Rhizosphere Concept and It’s Historical Background

The root system of higher plants is associated not only with soil environment
composed of inorganic and organic matter, but also with a vast community of
metabolically active microorganisms. As living plants create a unique habitat around
the roots, the microbial population on and around the roots is considerably higher
than that of root free soil environment and the differences may be both quantitative
and qualitative.

1. Rhizosphere: It is the zone/region of soil immediately surrounding the plant roots

together with root surfaces, or it is the region where soil and plant roots make contact,
or it is the soil region subjected to influence of plant roots and characterized by
increased microbial.

2. Rhizoplane: Root surface along with the closely adhering soil particles is termed as
rhizoplane.

Historical Background:

Term "Rhizosphere" was introduced for the first time by the German scientist Hiltner
(1904) to denote that region of soil which is subjected to the influence of plant roots.
The concept of "Rhizosphere Phenomenon" which shows the mutual interaction of
roots and microorganisms was came into existence with the work of Starkey et al
(1929), Clark (1939) and Rauath and Katznelson (1957).

N. V. Krassinikov (1934) found that free living nitrogen-fixing bacteria, Azotobacter

were unable to grow in the wheat rhizosphere.

Starkey (1938) examined the rhizosphere region of some plant species and
demonstrated the effect of root exudates on the predominance of bacterial population
in particular and other soil microorganisms in general in the rhizosphere region. Thus,
he put forth the concept of "Rhizosphere effect / phenomenon" for the first time.

F E Clark (1949) introduced / coined the term "Rhizoplane” to denote the root surface
together with the closely adhering soil particles.

R. I. Perotti (1925) suggested the boundaries of the rhizosphere region and showed
that it was bounded on one side by the general soil region (called as Edaphosphere)
and on the other side by the root tissues (called Histosphere).

G. Graf and S. Poschenrieder (1930) divided the rhizosphere region into two general
areas i.e. outer rhizosphere and inner rhizosphere for the purpose of describing the
same site of microbial action.
H. Katznelson (1946) suggested the R:S ratio i.e. the ratio between the microbial
population in the rhizosphere (R) and in the soil (S) to find out the degree or extent of
plant roots effect on soil microorganisms. R: S ratio gives a good picture of the relative
stimulation of the microorganisms in the rhizosphere of different plant species.

R: S ratio is defined as the ratio of microbial population per unit weight of rhizosphere
soil (R), to the microbial population per unit weight of the adjacent non-rhizosphere
soil (S)

A. G. Lochhead and H. Katznelson (1940) examined in detail the qualitative

differences between the microflora of the rhizosphere and microflora of the non-
rhizosphere region and reported that gram-negative, rod shaped and non-spore
forming bacteria are abundant in the rhizosphere than in the non-rhizosphere soil

C. Thom and H. Humfeld (1932) found that corn roots in acidic soils yielded
predominantly Trichoderma while roots from alkaline soils mainly contained
Penicillium.

M J. Timonin (1940) reported some differences in the fungal types and population in
the rhizosphere of cereals and legumes. R: S ratio of fungal population was believed
to be narrow in most of the plant species, usually not exceeding 10.

E. A. Peterson and others (1958) reported that the plant age and soil type influence
the nature of fungal flora in the rhizosphere, and the number of fungal population
gradually increases with the age of plant.

M. Adati (1932) studied many crops and found that though actinomycetes were
relatively less stimulated than bacteria, but in some cases the R: S ratio of
actinomycetes was as high as 62.

R. Venkatesan and G. Rangaswami (1965) studied the rhizosphere effect in rice plant
on bacteria, actinomycetes and fungi and reported that (i) for actinobacteris R: S was
more (ranging from 0 to 25) depending on the age of plant roots and the dominant
genera reported were Nocardia, (ii) R:S ratio reduced with the depth of soil.

E. A. Gonsalves and V. S. Yalavigi (1960) reported the presence of greater number of

algae in the rhizosphere

J. W. Rouatt et al reported positive rhizosphere effect on protozoa, but a negative

effect on algae in wheat plants.

