis lead to four haploid nuclei, two of the A mating type and two of the a mating type. One further mitotic division leads to four A and four a nucleus in each ascus. Meiosis is an essential part of the life cycle of all sexually reproducing organisms, and in its main features, meiosis in N. crassa seems typical of meiosis generally.

As the above events are occurring, the mycelial sheath that had enveloped the ascogonium develops as the wall of the perithecium, becomes impregnated with melanin, and blackens. The mature perithecium has a flask-shaped structure.

A mature perithecium may contain as many as 300 asci, each derived from identical fusion diploid nuclei. Ordinarily, in nature, when the perithecia mature the ascospores are ejected rather violently into the air. These ascospores are heat resistant and, in the lab, require heating at 60 °C for 30 minutes to induce germination. For normal strains, the entire sexual cycle takes 10 to 15 days. In a mature ascus containing eight ascospores, pairs of adjacent spores are identical in genetic constitution, since the last division is mitotic, and since the ascospores are contained in the ascus sac that holds them in a definite order determined by the direction of nuclear segregations during meiosis. Since the four primary products are also arranged in sequence, a first division segregation pattern of genetic markers can be distinguished from a second division segregation pattern.

Fine structure genetic analysis

Because of the above features N. crassa was found to be very useful for the study of genetic events occurring in individual meioses. Mature asci from a perithecium can be separated on a microscope slide and the spores experimentally manipulated. These studies usually involved the separate culture of individual ascospores resulting from a single meiotic event and determining the genotype of each spore. Studies of this type, carried out in several different laboratories, established the phenomenon of "gene conversion" (e.g. see references [11] [12] [13] ).

As an example of the gene conversion phenomenon, consider genetic crosses of two N. crassa mutant strains defective in gene pan-2. This gene is necessary for the synthesis of pantothenic acid (vitamin B5), and mutants defective in this gene can be experimentally identified by their requirement for pantothenic acid in their growth medium. The two pan-2 mutations B5 and B3 are located at different sites in the pan-2 gene, so that a cross of B5 ´ B3 yields wild-type recombinants at low frequency. [12] An analysis of 939 asci in which the genotypes of all meiotic products (ascospores) could be determined found 11 asci with an exceptional segregation pattern. These included six asci in which there was one wild-type meiotic product but no expected reciprocal double-mutant (B5B3) product. Furthermore, in three asci the ratio of meiotic products was 1B5:3B3, rather than in the expected 2:2 ratio. This study, as well as numerous additional studies in N. crassa and other fungi (reviewed by Whitehouse [14] ), led to an extensive characterization of gene conversion. It became clear from this work that gene conversion events arise when a molecular recombination event happens to occur near the genetic markers under study (e.g. pan-2 mutations in the above example). Thus studies of gene conversion allowed insight into the details of the molecular mechanism of recombination. Over the decades since the original observations of Mary Mitchell in 1955, [11] a sequence of molecular models of recombination have been proposed based on both emerging genetic data from gene conversion studies and studies of the reaction capabilities of DNA. Current understanding of the molecular mechanism of recombination is discussed in the Wikipedia articles Gene conversion and Genetic recombination. An understanding of recombination is relevant to several fundamental biologic problems, such the role of recombination and recombinational repair in cancer (see BRCA1) and the adaptive function of meiosis (see Meiosis).

Adaptive function of mating type

That mating in N. crassa can only occur between strains of different mating types suggests that some degree of outcrossing is favored by natural selection. In haploid multicellular fungi, such as N. crassa, meiosis occurring in the brief diploid stage is one of their most complex processes. Although physically much larger than the diploid stage, the haploid multicellular vegetative stage characteristically has a simple modular construction with little differentiation. In N. crassa, recessive mutations affecting the diploid stage of the life cycle are quite frequent in natural populations. [15] These mutations, when homozygous in the diploid stage, often cause spores to have maturation defects or to produce barren fruiting bodies with few ascospores (sexual spores). Most of these homozygous mutations cause abnormal meiosis (e.g., disturbed chromosome pairing or pachytene or diplotene). [16] The number of genes affecting the diploid stage was estimated to be at least 435 [15] (about 4% of the total number of 9,730 genes). Thus, outcrossing, promoted by the necessity for the union of opposite mating types, likely provides the benefit of masking recessive mutations that would otherwise be harmful to sexual spore formation (see Complementation (genetics)).

