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''Blakeslea trispora'' is a mould and member of the division
Zygomycota Zygomycota, or zygote fungi, is a former division or phylum of the kingdom Fungi. The members are now part of two phyla: the Mucoromycota and Zoopagomycota. Approximately 1060 species are known. They are mostly terrestrial in habitat, living ...
. This species has been well studied for its ability to produce carotenoids, particularly,
β-carotene β-Carotene is an organic, strongly coloured red-orange pigment abundant in fungi, plants, and fruits. It is a member of the carotenes, which are terpenoids (isoprenoids), synthesized biochemically from eight isoprene units and thus having 40 ...
and
lycopene Lycopene is an organic compound classified as a tetraterpene and a carotene. Lycopene (from the neo-Latin '' Lycopersicum'', the tomato species) is a bright red carotenoid hydrocarbon found in tomatoes and other red fruits and vegetables. Occu ...
. β-carotene is a vitamin A precursor and both of β-carotene and lycopene play a significant role in the inhibition of
oxidative stress Oxidative stress reflects an imbalance between the systemic manifestation of reactive oxygen species and a biological system's ability to readily detoxify the reactive intermediates or to repair the resulting damage. Disturbances in the normal ...
. ''Blakeslea trispora'' is commonly isolated from soil samples throughout the Southern United States and Southern Asia. ''B. trispora'' is a
pathogen In biology, a pathogen ( el, πάθος, "suffering", "passion" and , "producer of") in the oldest and broadest sense, is any organism or agent that can produce disease. A pathogen may also be referred to as an infectious agent, or simply a g ...
of tropical plants.
In vivo Studies that are ''in vivo'' (Latin for "within the living"; often not italicized in English) are those in which the effects of various biological entities are tested on whole, living organisms or cells, usually animals, including humans, and p ...
pathogenicity testing using
animal models A model organism (often shortened to model) is a non-human species that is extensively studied to understand particular biological phenomena, with the expectation that discoveries made in the model organism will provide insight into the working ...
suggests this fungus is not a cause of animal or human disease.


History

The genus ''Blakeslea'' was named in honor of American botanist
Albert Francis Blakeslee Albert Francis Blakeslee (November 9, 1874 – November 16, 1954) was an American botanist. He is best known for his research on the poisonous jimsonweed plant and the sexuality of fungi. He was the brother of the Far East scholar George Hubbar ...
based on ''B. trispora''. While A.F. Blakeslee was studying another fungus called
Botrytis rileyi
', he collaborated with
Roland Thaxter Roland Thaxter (August 28, 1858 – April 22, 1932) was an American mycologist, plant pathologist, botanist, and entomologist, renowned for his contribution to the insect parasitic fungi—Laboulbeniales. His college education was completed at Har ...
to further study this fungus. In 1914, while Thaxter was doing further research on ''Botrytis rileyi,'' ''Blakeslea trispora'' was accidentally isolated from a contaminated caterpillar infected with ''Botrytis rileyi''. ''Blakeslea trispora'' was first identified from the
larva A larva (; plural larvae ) is a distinct juvenile form many animals undergo before metamorphosis into adults. Animals with indirect development such as insects, amphibians, or cnidarians typically have a larval phase of their life cycle. ...
e of the caterpillar, which was growing on the
cowpea The cowpea (''Vigna unguiculata'') is an annual herbaceous legume from the genus ''Vigna''. Its tolerance for sandy soil and low rainfall have made it an important crop in the semiarid regions across Africa and Asia. It requires very few inputs, ...
plant. The caterpillar was infected by the fungus ''Botrytis rileyi;'' however, ''Blakeslea trispora'' was thought to be incidentally transferred to the diseased caterpillar feeding on a cowpea flower. When Thaxter first identified ''B. trispora'', he considered ''B. trispora'' to be very closely related to the genus ''
Choanephora ''Choanephora'' is a genus of Zygomycota fungi. ''Choanephora'' species are known as plant pathogens Plant pathology (also phytopathology) is the scientific study of diseases in plants caused by pathogens (infectious organisms) and envi ...
'' because of highly similar sporangiospore morphology. Both have a distinctive brown colour with faint, longitudinal striations on the sporangiole wall. The shape of the large spherical heads of their sporangiola are also similar. However, ''Choanephora'' and ''Blakeslea'' are considered to be distinct genera and can be distinguishable by the spore wall and its separation from the sporangiole wall. Species of ''Choanephora'' have highly adherent sporangiole wall in contrast to ''Blakeslea'' species where the sporangiole wall is readily separable from the underlying spore at maturity.


