Soil biology

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Soil biology is the study of microbial and faunal activity in the soil. This photo shows the activity of both. Worms in soil factory.jpg
Soil biology is the study of microbial and faunal activity in the soil. This photo shows the activity of both.

Soil biology is the study of microbial and faunal activity and ecology in soil. Soil life, soil biota, soil fauna, or edaphon is a collective term that encompasses all organisms that spend a significant portion of their life cycle within a soil profile, or at the soil-litter interface. These organisms include earthworms, nematodes, protozoa, fungi, bacteria, different arthropods, as well as some reptiles (such as snakes), and species of burrowing mammals like gophers, moles and prairie dogs. Soil biology plays a vital role in determining many soil characteristics. The decomposition of organic matter by soil organisms has an immense influence on soil fertility, plant growth, soil structure, and carbon storage. As a relatively new science, much remains unknown about soil biology and its effect on soil ecosystems.

Contents

Overview

The soil is home to a large proportion of the world's biodiversity. The links between soil organisms and soil functions are complex. The interconnectedness and complexity of this soil 'food web' means any appraisal of soil function must necessarily take into account interactions with the living communities that exist within the soil. We know that soil organisms break down organic matter, making nutrients available for uptake by plants and other organisms. The nutrients stored in the bodies of soil organisms prevent nutrient loss by leaching. Microbial exudates act to maintain soil structure, and earthworms are important in bioturbation. However, we find that we do not understand critical aspects about how these populations function and interact. The discovery of glomalin in 1995 indicates that we lack the knowledge to correctly answer some of the most basic questions about the biogeochemical cycle in soils. There is much work ahead to gain a better understanding of the ecological role of soil biological components in the biosphere.

In balanced soil, plants grow in an active and steady environment. The mineral content of the soil and its heartiful[ clarification needed ] structure are important for their well-being, but it is the life in the earth that powers its cycles and provides its fertility. Without the activities of soil organisms, organic materials would accumulate and litter the soil surface, and there would be no food for plants. The soil biota includes:

Of these, bacteria and fungi play key roles in maintaining a healthy soil. They act as decomposers that break down organic materials to produce detritus and other breakdown products. Soil detritivores, like earthworms, ingest detritus and decompose it. Saprotrophs, well represented by fungi and bacteria, extract soluble nutrients from delitro. The ants (macrofaunas) help by breaking down in the same way but they also provide the motion part as they move in their armies. Also the rodents, wood-eaters help the soil to be more absorbent.

Scope

Soil biology involves work in the following areas:

Complementary disciplinary approaches are necessarily utilized which involve molecular biology, genetics, ecophysiology, biogeography, ecology, soil processes, organic matter, nutrient dynamics [1] and landscape ecology.

Bacteria

Bacteria are single-cell organisms and the most numerous denizens of agriculture, with populations ranging from 100 million to 3 billion in a gram. They are capable of very rapid reproduction by binary fission (dividing into two) in favourable conditions. One bacterium is capable of producing 16 million more in just 24 hours. Most soil bacteria live close to plant roots and are often referred to as rhizobacteria. Bacteria live in soil water, including the film of moisture surrounding soil particles, and some are able to swim by means of flagella. The majority of the beneficial soil-dwelling bacteria need oxygen (and are thus termed aerobic bacteria), whilst those that do not require air are referred to as anaerobic, and tend to cause putrefaction of dead organic matter. Aerobic bacteria are most active in a soil that is moist (but not saturated, as this will deprive aerobic bacteria of the air that they require), and neutral soil pH, and where there is plenty of food (carbohydrates and micronutrients from organic matter) available. Hostile conditions will not completely kill bacteria; rather, the bacteria will stop growing and get into a dormant stage, and those individuals with pro-adaptive mutations may compete better in the new conditions. Some Gram-positive bacteria produce spores in order to wait for more favourable circumstances, and Gram-negative bacteria get into a "nonculturable" stage. Bacteria are colonized by persistent viral agents (bacteriophages) that determine gene word order in bacterial host.

From the organic gardener's point of view, the important roles that bacteria play are:

The nitrogen cycle Nitrogen Cycle.jpg
The nitrogen cycle

Nitrification

Nitrification is a vital part of the nitrogen cycle, wherein certain bacteria (which manufacture their own carbohydrate supply without using the process of photosynthesis) are able to transform nitrogen in the form of ammonium, which is produced by the decomposition of proteins, into nitrates, which are available to growing plants, and once again converted to proteins.

