Characterisation of seeds of a C4 phosphoenolpyruvate carboxylase-deficient mutant of Amaranthus edulis.

Seeds from the C(4) plant Amaranthus edulis were studied as part of the characterisation of a mutant (designated LaC(4) 2.16), which contains reduced amounts (5% of wild type) of the photosynthetic leaf form of phosphoenolpyruvate carboxylase (PEPC). On a per seed basis, the amount of PEPC activity was not significantly altered, while the weight and protein content of the mutant seeds were 34% lower than that of the wild type. Western gel blot analysis detected two PEPC polypeptides with molecular masses of 105 kDa (minor) and 100 kDa (major). The determination of in vitro phosphorylation in reconstituted assays revealed the presence of both calcium-dependent and calcium-independent PEPC-kinase activities in protein extracts of wild-type and mutant seeds. However, PEPC proteins were phosphorylated in dry seeds, and PEPC phosphorylation did not occur in vivo during seed imbibition in the presence of (32) P-phosphate. In contrast, (32) P-phosphate was incorporated into a range of proteins in wild-type seeds, but not in mutant seeds. In addition, ATP content was much reduced in germinating mutant seeds and this did not increase following the supply of phosphate. Collectively, these data suggest that the deficiency in C(4) PEPC in mutant A. edulis leaves has no effect on C(3) -type PEPC content and phosphorylation state in seeds, but causes impairment of energy production, thereby accounting for the reduced germination of the mutant.

Ecotypic variations in phosphoenolpyruvate carboxylase activity of the cordgrass Spartina densiflora throughout its latitudinal distribution range.

This study compared the specific activity of phosphoenolpyruvate carboxylase (PEPC) of Spartina densiflora Brongn., collected from four populations along its latitudinal distribution range. Spartina densiflora is a halophyte with C(4) photosynthesis that has a very wide latitudinal distribution, from Patagonia to the southwest Iberian Peninsula. The basis of intraspecific differences in PEPC activity were analysed by recording the phosphorylation state and amount of the enzyme, comparing leaf anatomy and evaluating leaf gas exchange. S. densiflora individuals from Patagonia had 60% higher PEPC specific activity than plants from the other three populations due to higher levels of PEPC protein that coincided with lower activation mediated by phosphorylation, yielding similar net photosynthesis rate (c. 29 micromol CO(2)xm(-2)xs(-1)). Patagonian plants had a higher area of photosynthetic mesophyll relative to total chlorophyll than plants from north Argentina and the southwest Iberian Peninsula. Ecotypic differentiation in PEPC activity and leaf anatomy were found, distinguishing a higher-latitude ecotype from lower-latitude populations. The higher PEPC protein levels of the Patagonian ecotype seemed to be a response to lower light activation level of the enzyme, as judged by the low PEPC phosphorylation state.