6-phosphogluconate Dehydrogenase Gene Research Articles

Fed-batch fermentation of Mucor circinelloides reveals significant improvement in biomass and lipid accumulation through performance evaluation, chemical analysis, and expression profiling

This study aimed to improve the lipid and biomass yields of Mucor circinelloides WJ11 by implementing four different fed-batch fermentation strategies, varied in time and glucose concentration (S1-S4). The S1 fermentation strategy yielded the highest biomass, lipid, and fatty acid content (22 ± 0.7 g/L, 53 ± 1.2 %, and 28 ± 1.6 %) after 120 and 144 h, respectively. The γ-linolenic acid titer of 0.75 ± 0.0 g/L was greatest in S3 after 48 h. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to analyze the transcription of key genes involved in lipid accumulation. The glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and ATP-citrate lyase genes showed increased expression levels. Fourier-transform infrared (FTIR) spectroscopy was used to analyze the biochemical profile during fermentation strategies. Optimal abiotic factors for production efficiency included pH 6.5, 25–26 °C, 15 % (v/v) inoculum, 500 rpm, 20 %-30 % dissolved oxygen, and 120 h fermentation. Glucose co-feeding offers valuable insights to develop effective fermentation strategies for lipid production.

Using Omics Techniques to Analyze the Effects of Gene Mutations and Culture Conditions on the Synthesis of β-Carotene in Pantoea dispersa

β-carotene possesses antioxidant properties and holds significant research value. In our study, we have successfully identified a strain of Pantoea dispersa MSC14 which has the capability to produce β-carotene. By incorporating corn steep liquor powder into culture medium and employing mutagenesis breeding techniques, we have successfully increased the production of β-carotene in the MSC14 strain by 13.97% and 29.22%, respectively. To gain further insights, we conducted genomic and transcriptomics analyses. These analyses revealed a significant mutation in the gndA (6-phosphogluconate dehydrogenase) gene of the mutant strain 14P9, resulting in a 33.74% decrease in 6-phosphogluconate dehydrogenase activity. Using transcriptomics analysis, we investigated the impact of this mutation on β-carotene production and explored the interconnectedness between carbon metabolism, fatty acid metabolism, amino acid metabolism, and β-carotene synthesis. The up-regulation of the trxC (Thioredoxin-2) gene, as observed in both transcriptomics results, prompted us to construct strains that overexpress trxC. This manipulation resulted in a notable 15.89% increase in β-carotene production, highlighting the significant impact of of the trxC gene on the β-carotene content of Pantoea dispersa. In conclusion, our study has successfully identified Pantoea dispersa MSC14 as a proficient producer of β-carotene. Furthermore, we have uncovered two genes implicated in the biosynthesis of β-carotene. These findings enhance our understanding of β-carotene synthesis and provide valuable guidance for carotenoid biosynthesis.

Transcriptomics-based analysis of the causes of sugar receding in Feizixiao litchi (Litchi chinensis Sonn.) pulp.