Growth and morphology

Blakeslea trispora undergoes both sexual and asexual reproduction. The asexual reproductive phase of ''Blakeslea trispora'' involves the production of sporangiospores produced in
sporangia A sporangium (; from Late Latin, ) is an enclosure in which spores are formed. It can be composed of a single cell or can be multicellular. Virtually all plants, fungi, and many other lineages form sporangia at some point in their life cy ...
. Once released, they can germinate in the presence of free water. Colonies of ''B. trispora'' grow rapidly on the agar growth media at 25 °C. They are white at first but become yellow to pale brown and very dark brown as they mature. The hyphae of ''B. trispora'' are aseptate, very dense, and highly branched. Sexual reproduction is by the formation of zygospores, which contain high concentrations of
triglycerol A triglyceride (TG, triacylglycerol, TAG, or triacylglyceride) is an ester derived from glycerol and three fatty acids (from '' tri-'' and ''glyceride''). Triglycerides are the main constituents of body fat in humans and other vertebrates, as w ...
-rich
lipid Lipids are a broad group of naturally-occurring molecules which includes fats, waxes, sterols, fat-soluble vitamins (such as vitamins A, D, E and K), monoglycerides, diglycerides, phospholipids, and others. The functions of lipids in ...
s and
phosphatidylcholine Phosphatidylcholines (PC) are a class of phospholipids that incorporate choline as a headgroup. They are a major component of biological membranes and can be easily obtained from a variety of readily available sources, such as egg yolk or soybea ...
. Zygospores can persist for long periods of time, and their germination is dependent on a cytoplasmic regulatory system that sustains dormancy and forestalls germination in the presence of unfavorable growth conditions. Zygospores range in size from 40-80μm. They are spherical or slightly flattened in shape. ''Blakeslea trispora'' has a
heterothallic Heterothallic species have sexes that reside in different individuals. The term is applied particularly to distinguish heterothallic fungi, which require two compatible partners to produce sexual spores, from homothallic ones, which are capable ...
mating system, having (+) and (-) mating types. Contact and interchange between the opposite mating types is a necessary precursor to induce sexual reproduction and development of zygospores. Extensions called gametangia are formed from each of the compatible
haploid Ploidy () is the number of complete sets of chromosomes in a cell, and hence the number of possible alleles for autosomal and pseudoautosomal genes. Sets of chromosomes refer to the number of maternal and paternal chromosome copies, respective ...
mycelia. Following anastomosis, a fertile
heterokaryotic A heterokaryon is a multinucleate cell that contains genetically different nuclei. Heterokaryotic and heterokaryosis are derived terms. This is a special type of syncytium. This can occur naturally, such as in the mycelium of fungi during sexual ...
zygosporangium is formed within which the zygospores develop. During sexual reproduction, carotenoid pigments are produced by both of mating type.
Carotenoid Carotenoids (), also called tetraterpenoids, are yellow, orange, and red organic pigments that are produced by plants and algae, as well as several bacteria, and fungi. Carotenoids give the characteristic color to pumpkins, carrots, parsnips, ...
s are precursors of many
apocarotenoid Apocarotenoids are organic compounds which occur widely in living organisms. They are derived from carotenoids by oxidative cleavage, catalyzed by carotenoid oxygenases. Examples include the vitamin A retinoids retinal, retinoic acid, and retinol; ...
s that contain very important sex-specific precursors, trisporic acid (TSA) for the sexual reproduction of ''Blakeslea trispora''.
Carotene The term carotene (also carotin, from the Latin ''carota'', "carrot") is used for many related unsaturated hydrocarbon substances having the formula C40Hx, which are synthesized by plants but in general cannot be made by animals (with the exc ...
s produced from carotenoids are further processed by carotene oxygenase to synthesize
trisporic acid Trisporic acids (TSAs) are C-18 terpenoid compounds synthesized via β-carotene and retinol pathways in the zygomycetes. They are pheromone compound responsible for sexual differentiation in those fungal species. TSAs and related compounds make u ...
(TSA). TSA produced from carotene stimulates both sexually complementary cells to make contact with each other. TSA is considered an important
signalling molecule In biology, cell signaling (cell signalling in British English) or cell communication is the ability of a cell to receive, process, and transmit signals with its environment and with itself. Cell signaling is a fundamental property of all cellula ...
for the initiation and control sexual reproduction.


