Fusion of pea pisum sativum cultivar little marvel root nodule protoplasts with tobacco nicotiana tabacum cultivar xanthi mesophyll protoplasts

Syono, K.; Nagata, T.; Suzuki, M.; Kajita, S.; Matsui, C.

Zeitschrift fuer Pflanzenphysiologie 95(5): 449-458


Accession: 005499623

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Protoplasts from pea root nodules were isolated enzymatically. Nodules from flowering pea were suitable material for the isolation of stable protoplasts. The isolated protoplasts were classified into 4 types ranging from small spheres from the early stage of infected [with Rhizobium leguminosarium] nodule cells (type 1) to large elongated protoplasts from the late stage of infected cells (type 4). In all these types, including the elongated ones, the lack of cell wall materials at the surface of the plasmalemma was confirmed by EM. The fine structure of vesicles around bacteria and the cytoplasm of the bacteroidal cells appeared well preserved during enzymatic isolation of protoplasts. The isolated nodule protoplasts, especially the spherical types, and tobacco mesophyll protoplasts were agglutinated with polyethylene glycol (PEG) and subsequently fused upon elution of the PEG to form completely spherical fusion products. [This research has application to studying symbiotic N2 fixation.].