PLANT MICROBE

INTERACTION
 INTRODUCTION

Microorganisms make available different types of nutrients to plants, which are

required for their growth. Following are the important roles of microorganisms in
plant growth.
 Certain microorganism produce growth promoting substances, eg auxin and
gibberellin which stimulates plant growth
 There may be a competition between microorganisms and plants for nutrients
available in soil
 Certain pathogenic microorganisms secrete some toxic substances, which may be
harmful to the plants
 INTRODUCTION

 Bacterial viruses show indirect harmful effect on plant growth by destroying

soil bacteria

 Most of the soil microorganisms are beneficial for plant growth

 INTRODUCTION

 The biological availability of N, P and K are considerable an economic

importance because they are the major plant nutrients derived from the soil.

 Of the three, nitrogen is the most important one because it is a key

building block of protein molecule upon which life is based

 The process of conversion of molecular nitrogen into ammonia is called

Nitrogen fixation
 MODES OF NITROGEN
FIXATION

1. Spontaneous Nitrogen Fixation

2. Industrial process

3. Biological Nitrogen Fixation

 A. SPONTANEOUS
NITROGEN FIXATION
B. INDUSTRIAL PROCESS

Production of ammonia by Haber process

 BIOLOGICAL NITROGEN
FIXATION

 About 78% of Nitrogen is present in gaseous state in the atmosphere.

 The microorganism utilize this gaseous nitrogen directly or indirectly in the soil
and convert into combined form of organic compound through biological reactions

 The phenomenon of fixation of atmospheric nitrogen by biological means is

called as diazotrophy or biological nitrogen fixation (BNF)

 The prokaryotes or microorganisms which fix the atmospheric are called

‘diazotrophs’ or nitrogen fixers
 BIOLOGICAL NITROGEN
FIXATION

 Beijerinck (1888) isolated Rhizobium root nodules for the first time

 Later Chlostridium pasteurianum (free living bacteria), frankia from root

nodules of non-legumes, Nostoc from lichens, Anabaena from Azolla leaves and
carroloid root from cycas were discovered which are natural nitrogen fixers

 Biological nitrogen fixation may be of two types

• Symbiotic
• Asymbiotic
 ASYMBIOTIC NITROGEN
FIXATION
 The process by which the microorganisms utilize the gaseous nitrogen directly
or independently in the soil is called as asymbiotic nitrogen fixation.
 The microorganisms involved in asymbiotic nitrogen fixation are called
asymbiotic or non-symbiotic nitrogen fixers.
 Some examples of such microorganisms are bactria like Azotobacter,
Rhodospirillum, Rhodopseudomona, Chlorobium, Diplococcus pneumoniae,
Micrococcus sulfurens etc.
 These microorganisms are known to fix atmospheric nitrogen asymbiotically
 MECHANISM OF NITROGEN
FIXATION

 The mechanism of nitrogen fixation is explained on the basis of physiological

basis
 The protoplasm of Azotobacter produces special catalysts which take part in
fixing the free nitrogen
 The catalyst contains carboxyl and amino group as an important part which do not
take any active part in fixing atmospheric nitrogen
 Phenyl hydrogen and hydroxylamine are also present in the composition of the
catalyst.
 MECHANISM
 It was presumed that molecular nitrogen reacts with catalyst of nitrogen fixation
through oxygen of carboxyl group and produces hydrazine as the first product of the
reaction
 The hydrazine is then further reduced by active hydrogen and is transformed into
amino acids.
 They finally enter the proteins of cell protoplasm.
 This incorporation of amino acid into protoplasm helps into the growth of
bacterium.
 REQUIREMENTS OF
NITROGEN FIXATION

