''Azotobacter salinestris''
is a
Gram-negative
Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. They are characterized by their cell envelopes, which are composed of a thin peptidoglycan cell wa ...
,
nitrogen-fixing
Nitrogen fixation is a chemical process by which molecular nitrogen (), with a strong triple covalent bond, in the air is converted into ammonia () or related nitrogenous compounds, typically in soil or aquatic systems but also in industry. At ...
bacterium
Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, bacteria were a ...
; its specific name, ''salinestris'', comes from the Latin words ''salinus'' meaning saline and ''estris'' which means "living in".
[Page, and Shivprasad. "ITIS Standard Report Page: Azotobacter Salinestris." ITIS Standard Report Page: Azotobacter Salinestris. N.p., 1991. Web. 8 Feb. 2016. ] It can be found living in soil or marine habitats as single cells or in chains of six to eight cells.
This organism is
motile
Motility is the ability of an organism to move independently, using metabolic energy.
Definitions
Motility, the ability of an organism to move independently, using metabolic energy, can be contrasted with sessility, the state of organisms th ...
at younger stages, but loses its
flagella
A flagellum (; ) is a hairlike appendage that protrudes from certain plant and animal sperm cells, and from a wide range of microorganisms to provide motility. Many protists with flagella are termed as flagellates.
A microorganism may have f ...
at older stages.
This species is known for its potential use in
bioremediation
Bioremediation broadly refers to any process wherein a biological system (typically bacteria, microalgae, fungi, and plants), living or dead, is employed for removing environmental pollutants from air, water, soil, flue gasses, industrial effluent ...
.
Isolation
William J. Page and Shailaja Shivprasad isolated ''A. salinestris'' from
saline soils.
The colonies used for their study were first taken from air-dried surface soil from
Alberta, Canada
Alberta ( ) is one of the thirteen provinces and territories of Canada. It is part of Western Canada and is one of the three prairie provinces. Alberta is bordered by British Columbia to the west, Saskatchewan to the east, the Northwest Territo ...
.
The soil was inoculated into a Burk nitrogen-free mineral-salt
medium
Medium may refer to:
Science and technology
Aviation
*Medium bomber, a class of war plane
* Tecma Medium, a French hang glider design
Communication
* Media (communication), tools used to store and deliver information or data
* Medium of ...
, which contained 1% glucose and 0.25
micrograms
In the metric system, a microgram or microgramme is a unit of mass equal to one millionth () of a gram. The unit symbol is μg according to the International System of Units (SI); the recommended symbol in the United States and United Kingdom wh ...
of
copper chloride per milliliter of solution.
The medium was incubated at 30 °C and stored at 4 °C.
Colony formation was noted after 2–3 days of incubation.
The same soil samples were also used to inoculate slant cultures, which were stored at room temperature.
''A. salinestris'' was found to lose
viability
Viability is the ability of a thing (a living organism, an artificial system, an idea, etc.) to maintain itself or recover its potentialities.
Viability or viable may refer to:
Biology, medicine or ecology
* Viability selection, the selection of ...
in the slant cultures when stored at 4 °C.
Through these cultures and characterization tests, ''A. salinestris'' was found to share many of the general characteristics specific to the species ''
Azotobacter
''Azotobacter'' is a genus of usually motile, oval or spherical bacteria that form thick-walled cysts (and also has hard crust) and may produce large quantities of capsular slime. They are aerobic, free-living soil microbes that play an im ...
.''
Originally, ''A. salinestris'' colonies were classified as ''
Azotobacter chroococcum
''Azotobacter chroococcum'' is a bacterium that has the ability to fix atmospheric nitrogen. It was discovered by Martinus Beijerinck in 1901, and was the first aerobic, free-living nitrogen fixer discovered. ''A. chroococcum'' could be useful ...
'', but were later identified as a separate species based on their salt-dependent growth.