Current research

Neurospora crassa is not only a model organism for the study of phenotypic types in knock-out variants, but a particularly useful organism widely used in computational biology and the circadian clock. It has a natural reproductive cycle of 22 hours and is influenced by external factors such as light and temperature. Knock out variants of wild type N. crassa are widely studied to determine the influence of particular genes (see Frequency (gene) ).

See also

Notes and references

  1. Davis, Perkins (2002). "Neurospora: a model of model microbes". Nature Reviews Genetics. 3 (5): 397–403. doi:10.1038/nrg797. PMID   11988765. S2CID   15642417.
  2. Trans-NIH Neurospora Initiative
  3. Horowitz NH, Berg P, Singer M, et al. (January 2004). "A centennial: George W. Beadle, 1903–1989". Genetics. 166 (1): 1–10. doi:10.1534/genetics.166.1.1. PMC   1470705 . PMID   15020400.
  4. Galagan J.; Calvo S.; Borkovich K.; Selker E.; Read N. D.; et al. (2003). "The genome sequence of the filamentous fungus Neurospora crassa". Nature. 422 (6934): 859–868. Bibcode:2003Natur.422..859G. doi: 10.1038/nature01554 . PMID   12712197.
  5. Colot H.V.; Park G.; Turner G.E.; Ringleberg C.; Crew C.M.; Litvinkova L.; Weiss R.L.; Borkovitch K.A.; Dunlap J.C.; et al. (2006). "A high-throughput gene knockout procedure for Neurospora reveals functions for multiple transcription factors". Proceedings of the National Academy of Sciences, USA. 103 (27): 10352–10357. Bibcode:2006PNAS..10310352C. doi: 10.1073/pnas.0601456103 . PMC   1482798 . PMID   16801547.
  6. Perkins D. D.; Turner B. C. (1988). "Neurospora from natural populations: Toward the population biology of a haploid eukaryote". Experimental Mycology. 12 (2): 91–131. doi:10.1016/0147-5975(88)90001-1.
  7. Westergaard M, Mitchell HK (1947). "Neurospora. V. "A synthetic medium favoring sexual reproduction". Am J Bot. 34 (10): 573–577. doi:10.2307/2437339. JSTOR   2437339.
  8. Nelson MA, Metzenberg RL (September 1992). "Sexual development genes of Neurospora crassa". Genetics. 132 (1): 149–62. doi:10.1093/genetics/132.1.149. PMC   1205113 . PMID   1356883.
  9. Fincham J RS, Day PR (1963). Fungal Genetics. Blackwell Scientific Publications, Oxford, UK. ASIN: B000W851KO
  10. Wagner RP, Mitchell HK. (1964). Genetics and Metabolism. John Wiley and Sons, Inc., New York ASIN: B00BXTC5BO
  11. 1 2 Mitchell MB (April 1955). "ABERRANT RECOMBINATION OF PYRIDOXINE MUTANTS OF Neurospora". Proc. Natl. Acad. Sci. U.S.A. 41 (4): 215–20. Bibcode:1955PNAS...41..215M. doi: 10.1073/pnas.41.4.215 . PMC   528059 . PMID   16589648.
  12. 1 2 Case ME, Giles NH (May 1958). "EVIDENCE FROM TETRAD ANALYSIS FOR BOTH NORMAL AND ABERRANT RECOMBINATION BETWEEN ALLELIC MUTANTS IN Neurospora Crassa". Proc. Natl. Acad. Sci. U.S.A. 44 (5): 378–90. Bibcode:1958PNAS...44..378C. doi: 10.1073/pnas.44.5.378 . PMC   335434 . PMID   16590210.
  13. Stadler DR (July 1959). "Gene Conversion of Cysteine Mutants in Neurospora". Genetics. 44 (4): 647–56. doi:10.1093/genetics/44.4.647. PMC   1209971 . PMID   17247847.
  14. Whitehouse, HLK. (1982). Genetic Recombination. Wiley, New York ISBN   978-0471102052
  15. 1 2 Leslie JF, Raju NB (December 1985). "Recessive mutations from natural populations of Neurospora crassa that are expressed in the sexual diplophase". Genetics. 111 (4): 759–77. doi:10.1093/genetics/111.4.759. PMC   1202670 . PMID   2933298.
  16. Raju NB, Leslie JF (October 1992). "Cytology of recessive sexual-phase mutants from wild strains of Neurospora crassa". Genome. 35 (5): 815–26. doi:10.1139/g92-124. PMID   1427061.