Nitrogen fixation

In another part of the cycle, the process of nitrogen fixation constantly puts additional nitrogen into biological circulation. This is carried out by free-living nitrogen-fixing bacteria in the soil or water such as Azotobacter , or by those that live in close symbiosis with leguminous plants, such as rhizobia. These bacteria form colonies in nodules they create on the roots of peas, beans, and related species. These are able to convert nitrogen from the atmosphere into nitrogen-containing organic substances. [2]

Denitrification

While nitrogen fixation converts nitrogen from the atmosphere into organic compounds, a series of processes called denitrification returns an approximately equal amount of nitrogen to the atmosphere. Denitrifying bacteria tend to be anaerobes, or facultatively anaerobes (can alter between the oxygen dependent and oxygen independent types of metabolisms), including Achromobacter and Pseudomonas . The purification process caused by oxygen-free conditions converts nitrates and nitrites in soil into nitrogen gas or into gaseous compounds such as nitrous oxide or nitric oxide. In excess, denitrification can lead to overall losses of available soil nitrogen and subsequent loss of soil fertility. However, fixed nitrogen may circulate many times between organisms and the soil before denitrification returns it to the atmosphere. The diagram above illustrates the nitrogen cycle.

Actinomycetota

Actinomycetota are critical in the decomposition of organic matter and in humus formation. They specialize in breaking down cellulose and lignin along with the tough chitin found on the exoskeletons of insects. Their presence is responsible for the sweet "earthy" aroma associated with a good healthy soil. They require plenty of air and a pH between 6.0 and 7.5, but are more tolerant of dry conditions than most other bacteria and fungi. [3]

Fungi

A gram of garden soil can contain around one million fungi, such as yeasts and moulds. Fungi have no chlorophyll, and are not able to photosynthesise. They cannot use atmospheric carbon dioxide as a source of carbon, therefore they are chemo-heterotrophic, meaning that, like animals, they require a chemical source of energy rather than being able to use light as an energy source, as well as organic substrates to get carbon for growth and development.

Many fungi are parasitic, often causing disease to their living host plant, although some have beneficial relationships with living plants, as illustrated below. In terms of soil and humus creation, the most important fungi tend to be saprotrophic; that is, they live on dead or decaying organic matter, thus breaking it down and converting it to forms that are available to the higher plants. A succession of fungi species will colonise the dead matter, beginning with those that use sugars and starches, which are succeeded by those that are able to break down cellulose and lignins.

Fungi spread underground by sending long thin threads known as mycelium throughout the soil; these threads can be observed throughout many soils and compost heaps. From the mycelia the fungi is able to throw up its fruiting bodies, the visible part above the soil (e.g., mushrooms, toadstools, and puffballs), which may contain millions of spores. When the fruiting body bursts, these spores are dispersed through the air to settle in fresh environments, and are able to lie dormant for up to years until the right conditions for their activation arise or the right food is made available.

Mycorrhizae

Those fungi that are able to live symbiotically with living plants, creating a relationship that is beneficial to both, are known as mycorrhizae (from myco meaning fungal and rhiza meaning root). Plant root hairs are invaded by the mycelia of the mycorrhiza, which lives partly in the soil and partly in the root, and may either cover the length of the root hair as a sheath or be concentrated around its tip. The mycorrhiza obtains the carbohydrates that it requires from the root, in return providing the plant with nutrients including nitrogen and moisture. Later the plant roots will also absorb the mycelium into its own tissues.

Beneficial mycorrhizal associations are to be found in many of our edible and flowering crops. Shewell Cooper suggests that these include at least 80% of the Brassica and Solanum families (including tomatoes and potatoes), as well as the majority of tree species, especially in forest and woodlands. Here the mycorrhizae create a fine underground mesh that extends greatly beyond the limits of the tree's roots, greatly increasing their feeding range and actually causing neighbouring trees to become physically interconnected. The benefits of mycorrhizal relations to their plant partners are not limited to nutrients, but can be essential for plant reproduction. In situations where little light is able to reach the forest floor, such as the North American pine forests, a young seedling cannot obtain sufficient light to photosynthesise for itself and will not grow properly in a sterile soil. But, if the ground is underlain by a mycorrhizal mat, then the developing seedling will throw down roots that can link with the fungal threads and through them obtain the nutrients it needs, often indirectly obtained from its parents or neighbouring trees.