Characteristics
Morphology
''Azotobacter salinestris'' is a
Gram-negative
Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. They are characterized by their cell envelopes, which are composed of a thin peptidoglycan cell wa ...
, rod-shaped organism.
This organism's cells are about 2 x 4
μm
The micrometre ( international spelling as used by the International Bureau of Weights and Measures; SI symbol: μm) or micrometer (American spelling), also commonly known as a micron, is a unit of length in the International System of Uni ...
in size when they are 18 hours old and can grow up to 5 μm in diameter.
Older cells can also form cysts.
''A. salinestris'' colonies appear to be brown-black in color because they produce
water-soluble
In chemistry, solubility is the ability of a substance
Substance may refer to:
* Matter, anything that has mass and takes up space
Chemistry
* Chemical substance, a material with a definite chemical composition
* Drug substance
** Substan ...
catechol melanin
Melanin (; from el, μέλας, melas, black, dark) is a broad term for a group of natural pigments found in most organisms. Eumelanin is produced through a multistage chemical process known as melanogenesis, where the oxidation of the amino ...
.
Physiology
The bacteria that performed the most efficient atmospheric nitrogen fixation were from samples grown in 0.05% to 0.10% saline concentration soils.
Nitrogen fixation rates were not affected by the presence of oxygen.
''A. salinestris'' that grows in soils is a
facultative anaerobe
A facultative anaerobic organism is an organism that makes ATP by aerobic respiration if oxygen is present, but is capable of switching to fermentation if oxygen is absent.
Some examples of facultatively anaerobic bacteria are ''Staphylococcus' ...
.
Colonies growing in aquatic habitats were determined to be
microaerophilic
A microaerophile is a microorganism that requires environments containing lower levels of dioxygen than that are present in the atmosphere (i.e. < 21% O2; typically 2–10% O2) for optimal growth.[ A more re ...]
and very sensitive to the presence of
hydrogen peroxide
Hydrogen peroxide is a chemical compound with the formula . In its pure form, it is a very pale blue liquid that is slightly more viscous than water. It is used as an oxidizer, bleaching agent, and antiseptic, usually as a dilute solution (3% ...
since they do not produce a
catalase
Catalase is a common enzyme found in nearly all living organisms exposed to oxygen (such as bacteria, plants, and animals) which catalyzes the decomposition of hydrogen peroxide to water and oxygen. It is a very important enzyme in protecting t ...
enzyme.
Metabolism
''A. salinestris'' can use
melibiose
Melibiose is a reducing disaccharide formed by an α-1,6 linkage between galactose and glucose (D-Gal-α(1→6)-D-Glc). It differs from lactose in the chirality of the carbon where the galactose ring is closed and that the galactose is linked to a ...
,
galactose
Galactose (, '' galacto-'' + '' -ose'', "milk sugar"), sometimes abbreviated Gal, is a monosaccharide sugar that is about as sweet as glucose, and about 65% as sweet as sucrose. It is an aldohexose and a C-4 epimer of glucose. A galactose molec ...
,
mannitol
Mannitol is a type of sugar alcohol used as a sweetener and medication. It is used as a low calorie sweetener as it is poorly absorbed by the intestines. As a medication, it is used to decrease pressure in the eyes, as in glaucoma, and to low ...
,
sucrose
Sucrose, a disaccharide, is a sugar composed of glucose and fructose subunits. It is produced naturally in plants and is the main constituent of white sugar. It has the molecular formula .
For human consumption, sucrose is extracted and refi ...
,
glucose
Glucose is a simple sugar with the molecular formula . Glucose is overall the most abundant monosaccharide, a subcategory of carbohydrates. Glucose is mainly made by plants and most algae during photosynthesis from water and carbon dioxide, usi ...
, and
fructose
Fructose, or fruit sugar, is a ketonic simple sugar found in many plants, where it is often bonded to glucose to form the disaccharide sucrose. It is one of the three dietary monosaccharides, along with glucose and galactose, that are absorb ...
as primary carbon sources.