To investigate the causes of the "sugar receding" in 'Feizixiao' litchi (Litchi chinensis Sonn.) pulp, the main sugar contents and sucrose metabolism enzyme activities were measured in pulp obtained in 2020 and 2021. Pulp RNA obtained in 2020 was extracted at 35, 63, and 69 days after anthesis (DAA) for transcriptome sequencing analysis. The differential expression of genes was verified by real-time PCR for both years. The results showed that after 63 DAA, the contents of soluble sugars and sucrose decreased, and the contents of fructose and glucose increased in both years. The dynamic changes in sucrose metabolism enzyme activities were similar in both years. After 63 DAA, except for acid invertase (AI) in 2021, the activities of other enzymes decreased significantly, and the net activity of sucrose metabolism enzymes showed a strong sucrose cleavage activity. There were 18061, 19575, and 985 differentially expressed genes in 35d vs. 63d, 35d vs. 69d, and 63d vs. 69d, respectively. Ninety-one sugar metabolism genes were screened out, including sucrose synthase (SS), sucrose phosphate synthase (SPS), AI, neutral invertase (NI), hexokinase (HK), glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), phosphofructokinase (PFK), and pyruvate kinase (PK) genes. In 63d vs. 69d, seventy-five percent of sucrose metabolism genes were downregulated, seventy-seven percent of genes in glycolysis (EMP) were upregulated and the PFK genes were significantly upregulated. There was a significant linear correlation between the expression of 15 genes detected by real-time PCR and the transcriptome sequencing results (r2020 = 0.9139, r2021 = 0.8912). These results suggest that the upregulated expression of PFK genes at maturity may enhance PFK activity and promote the degradation of soluble sugar in pulp through the EMP pathway, resulting in decreased soluble sugar and sucrose contents and "sugar receding" in pulp. Moreover, the downregulated expression of sucrose metabolism genes in pulp decreased the activities of these enzymes, but the net activity of these enzymes resulted in cleaved sucrose and replenished levels of reducing sugars, resulting in a stable reducing sugar content.

Glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase genes of winter wheat enhance the cold tolerance of transgenic Arabidopsis

In this study, winter wheat G6PDH (TaG6PDH) and 6PGDH (Ta6PGDH) were investigated. Both their expression and their activity were upregulated under cold stress, suggesting that TaG6PDH and Ta6PGDH positively respond to cold stress in winter wheat. Exogenous abscisic acid (ABA) treatment markedly increased the expression and activity levels of TaG6PDH and Ta6PGDH in winter wheat under cold stress. Subsequently, TaG6PDH-and Ta6PGDH were overexpressed in Arabidopsis, and showed stronger reactive oxygen species (ROS)-scavenging ability and higher survival rate compared with wild-type (WT) plants under cold stress. In addition, we found that TaG6PDH and Ta6PGDH overexpression can promote the oxidative pentose phosphate pathway (OPPP) in the cytoplasm and peroxisomes of Arabidopsis. In summary, Arabidopsis overexpressing TaG6PDH and Ta6PGDH showed improved cold tolerance.

Overexpression of a Cytosolic 6-Phosphogluconate Dehydrogenase Gene Enhances the Resistance of Rice to Nilaparvata lugens.

The pentose phosphate pathway (PPP) plays an important role in plant growth and development, and plant responses to biotic and abiotic stresses. Yet, whether the PPP regulates plant defenses against herbivorous insects remains unclear. In this study, we cloned a rice cytosolic 6-phosphogluconate dehydrogenase gene, Os6PGDH1, which encodes the key enzyme catalyzing the third step in the reaction involving the oxidative phase of the PPP, and explored its role in rice defenses induced by brown planthopper (BPH) Nilaparvata lugens. Levels of Os6PGDH1 transcripts were detected in all five examined tissues, with the highest in outer leaf sheaths and lowest in inner leaf sheaths. Os6PGDH1 expression was strongly induced by mechanical wounding, infestation of gravid BPH females, and jasmonic acid (JA) treatment. Overexpressing Os6PGDH1 (oe6PGDH) decreased the height of rice plants and the mass of the aboveground part of plants, but slightly increased the length of plant roots. In addition, the overexpression of Os6PGDH1 enhanced levels of BPH-induced JA, jasmonoyl-isoleucine (JA-Ile), and H2O2, but decreased BPH-induced levels of ethylene. Bioassays revealed that gravid BPH females preferred to feed and lay eggs on wild-type (WT) plants over oe6PGDH plants; moreover, the hatching rate of BPH eggs raised on oe6PGDH plants and the fecundity of BPH females fed on these were significantly lower than the eggs and the females raised and fed on WT plants. Taken together, these results indicate that Os6PGDH1 plays a pivotal role not only in rice growth but also in the resistance of rice to BPH by modulating JA, ethylene, and H2O2 pathways.