Microorganisms in the Rhizosphere and Rhizosphere Effect

The rhizosphere region is a highly favorable habitat for the proliferation, activity and
metabolism of numerous microorganisms. The rhizosphere microflora can be
enumerated intensively by microscopic, cultural and biochemical techniques.
Microscopic techniques reveal the types of organisms present and their physical
association with the outer root tissue surface / root hairs. The cultural technique most
commonly followed is "serial dilution and plate count method" which reveal the
quantitative and qualitative population of microflora. At the same time, a cultural
method shows the selective enhancement of certain categories of bacteria. The
biochemical techniques used are designed to measure a specific change brought about
by the plant or by the microflora. The rhizosphere effect on most commonly found
microorganisms viz. bacteria, actinomycetes, fungi, algae and protozoa is being
discussed herewith in the following paragraphs.

A. Bacteria:

The greater rhizosphere effect is observed with bacteria (R: S values ranging from 10-
20 or more) than with actinomycetes and fungi. Gram-negative, rod shaped, non-
sporulating bacteria which respond to root exudates are predominant in the
rhizosphere (Pseudomonas, Agrobacterium). While Gram-positive, rods, Cocci and
aerobic spore forming (Bacillus, Clostridium) are comparatively rare in the rhizosphere.
The most common genera of bacteria are: Pseudomonas, Arthrobacter, Agrobacterium,
Alcaligenes, Azotobacter, Mycobacterium, Flavobacter, Cellulomonas, Micrococcus and
others have been reported to be either abundant or sparse in the rhizosphere. From
the agronomic point of view, the abundance of nitrogen fixing and phosphate
solubilizing bacteria in the rhizosphere assumes a great importance. The aerobic
bacteria are relatively less in the rhizosphere because of the reduced oxygen levels due
to root respiration. The bacterial population in the rhizosphere is enormous in the
ranging form 10^8 to 10^9 per gram of rhizosphere soil. They cover about 4-10% of
the total root area occurring profusely on the root hair region and rarely in the root
tips. There is predominance of amino acids and growth factors required by bacteria,
are readily provided by the root exudates in the region of rhizosphere.

B. Fungi:

In contrast to their effects on bacteria, plant roots do not alter / enhance the total count
of fungi in the rhizosphere. However, rhizosphere effect is selective and significant on
specific fungal genera (Fusarium, Verticillium, Aspergillus and Penicillium) which are
stimulated. The R:S ratio of fungal population is believed to be narrow in most of the
plants, usually not exceeding to 10. The soil / serial dilution and plating technique
used for the enumeration of rhizosphere fungi may often give erratic results as most
of the spore formers produce abundant colonies in culture media giving a wrong
picture / estimate (eg Aspergilli and Penicillia). In fact the mycelial forms are more
dominant in the field. The zoospore / forming lower fungi such as Phytophthora,
Pythium, Aphanomyces are strongly attracted to the roots in response to particular
chemical compounds excreted by the roots and cause diseases under favorable
conditions. Several fungi eg Gibberella and fujikurio produces phytohormones and
influence the plant growth.

C. Actinobacteria, Protozoa and Algae:

Stimulation of actinobacteria in the rhizosphere has not been studied in much detail
so far. It is generally understood that the actinomycetes are less stimulated in the
rhizosphere than bacteria. However, when antagonistic actinobacteria increase in
number they suppress bacteria. Actinobacteria may also increase in number when
antibacterial agents are sprayed on the crop. Among the actinobacteria, the phosphate
solublizers (eg. Nocardia, Streptomyces) have a dominant role to play.

As rule actinobacteria, protozoa and algae are not significantly influenced by their
proximity to the plant roots and their R: S ratios rarely exceed 2 to 3: 1 and around
roots of plants, R: S ratio for these microorganisms may go to high. Because of large
bacterial community, an increase in the number or activity of protozoa is expected in
the rhizosphere. Flagellates and amoebae are dominant and ciliates are rare in the
region.

Factors affecting microbial flora of the Rhizosphere / Rhizosphere Effect

The most important factors which affect / influence the microbial flora of the
rhizosphere or rhizosphere effect are: soil type & its moisture, soil amendments, soil
PH, proximity of root with soil, plant species, and age of plant and root exudates.