They prefer to use
sodium ion
Sodium is a chemical element with the symbol Na (from Latin ''natrium'') and atomic number 11. It is a soft, silvery-white, highly reactive metal. Sodium is an alkali metal, being in group 1 of the periodic table. Its only stable iso ...
s as their
electron acceptor
An electron acceptor is a chemical entity that accepts electrons transferred to it from another compound. It is an oxidizing agent that, by virtue of its accepting electrons, is itself reduced in the process. Electron acceptors are sometimes mist ...
, but also use
rubidium
Rubidium is the chemical element with the symbol Rb and atomic number 37. It is a very soft, whitish-grey solid in the alkali metal group, similar to potassium and caesium. Rubidium is the first alkali metal in the group to have a density hig ...
.
Strains that do not have access to sodium ions produce acid as a product of the metabolism of their growth-promoting carbon
substrate
Substrate may refer to:
Physical layers
*Substrate (biology), the natural environment in which an organism lives, or the surface or medium on which an organism grows or is attached
** Substrate (locomotion), the surface over which an organism lo ...
.
Ecology
All known ''A. salinestris'' samples were isolated from soils that had a
neutral or slightly
basic
BASIC (Beginners' All-purpose Symbolic Instruction Code) is a family of general-purpose, high-level programming languages designed for ease of use. The original version was created by John G. Kemeny and Thomas E. Kurtz at Dartmouth College ...
pH.
The optimal growth pH of this species is 7.2-7.5.
Soils must have at least one
millimolar
Molar concentration (also called molarity, amount concentration or substance concentration) is a measure of the concentration of a chemical species, in particular of a solute in a solution, in terms of amount of substance per unit volume of solu ...
concentration of saline for this organism to grow.
Unlike other ''Azotobacter'' species, iron was absolutely required for growth.
While most ''Azotobacter'' species are commonly found in soil, ''A. salinestris'' is unique due to being found in soils with high salt content and requiring the presence of iron to grow.
These organisms can survive in
aerobic
Aerobic means "requiring air," in which "air" usually means oxygen.
Aerobic may also refer to
* Aerobic exercise, prolonged exercise of moderate intensity
* Aerobics, a form of aerobic exercise
* Aerobic respiration, the aerobic process of cell ...
and
anaerobic conditions.
It is dependent on, so can also be found in
marine environment
Marine habitats are habitats that support marine life. Marine life depends in some way on the saltwater that is in the sea (the term ''marine'' comes from the Latin ''mare'', meaning sea or ocean). A habitat is an ecological or environmen ...
s.
''A. chroococcum'' is the most common species from ''Azotobacter'' to be isolated from soil samples.
It is also a close relative to ''A. salinestris''.
All growth conditions used to isolate and determine optimum living conditions for ''A. salinestris'' were based on the optimal living conditions for ''A.'' ''chroococcum''.
The defining factor between these two species was the dependence on sodium ions to live. ''A. salinestris'' displayed a stronger dependence on sodium to live than ''A. chroococcum''.
Genetics
While Page and Shivprasad are credited with the discovery and characterization of ''A. salinestris,'' Eydne and Wachter are credited with the sequencing of the bacterium’s
5S rRNA
The 5S ribosomal RNA (5S rRNA) is an approximately 120 nucleotide-long ribosomal RNA molecule with a mass of 40 kDa. It is a structural and functional component of the large subunit of the ribosome in all domains of life (bacteria, archaea, an ...
in 1987.
Although the results were never published,
sequencing data placed this bacterial strain in the genus'' Azotobacter''.
The bacterium’s DNA has a melting point of 96.68 to 97.08 °C and the
GC content
In molecular biology and genetics, GC-content (or guanine-cytosine content) is the percentage of nitrogenous bases in a DNA or RNA molecule that are either guanine (G) or cytosine (C). This measure indicates the proportion of G and C bases out of ...
was 67.73-67.8%.