Establishing an innovative carbohydrate metabolic pathway for efficient production of 2-keto-l-gulonic acid in Ketogulonicigenium robustum initiated by intronic promoters

Background2-Keto-l-gulonic acid (2-KGA), the precursor of vitamin C, is currently produced by two-step fermentation. In the second step, l-sorbose is transformed into 2-KGA by the symbiosis system composed of Ketogulonicigenium vulgare and Bacillus megaterium. Due to the different nutrient requirements and the uncertain ratio of the two strains, the symbiosis system significantly limits strain improvement and fermentation optimization.ResultsIn this study, Ketogulonicigenium robustum SPU_B003 was reported for its capability to grow well independently and to produce more 2-KGA than that of K. vulgare in a mono-culture system. The complete genome of K. robustum SPU_B003 was sequenced, and the metabolic characteristics were analyzed. Compared to the four reported K. vulgare genomes, K. robustum SPU_B003 contained more tRNAs, rRNAs, NAD and NADP biosynthetic genes, as well as regulation- and cell signaling-related genes. Moreover, the amino acid biosynthesis pathways were more complete. Two species-specific internal promoters, P1 (orf_01408 promoter) and P2 (orf_02221 promoter), were predicted and validated by detecting their initiation activity. To efficiently produce 2-KGA with decreased CO2 release, an innovative acetyl-CoA biosynthetic pathway (XFP-PTA pathway) was introduced into K. robustum SPU_B003 by expressing heterologous phosphoketolase (xfp) and phosphotransacetylase (pta) initiated by internal promoters. After gene optimization, the recombinant strain K. robustum/pBBR-P1_xfp2502-P2_pta2145 enhanced acetyl-CoA approximately 2.4-fold and increased 2-KGA production by 22.27% compared to the control strain K. robustum/pBBR1MCS-2. Accordingly, the transcriptional level of the 6-phosphogluconate dehydrogenase (pgd) and pyruvate dehydrogenase genes (pdh) decreased by 24.33 ± 6.67 and 8.67 ± 5.51%, respectively. The key genes responsible for 2-KGA biosynthesis, sorbose dehydrogenase gene (sdh) and sorbosone dehydrogenase gene (sndh), were up-regulated to different degrees in the recombinant strain.ConclusionsThe genome-based functional analysis of K. robustum SPU_B003 provided a new understanding of the specific metabolic characteristics. The new XFP-PTA pathway was an efficient route to enhance acetyl-CoA levels and to therefore promote 2-KGA production.

Live attenuated Salmonella enterica serovar Choleraesuis vaccine vector displaying regulated delayed attenuation and regulated delayed antigen synthesis to confer protection against Streptococcus suis in mice

Salmonella enterica serotype Choleraesuis (S. Choleraesuis) and Streptococcus suis (S. suis) are important swine pathogens. Development of a safe and effective attenuated S. Choleraesuis vaccine vector would open a new window to prevent and control pig diseases. To achieve this goal, the mannose and arabinose regulated delayed attenuated systems (RDAS), Δpmi and ΔPcrp::TT araC PBADcrp, were introduced into the wild type S. Choleraesuis strain C78-3. We also introduced ΔrelA::araC PBADlacI TT to achieve regulated delayed antigen synthesis and ΔasdA to constitute a balanced-lethal plasmid system. The safety and immunogenicity of the resulted RDAS S. Choleraesuis strain rSC0011 carrying 6-phosphogluconate dehydrogenase (6-PGD) of S. suis serotype 2 (SS2) were evaluated in vitro and in vivo. Compared with the wild type parent strain C78-3 and vaccine strain C500, a live attenuated S. Choleraesuis vaccine licensed for piglet in China, the results showed that the survival curves of the vaccine strain rSC0011 were similar to those of strains C78-3 and C500 at the early stage of infection, but lower than those of C78-3 and higher than those of C500 at the later stage in both porcine alveolar macrophages and peripheral porcine monocytes. The LD50 of the RDAS strains rSC0011 by oral route in mice was close to that of C500 and 10,000-fold higher than that of C78-3. Similar results were achieved by intraperitoneal (i.p.) route, suggesting that the RDAS strains rSC0011 achieved similar attenuation as C500. However, the RDAS strain rSC0011 was superior to C500 in colonization of Peyer's patches. Adult mice orally immunized with strain rSC0011 carrying a plasmid expression 6-phosphogluconate dehydrogenase (6-PGD) gene from SS2 developed strong immune responses against 6-PGD and Salmonella antigens, and conferred high protection against i.p. challenge with SS2.

