Ribulose bisphosphate carboxylase activity was examined in Pinus silvestris L. during successive seasons. The enzyme activities were studied both in seedlings, kept under controlled conditions in a climate chamber, and in needles from a 15‐year‐old tree in a natural stand. The enzyme activities were analysed in cell‐free extracts prepared with Tween 80 as protective agent. The carboxylase activity fluctuated periodically both in the seedlings and in the natural stand. In the seedlings, the weight‐related activity in the older needles increased 50–100% (in the cotyledons c. 200%) in the beginning of the “summer”. It decreased as the new shoot developed. The specific activity increased c. 100%. With chlorophyll as base, the activity usually decreased during “summer”. In the developing current needles the carboxylase activity increased when expressed on a weight or on a protein basis. The decrease in weight‐related carboxylase activity in the older needles was preceded by, or simultaneous with, loss of total protein. It is suggested that protein, including the carboxylase, is utilized as nitrogen reserve for the new shoot. During hardening by combined photoperiod and thermoperiod, the carboxylase activity decreased when expressed relative to dry weight and protein. Calculated on a chlorophyll basis, the activity was rather constant. In the natural stand the activity in the one‐ and two‐year‐old needles increased during spring and summer and decreased during autumn and winter. Even at severe winter stress substantial carboxylase activity remained in the needles. The activity of the enzyme in vivo is discussed with respect to electron transport and net photosynthesis.
Net photosynthesis of seedlings of Pinus silvestris has been measured and compared with the activities of photosynthetic electron transport and extracted RuBP carboxylase. The effects of prolonged frost hardening (photoperiod 8 h, + 3°C) followed by winter stress at subzero temperatures were analysed. There was a parallel effect of frost hardening and winter stress on the photosynthetic properties of both intact seedlings and isolated chloroplast thylakoids. The activity of extracted RuBP carboxylase was less affected by the treatments. In relation to earlier works we conclude that the decay of net photosynthesis in winter climate is determined by the electron transport properties of the chloroplast thylakoids, i.e. by the pool sizes of photosynthetically active plastoquinone. The results of this work justify the definition of two phases in the response of conifers towards autumn and winter climates: I. Frost hardening occurs at temperatures slightly above zero and it does not affect the efficiency of photosynthesis as defined by the quantum yield at rate limiting light absorption. II. Winter stress occurs at subzero temperatures and it is characterized by a suppression of the photosynthetic efficiency as a result of damage within the photosynthetic apparatus.
Scots pine (Pinus sylvestris L.) seedlings of a provenance from northern Sweden were cultivated hydroponically for 7 weeks in a climate chamber. The nutrient solution contained either 2.5 (low-N) or 50 (high-N) mg N l(-1) with other essential elements added in a fixed optimal proportion to the nitrogen. After 5 and 7 weeks, the seedlings were analyzed for growth, total nitrogen and other essential nutrients, protein and free amino acids. Low-N seedlings grew more slowly and had higher root/shoot ratios than high-N seedlings. With respect to total nitrogen, the effect of the lower nutrient supply was mainly on the nitrogen content of the whole plant and the allocation of nitrogen among tissues, not on tissue nitrogen concentration. This was also the case for potassium, phosphorus, calcium and magnesium. The proportions by weight among these macronutrients in the whole seedlings were similar in both nutrient regimes. The proportion and concentration of sulfur were significantly lower in low-N seedlings than in high-N seedlings, because of a lower net uptake of sulfur than of other macronutrients. The shoot, needles and stem of low-N seedlings had higher concentrations of free amino acids and lower concentrations of protein than the shoot, needles and stem of high-N seedlings. Arginine dominated the pool of free amino acids in the low-N seedlings, whereas glutamine predominated in the high-N seedlings. We conclude that Scots pine seedlings accumulated soluble nitrogen as arginine when net protein synthesis was limited by factors other than nitrogen availability. Nutritional imbalance, as revealed by growth characteristics and a suboptimal proportion and concentration of sulfur in the seedlings, probably affected synthesis of S-amino acids, resulting in the diversion of assimilated nitrogen to arginine instead of protein.
The concentration of free amino acids and total nitrogen was studied in needles, stems and roots of seedlings of Pinus sylvestris L. for five weeks during the second growth period (“summer”). In one group of seedlings the source/sink relation was disturbed through removal of the terminal buds. The seedlings were cultivated in artificial year‐cycles in a climate chamber. Total nitrogen increased in needles and sterns of intact seedlings in the beginning of the “summer” and decreased during shoot growth. In seedlings, from which the buds had been removed, nitrogen remained at high levels in the primary needles and accumulated in steins and roots. The results are consistent with utilization of nitrogen in older needles and in the stem during shoot elongation. The pool of free amino acids increased in the beginning of the “summer” and decreased after bud break in primary needles, stems and roots. Arginine and glutamine, in the roots also asparagine, were the dominating amino acids (amides included). Together, these compounds (plus glutamate and aspartate) contributed about 90% of the nitrogen in the amino acid pool in all organs. In primary needles and in the stem, arginine predominated at the end of hardening (75–85% of the amino acid nitrogen). Free amino acids contributed at most ca 10% of the total nitrogen in primary needles, where the ratio of free amino acid nitrogen: total nitrogen was highest at the end of dormancy and in the early “summer”. Free amino acids accumulated after bud removal in primary needles and especially in stems and roots. Glutamine became relatively more dominant than arginine in the different organs. The observations are consistent with the role of arginine and glutamine for storage and transport of nitrogen in conifers. Because of the low concentrations of amino acid nitrogen in the primary needles, arginine is not considered a major nitrogen reserve in needles of Scots pine seedlings.
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