1. Oxygen and carboxyl group of the catalysts play an important

role in nitrogen fixation
2. A very small amount of chemical energy (ATP) is used up during
nitrogen fixation (about 1 kilo calories to fix 2mg of atmospheric
nitrogen.
3. It also gives the relationship between respiration and nitrogen
fixation.
 SYMBIOTIC NITROGEN
FIXATION
 The process of nitrogen fixation where the microorganism utilize the gaseous
nitrogen indirectly through the mediation of other living organisms in the soil is
called symbiotic nitrogen fixation
 The microorganism involved are called symbiotic nitrogen fixing bacteria
 The microorganism develop symbiotic relationship with different parts of plants
and may develop special structures as the site of nitrogen fixation
 Non nodule forming diazotrophs for example Azotobacter, Beijernickia, Derxia
are associated with the roots of certain plants
 Azotobacter paspali is restricted to the roots of tropical grasses
 SYMBIOTIC NITROGEN
FIXATION

 Beijerinckia shows host specificity with sugarcane root

 Azospirilum is associated with the roots of corn, wheat, Sorghum etc

 In addition to these bacteria, Frankia, Rhizobium sp and cyanobacteria

undergo symbiosis by getting established inside the plant tissues and may or may
not develop special symbiotic structures.
 SOIL FERTILITY
 The ability of soil to provide all essential nutrients in available forms and in a
suitable balance is called soil fertility.
 A fertile soil is considered to be one that produces abundant crops under suitable
environmental conditions.
 Microorganisms play a significant role in maintaining the soil fertility.
 Nitrogen fixing microorganisms are now employed in soil for improving soil
fertility.
 The atmosphere contains about 1015 tons of nitrogen gas and the nitrogen cycle
involves the transformation of some 3x10 tons of nitrogen per year on a global basis
 However, transformations are not exclusively biological.

 Lightening probably accounts for about 10% of the world's supply of

fixed nitrogen.

 Out of the total quantity of nitrogen reaching the soil, only an estimated
60% is available for plant growth.

 The rest 40% can be easily achieved with the use of bio-fertilizers or any
method of biological nitrogen fixation.
 Biofertilizers are the cultures of microorganisms used for inoculating seed
or soil or both under ideal conditions to increase the availability of plant
nutrients.
 Their purpose is to supplement chemical fertilizers and not to replace
them.
 It has been reported that the microorganisms have the beneficial role of
biological nitrogen fixation to supply nitrogen to crops, solubilizing insoluble
phosphates to soluble forms to make them available to crops
 If these specific organisms are not present in the soil, they have to be
inoculated to initiate and accelerate biological activity.
 Such microbial inoculants have been called bio-fertilizers.
 Bio-fertilizers can be put broadly into two groups ie
• Inoculants of specific organisms such as Rhizobia, Azotobacter, blue-green algae,
phosphate solubilizers, cellulolytic microorganisms
• and biomass producing organisms, e.g. Azolla.
 Microorganisms are abundant in upper six inches or surface layer of soil.
 All of them play an important role in improving soil fertility.
 Bacteria, Azotobacter, Rhizobium, Actinomycete, Frankia etc. fix atmospheric
nitrogen and increase soil fertility.
 Fungi like Chaetomium globosum, Polyporus abietinus etc. help in recovering
soil nutrients.
 An increased yield to the extent of 10 to 15 percent could be obtained through
blue- green algae application
 A conservative estimate indicates that blue-green algae contribute about
25 to 35 kg of nitrogen per hectare per cropping season.

 Phosphate solubilizing microorganisms, particularly the soil bacteria

belonging to the genera Pseudomonas and Bacillus, and fungi belonging to
the genera Penicillium and Aspergillus possesses the ability to transform
insoluble phosphates into soluble forms.
 SYMBIOTIC ASSOCIATIONS
 later, another bacterium plant association was found to fix nitrogen
symbiotically.

 The bacterium Spirillum lipoferum can live symbiotically in the roots of

several non-legume tropical grasses and was reported to fix nitrogen at a rate
comparable to the Rhizobium legume system.

 However, the anatomical relationship appears to be somewhat different.

Plants infected with Spirillum lipoferum do not form nodules on their roots.

 The bacteria in the infected root live within the cells of the root cortex,
the layer of cells immediately under the epidermis.
 There are several plant species which grow as weeds and have well
established microbial symbiotic association, e.g. grasses and Pteriodphytes.