Abnormal Dosage Compensation of Reporter Genes Driven by the Drosophila Glass Multiple Reporter (GMR) Enhancer-Promoter

In Drosophila melanogaster the male specific lethal (MSL) complex is required for upregulation of expression of most X-linked genes in males, thereby achieving X chromosome dosage compensation. The MSL complex is highly enriched across most active X-linked genes with a bias towards the 3′ end. Previous studies have shown that gene transcription facilitates MSL complex binding but the type of promoter did not appear to be important. We have made the surprising observation that genes driven by the glass multiple reporter (GMR) enhancer-promoter are not dosage compensated at X-linked sites. The GMR promoter is active in all cells in, and posterior to, the morphogenetic furrow of the developing eye disc. Using phiC31 integrase-mediated targeted integration, we measured expression of lacZ reporter genes driven by either the GMR or armadillo (arm) promoters at each of three X-linked sites. At all sites, the arm-lacZ reporter gene was dosage compensated but GMR-lacZ was not. We have investigated why GMR-driven genes are not dosage compensated. Earlier or constitutive expression of GMR-lacZ did not affect the level of compensation. Neither did proximity to a strong MSL binding site. However, replacement of the hsp70 minimal promoter with a minimal promoter from the X-linked 6-Phosphogluconate dehydrogenase gene did restore partial dosage compensation. Similarly, insertion of binding sites for the GAGA and DREF factors upstream of the GMR promoter led to significantly higher lacZ expression in males than females. GAGA and DREF have been implicated to play a role in dosage compensation. We conclude that the gene promoter can affect MSL complex-mediated upregulation and dosage compensation. Further, it appears that the nature of the basal promoter and the presence of binding sites for specific factors influence the ability of a gene promoter to respond to the MSL complex.

Enhancement of riboflavin production with Bacillus subtilis by expression and site-directed mutagenesis of zwf and gnd gene from Corynebacterium glutamicum

Zwf (code for glucose-6-phosphate dehydrogenase) and gnd (code for 6-phosphogluconate dehydrogenase) genes from Corynebacterium glutamicum were firstly cloned, and then site-directed mutagenesis was successfully introduced to remove allosteric inhibition by intracellular metabolites. Expression of the mutant zwf and gnd in Bacillus subtilis RH33 resulted in significant enhancement of riboflavin productivity, while the specific growth rate decreased slightly and the specific glucose uptake rate was unchanged. Introduction of the mutant zwf and gnd led to approximately 18% and 22% increased riboflavin production, respectively. An improvement by 31% and 39% of the riboflavin production was obtained by co-expression of the mutated dehydrogenases in shaker flask and fed-batch cultivation. Intracellular metabolites analysis indicated that metabolites detected in pentose phosphate pathway or riboflavin synthesis pathway of engineered strains showed higher concentration, while TCA cycle and glycolysis metabolites detected were lower abundance than that of parent strain.

Relationship between glucose catabolism and xanthan production in Xanthomonas oryzae pv. oryzae

Two genes involved in central carbon metabolism were inactivated to modulate intracellular glucose 6-phosphate and to evaluate its effects on xanthan production in wild-type Xanthomonas oryzae pv. oryzae. Upon the inactivation of the phosphogluconate dehydratase gene (edd), intracellular glucose 6-phosphate increased from 0.05 to 1.17 mmol/g (dry cell wt). This was accompanied by increased xanthan production of up to 2.55 g/l (culture medium). In contrast, inactivation of 6-phosphogluconate dehydrogenase gene (gndA) did not influence intracellular glucose 6-phosphate nor xanthan production. The intracellular availability of glucose 6-phosphate is proposed as a rate-limiting factor in xanthan production, and it may be possible to increases production of xanthan by modulating the activities of enzymes in central carbon metabolism.