David Attenborough points out the plant, fungi, animal relationship that creates a "three way harmonious trio" to be found in forest ecosystems, wherein the plant/fungi symbiosis is enhanced by animals such as the wild boar, deer, mice, or flying squirrel, which feed upon the fungi's fruiting bodies, including truffles, and cause their further spread (Private Life Of Plants, 1995). A greater understanding of the complex relationships that pervade natural systems is one of the major justifications of the organic gardener, in refraining from the use of artificial chemicals and the damage these might cause.[ citation needed ]

Recent research has shown that arbuscular mycorrhizal fungi produce glomalin, a protein that binds soil particles and stores both carbon and nitrogen. These glomalin-related soil proteins are an important part of soil organic matter. [4]

Invertebrates

Soil fauna affect soil formation and soil organic matter dynamically on many spatiotemporal scales. [5] Earthworms, ants and termites mix the soil as they burrow, significantly affecting soil formation. Earthworms ingest soil particles and organic residues, enhancing the availability of plant nutrients in the material that passes through and out of their bodies. By aerating and stirring the soil, and by increasing the stability of soil aggregates, these organisms help to assure the ready infiltration of water. These organisms in the soil also help improve pH levels.

Ants and termites are often referred to as "Soil engineers" because, when they create their nests, there are several chemical and physical changes made to the soil. Among these changes are increasing the presence of the most essential elements like carbon, nitrogen, and phosphorus—elements needed for plant growth. [6] They also can gather soil particles from differing depths of soil and deposit them in other places, leading to the mixing of soil so it is richer with nutrients and other elements.

Vertebrates

Gopher sticking out of burrow Botta's pocket gopher (Thomomys bottae)DSC2908vv.jpg
Gopher sticking out of burrow

The soil is also important to many mammals. Gophers, moles, prairie dogs, and other burrowing animals rely on this soil for protection and food. The animals even give back to the soil as their burrowing allows more rain, snow and water from ice to enter the soil instead of creating erosion. [7]

Table of soil life

This table includes some familiar types of soil life of soil life, [8] coherent with prevalent taxonomy as used in the linked Wikipedia articles.

DomainKingdomPhylumClassOrderFamilyGenus
Prokaryote Bacteria Pseudomonadota Betaproteobacteria Nitrosomonadales Nitrosomonadaceae Nitrosomonas
Prokaryote Bacteria Pseudomonadota Alphaproteobacteria Hyphomicrobiales Nitrobacteraceae Nitrobacter
Prokaryote Bacteria Pseudomonadota Alphaproteobacteria Hyphomicrobiales Rhizobiaceae Rhizobium [lower-alpha 1]
Prokaryote Bacteria Pseudomonadota Gammaproteobacteria Pseudomonadales Azotobacteraceae Azotobacter
Prokaryote Bacteria Actinomycetota Actinomycetia
Prokaryote Bacteria "Cyanobacteria (Blue-green algae)
Prokaryote Bacteria Bacillota Clostridia Clostridiales Clostridiaceae Clostridium
Eukaryote Fungi Ascomycota Eurotiomycetes Eurotiales Trichocomaceae Penicillium
Eukaryote Fungi Ascomycota Eurotiomycetes Eurotiales Trichocomaceae Aspergillus
Eukaryote Fungi Ascomycota Sordariomycetes Hypocreales Nectriaceae Fusarium
Eukaryote Fungi Ascomycota Sordariomycetes Hypocreales Hypocreaceae Trichoderma
Eukaryote Fungi Basidiomycota Agaricomycetes Cantharellales Ceratobasidiaceae Rhizoctonia
Eukaryote Fungi Zygomycota Zygomycetes Mucorales Mucoraceae Mucor
Eukaryote SAR (clade) Heterokontophyta Bacillariophyceae (Diatomea algae)
Eukaryote SAR (clade) Heterokontophyta Xanthophyceae (Yellow-green algae)
Eukaryote Alveolata (clade) Ciliophora
Eukaryote Amoebozoa (clade)
Eukaryote Plantae Chlorophyta (green algae) Chlorophyceae
Eukaryote Animalia Nematoda
Eukaryote Animalia Rotifer
Eukaryote Animalia Tardigrada
Eukaryote Animalia Arthropoda Entognatha Collembola
Eukaryote Animalia Arthropoda Arachnida Acarina
Eukaryote Animalia Arthropoda Arachnida Pseudoscorpionida
Eukaryote Animalia Arthropoda Insecta Choleoptera (larvae)
Eukaryote Animalia Arthropoda Insecta Coleoptera Carabidae (Ground beetles)
Eukaryote Animalia Arthropoda Insecta Coleoptera Staphylinidae (Rove beetle)
Eukaryote Animalia Arthropoda Insecta Diptera (larvae)
Eukaryote Animalia Arthropoda Insecta Hymenoptera Formicidae (Ant)
Eukaryote Animalia Arthropoda Chilopoda (Centipede)
Eukaryote Animalia Arthropoda Diplopoda (Millipede)
Eukaryote Animalia Arthropoda Malacostraca Isopoda (woodlouse)
Eukaryote Animalia Annelida Clitellata Haplotaxida Enchytraeidae
Eukaryote Animalia Annelida Clitellata Haplotaxida Lumbricidae
Eukaryote Animalia Mollusca Gastropoda

