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The interaction of the chloroplast precursor protein of ferredoxin with mixed model membranes composed of 2H chain labeled monogalactosyl diacylglycerol and phosphatidylcholine was studied by 2H and 31P NMR. The bilayers were found to have special chain packing properties which most likely are the result of a specific arrangement of head groups at the interface. The precursor and not the corresponding apoprotein induced a bilayer → isotropic transition in lipid organization as a result of the transit sequence—lipid interaction. The implications of these observations for proteins import into chloroplasts are indicated.

During greening of detached leaves from dark-grown barley seedlings, the linolenic acid content of the lipids increases in the early stages of the formation of the chloroplast lamellar system. Primarily the fraction containing monogalactosyl diglyceride is enriched with linolenic acid. Incorporation of 14C-labeled acetate into the leaf lipids of detached whole leaves is low, but increases 10- to 20-fold during greening. Increasing percentages of label appear in linolenic acid during the first 15 hr of greening, whereafter they remain constant. A constant, relatively high amount of acetate is incorporated into lipids when slices of leaves at various stages of greening are incubated by submersion in acetate solution, a treatment that blocks further chlorophyll synthesis during incubation. At the initial greening stages 75% of the label is channeled into steroids and other unsaponifiable lipids, but in advanced stages of chloroplast development 75% of the incorporated acetate is built into phospho-, sulfo- and galacto-lipids, and only 25% is channeled into unsaponifiable lipids. Experimental variation of the physiological conditions of the tissue during incubation resulted in differences in the amount of label found in the various phospho- and galacto-lipids. The amounts of labeling of the individual fatty acids in the lipid classes studied differ markedly and could be changed by varying the conditions of incubation. Labeling of linolenic acid was found to be highest in the monogalactosyl diglyceride fraction at all stages of greening.

The capacity of leaf slices from light-grown seedlings of wild type barley and 10 xantha mutants at six different gene loci to incorporate acetate-(14)C into various lipids has been investigated. The fatty acid composition of the leaf lipids in these lethal mutants was similar to that of the wild type, but the fatty acid labeling pattern in the individual lipid classes can be drastically altered by these mutations, which affect chloroplast differentiation. A genetic block in chlorophyll synthesis, caused by mutations in the xan-f locus, leads to a repression of the formation of chloroplast membranes and of acetate incorporation into phospho-, sulfo-, and galacto-lipids (the acetate being preferentially channeled into a lipid fraction containing steroids and free fatty acids). Two leucine "auxotrophs" at different loci, which in the absence of leucine in the growth medium produce giant grana and accumulate some chlorophyll, differed considerably in the amount of labeling of their polar lipids during incubation. Leaves of xan-a(11), containing plastids with little chlorophyll, highly disorganized membrane systems, and large bodies with osmiophilic deposits, were nonetheless equal to wild type in their capacity to incorporate acetate-(14)C into phospho-, sulfo-, and galacto-lipids. The mutants at the xan-m locus have plastids with undispersed prolamellar bodies and osmiophilic packages of grana-like membranes associations. Leaf slices of these mutants synthesized considerably more linolenic acid-(14)C, which was incorporated into monogalactosyl diglycerides, than did slices of the wild type. This led to a labeling pattern of the fatty acids in the monogalactolipids which was remarkably similar to their endogenous fatty acid composition.