PGD variants in several wild pig populations of East Asia

PGD (6-phosphogluconate dehydrogenase) gene frequencies were reported in wild pigs, Sus scrofa, of three subspecies, i.e. Japanese wild pig, S.s. leucomystax, Ryukyu wild pig, S.s. riukiuanus, and Formosan wild pig, S.s. taivanus. Five phenotypes (A, AB, B, AC' and C') were observed. The C' variant was found only in the S.s. leucomystax, and may be identical to PGD-C reported by Archibald & McTeir (1988). PGD-A was a common variant in all the species in the genus Sus including wild pig, Sus scrofa, Javan pig, Sus verrucosus, and Bearded pig, Sus barbatus, and predominated in the whole populations examined except some of those of the S.s. riukiuanus. This suggested that the PGDA appeared before the other two alleles (B and C') during the evolution of the genus Sus.

The porcine PGD gene is preferentially lost from chromosome 6 in pig × rodent somatic cell hybrids

The 6-phosphogluconate dehydrogenase (PGD) and glucose phosphate isomerase (GPI) genes are both located on chromosome 6 in the pig (Sus scrofa domestica). Nonetheless, the PGD gene was absent in a total of 17 GPI-positive cell lines found in three independently derived panels of pig x rodent somatic cell hybrids. In most of these cell lines we found an apparently normal pig chromosome 6 at cytogenetic analysis. These results suggest instability of the porcine PGD gene region in interspecies hybrid cells.

Localization of the 6-phosphogluconate dehydrogenase (PGD) gene in horses by in situ hybridization.

Localization of the 6-phosphogluconate dehydrogenase (PGD) gene in horses by in situ hybridization.

Sequence-indexed mutations in maize using the UniformMu transposon-tagging population

BackgroundGene knockouts are a critical resource for functional genomics. In Arabidopsis, comprehensive knockout collections were generated by amplifying and sequencing genomic DNA flanking insertion mutants. These Flanking Sequence Tags (FSTs) map each mutant to a specific locus within the genome. In maize, FSTs have been generated using DNA transposons. Transposable elements can generate unstable insertions that are difficult to analyze for simple knockout phenotypes. Transposons can also generate somatic insertions that fail to segregate in subsequent generations.ResultsTransposon insertion sites from 106 UniformMu FSTs were tested for inheritance by locus-specific PCR. We confirmed 89% of the FSTs to be germinal transposon insertions. We found no evidence for somatic insertions within the 11% of insertion sites that were not confirmed. Instead, this subset of insertion sites had errors in locus-specific primer design due to incomplete or low-quality genomic sequences. The locus-specific PCR assays identified a knockout of a 6-phosphogluconate dehydrogenase gene that co-segregates with a seed mutant phenotype. The mutant phenotype linked to this knockout generates novel hypotheses about the role for the plastid-localized oxidative pentose phosphate pathway during grain-fill.ConclusionWe show that FSTs from the UniformMu population identify stable, germinal insertion sites in maize. Moreover, we show that these sequence-indexed mutations can be readily used for reverse genetic analysis. We conclude from these data that the current collection of 1,882 non-redundant insertion sites from UniformMu provide a genome-wide resource for reverse genetics.