See also

Notes

  1. See Rhizobia for a list of other nitrogen-fixing bacteria occupying the similar niche of root nodules.

Related Research Articles

<span class="mw-page-title-main">Ecosystem</span> Community of living organisms together with the nonliving components of their environment

An ecosystem is a system that environments and their organisms form through their interaction. The biotic and abiotic components are linked together through nutrient cycles and energy flows.

<span class="mw-page-title-main">Humus</span> Organic matter in soils resulting from decay of plant and animal materials

In classical soil science, humus is the dark organic matter in soil that is formed by the decomposition of plant and animal matter. It is a kind of soil organic matter. It is rich in nutrients and retains moisture in the soil. Humus is the Latin word for "earth" or "ground".

<span class="mw-page-title-main">Mycorrhiza</span> Fungus-plant symbiotic association

A mycorrhiza is a symbiotic association between a fungus and a plant. The term mycorrhiza refers to the role of the fungus in the plant's rhizosphere, its root system. Mycorrhizae play important roles in plant nutrition, soil biology, and soil chemistry.

<span class="mw-page-title-main">Decomposer</span> Organism that breaks down dead or decaying organisms

Decomposers are organisms that break down dead or decaying organisms; they carry out decomposition, a process possible by only certain kingdoms, such as fungi. Like herbivores and predators, decomposers are heterotrophic, meaning that they use organic substrates to get their energy, carbon and nutrients for growth and development. While the terms decomposer and detritivore are often interchangeably used, detritivores ingest and digest dead matter internally, while decomposers directly absorb nutrients through external chemical and biological processes. Thus, invertebrates such as earthworms, woodlice, and sea cucumbers are technically detritivores, not decomposers, since they are unable to absorb nutrients without ingesting them.

<span class="mw-page-title-main">Detritivore</span> Animal that feeds on decomposing plant and animal parts as well as faeces

Detritivores are heterotrophs that obtain nutrients by consuming detritus. There are many kinds of invertebrates, vertebrates, and plants that carry out coprophagy. By doing so, all these detritivores contribute to decomposition and the nutrient cycles. Detritivores should be distinguished from other decomposers, such as many species of bacteria, fungi and protists, which are unable to ingest discrete lumps of matter. Instead, these other decomposers live by absorbing and metabolizing on a molecular scale. The terms detritivore and decomposer are often used interchangeably, but they describe different organisms. Detritivores are usually arthropods and help in the process of remineralization. Detritivores perform the first stage of remineralization, by fragmenting the dead plant matter, allowing decomposers to perform the second stage of remineralization.

<span class="mw-page-title-main">Arbuscular mycorrhiza</span> Symbiotic penetrative association between a fungus and the roots of a vascular plant

An arbuscular mycorrhiza (AM) is a type of mycorrhiza in which the symbiont fungus penetrates the cortical cells of the roots of a vascular plant forming arbuscules. Arbuscular mycorrhiza is a type of endomycorrhiza along with ericoid mycorrhiza and orchid mycorrhiza .They are characterized by the formation of unique tree-like structures, the arbuscules. In addition, globular storage structures called vesicles are often encountered.