Physiology

At the beginning of the sexual reproduction cycle of ''B. trispora,'' the initial step is the production of carotenes from carotenoids. Carotenes are further processed by carotene oxygenase, which is encoded in the tsp3 gene of the ''B. trispora'', to produce TSA. TSA is produced by both of the mating types: (+) and (-) strains, and it is copiously produced especially when compatible mycelia are grown together. As these two different sex types produce TSA, they sense sexually complementary cells and form gametangia. Eventually, those gametangia merge and zygosporangia form. As these two different mating types meet each other, each mating type transfers the sex-specific precursor of the trisporoid TSA, and acts as a signal for the synthesis of the surface protein
agglutinin An agglutinin is a substance in the blood that causes particles to coagulate and aggregate; that is, to change from fluid-like state to a thickened-mass (solid) state. Agglutinins can be antibodies that cause antigens to aggregate by binding t ...
. Agglutinin allows the two TSAs to recognize each other. It then causes rapid contact and efficient interactions between those two different mating types. Additionally, stimulating both of the mating types by TSA promotes synthesis of
β-carotene β-Carotene is an organic, strongly coloured red-orange pigment abundant in fungi, plants, and fruits. It is a member of the carotenes, which are terpenoids (isoprenoids), synthesized biochemically from eight isoprene units and thus having 40 ...
. As β-carotene is produced, it becomes a precursor of trisporoid, which is a pheromone for ''B. trispora''. Production of β-carotene promotes a
positive feedback Positive feedback (exacerbating feedback, self-reinforcing feedback) is a process that occurs in a feedback loop which exacerbates the effects of a small disturbance. That is, the effects of a perturbation on a system include an increase in the ...
process that further stimulates carotenogenesis and the production of trisporoid which serves as a β-carotene increasing substance. Furthermore, it act as a hormone stimulator of its
biosynthesis Biosynthesis is a multi-step, enzyme-catalyzed process where substrates are converted into more complex products in living organisms. In biosynthesis, simple compounds are modified, converted into other compounds, or joined to form macromolecules. ...
. Thus, ''Blakeslea trispora'' requires certain concentrations of TSA to activate carotenogenesis and produce more carotenoids (about 0.5% of its
dry weight Vehicle weight is a measurement of wheeled motor vehicles; either an actual measured weight of the vehicle under defined conditions or a gross weight rating for its weight carrying capacity. Curb or kerb weight Curb weight (U.S. English) or kerb ...
) which can be accumulated in the zygospores of ''B. trispora''. Therefore, both TSA and trisporoid acts as sex hormones in ''Blakeslea trispora'', which triggers the sexual reproduction and controls intimate contact between heterothallic strains, further governing the formation of sex structures, zygospores. Carotenoids are absolutely necessary not only for the production of trisporic acid, but also for the process of zygote formation, as significant factors for the production of
sporopollenin 270px, SEM image of pollen grains Sporopollenin is one of the most chemically inert biological polymers. It is a major component of the tough outer (exine) walls of plant spores and pollen grains. It is chemically very stable and is usually well ...
, a structural component of the zygospore cell wall. It is consequential to regulate this feedback-type synthesis with carotenoids and further synthesis of TSA. Therefore, the formation of the zygospores can be prevented by the inhibition of carotenogenesis in ''Blakeslea trispora''.


Applications

''Blakeslea trispora'' is useful as sources of β-carotene and its precursor molecule, lycopene, in industrial production. These molecules are useful
food coloring Food coloring, or color additive, is any dye, pigment, or substance that imparts color when it is added to food or drink. They come in many forms consisting of liquids, powders, gels, and pastes. Food coloring is used in both commercial food ...
agents and may have beneficial effects for human health as
antioxidant Antioxidants are compounds that inhibit oxidation, a chemical reaction that can produce free radicals. This can lead to polymerization and other chain reactions. They are frequently added to industrial products, such as fuels and lubricant ...
s. The production of lycopene primarily requires some interaction between the mating strains. ''Blakeslea trispora'' needs both mating types to synthesize lycopene on a commercially applicable scale. The (-) strain is twice proportionally as important as the (+) strain in determining the productivity of the synthesis of the lycopene. To produce an optimal amount of lycopene, excess (-) mating type at a 1:2 (+/-) ratio with
inoculum In biology, inoculum refers to the source material used for inoculation. ''Inoculum'' may refer to: * In medicine, material that is the source of the inoculation in a vaccine * In microbiology, propagules: cells, tissue, or viruses that are used t ...
ages 36 and 48 hours respectively is favorable.