A separate sequencing of the
16S rRNA 16S rRNA may refer to:
* 16S ribosomal RNA
16 S ribosomal RNA (or 16 S rRNA) is the RNA component of the 30S subunit of a prokaryotic ribosome ( SSU rRNA). It binds to the Shine-Dalgarno sequence and provides most of the SSU structure.
The ...
sequence, conducted by Moore ''et al.'' confirmed that ''A. salinestris'' was indeed a separate species of that genus.
Importance
''A. salinestris'' was the first
prokaryote
A prokaryote () is a single-celled organism that lacks a nucleus and other membrane-bound organelles. The word ''prokaryote'' comes from the Greek πρό (, 'before') and κάρυον (, 'nut' or 'kernel').Campbell, N. "Biology:Concepts & Con ...
to show Na
+/succinic acid
efflux.
It can tolerate up to 5%
glyphosate
Glyphosate (IUPAC name: ''N''-(phosphonomethyl)glycine) is a broad-spectrum systemic herbicide and crop desiccant. It is an organophosphorus compound, specifically a phosphonate, which acts by inhibiting the plant enzyme 5-enolpyruvylshik ...
, which is a
pesticide used to kill weeds that compete with crops.
Because the species is a common nitrogen fixer, it is important to the agricultural industry for the species to be able to survive in the presence of such a common pesticide.
It can degrade
endosulfan, which is an
insecticide that is highly hazardous to human,
mammal, and fish health.
Endosulfan use was banned in 2012 by the United States, following a precedent established by
New Zealand
New Zealand ( mi, Aotearoa ) is an island country in the southwestern Pacific Ocean. It consists of two main landmasses—the North Island () and the South Island ()—and over 700 List of islands of New Zealand, smaller islands. It is the ...
and the
European Union
The European Union (EU) is a supranational political and economic union of member states that are located primarily in Europe. The union has a total area of and an estimated total population of about 447million. The EU has often been ...
.
[Martin, David S. EPA moves to ban DDT cousin. CNN. June 10, 2010. ] The decision to ban endosulfan use came after a study that showed the health risks to humans and wildlife were much higher than expected.
[Cone, M. (2010, June 10). Endosulfan to Be Banned, Pesticide Poses "Unacceptable Risks," EPA Says. Retrieved April 27, 2016, from http://www.scientificamerican.com/article/endosulfan-banned-epa/] It is similar to
dichlorodiphenyltrichloroethane
Dichlorodiphenyltrichloroethane, commonly known as DDT, is a colorless, tasteless, and almost odorless crystalline chemical compound, an organochloride. Originally developed as an insecticide, it became infamous for its environmental impacts. ...
, causes birth defects, and is an
estrogen
Estrogen or oestrogen is a category of sex hormone responsible for the development and regulation of the female reproductive system and secondary sex characteristics. There are three major endogenous estrogens that have estrogenic hormonal a ...
analog.
Therefore, the ability of ''A. salinestris'' to break down endosulfan is important for
bioremediation
Bioremediation broadly refers to any process wherein a biological system (typically bacteria, microalgae, fungi, and plants), living or dead, is employed for removing environmental pollutants from air, water, soil, flue gasses, industrial effluent ...
to the environments where the substance was used.
See also
*
Prokaryote
A prokaryote () is a single-celled organism that lacks a nucleus and other membrane-bound organelles. The word ''prokaryote'' comes from the Greek πρό (, 'before') and κάρυον (, 'nut' or 'kernel').Campbell, N. "Biology:Concepts & Con ...
References
External links
Type strain of ''Azotobacter salinestris'' at Bac''Dive'' - the Bacterial Diversity Metadatabase
{{Taxonbar, from=Q24976883
Gram-negative bacteria
Pseudomonadales
Bacteria described in 1991