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<span class="mw-page-title-main">Meiosis</span> Cell division producing haploid gametes

Meiosis is a special type of cell division of germ cells and apicomplexans in sexually-reproducing organisms that produces the gametes, the sperm or egg cells. It involves two rounds of division that ultimately result in four cells, each with only one copy of each chromosome (haploid). Additionally, prior to the division, genetic material from the paternal and maternal copies of each chromosome is crossed over, creating new combinations of code on each chromosome. Later on, during fertilisation, the haploid cells produced by meiosis from a male and a female will fuse to create a zygote, a cell with two copies of each chromosome again.

<span class="mw-page-title-main">Ascomycota</span> Division or phylum of fungi

Ascomycota is a phylum of the kingdom Fungi that, together with the Basidiomycota, forms the subkingdom Dikarya. Its members are commonly known as the sac fungi or ascomycetes. It is the largest phylum of Fungi, with over 64,000 species. The defining feature of this fungal group is the "ascus", a microscopic sexual structure in which nonmotile spores, called ascospores, are formed. However, some species of Ascomycota are asexual and thus do not form asci or ascospores. Familiar examples of sac fungi include morels, truffles, brewers' and bakers' yeast, dead man's fingers, and cup fungi. The fungal symbionts in the majority of lichens such as Cladonia belong to the Ascomycota.

<span class="mw-page-title-main">Ascus</span> Spore-bearing cell in ascomycete fungi

An ascus is the sexual spore-bearing cell produced in ascomycete fungi. Each ascus usually contains eight ascospores, produced by meiosis followed, in most species, by a mitotic cell division. However, asci in some genera or species can occur in numbers of one, two, four, or multiples of four. In a few cases, the ascospores can bud off conidia that may fill the asci with hundreds of conidia, or the ascospores may fragment, e.g. some Cordyceps, also filling the asci with smaller cells. Ascospores are nonmotile, usually single celled, but not infrequently may be coenocytic, and in some cases coenocytic in multiple planes. Mitotic divisions within the developing spores populate each resulting cell in septate ascospores with nuclei. The term ocular chamber, or oculus, refers to the epiplasm that is surrounded by the "bourrelet".

<i>Saccharomyces cerevisiae</i> Species of yeast

Saccharomyces cerevisiae is a species of yeast. The species has been instrumental in winemaking, baking, and brewing since ancient times. It is believed to have been originally isolated from the skin of grapes. It is one of the most intensively studied eukaryotic model organisms in molecular and cell biology, much like Escherichia coli as the model bacterium. It is the microorganism behind the most common type of fermentation. S. cerevisiae cells are round to ovoid, 5–10 μm in diameter. It reproduces by budding.

<i>Schizosaccharomyces pombe</i> Species of yeast

Schizosaccharomyces pombe, also called "fission yeast", is a species of yeast used in traditional brewing and as a model organism in molecular and cell biology. It is a unicellular eukaryote, whose cells are rod-shaped. Cells typically measure 3 to 4 micrometres in diameter and 7 to 14 micrometres in length. Its genome, which is approximately 14.1 million base pairs, is estimated to contain 4,970 protein-coding genes and at least 450 non-coding RNAs.

<span class="mw-page-title-main">Karyogamy</span> Fusion of the nuclei of two haploid eukaryotic cells

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<span class="mw-page-title-main">Evolution of sexual reproduction</span> How sexually reproducing multicellular organisms could have evolved from a common ancestor species

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<i>Sordaria fimicola</i> Species of fungus

Sordaria fimicola is a species of microscopic fungus. It is commonly found in the feces of herbivores. Sordaria fimicola is often used in introductory biology and mycology labs because it is easy to grow on nutrient agar in dish cultures. The genus Sordaria, closely related to Neurospora and Podospora, is a member of the large class Sordariomycetes, or flask-fungi. The natural habitat of the three species of Sordaria that have been the principal subjects in genetic studies is dung of herbivorous animals. The species S. fimicola is common and worldwide in distribution. The species of Sordaria are similar morphologically, producing black perithecia containing asci with eight dark ascospores in a linear arrangement. These species share a number of characteristics that are advantageous for genetic studies. They all have a short life cycle, usually 7–12 days, and are easily grown in culture. Most species are self-fertile and each strain is isogenic. All kinds of mutants are easily induced and readily obtainable with particular ascospore color mutants. These visual mutants aid in tetrad analysis, especially in analysis of intragenic recombination.

