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Q8:
What are the conditions necessary for fixation of atmospheric nitrogen by Rhizobium. What is their role in $N_2$-fixation?

Solution :

1. Physiological Conditions for Nitrogen Fixation by Rhizobium

The symbiotic nitrogen fixation process, mediated by the association between the bacterium Rhizobium and the roots of leguminous plants, requires a highly regulated micro-environment. The necessary conditions are:

  • Anaerobic Environment: The enzyme nitrogenase is highly sensitive to molecular oxygen ($O_2$), which irreversibly inactivates the catalytic iron-molybdenum protein complex [Per the kinetics of the Nitrogenase complex].
  • Presence of Leghaemoglobin: An oxygen-scavenging pigment is required to maintain a low $O_2$ tension while simultaneously supplying oxygen for bacterial respiration.
  • ATP Supply: Nitrogen fixation is an energetically expensive process requiring significant chemical energy [Per the stoichiometry of the nitrogenase reaction: $N_2 + 8e^- + 8H^+ + 16ATP \rightarrow 2NH_3 + H_2 + 16ADP + 16Pi$].
  • Presence of Nitrogenase Enzyme: The specific biological catalyst (dinitrogenase) containing Mo-Fe (Molybdenum-Iron) cofactor must be present.
  • Reducing Environment: A steady supply of reductants (electrons) typically derived from ferredoxin is essential for the reduction of atmospheric nitrogen to ammonia.

2. Role of Rhizobium in $N_2$-Fixation

The role of Rhizobium is multifaceted, involving structural, enzymatic, and biochemical coordination within the root nodule:

Role Component Biological Function
Nod Factor Secretion Bacterial signaling molecules trigger root hair curling and the initiation of nodule primordia development [By chemo-attraction and signal transduction pathways].
Nitrogenase Synthesis The bacteria synthesize the nitrogenase complex, which reduces atmospheric $N_2$ into the plant-assimilable form, ammonia ($NH_3$).
Leghaemoglobin Production The bacterium contributes to the synthesis of the apoprotein part of leghaemoglobin, which acts as an "$O_2$ buffer" to protect nitrogenase from inhibition.
Ammonia Translocation The bacteria export the synthesized $NH_3$ to the host plant cytosol, where it is promptly protonated to $NH_4^+$ to avoid toxicity, and incorporated into amino acids via the GS-GOGAT pathway.

Visual Representation of the Nodule Environment

Leghaemoglobin $N_2$ Gas $NH_3$ Product

Final Solution: The primary conditions for Rhizobium-mediated nitrogen fixation include the maintenance of a low oxygen tension via leghaemoglobin, an ample supply of ATP, and the presence of the nitrogenase enzyme. Rhizobium acts by providing the metabolic machinery (enzymes) for the reduction of inert $N_2$ into ammonia, which is then utilized by the host plant for protein synthesis and developmental growth.


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