 It has been reported that most of the C4 plants (e.g. tropical grasses like
Cyperus, Saccharum, Cyanodon, Andropogon, Spartina etc. harbour
microbial populations in their root regions.

 These microbes can also fix atmosphere nitrogen in the root region
(rhizosphere).

 All tropical grasses and plants so far found to stimulate much activity in
nitrogen fixation.
 The important nitrogen fixing bacterium associated with these weeds is
Spirillum lipoferum.
1. Rhizobium
 The bacterial-plant, Rhizobium-legume, mutualistic association is a widely
known classical type of symbiosis.
 The bacterial partners, one of several species of Rhizobium, are common
types of bacteria that live in the soil, and the other partner is one of several
species of pants called legumes-soybeans, beans, peas, alfalfa and clovers are
common examples.
 When roots of a susceptible legume become infected with an appropriate
strain of Rhizobium, the roots undergo a modification and form structures called
nodules.
 Subsequently, specialized cells with the bacterial infected nodules convert
gaseous nitrogen (a form in the atmosphere that cannot be used by either the
bacteria or plant directly) into a combined form of nitrogen that can be used
by both types of organisms.

 The process is called symbiotic nitrogen fixation. Both the partners in the
association (the legume and the bacterium) can grow separately in the soil as
independent free-living organisms

 Although the association is facultative, some strains of Rhizobium exhibit

a high degree or specificity for the roots of certain kinds of leguminous
plants.
 In other words, a specific strain of Rhizobium can only infect specific
susceptible species of legumes. For example strains of Rhizobium that cause
nodules formation in soybeans are unable to stimulate nodule formation in
other legumes, such as clover, alfalfa etc

 It is thought that lectins present on the surface of soybeans are

responsible for the selectivity.
 FORMATION OF NODULE
 Rhizobia have the capacity to fix free atmospheric nitrogen of soil in
nodule.

 They infect or get their entrance in root through the soft-hair or other
epidermal cells by damaging them.

 In the first stage of the infection, bacteria grow very profusely at the tip of
the root hair and form a long filament in the root hair called infection thread.

 This thread reaches the endodermis and pericycle area through cortex
tissue. Cells of this area (cortex) go on dividing and form a young nodule.
 The newly formed young nodule pushes the overlying parenchyma and the
epidermal tissue towards outside and produces a small swelling on the surface
of the root.

 Anatomically, a nodule is made of thin walled parenchymatous cells which

are filled up with the nitrogen fixing organisms.

 The size and shape of the root nodule varies according, to plants in which
it occurs. It is not necessary that all the bacteria, which infect the root produce
nodule.

 According to Wipf and Cooper (1939) the root nodule always contains
double number of chromosomes as against of normal somatic tissue.
 If the root lacks such cells (cells with double chromosome number), there will
be no formation of root nodule, even after the formation of infection thread.
 Those plants which do not bear root-nodules will never be able to fix
atmospheric nitrogen in the plants.
 It has been observed by Allen and Allen (1947) that out of 429 genera of family
Fabaceae about 179 genera containing 949 species form nodules.\
Nitrogen Fixing Organisms found in Nodules
Rhizobium:
 The presence of these bacteria in the root nodule can be seen under high power
microscope by crushing a washed nodule between two glass slider.
 Frank (1877) and Beijerinck (1888) discovered Rhizobium a free living or
symbiotic bacteria of leguminous as well as non-leguminous plants.
 This kind of bacteria, preferentially, infect legumes as compared to non-
legumes.
 Structurally, Rhizobia are rod shaped but great variation can be observed
during their life cycle.
 These may be coccoid, very small, highly motile or ellipsoidal forms.
 The bacteroids are usually irregular with X, Y, star and club-Shaped forms.
 They are 0.5 to 0.9 micron by 1.2 to 3.0 micron in size.
 All the species are non-spore forming, acid producing and fast growing.
 These symbiotic bacteria (Rhizobium) are difficult to cultivate in ordinary
culture media but they grow luxuriantly on mannitol agar.