<span class="mw-page-title-main">Soil food web</span>

The soil food web is the community of organisms living all or part of their lives in the soil. It describes a complex living system in the soil and how it interacts with the environment, plants, and animals.

<span class="mw-page-title-main">Rhizosphere</span> Region of soil or substrate comprising the root microbiome

The rhizosphere is the narrow region of soil or substrate that is directly influenced by root secretions and associated soil microorganisms known as the root microbiome. Soil pores in the rhizosphere can contain many bacteria and other microorganisms that feed on sloughed-off plant cells, termed rhizodeposition, and the proteins and sugars released by roots, termed root exudates. This symbiosis leads to more complex interactions, influencing plant growth and competition for resources. Much of the nutrient cycling and disease suppression by antibiotics required by plants occurs immediately adjacent to roots due to root exudates and metabolic products of symbiotic and pathogenic communities of microorganisms. The rhizosphere also provides space to produce allelochemicals to control neighbours and relatives.

Soil ecology is the study of the interactions among soil organisms, and between biotic and abiotic aspects of the soil environment. It is particularly concerned with the cycling of nutrients, formation and stabilization of the pore structure, the spread and vitality of pathogens, and the biodiversity of this rich biological community.

<span class="mw-page-title-main">Microbial loop</span> Trophic pathway in marine microbial ecosystems

The microbial loop describes a trophic pathway where, in aquatic systems, dissolved organic carbon (DOC) is returned to higher trophic levels via its incorporation into bacterial biomass, and then coupled with the classic food chain formed by phytoplankton-zooplankton-nekton. In soil systems, the microbial loop refers to soil carbon. The term microbial loop was coined by Farooq Azam, Tom Fenchel et al. in 1983 to include the role played by bacteria in the carbon and nutrient cycles of the marine environment.

<span class="mw-page-title-main">Soil respiration</span> Chemical process produced by soil and the organisms within it

Soil respiration refers to the production of carbon dioxide when soil organisms respire. This includes respiration of plant roots, the rhizosphere, microbes and fauna.

Soil organic matter (SOM) is the organic matter component of soil, consisting of plant and animal detritus at various stages of decomposition, cells and tissues of soil microbes, and substances that soil microbes synthesize. SOM provides numerous benefits to the physical and chemical properties of soil and its capacity to provide regulatory ecosystem services. SOM is especially critical for soil functions and quality.

<span class="mw-page-title-main">Microbiology of decomposition</span>

Microbiology of decomposition is the study of all microorganisms involved in decomposition, the chemical and physical processes during which organic matter is broken down and reduced to its original elements.

Soil microbiology is the study of microorganisms in soil, their functions, and how they affect soil properties. It is believed that between two and four billion years ago, the first ancient bacteria and microorganisms came about on Earth's oceans. These bacteria could fix nitrogen, in time multiplied, and as a result released oxygen into the atmosphere. This led to more advanced microorganisms, which are important because they affect soil structure and fertility. Soil microorganisms can be classified as bacteria, actinomycetes, fungi, algae and protozoa. Each of these groups has characteristics that define them and their functions in soil.

<span class="mw-page-title-main">Mycorrhizal fungi and soil carbon storage</span>

Soil carbon storage is an important function of terrestrial ecosystems. Soil contains more carbon than plants and the atmosphere combined. Understanding what maintains the soil carbon pool is important to understand the current distribution of carbon on Earth, and how it will respond to environmental change. While much research has been done on how plants, free-living microbial decomposers, and soil minerals affect this pool of carbon, it is recently coming to light that mycorrhizal fungi—symbiotic fungi that associate with roots of almost all living plants—may play an important role in maintaining this pool as well. Measurements of plant carbon allocation to mycorrhizal fungi have been estimated to be 5 to 20% of total plant carbon uptake, and in some ecosystems the biomass of mycorrhizal fungi can be comparable to the biomass of fine roots. Recent research has shown that mycorrhizal fungi hold 50 to 70 percent of the total carbon stored in leaf litter and soil on forested islands in Sweden. Turnover of mycorrhizal biomass into the soil carbon pool is thought to be rapid and has been shown in some ecosystems to be the dominant pathway by which living carbon enters the soil carbon pool.