Lycopene

''Blakeslea trispora'' is known to be the most effective producer of lycopene. Lycopene is processed by lycopene cyclase which leads to the production of β-carotene. For industrial production of lycopene, ''Blakeslea trispora'' is grown with an lycopene cyclase inhibitor which can be introduced into the
fermentation Fermentation is a metabolic process that produces chemical changes in organic substrates through the action of enzymes. In biochemistry, it is narrowly defined as the extraction of energy from carbohydrates in the absence of oxygen. In food ...
process. The zygospores of ''Blakeslea trispora'' tend to contain a maximum amount of lycopene. Lycopene is an intermediate in the biosynthesis of all dicyclic carotenoids including β-carotene. Lycopene is one of the most important carotene molecules because it is capable of producing both β-carotene and other carotenoids, well known for their potent
anti-oxidant Antioxidants are compounds that inhibit oxidation, a chemical reaction that can produce free radicals. This can lead to polymerization and other chain reactions. They are frequently added to industrial products, such as fuels and lubricants, ...
activities. As such, β-carotene and other carotenoids play crucial roles for
oxidative stress Oxidative stress reflects an imbalance between the systemic manifestation of reactive oxygen species and a biological system's ability to readily detoxify the reactive intermediates or to repair the resulting damage. Disturbances in the normal ...
reduction and cardiovascular protection. Carotenoids have highly efficient antioxidant scavenging activities against ROS (
reactive oxygen species In chemistry, reactive oxygen species (ROS) are highly reactive chemicals formed from diatomic oxygen (). Examples of ROS include peroxides, superoxide, hydroxyl radical, singlet oxygen, and alpha-oxygen. The reduction of molecular oxygen () p ...
), such as singlet-oxygen and
free radicals In chemistry, a radical, also known as a free radical, is an atom, molecule, or ion that has at least one unpaired valence electron. With some exceptions, these unpaired electrons make radicals highly chemically reactive. Many radicals spont ...
. Therefore, they have the ability to prevent
chronic diseases A chronic condition is a health condition or disease that is persistent or otherwise long-lasting in its effects or a disease that comes with time. The term ''chronic'' is often applied when the course of the disease lasts for more than three mo ...
such as cancer,
cerebrovascular Cerebrovascular disease includes a variety of Medicine, medical conditions that affect the blood vessels of the brain and the cerebral circulation. Arteries supplying oxygen and nutrients to the brain are often Angiopathy, damaged or deformed in ...
and cardiovascular diseases and
myocardial infarction A myocardial infarction (MI), commonly known as a heart attack, occurs when blood flow decreases or stops to the coronary artery of the heart, causing damage to the heart muscle. The most common symptom is chest pain or discomfort which may ...
. Lycopene is considered a very important and relevant source to human health. A case study by Weilian Hu and his colleagues in 2013 showed that the administration of lycopene in adult mice appeared to improve the activity of antioxidant enzyme. They have reported that, the administration of ''Blakeslea trispora'' powder, which contains high amounts of lycopene has the potential to protect the liver, brain, kidney and skin against oxidative stress. This is done by reducing the concentration of ROS and by enhancing the activities of the antioxidant enzyme. Furthermore, they are further investigating whether the fungus ''Blakeslea trispora'' could be a potent effector of anti-aging because of its ability to efficiently mass-produce amounts of lycopene.


β-carotene

β-carotene is a molecule which displays a red-orange pigment. Therefore, it is used as a coloring agent for food products. β-carotene is a member of carotenes which are highly unsaturated
isoprene Isoprene, or 2-methyl-1,3-butadiene, is a common volatile organic compound with the formula CH2=C(CH3)−CH=CH2. In its pure form it is a colorless volatile liquid. Isoprene is an unsaturated hydrocarbon. It is produced by many plants and animals ...
derivatives. Because ''Blakeslea trispora'' has an effective ability to produce great amount of β-carotene from lycopene, ''Blakeslea trispora'' is the main organism used for its production on an industrial scale. β-carotenes are known to be a powerful stimulant of the human
immune system The immune system is a network of biological processes that protects an organism from diseases. It detects and responds to a wide variety of pathogens, from viruses to parasitic worms, as well as cancer cells and objects such as wood splinte ...
and play significant roles in the prevention of
degenerative disease Degenerative disease is the result of a continuous process based on degenerative cell changes, affecting tissues or organs, which will increasingly deteriorate over time. In neurodegenerative diseases, cells of the central nervous system stop wor ...
s and cancers. All cells are capable of producing and regulating ROS. However, dysregulation of ROS can lead to DNA damaging, inactivation of enzymes and proteins, disruption of membranes. This ultimately causes cell death, becoming very toxic to the individuals. Further investigations of β-carotene usage collected from ''Blakeslea trispora'' may lead to great improvements to human health in the treatment and prevention of certain chronic diseases such as cancer.


References

{{DEFAULTSORT:Blakeslea Trispora Fungi described in 1914 Fungal plant pathogens and diseases Zygomycota