Nitrogen (N) is one of the most required mineral elements for plant growth, and potassium (K) plays a vital role in nitrogen metabolism, both elements being widely applied as fertilizers in agricultural production. However, the exact relationship between K and nitrogen use efficiency (NUE) remains unclear. Apple dwarf rootstock seedlings (M9T337) were used to study the impacts of different K levels on plant growth, nitrogen metabolism, and carbon (C) assimilation in water culture experiments for 2 years. The results showed that both deficiency and excess K inhibited the growth and root development of M9T337 seedlings. When the K supply concentration was 0 mM and 12 mM, the biomass of each organ, root-shoot ratio, root activity and NO 3 − ion flow rate decreased significantly, net photosynthetic rate (P n) and photochemical efficiency (F v /F m) being lower. Meanwhile, seedlings treated with 6 mM K + had higher N and C metabolizing enzyme activities and higher nitrate transporter gene expression levels (NRT1.1; NRT2.1). 13 C and 15 N labeling results showed that deficiency and excess K could not only reduce 15 N absorption and 13 C assimilation accumulation of M9T337 seedlings, but also reduced the 15 N distribution ratio in leaves and 13 C distribution ratio in roots. These results suggest that appropriate K supply (6 mM) was optimal as it enhanced photoassimilate transport from leaves to roots and increased NUE by influencing photosynthesis, C and N metabolizing enzyme activities, nitrate assimilation gene activities, and nitrate transport.

The fibroblast growth factor (FGF) receptor complex is a regulator of adult organ homeostasis in addition to its central role in embryonic development and wound healing. FGF receptor 4 (FGFR4) is the sole FGFR receptor kinase that is significantly expressed in mature hepatocytes. Previously, we showed that mice lacking mouse FGFR4 (mR4 ؊/؊ ) exhibited elevated fecal bile acids, bile acid pool size, and expression of liver cholesterol 7␣-hydroxylase (CYP7A1), the rate-limiting enzyme for canonical neutral bile acid synthesis. To prove that hepatocyte FGFR4 was a negative regulator of cholesterol metabolism and bile acid synthesis independent of background, we generated transgenic mice overexpressing a constitutively active human FGFR4 (CahR4) in hepatocytes and crossed them with the FGFR4-deficient mice to generate CahR4/mR4 ؊/؊ mice. In mice expressing active FGFR4 in liver, fecal bile acid excretion was 64%, bile acid pool size was 47%, and Cyp7a1 expression was 10 -30% of wild-type mice. The repressed level of Cyp7a1 expression was resistant to induction by a high cholesterol diet relative to wild-type mice. Expression of CahR4 in mR4 ؊/؊ mouse livers depressed bile acid synthesis below wild-type levels from the elevated levels observed in mR4 ؊/؊ . Levels of phosphorylated cJun N-terminal kinase (JNK), which is part of a pathway implicated in bile acid-mediated repression of synthesis, was 30% of wild-type levels in mR4 ؊/؊ livers, whereas CahR4 livers exhibited an average 2-fold increase. However, cholate still strongly induced phospho-JNK in mR4 ؊/؊ livers. These results confirm that hepatocyte FGFR4 regulates bile acid synthesis by repression of Cyp7a1 expression. Hepatocyte FGFR4 may contribute to the repression of bile acid synthesis through JNK signaling but is not required for activation of JNK signaling by bile acids.

An 8-week growth trial was carried out in a semirecirculation system to investigate the e¡ect of high dietary starch levels on the growth performance, blood chemistry, starch utilization and body composition of gibel carp (Carassius auratus var. gibelio). Five isonitrogenous and isocarloric experimental diets were formulated to contain di¡erent starch levels (24%, 28%, 32%, 36% and 40% respectively). Triplicate groups of ¢sh (24 ¢sh per tank with an average body weight, of 8.5 g) were assigned to each diet. The results showed that dietary carbohydrate levels signi¢cantly a¡ected the growth performance, hepatopancreatic lipid content, pyruvate kinase (PK) activity and whole-body lipid content. Growth performance, body crude lipid and plasma glucose concentrations showed a decreasing trend with an increase in dietary starch from 24% to 40%. Pyruvate kinase activities and hepatopancreatic lipid content showed an increasing trend with the dietary starch increasing from 24% to 32%, and then a decreasing trend with the dietary starch increasing from 32% to 40%. No signi¢cant di¡erence in the hepatopancreatic hexokinase (HK) activity, plasma triglyceride contents, body crude protein, ash and calcium (Ca) and phosphorus (P) contents was observed between di¡erent treatments. In conclusion, higher dietary starch levels (32^40%) signi¢-cantly (Po0.05) decreased the growth of gibel carp in the present study.

