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In addition to the GBSSI isoform of starch synthase described previously, the pea plant contains a second, granule-bound isoform, GBSSIb. GBSSI is abundant in pea embryos and Rhizobium root nodules, is present at low levels in pods and is absent from leaves. Mutations at the lam locus eliminate GBSSI from all of these organs. GBSSIb is present in pods, leaves and nodules and is unaffected by mutations at the lam locus. GBSSI and GBSSIb are very similar in molecular mass, primary sequence, activity and antigenic properties. GBSSIb, like GBSSI, can synthesize amylose in the presence of malto-oligosaccharides in isolated starch granules. However, its role in vivo is unclear. The lam mutation eliminates amylose from the starch of embryos but does not affect the relatively small amounts of amylose-like material in the starch of pods, leaves and nodules. The significance of these results for understanding of the regulation of amylose synthesis is discussed.

Mutants of Pisum sativum L. with seeds containing low-amylose starch were isolated by screening a population derived from chemically mutagenized material. In all of the mutant lines selected, the low-amylose phenotype was caused by a recessive mutation at a single locus designated lam. In embryos of all but one mutant line, the 59 kDa granule-bound starch synthase (GBSSI) was absent or greatly reduced in amount. The granule-bound starch synthase activity in developing embryos of the mutants was reduced but not eliminated. These results provide further evidence that amylose synthesis is unique to GBSSI. Other granule-bound isoforms of starch synthase cannot substitute for this protein in amylose synthesis. Examination of iodine-stained starch granules from mutant embryos by light microscopy revealed large, blue-staining cores surrounded by a pale-staining periphery. In this respect, the low-amylose mutants of pea differ from those of other species. The differential staining may indicate that the structure of amylopectin varies between the core and peripheral regions.

Analyses of seeds from a wide range of pea lines have revealed extensive variation for all of the major fatty acids normally found in pisum. Unlike total lipid content, this variation was not affected by the presence of mutant alleles at the r and rb loci. To allow chemically analysed seed to be used in crosses, which is of particular importance for F 2 seed, a method has been developed by which individual seeds can be analysed non-destructively. This system entails drilling a small hole in the seed, away from the embryonic axis. The small quantity of powder produced by this process can be used for the chemical analysis but does not prevent the seed from being grown to produce progeny. Reducing the sample size had no significant effect upon the compositional analysis of the fatty acids.