Search

Aeonium domesticum cv. variegatum is a mesochimera of the constitution green/white/green with normal proplastids and chloroplasts in the unaffected tissues and ribosome-deficient colourless mutant plastids in the white leaf tissues. All the different plastid types contain 'succulent protein crystalloids' (SPC). For more detailed characterization, the SPC elements were freed from the plastids and purified by gel filtration. Electron microscopy of different fractions revealed five levels of structural organization. Beginning with the most complex state, the levels are designated as 'succulent protein (SP) organizational state' V (hexagonally arranged and closely packed tubules in the stroma of intact plastids) to I (globular protomers of 5 nm diameter as the basic structure of SPCs). Highly purified SP-fractions were shown by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to consist of two or three proteins of Mr 56 kdalton, 58 kdalton and 60 kdalton, depending on the buffer medium used for SP isolation and the duration of storage of leaves in the frozen state. In the urea/SDS-PAGE system, these proteins show similar mobilities to α- and β-tubulin, but no immunoreaction against antitubulin. The proteolytic cleavage pattern of tubulin subunits and SP proteins are different. Their locations on two-dimensional isoelectric focusing-SDS gels show some overlappings because of microheterogeneities in both proteins in the pH gradient from pH 4.5 to 6.5. Malatedehydrogenase activity could not be detected in the purified SP fractions.

Ripe, deep-red fruits of Palisota barteri contain tubulous chromoplasts which develop from unpigmented leucoplasts. These plastids contain, besides large spherical inclusion bodies, numerous osmiophilic globules which, in the course of ripening, frequently show transition states to tubular structures. The tubules contain, in all stages of their development, acylated β-citraurin, which is also the main pigment of Citrus fruits. The tubular structures have been isolated, fragmented by French-pressure treatment, and separated into three fractions on sucrose gradients. The lightest fraction (1.044 g·cm(-3)) contained thick fragments ('saccules') with diameters of 50-60 nm, whereas the heaviest (1.083 g·cm(-3)) consisted of tubules 20-25 nm in diameter. The relative amounts of polar lipids, proteins, and carotenoids of the different fractions are consistent with a molecular structure model of tubules and saccules, according to which a wick of longitudinally oriented carotenoid molecules of variable thickness is coated by a monolayer of polar lipids and proteins. High-resolution 'negative-stainings' showed the surface of the tubules to be covered with globular particles of about 6 nm diameter. The main protein of all fractions is a 30-kDa polypeptide; it is assumed that the particles are oligomers of this specific protein.

Protein crystalloids are typical constituents of Aeonium domesticum plastids. They are composed of hexagonally arranged tube-like elements situated in the stroma without a bordering membrane. The single tubule has an external diameter of about 20 nm and an internal one of about 10 nm. The green-white-green mesochimera Ae. domesticum cv. variegatum contains normal chloroplasts in the green tissue and colourless plastids in the pale tissue. The defective plastids have a double-layered envelope, scarce internal membrane structures and contain, in the mature stage, a large vacuole. Plastid ribosomes can be detected only rarely in proplastids. They lose their ribosome complement entirely in the course of development. Polyacrylamide gel electrophoresis of total nucleic acids extracted from white tissue revealed the absence of the 23S and 16S rRNA normally present in plastids. Despite the loss of ribosomes, the plastids contain large protein crystalloids, which are structurally identical with those of normal green chloroplasts. Consequences concerning problems of encoding and transport of crystalloid protein(s) are briefly discussed.