Gender- and age-related changes in 6-phosphogluconate dehydrogenase gene expression in white adipose tissue of rats ( Rattus norvegicus) are not related to serum testosterone concentration

Previously, we have shown that the age-related changes in 6-phosphogluconate dehydrogenase (6PGDH) activity depend on sex, and that oestradiol is playing a crucial role in the regulation of 6PGDH gene expression in rat liver, but not in other tissues [Pankiewicz, A., Sledzinski, T., Nogalska, A., Swierczynski, J., 2003. Tissue specific, sex and age-related differences in the 6-phosphogluconate dehydrogenase gene expression. Int. J. Biochem. Cell Biol. 35, 235–245.]. To complete the knowledge on the influence of sex hormones on 6PGDH activity, experiments have been performed on the effect of testosterone on 6PGDH gene expression in rat white adipose tissue and liver. The results presented here disclosed that in young male rats high serum testosterone concentration was associated with high white adipose tissue 6PGDH activity. After orchidectomy, a decrease in serum testosterone concentration (both in young and old rats) was observed. In contrast, no changes in white adipose tissue and liver 6PGDH activity were found. In female rats, both young and old, serum testosterone concentration was below the limit of detection, whereas 6PGDH activity was much higher in young than in old animals. Moreover, the testosterone administration to 9-month old male rats (which displayed much lower serum testosterone concentration that young animals) resulted in no effect on 6PGDH activity either in WAT or in the liver. In conclusion, the results presented in this paper indicate that testosterone does not play any role in the age- and gender-related differences in 6PGDH gene expression in white adipose tissue.

Variation at isozyme loci in culturedin vitro sugar beet regenerants of gynogenetic origin

Assessment of alcohol dehydrogenase, isocitrate dehydrogenase and 6-phosphogluconate dehydrogenase genes expression has been conducted in culturedin vitro sugar beet vegetative shoots of gynogenetic origin characterized by different level of mixoploidy (x+xx). It was shown that in all 12 originally haploid lines under investigation at alcohol dehydrogenase (ADH1) locus only homozygous phenotypes were observed. At the same time, some of cultured gynogenetic lines demonstrated polymorphism at isocitrate dehydrogenase (IDH1) and 6-phosphogluconate dehydrogenase (PGD1) loci. The conclusion has been done that such a variation in mixoploid tissues of regenerants is associated with spontaneous polyploidization caused by their prolonged culturing underin vitro conditions. Instability of Idh1 and Pgd1 loci, encoding IDH1 and PGD1, together with stability of Adh1 locus, encoding ADH1, testifies that change in ploidy level observed in somatic cells of cultured sugar beet regenerants is unlikely the only factor determining different gene expression duringin vitro culturing.

Genetic and biological diversity among populations of Leishmania major from Central Asia, the Middle East and Africa

Evidence is provided for genetic and biological variation among Leishmania major strains that correlates with their geographical origin. The host–parasite relationship also appears to be specific. Great gerbils, Rhombomys opimus, and fat sand rats, Psammomys obesus, are the main reservoir hosts in Central Asia and the Middle East, respectively. However, the Central Asian parasite failed to infect the Middle Eastern rodent host in the laboratory, and vice versa. A permissively primed intergenic polymorphic (PPIP)-PCR and a single-stranded conformation polymorphism (SSCP)-PCR exposed genetic polymorphism among 30 strains of L. major from different geographical regions. This was verified by subsequent sequencing of DNA from the same strains using four genomic targets: (a) the NADH-dehydrogenase (NADH-DH) gene, (b) the 6-phosphogluconate dehydrogenase (6PGD) gene, (c) the ribosomal internal transcribed spacers, and (d) an anonymous DNA sequence originally amplified with random primers. All the genetic markers indicated that the nine Central Asian strains were a separate homogenous genetic group. The Middle Eastern strains formed another geographical group that displayed heterogeneity corresponding with their different Middle Eastern locations. Molecular markers and host–parasite relationships confirmed that Central Asian and Middle Eastern strains are genetically and biologically distinct sub-populations of L. major. Three African strains of L. major were genetically closer to the Middle Eastern strains, and a representative one did infect fat sand rats, but they had distinct permissively primed inter-genic polymorphic PCR patterns and internal transcribed spacer 2 types.

