BOTANY:BOT-409

Microbes, man and environment

Dr. Shahbaz ali

Associate professor
Govt. mao college Lahore
Course contents:
 Nitrogen-Fixing Bacteria

• The conversion of gaseous nitrogen (N2) to ammonia (NH3) often occurs

as part of a symbiotic relationship between bacteria and plants.
• These symbioses produce more than 100 million metric tons of fixed
nitrogen annually and are a vital part of the global nitrogen cycle, and
without them life on Earth could not have evolved.
• Currently, symbiotic nitrogen fixation accounts for more than half of
the nitrogen used in agriculture (the remainder is applied as fertilizer).
• The provision of fixed nitrogen enables the growth of host plants in
soils that would otherwise be nitrogen limiting.
• At the same time, it reduces loss of nitrogen by denitrification and
leaching.
 Root Nodules
• Several bacterial genera form nitrogen-fixing nodules with legumes.
• These include Allorhizobium, Azorhizobium, Bradyrhizobium, Mesorhizobium,
Sinorhizobium, and Rhizobium. Collectively these bacteria are often called
rhizobia.
• Here we discuss the general process of nodulation, as revealed through studies of
bacteria in the genus Rhizobium. Rhizobia live freely in the soil.
• In nitrogen-sufficient soils, plants secrete ethylene-mediated compounds that
inhibit nodulation. But even in nitrogen-starved soils, when rhizobia approach the
plant root, they are assumed to be invaders.
• The plant responds with an oxidative burst, producing a mixture of compounds
that can contain superoxide radicals, hydrogen peroxide, and N2O.
• This oxidative burst is critical in determining the fate of the infection process.
Rhizobia that become effective colonizers mount antioxidant defenses.
• Only rhizobia with sufficient antioxidant abilities proceed to the next step in the
infection process.
• Infection is initiated by the exchange of signaling molecules between the plant and
rhizobia in the rhizosphere. Plant roots release flavonoid inducer molecules (2-
phenyl-1,4-benzopyrone derivatives) that stimulate rhizobial colonization of the root
surfaces (figure 31.9a).
• Flavonoids bind the bacterial protein NodD, which can then function as a
transcriptional regulator.
• NodD activates transcription of nod genes, which encode the biosynthetic enzymes
needed for the production of Nod factor signaling compounds (figure 31.9b).
Conversely flavonoids from non-host plants inhibit the production of Nod factors.
• Upon receipt of the Nod factor signal, gene expression in the outer (epidermal) cells
of the roots is altered, resulting in changes in the levels of intracellular calcium. This
“calcium spiking” triggers the root hairs to curl, thereby entrapping bacteria (figure
31.9c,d).
• Nod factors trigger root cortex cells to initiate cell division, and these cells will
eventually form the nodule primordium that accepts the invading rhizobium.
• In addition to Nod factors, the bacteria now produce an exopolysaccharide
that induces changes in the plant cell wall, and the plant plasma membrane
invaginates.
• This leads to the development of a bacteria-filled, tube-like structure called
the infection thread (figure 31.9e,f).
• When bacteria penetrate the base of a root hair cell, they stimulate growth
of the infection thread through additional plant cell walls and membranes.
• This is dependent on the plant hormone cytokinin in concert with Nod
factor stimulation of plant cell mitosis.
• Finally, the infection thread filled with bacteria reaches the inner plant
cortex, where each bacterial cell is endocytosed by a plant cell in a discrete,
un-walled membrane compartment that arises from the infection thread.
• This unit, consisting of an individual bacterium and the surrounding
endocytic membrane, is called the symbiosome (figure 31.9g).
• It is here that each bacterial cell differentiates into the nitrogen-fixing form called
a bacteroid.
• The low levels of oxygen within the symbiosome triggers nitrogen fixation and the
switch to microaerobic respiration needed to provide ATP and reducing
equivalents to the nitrogenase enzyme.
• Overall, most metabolic processes within bacteroids stop but nitrogen fixation
and respiration, are upregulated.
• The assembly of many symbiosomes gives rise to the root nodule (figure 31.9h,i).
• The multistep conversion of N2 to NH3 by nitrogen fixation can be summarized as
follows:
• The reaction is driven by the enzyme complex nitrogenase.
 Leghemoglobin
• The reduction of atmospheric N2 to ammonia by the differentiated bacteroids depends on the
production of a protein called leghemoglobin.
• Recall that the nitrogenase enzyme is disabled by oxygen. To help protect the nitrogenase,
leghemoglobin binds oxygen and helps maintain microaerobic conditions within the mature
nodule.
• Leghemoglobin is similar in structure to the hemoglobin found in animals; however, it has a
higher affinity for oxygen. Interestingly, the protein part of leghemoglobin is encoded by plant
genes, whereas the heme group is a bacterial product.
• The transfer of ammonia from the rhizobium to the host plant involves a complex cycling of
amino acids. The plant provides certain amino acids to the bacteroids so that they do not need
to assimilate ammonia.
• In return, the bacteroids shuttle amino acids (which bear the newly fixed nitrogen) back to the
plant.
• This creates an interdependent relationship, providing selective pressure for the evolution of
mutualism. The bacteria also receive carbon and energy in the form of dicarboxylic acids from
• The molecular mechanisms by which both the legume host and the
rhizobial symbionts establish productive nitrogen-fixing bacteroids within
nodules is an intense area of research.
• A major goal of biotechnology is to enable nitrogen fixation within cereal
plants such as rice, wheat, and corn. Three approaches are being
pursued:
• (1) modifying rhizobia to form productive nodules on these non-
leguminous plants;
• (2) creating strains of non-nodulating nitrogen-fixing bacteria that will
colonize cereal plants; and
• (3) introducing nitrogen-fixation genes into plants that are not rhizobium
hosts.
• Although progress has been made using each approach, nitrogen fixation
on these important crops has yet to be realized, and the use of human-