The sequence NYKKPKL in the NH 2 terminus of fibroblast growth factor (FGF)-1 has been proposed to affect the long term activities of FGF-1 through its function as a nuclear translocation signal or its role in stabilization of the structure required to sustain binding and activation of the transmembrane receptor kinase. A dynamic molecular model of FGF-1 docked into a duplex of the FGF receptor ectodomain and a hexadecameric heparin chain suggests that the NYKKPKL sequence does not directly interact with heparin or the receptor, but rather the lysine-leucine residues within the sequence indirectly stabilize a major receptor-binding domain. Concurrent with a marked increase in dependence on exogenous heparin for optimal activity, sequential deletion of residues in the NYKKPKL sequence in FGF-1 resulted in a progressive loss of thermal stability, resistance to protease, mitogenic activity, and affinity for the transmembrane receptor. The largest change resulted from deletion of the entire sequence through the lysineleucine residues. In the presence of sufficiently high concentrations of heparin, the deletion mutants exhibited mitogenic activity equal to wild-type FGF-1. The results confirm that a primary role of the NYKKPKL sequence domain is to maintain the structural integrity of FGF-1 required for optimal binding to and activation of the heparan sulfate-transmembrane receptor complex.Sequence subdomains within the NH 2 -terminal sequence of FGF-1 and FGF-2 that are members of the heparin-binding fibroblast growth factor (FGF) 1 family have been proposed to signal the nuclear accumulation that occurs during sustained exposure of cells to the ligands (1). FGF-(28 -154), a form in which the nuclear translocation sequence 21 NYKKPKL 27 had been deleted, bound to the tyrosine kinase receptor and activated kinase activity and immediate early gene expression but failed to elicit a mitogenic response in cultured fibroblasts equal to that of a wild-type isoform, FGF-(21-154), containing the sequence (1). Subsequent results revealed that FGF-(28 -154) and several mutants with substitutions in the 21 NYKKPKL 27 sequence of FGF-(21-154) promoted levels of mitogenic activity much more similar to wild-type FGF-(21-154) when they were purified by cation exchange rather than reversed phase chromatography (2). The results suggested that deletion of the 21 NYKKPKL 27 sequence may affect the structure and stability of FGF-1 that affects its interaction with the transmembrane receptor rather than, or in addition to, having a role in the intracellular trafficking of the ligand.In this report, we show that a model of FGF-1 that results after performance of molecular dynamics calculations on the ligand docked into a complex of heparin and the FGF receptor kinase ectodomain (hereafter referred to as FGFR1␤) predicts that the nuclear translocation sequence 21 NYKKPKL 27 , particularly Lys 26 and Leu 27 , play a role in stability and structure of a key domain of FGF-1 that directly interacts with the receptor. Sequential deletion of res...

The human hepatoblastoma cell line, HepG2, exhibits an array of stable properties in culture that have made it a popular cell culture model for studies on regulation of liver-specific gene expression and properties of hepatoma cells. In contrast to other hepatoma cell lines, HepG2 cells overexpress a characteristic detergent-extractable, wheat germ lectin-binding protein with apparent molecular mass of 130 kDa. Using an antibody to screen a phage expression library of HepG2 complementary DNA (cDNA), we identified and cloned a 4734 base pair cDNA which codes for a 130-kDa leucine-rich protein (lrp 130) when expressed in transfected cells. The deduced sequence of lrp130 exhibits sequences weakly homologous to the consensus sequence for the ATP binding site in ATP-dependent kinases and the protein kinase C phosphorylation site of the epidermal growth factor receptor. Consistent with the higher levels of expression of lrp130 antigen, Northern hybridization analysis indicated that HepG2 cells express high levels of the major 4.8 kilobase lrp130 mRNA relative to other hepatoma cells. Although currently of unknown function, lrp130 may be of utility as a marker for liver cell lineages represented by the HepG2 cell line.

Calcarisporium cordycipiticola is the pathogen in the white mildew disease of Cordyceps militaris, one of the popular mushrooms. This disease frequently occurs and there is no effective method for disease prevention and control. In the present study, C. militaris is found to be the only host of C. cordycipiticola, indicating strict host specificity. The infection process was monitored by fluorescent labeling and scanning and transmission electron microscopes. C. cordycipiticola can invade into the gaps among hyphae of the fruiting bodies of the host and fill them gradually. It can degrade the hyphae of the host by both direct contact and noncontact. The parasitism is initially biotrophic, and then necrotrophic as mycoparasitic interaction progresses. The approximate chromosome-level genome assembly of C. cordycipiticola yielded an N50 length of 5.45 Mbp and a total size of 34.51 Mbp, encoding 10,443 proteins. Phylogenomic analysis revealed that C. cordycipiticola is phylogenetically close to its specific host, C. militaris. A comparative genomic analysis showed that the number of CAZymes of C. cordycipiticola was much less than in other mycoparasites, which might be attributed to its host specificity. Secondary metabolite cluster analysis disclosed the great biosynthetic capabilities and potential mycotoxin production capability. This study provides insights into the potential pathogenesis and interaction between mycoparasite and its host.

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Albinism has been used for new variety screening in some edible mushrooms and the underlying mechanisms are fascinating. Albino fruiting body of Cordyceps militaris, a well-known edible fungus and model organism for Cordyceps, has the potential to be a nutraceutical or functional food due to its high content of metabolites and antioxidant activities. In this study, a spontaneous albino mutant strain (505) of C. militaris was obtained. In comparison to its normal sibling strain (498), the albino strain stably remained white in response to light and had significantly decreased conidia and carotenoid production but accumulated more cordycepin. Transcriptome analysis of both strains revealed that all the seven photoreceptors were expressed similarly in response to light. However, many more genes in the albino strain were differentially expressed in response to light than its sibling strain. The significantly enriched pathways in 498L vs. 505L were mainly associated with replication and repair. Some secondary metabolite backbone genes including those encoding DMAT, two NRPS-like proteins, three NPRS, and lanosterol synthase were differentially expressed in the albino when compared with that of the normal strains. Transcriptome and real-time quantitative PCR analyses indicated that some cytochrome P450s and methyltransferases might be related to the phenotypic differences observed between the two strains. This study compared the genome-wide transcriptional responses to light irradiation in a spontaneous albino mutant and its normal sibling strain of an edible fungus, and these findings potentially pave the way for further investigation of the pigment biosynthetic pathway.