A novel gnd mutation leading to increased l-lysine production in Corynebacterium glutamicum

Toward more efficient l-lysine production, we have been challenging genome-based strain breeding by the approach of assembling only relevant mutations in a single wild-type background. Following the creation of a new l-lysine producer Corynebacterium glutamicum AHP-3 that carried three useful mutations ( lysC311, hom59, and pyc458) on the relevant downstream pathways, we shifted our target to the pentose phosphate pathway. Comparative genomic analysis for the pathway between a classically derived l-lysine producer and its parental wild-type identified several mutations. Among these mutations, a Ser-361 → Phe mutation in the 6-phosphogluconate dehydrogenase gene ( gnd) was defined as a useful mutation for l-lysine production. Introduction of the gnd mutation into strain AHP-3 by allelic replacement led to approximately 15% increased l-lysine production. Enzymatic analysis revealed that the mutant enzyme was less sensitive than the wild-type enzyme to allosteric inhibition by intracellular metabolites, such as fructose 1,6-bisphosphate, d-glyceraldehyde 3-phosphate, phosphoribosyl pyrophosphate, ATP, and NADPH, which were known to inhibit this enzyme. Isotope-based metabolic flux analysis demonstrated that the gnd mutation resulted in 8% increased carbon flux through the pentose phosphate pathway during l-lysine production. These results indicate that the gnd mutation is responsible for diminished allosteric regulation and contributes to redirection of more carbon to the pentose phosphate pathway that was identified as the primary source for NADPH essential for l-lysine biosynthesis, thereby leading to improved product formation.

Molecular cloning and characterization of rice 6-phosphogluconate dehydrogenase gene that is up-regulated by salt stress.

The pentose phosphate pathway (PPP) is the important metabolism pathway in plant. In the present study, a cDNA encoding one of the key enzymes of PPP, 6-phosphogluconate dehydrogenase(6PGDH), was isolated from rice and designated as Os6PGDH. The Os6PGDH encoding protein is a cytosolic isoenzyme according to the absence of plastid transit peptide at the N-terminus. The full-length cDNA of 1751 bp encodes 480 amino acids and its putative protein sequence is 94%, 84% and 83% identical to maize, spinach and alfalfa 6PGDHs respectively. Comparison of the cloned mRNA sequence with that of the genomic sequence from the Rice Genome Project showed a simple genomic organization devoid of introns in the translated region of the gene. RT-PCR experiments revealed that Os6PGDH expression was high in inflorescence, low in root and embryos but almost absent in leaves. Furthermore, Os6PGDH was up-regulated in the shoots under salt stress. It is suggested that 6PGDH in plant may play an important role in cell division and salt response.

Tissue specific, sex and age—related differences in the 6-phosphogluconate dehydrogenase gene expression

Data presented in this paper indicate that: (1) the age-related changes in 6-phosphogluconate dehydrogenase (6PGDH) activity depend on sex and tissue. No differences in the liver 6PGDH activity between young (1-month-old) males and females were found. In adult males, the activity was the same as in young animals but, in adult females, it reached the value twice as high as in the young. In adipose tissue (both white and brown) and kidney cortex, the enzyme activity decreased with age both in males and females. There were no differences between males and females 6PGDH activity in brain, heart and skeletal muscle. (2) The sex and age-related changes in the liver 6PGDH activity occur predominantly at the level of mRNA cellular concentration. (3) In the liver of ovariectomized rats decrease of 6PGDH activity and mRNA level was observed. Oestradiol administration to ovariectomized rats restored liver 6PGDH activity and liver 6PGDH mRNA levels to that observed in controls. No changes in 6PGDH activity and mRNA levels were found in white adipose tissue (WAT) of ovariectomized adult rats and in ovariectomized rats treated with oestradiol. (4) Oestradiol administration to males caused an increase of liver 6PGDH activity and mRNA levels to values observed in females, but was without an effect on WAT 6PGDH activity and mRNA level. (5) These results suggest that 6PGDH activity in different tissues is not regulated in coordinate fashion and that oestradiol plays an important role in the liver enzyme activity regulation.