Reproductive growth of intracellular bacteria from isolated protoplasts in nodules of clover and soybean was directly investigated using a microchamber with visual and video recording. Differentiated bacteriods from clover nodules uniformly failed to reproduce. Such growth as occurred came from undifferentiated rhizobia from within the protoplast or extracellularly in the nodule. Plating investigation gave results in agreement with this conclusion. Osmoprotective media failed to secure the reproduction of differentiated clover bacteroids. Reproductive growth of bacteroids from protoplasts and crushed nodules of soybean was regularly observed in the microchamber and determined as proportionate colony-forming ability (CFA) on laboratory media. The CFA markedly increased with age of nodule and with the addition of nodule or root extract. The promoting effect of such extracts was reduced after heating for 60 min at 100°C, and lost completely after 20 min at 121°C. High osmolarity in the suspending and culture media was detrimental to bacteroid recovery.
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Bergersen's modified medium
- B+m :
BMM with additional mannitol
Chlamydomonas dilution buffer
protoplast dilution buffer
PDB without mannitol or sorbitol
Rhizobium minimal medium
- R+m :
RMM with mannitol instead of sucrose
- YMA, YMB:
yeast mannitol agar and broth, respectively. For details, see Materials and methods
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Zhou, J.C., Tchan, Y.T. & Vincent, J.M. Reproductive capacity of bacteroids in nodules of Trifolium repens L. and Glycine max (L.) Merr.. Planta 163, 473–482 (1985). https://doi.org/10.1007/BF00392704
- Bacteroid (reproductive capacity)
- Glycine (bacteroids)
- Nodule extract
- Nodule protoplast