A. Soil type and its moisture: In general, microbial activity and population is high in
the rhizosphere region of the plants grown in sandy soils and least in the high humus
soils, and rhizosphere organisms are more when the soil moisture is low. Thus, the
rhizosphere effect is more in the sandy soils with low moisture content.

B. Soil amendments and fertilizers: Crop residues, animal manure and chemical
fertilizers applied to the soil cause no appreciable effect on the quantitative or
qualitative differences in the microflora of rhizosphere. In general, the character of
vegetation is more important than the fertility level of the soil.
C. Soil pH/ Rhizosphere pH: Respiration by the rhizosphere microflora may lead to
the change in soil rhizosphere pH. If the activity and population of the rhizosphere
microflora is more, then the pH of rhizosphere region is lower than that of
surrounding soil or non-rhizosphere soil. Rhizosphere effect for bacteria and protozoa
is more in slightly alkaline soil and for that of fungi is more in acidic soils.

D. Proximity of root with Soil: Soil samples taken progressively closer to the root
system have increasingly greater population of bacteria, and actinomycetes and
decreases with the distance and depth from the root system. Rhizosphere effect
decline sharply with increasing distance between plant root and soil.

E. Plant Species: Different plant species inhabit often somewhat variable microflora
in the rhizosphere region. The qualitative and quantitative differences are attributed
to variations in the rooting habits, tissue composition and excretion products. In
general, legumes show / produce a more pronounced rhizosphere effect than grasses
or cereals. Biennials, due to their long growth period exert more prolonged
stimulation on rhizosphere effect than annuals.

F. Age of Plant: The age of plant also alters the rhizosphere microflora and the stage
of plant maturity controls the magnitude of rhizosphere effect and degree of response
to specific microorganisms. The rhizosphere microflora increases in number with the
age of the plant and reaching at peak during flowering which is the most active period
of plant growth and metabolism. Hence, the rhizosphere effect was found to be more
at the time of flowering than in the seedling or full maturity stage of the plants. The
fungal flora (especially, Cellulolytic and Amylolytic) of the rhizosphere usually
increases even after fruiting and the onset of senescence due to accumulation of
moribund tissue and sloughed off root parts / tissues: whereas, bacterial flora of the
rhizosphere decreases after the flowering period and fruit setting.

G. Root / exudates /excretion: One of the most important factors responsible for
rhizosphere effect is the availability of a great variety of organic substances at the root
region by way of root exudates/excretions. The quantitative and qualitative
differences in the microflora of the rhizosphere from that of general soil are mainly
due to influences of root exudates. The spectrum of chemical composition root
exudates varies widely, and hence their influence on the microflora also varies widely.
S. No Root Executes Chemical Substances

1 Amino Acids All naturally occurring amino acids.

Acetic, butyric, citric, fumaric, lactic, malic, propionic,

2 Organic acids
succinic etc.

Arabinose, fructose, galactose, glucose, maltose,

Carbohydrates /
3 mannose, oligosaccharides, raffinose, ribose, sucrose,
sugars
xylose etc.

Nucleic acid
4 Adenine, cystidine, guanine, undine
derivatives

Growth factors
5 Biotin, choline, inositol, pyridoxine etc
(phytohormones)

6 Vitamins Thiamine, nicotinic acid, biotin etc

7 Enzymes Amylase, invertase, protease, phosphatase etc.

Auxins, glutamine, glycosides, hydrocyanic acid

peptides, Uv-absorbing compounds, nematode
8 Other compounds
attracting factors, spore germination stimulators, spore
inhibitors etc.

The nature and amount of chemical substances thus exuded are dependent on the
species of plant, plant age, inorganic nutrients, and temperature, light intercity, O2 /
CO2 level, root injury etc. Another source of nutrients for the microorganisms in the
rhizosphere region is the sloughed off root epidermis which exert selective
stimulation effect on some specific groups of microorganisms. For instance, glucose
and amino acids in the exudates readily attract Gram-negative rods which
predominantly colonize the roots. Sugars and amino acids in the root exudates
stimulate the germination of chlamydospores and other resting spores of fungi;
stimulation effect of root exudates on plant pathogenic fungi, nematodes is also well
known.