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<span class="mw-page-title-main">Mating of yeast</span> Biological process

The yeast Saccharomyces cerevisiae is a simple single-celled eukaryote with both a diploid and haploid mode of existence. The mating of yeast only occurs between haploids, which can be either the a or α (alpha) mating type and thus display simple sexual differentiation. Mating type is determined by a single locus, MAT, which in turn governs the sexual behaviour of both haploid and diploid cells. Through a form of genetic recombination, haploid yeast can switch mating type as often as every cell cycle.

<i>Neurospora</i> Genus of fungi

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<span class="mw-page-title-main">Mating in fungi</span> Combination of genetic material between compatible mating types

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<span class="mw-page-title-main">David Perkins (geneticist)</span> American geneticist

David Dexter Perkins was an American geneticist, a member of the faculty of the Department of Biology at Stanford University for more than 58 years, from 1948 until his death in 2007. He received his PhD in Zoology in 1949 from Columbia University. A member of the National Academy of Sciences, he served as president of the Genetics Society of America in 1977. In a scientific career that spanned more than six decades, Perkins collaborated on more than 300 papers. His associates included many graduate students and postdoctoral fellows who went on to scientific careers throughout the world.

The mating-type locus is a specialized region in the genomes of some yeast and other fungi, usually organized into heterochromatin and possessing unique histone methylation patterns. The genes in this region regulate the mating type of the organism and therefore determine key events in its life cycle, such as whether it will reproduce sexually or asexually. In fission yeast such as S. pombe, the formation and maintenance of the heterochromatin organization is regulated by RNA-induced transcriptional silencing, a form of RNA interference responsible for genomic maintenance in many organisms. Mating type regions have also been well studied in budding yeast S. cerevisiae and in the fungus Neurospora crassa.

<span class="mw-page-title-main">Tetrad (meiosis)</span> Product of meiosis in spore-producing organisms

The tetrad is the four spores produced after meiosis of a yeast or other Ascomycota, Chlamydomonas or other alga, or a plant. After parent haploids mate, they produce diploids. Under appropriate environmental conditions, diploids sporulate and undergo meiosis. The meiotic products, spores, remain packaged in the parental cell body to produce the tetrad.

The parasexual cycle, a process restricted to fungi and single-celled organisms, is a nonsexual mechanism of parasexuality for transferring genetic material without meiosis or the development of sexual structures. It was first described by Italian geneticist Guido Pontecorvo in 1956 during studies on Aspergillus nidulans. A parasexual cycle is initiated by the fusion of hyphae (anastomosis) during which nuclei and other cytoplasmic components occupy the same cell. Fusion of the unlike nuclei in the cell of the heterokaryon results in formation of a diploid nucleus (karyogamy), which is believed to be unstable and can produce segregants by recombination involving mitotic crossing-over and haploidization. Mitotic crossing-over can lead to the exchange of genes on chromosomes; while haploidization probably involves mitotic nondisjunctions which randomly reassort the chromosomes and result in the production of aneuploid and haploid cells. Like a sexual cycle, parasexuality gives the species the opportunity to recombine the genome and produce new genotypes in their offspring. Unlike a sexual cycle, the process lacks coordination and is exclusively mitotic.

<i>Podospora anserina</i> Species of fungus

Podospora anserina is a filamentous ascomycete fungus from the order Sordariales. It is considered a model organism for the study of molecular biology of senescence (aging), prions, sexual reproduction, and meiotic drive. It has an obligate sexual and pseudohomothallic life cycle. It is a non-pathogenic coprophilous fungus that colonizes the dung of herbivorous animals such as horses, rabbits, cows and sheep.

References