Tyrosinemia type II (Richner-Hanhart syndrome, RHS) is a disorder of autosomal recessive inheritance characterized by keratitis, palmoplantar hyperkeratosis, mental retardation, and elevated blood tyrosine levels. The disease results from deficiency in hepatic tyrosine aminotransferase (TAT). We have previously described one deletion and six different point mutations in four RHS patients. We have now analyzed the TAT genes in a further seven unrelated RHS families from Italy, France, the United Kingdom, and the United States. We have established PCR conditions for the amplification of all twelve TAT exons and have screened the products for mutations by direct sequence analysis or by first performing single-strand conformation polymorphism analysis. We have thus identified the presumably pathological mutations in eight RHS alleles, including two nonsense mutations (R57X, E411X) and four amino acid substitutions (R119W, L201R, R433Q, R433W). Only the R57X mutation, which was found in one Scottish and two Italian families, has been previously reported in another Italian family. Haplotype analysis indicates that this mutation, which involves a CpG dinucleotide hot spot, has a common origin in the three Italian families but arose independently in the Scottish family. Two polymorphisms have also been detected, viz., a protein polymorphism, P15S, and a silent substitution S103S (TCG-->TCA). Expression of R433Q and R433W demonstrate reduced activity of the mutant proteins. In all, twelve different TAT gene mutations have now been identified in tyrosinemia type II.

From laboratory experiments with seedlings and young trees of Norway spruce (Picea abies L. Karst.), a cycling pool of soluble non‐protein N compounds is thought to be indicative of the N‐nutritional status of trees. In order to test whether this assumption can be transferred to mature trees grown in the field, xylem sap and phloem exudate were collected from spruce trees in two remote forest stands: (1) a N‐limited stand (Villingen site), and (2) a stand where trees are sufficiently supplied with N from the soil (Schluchsee site). Trees at these sites were c. 80–100 (Villingen site) and c. 40–60 (Schluchsee site) yr old. In addition to untreated control areas, one entire watershed area at both sites was subjected to (NH2)2SO4 fertilization to the soil. In the xylem sap of the spruce roots at both sites Gln, Asp and Arg were the dominant total soluble non‐protein nitrogen (TSNN) compounds. In the xylem sap of the trunk and the twigs Arg was virtually absent and Gln plus Asp dominated TSNN. On average, TSNN in the xylem sap of trees at the Schluchsee site was 1·5–2‐fold higher than those at Villengen. Highest TSNN contents in the xylem sap were found during growth and development of current year tissues, while the lowest TSNN contents were found in summer. At the Villingen site (NH4)2SO4 fertilization caused an increase in the Gln content in the xylem sap of all tree sections analysed as well as an increase in the Arg content in the xylem sap of the roots. At the Schluchsee site only a small increase in TSNN contents of the xylem was observed, mostly in the xylem sap of the roots. In phloem exudates TSNN contents were much higher in trees on the Schluchsee than on the Villingen site. The seasonal pattern of TSNN in phloem exudates was similar to the seasonal pattern found in the xylem sap. During spring and early summer Gln was the predominant TSNN compound in phloem exudates, but during late summer and autumn Arg became predominant. At the Villingen site (NH4)2SO4 fertilization caused a significant increase in TSNN contents in phloem exudates of twigs and roots, but at Schluchsee an increase in TSNN was found only in phloem exudates of the roots. At both field sites Arg that was not transported to the shoot by xylem transport, was allocated from the leaves to the roots by phloem transport and was cycled within the root system by both xylem and phloem transport. From these results it is calculated that shoot‐to‐root signalling by long‐distance transport of amino compounds can also contribute to the regulation of N‐nutrition of mature spruce trees. Apparently, the internal cycling of individual N compounds within spruce trees differs considerably.

and Rennenberg, H, 1997, Nitrogen distribution in young Norway spmce (Picea abies) trees as affected by pedospheric nitrogen supply. -Physiol. Plant. 101: 764-769.In numerous locations in Europe spmce trees are exposed to high loads of nitrogen. The present study was performed to characterize the distribution of nitrogen compounds under these conditions. For this purpose Norway spruce (Picea abies [L.] Karst.) trees were cultivated under close-to-natural conditions of a forest understory in soil from an apparently nitrogen-limited field site in the Black Forest either with, or without supplementation of nitrogen as ammonium nitrate. After 11 and 20 months, growth, total nitrogen contents of the biomass, and total soluble non-proteinogenic nitrogen compounds (TSNN, i.e. nitrate, ammonium, soluble proteinogenic and nonproteinogenic amino compounds) in needles, xylem sap and phloem exudate were analysed. After 20 months of growth, N-fertilization had slightly enhanced the biomass of current-, but not of 1-year-old shoots. At both harvests, total N-content of 1year-old needles was increased by N-fertilization, whereas current-year needles were not significantly affected. By contrast, TSNN was elevated by N-fertilization in both current-year and 1-year-old needles. The increase in TSNN was mainly attributed to an accumulation of arginine. Xylem sap analysis showed that the increase in TSNN of the needles was a consequence of enhanced nitrogen assimilation of the roots rather than the shoot. Since also TSNN in phloem exudates was enhanced, it appears that Nfertilization elevates the cycling pool of amino compounds in young Norway spruce trees. However, this pool seems to be subject to metabolic interconversion, since mainly glutamine and aspartate are transported in the xylem from the roots to the shoot, but arginine accumulated in the needles and the phloem.

In numerous locations in Europe spruce trees are exposed to high loads of nitrogen. The present study was performed to characterize the distribution of nitrogen compounds under these conditions. For this purpose Norway spruce (Picea abies [L.] Karst.) trees were cultivated under close‐to‐natural conditions of a forest understory in soil from an apparently nitrogen‐limited field site in the Black Forest either with, or without supplementation of nitrogen as ammonium nitrate. After 11 and 20 months, growth, total nitrogen contents of the biomass, and total soluble non‐proteinogenic nitrogen compounds (TSNN, i.e. nitrate, ammonium, soluble proteinogenic and non‐proteinogenic amino compounds) in needles, xylem sap and phloem exudate were analysed. After 20 months of growth, N‐fertilization had slightly enhanced the biomass of current‐, but not of 1‐year‐old shoots. At both harvests, total N‐content of 1‐year‐old needles was increased by N‐fertilization, whereas current‐year needles were not significantly affected. By contrast, TSNN was elevated by N‐fertilization in both current‐year and 1‐year‐old needles. The increase in TSNN was mainly attributed to an accumulation of arginine. Xylem sap analysis showed that the increase in TSNN of the needles was a consequence of enhanced nitrogen assimilation of the roots rather than the shoot. Since also TSNN in phloem exudates was enhanced, it appears that N‐fertilization elevates the cycling pool of amino compounds in young Norway spruce trees. However, this pool seems to be subject to metabolic interconversion, since mainly glutamine and aspartate are transported in the xylem from the roots to the shoot, but arginine accumulated in the needles and the phloem.