<span class="mw-page-title-main">Root microbiome</span> Microbe community of plant roots

The root microbiome is the dynamic community of microorganisms associated with plant roots. Because they are rich in a variety of carbon compounds, plant roots provide unique environments for a diverse assemblage of soil microorganisms, including bacteria, fungi, and archaea. The microbial communities inside the root and in the rhizosphere are distinct from each other, and from the microbial communities of bulk soil, although there is some overlap in species composition.

The fungal loop hypothesis suggests that soil fungi in arid ecosystems connect the metabolic activity of plants and biological soil crusts which respond to different soil moisture levels. Compiling diverse evidence such as limited accumulation of soil organic matter, high phenol oxidative and proteolytic enzyme potentials due to microbial activity, and symbioses between plants and fungi, the fungal loop hypothesis suggests that carbon and nutrients are cycled in biotic pools rather than leached or effluxed to the atmosphere during and between pulses of precipitation.

<span class="mw-page-title-main">Viral shunt</span>

The viral shunt is a mechanism that prevents marine microbial particulate organic matter (POM) from migrating up trophic levels by recycling them into dissolved organic matter (DOM), which can be readily taken up by microorganisms. The DOM recycled by the viral shunt pathway is comparable to the amount generated by the other main sources of marine DOM.

Seventeen elements or nutrients are essential for plant growth and reproduction. They are carbon (C), hydrogen (H), oxygen (O), nitrogen (N), phosphorus (P), potassium (K), sulfur (S), calcium (Ca), magnesium (Mg), iron (Fe), boron (B), manganese (Mn), copper (Cu), zinc (Zn), molybdenum (Mo), nickel (Ni) and chlorine (Cl). Nutrients required for plants to complete their life cycle are considered essential nutrients. Nutrients that enhance the growth of plants but are not necessary to complete the plant's life cycle are considered non-essential, although some of them, such as silicon (Si), have been shown to improve nutrent availability, hence the use of stinging nettle and horsetail macerations in Biodynamic agriculture. With the exception of carbon, hydrogen and oxygen, which are supplied by carbon dioxide and water, and nitrogen, provided through nitrogen fixation, the nutrients derive originally from the mineral component of the soil. The Law of the Minimum expresses that when the available form of a nutrient is not in enough proportion in the soil solution, then other nutrients cannot be taken up at an optimum rate by a plant. A particular nutrient ratio of the soil solution is thus mandatory for optimizing plant growth, a value which might differ from nutrient ratios calculated from plant composition.

Saprotrophic bacteria are bacteria that are typically soil-dwelling and utilize saprotrophic nutrition as their primary energy source. They are often associated with soil fungi that also use saprotrophic nutrition and both are classified as saprotrophs.

References

  1. Ochoa-Hueso, R; Delgado-Baquerizo, M; King, PTA; Benham, M; Arca, V; Power, SA (February 2019). "Ecosystem type and resource quality are more important than global change drivers in regulating early stages of litter decomposition". Soil Biology and Biochemistry. 129: 144–152. doi:10.1016/j.soilbio.2018.11.009. hdl: 10261/336676 . S2CID   92606851.
  2. Nitrogen cycle diagram: http://www.epa.gov/maia/html/nitrogen.html (broken)
  3. "Actinomycetes - Remarkable Antibiotic, Nitrogen Fixing, Decomposer Bacteria". www.the-compost-gardener.com. Retrieved 2019-05-08.
  4. Comis, Don (September 2002). "Glomalin: Hiding Place for a Third of the World's Stored Soil Carbon". Agricultural Research: 4–7.
  5. Frouz, Jan (2018-12-15). "Effects of soil macro- and mesofauna on litter decomposition and soil organic matter stabilization". Geoderma. 332: 161–172. Bibcode:2018Geode.332..161F. doi:10.1016/j.geoderma.2017.08.039. ISSN   0016-7061. S2CID   135319222.
  6. "Impact of termite activity and its effect on soil composition". ResearchGate. Retrieved 2019-05-08.
  7. "What types of animals live in the soil? Why is soil condition important to them?". Soils Matter, Get the Scoop!. 2015-06-30. Retrieved 2019-05-08.
  8. (in French) Dominique Soltner  [ fr ], Les Bases de la Production Végetal, tome I: Le Sol et son amélioration, Collection Sciences et Téchniques Agricoles, 2003

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