STEM

NODULATION
By
Group Agronomy
Nodulation
• Nodulation involves the production of a special organ, the nodule,
and also what has been called a novel organelle, the symbiosome,
consisting of nitrogen-fixing bacteroids enclosed in a primarily
host-derived peribacteroid membrane.
Types of Nodulation

Root

Stem

Leaves
 Types of Stem nodulation
Aquatic legumes using
Nodulation stem primordia.

Leaves nodulation
Leaves glands of some
species of Leguminosae
family.

Root nodulation
Occurs in roots in nodule
forming legumes.

We will discuss Stem nodulation

Stem Nodulation
The symbiotic relationship between legume plant and nitrogen fixing
bacteria forming nitrogen fixing nodule at stem located root
primordia is called as stem nodule and the process of forming
nodules on stem is called stem nodulation
• First time this phenomena was reported in 1928, by Hagerup in
Aeschynomene aspera L.
Genra of Stem nodulated
Legumes

Aeschynomene

Discolobium Sesbania

Neptunia
Characteristics of Stem-nodulating
legumes
• Mostly aquatic annual or perennial herbs or shrubs and small trees.
• Have the ability to grow in waterlogged soils, swamps, or riverbanks.
• Used as green manures in lowland rice-growing tropical regions of the world.
• The presence of predetermined nodulation sites on stem.
• Comprises of primordia of adventious roots.
• These primordia appearance on whole length of stem or on small portion of stem.
• Arranged in straight vertical rows or in spiral shape rows winding around the stem.
Inoculation process
• Shoots are sprayed with suspension containing 10-8 bacteria/ml.
• Using either a liquid culture of rhizobia, a colloidal suspension obtained by
mixing entrapped rhizobia in a phosphate buffer (0.06m, pH 6.8), or a
suspension of crushed stem nodules with water passed through a filter.
• Two sources are there for survival of plant nitrogen fixing bacteria as they are
true soil living bacteria's using soil organic matter as source of energy to
survive but they use phyllosphere of the host plant to be an alternate source for
non-symbiotic growth of stem-nodule rhizobia.
Infection Process and Stem nodule
Development
• In root-nodulated legumes, there are two main types of nodules: indeterminate and
determinate.
• Indeterminate nodules generally develop on temperate legumes and are characterized by the
formation of infection threads through root hairs and by a cylindrical shape due to the
persistent apical activity of the nodule meristem.
• Determinate nodules occur on most tropical legumes. The rhizobia generally infect their host
plant intercellularly by direct “crack entry” without formation of infection threads.
• In this case, the nodule meristem activity is transient and the nodule displays the round shape
typical of determinate nodules.
Continued…
• Stem and root nodule formation share a number of similarities:
• Nodulation sites are predetermined and restricted to the cavities formed, either at the base of
the stem located primordia or at the base of lateral roots emerging from the tap root.
• The absence of root hair induction at the stem nodulation site, compared with that observed at
the base of the lateral roots.
• In both cases, however, the nitrogen fixing bacteria pentrate by direct intercellular infection
(“crack entry” of determinate nodules) between the basal cells of the primordium or the lateral
root.
• Simultaneously, dedifferentiation of cortical cells starts to form the nodule primordium.
Continued…
• The large intercellular spaces then extend toward the cortex progressively forming narrow,
branched, intercellular infection threads that spread into the meristematic cells induced in the
cortex.
• Nitrogen fixing bacteria are then released from the infection threads into the cell cytoplasm
and are surrounded by the peri-bacteroid membrane.
• At this early stage of nodule development (4 to 5 d after inoculation), the central infected
zone starts to develop the red color characteristic of leghemoglobin and the nitrogen fixing
bacteria begin to fix N, symbiotically while all stages of symbiont differentiation can still be
observed in the same section of a growing nodule.
• Interestingly, these last features are typical of indeterminate nodules.
Continued…
• The meristematic activity, that normally continues for many weeks in indeterminate nodules,
abruptly comes to an end after 1 or 2 weeks. Then they display the round shape of determinate
legumes typical of most tropical legumes.
• Differentiation of stem and root nodules thus appears as intermediate between indeterminate
and determinate types of nodule development.
• Stem nodules, and not the dark-developed root nodules, show a green cortex containing
chloroplasts closely surrounding the nitrogen-fixing cells. Proteins involved with, evolution in
photosystem-II have been shown to be present in the inner and mid cortex of the stem nodule.
It is suggested that the higher level of intercellular glycoprotein observed in the cortex of
aerial nodules could prevent oxy- gen damage to the nitrogen-fixing zone.
Continued…
• Morphologically, the stem nodules are root nodules present on the stem. The main differences
between stem and root nodules result from the aerial stem environment, which could involve
the adaptation of the microsymbionts more than that of the host plant itself.
Factors Affecting inoculation process
• As the nitrogen fixing bacteria comes form two sources viz, soil and phyllo-sphere therefore
affected by a number of external factors.
• Soil splash due to rain brings rhizobia to lower stem.
• Ants are also possible vectors to inoculate primordia on stem.
• Wind is often responsible for higher inoculation.
• Wind transport contaminated soil particles on above ground plant parts favoring inoculation.
• A carbon source is critical for their survival in the soil.
• Phyllo-spheric or Epiphytic environment must contain nutritional substances like amino acids
and carbohydrates.
References
• Boivin, C. et al. (1997) ‘Stem Nodulation in Legumes: Diversity, Mechanisms, and Unusual
Characteristics’, Critical Reviews in Plant Sciences, 16(1), pp. 1–30. doi: 10.1080/07352689709701944.
• Suzuki, S. et al. (2007) ‘Rhizobial factors required for stem nodule maturation and maintenance in
Sesbania rostrata-Azorhizobium caulinodans ORS571 symbiosis’, Applied and Environmental
Microbiology, 73(20), pp. 6650–6659. doi: 10.1128/AEM.01514-07.
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