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PROPERTIES OF RuBP carboxylase/OXYGENASE IN A MUTANT TOBACCO

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Chinese Science Bulletin 1987年第20期32卷 1420-1422页

作者：李立人 S.D.KUNG Shanghai Institute of Plant Physiology Academia Sinica University of Maryland USA

The aurea mutant （Su/su）of tobacco is a nuclear mutation found in Nicotiana tobacum Var. John Williams Broadleaf （Su/su）. Under greenhouse conditions and atmosphere CO2 levels, the yellow mutant tobacco （Su/su） grows much slower than the dark green wild type （su/su）. Zelitch reported that the yellow mutant has a higher photorespiratory rate and a lower net rate of photosynthesis, compared with the wild type. RuBP carboxylase/oxygenase is a key enzyme in photosynthesis and photorespiration, and the small subunit of the enzyme is nuclear-encoded. This study was designed to examine if the nuclear gene mutation influenced the structure and properties of the enzyme.

关键词： tobacco mutant Nicotiana photosynthesis carboxylase greenhouse yellow oxygenase slower subunit

Concomitant Increases of the Developing Seed Phosph<i>oenol</i>pyruvate carboxylase Ac-tivity and the Seed Protein Content of Field-Grown Wheat with Nitrogen Supply

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Agricultural Sciences 2014年第14期5卷 1558-1565页

作者： Naoki Yamamoto Toshio Sugimoto Takehiro Masumura Plant Nutrition Laboratory Department of Biological and Environmental Science Faculty of Agriculture Graduate School of Agricultural Science Kobe University Kobe Japan Laboratory of Genetic Engineering Graduate School of Life and Environmental Sciences Kyoto Prefectural University Kyoto Japan Kyoto Prefectural Institute of Agricultural Biotechnology Kyoto Japan

Wheat seed storage protein is of great importance for human food. To increase the contents of storage proteins effectively, nitrogen fertilizer at flowering stages is commonly applied. In our previous study, rice phosphoenolpyruvate carboxylase (PEPCase) activity in developing seeds was observed in response to nitrogen application at a flowering stage and was positively correlated to the response of the protein content in seeds of six cultivars. This observation might indicate that the seeds have a biological system for accepting nitrogen in seeds by using PEPCase. To test whether this physiological event occurs in wheat, we examined the PEPCase activity and protein content in field-grown wheat seeds under different nitrogen supply conditions. With only basal dressing, seeds showed lower PEPCase activity and protein content (both 0.90-fold) compared to seeds without basal fertilizer. With ammonium sulfate application at 8.3 and 25 g/m2 at a flowering stage, seeds showed higher PEPCase activity (1.08- and 1.17-fold, respectively) and protein content (1.15- and 1.42-fold, respectively), depending on the nitrogen level. We investigated the relationship between PEPCase activity and protein content in the seeds among four conditions. The effect of the nitrogen supply on PEPCase activity during grain-filling stages was validated by the results of a hydroponic culture experiment. Together the results demonstrate that our hypothesis seems to apply to field-grown wheat.

关键词： Nitrogen Application Phosphoenolpyruvate carboxylase Seed Storage Protein Wheat

Comparison of Five Endogenous Reference Genes for Specific PCR Detection and Quantification of Rice

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Rice science 2019年第4期26卷 248-256,I0006,I0007页

作者： ZHANG Xiujie JIN Wujun XU Wentao LI Xiaying SHANG Ying LI Sha OUYANG Hongsheng College of Animal Sciences Jilin University Changchun 130062 China Development Center of Science and Technology Ministry of Agriculture and Rural Affairs Beijing 100176 China Biotechnology Research Institute Chinese Academy of Agricultural Sciences Beijing 100081China College of Food Science and Nutritional Engineering China Agricultural University Beijing 100083China

Endogenous reference genes (ERGs) provide vital information regarding genetically modified organisms (GMOs). The successful detection of ERGs can identity GMOs and the source of genes, verify stability and reliability of the detection system, and calculate the level of genetically modified (GM) ingredients in mixtures. The reported ERGs in rice include sucrose-phosphate synthase (SPS), phospholipase D (PLD), RBE4 and rice root-specific GOS9 genes. Based on the characteristics of ERGs, a new ERG gene, phosphoenolpyruvate carboxylase (PEPC), was selected, and further compared with the four existing genes. A total of 18 rice varieties and 29 non-rice crops were used to verify the interspecies specificity, intraspecies consistency, sensitivity, stability and reliability of these five ERGs using qualitative and quantitative PCR. Qualitative detection indicated that SPS and PEPC displayed sufficient specificity, and the detection sensitivity was 0.05% and 0.005%, respectively. Although the specificity of both RBE4 and GOS9 were adequate, the amplicons were small and easily confused with primer dimers. Non-specific amplification of the PLD gene was present in maize and potato. Real-time quantitative PCR detection indicated that PLD, SPS and PEPC displayed good specificity, with R2 of the standard curve greater than 0.98, while the amplification efficiency ranged between 90% and 110%. Both the detection sensitivities of PLD and PEPC were five copies and that of SPS was ten copies. RBE4 showed typical amplification in maize, beet and Arabidopsis, while GOS9 was found in maize, tobacco and oats. PEPC exhibited excellent detection sensitivity and species specificity, which made it a potentially useful application in GM-rice supervision and administration. Additionally, SPS and PLD are also suitable for GM-rice detection. This study effectively established a foundation for GMO detection, which not only provides vital technical support for GMO identification, but also is of grea

关键词： endogenous reference gene rice genetically modified crop phosphoenolpyruvate carboxylase gene sucrose-phosphate synthase gene phospholipase D gene starch branching enzyme 4 gene rice root-specific GOS9 gene

Overexpression of PtPEPCK1 gene promotes nitrogen metabolism in poplar

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Journal of Forestry Research 2019年第6期30卷 2289-2303页

作者： Lina Wang Miao He Song Chen Kean Wang Donghai Cui Xin Huang Lijie Liu State Key Laboratory of Tree Genetics and Breeding Northeast Forestry University51 Hexing RoadHarbin 150040People’s Republic of China College of Life Sciences and Agriculture and Forestry Qiqihar UniversityQiqihar 161006People’s Republic of China

To understand the function of phosphoenolpyruvate carboxylase kinase,we introduced PtPEPCK1 gene under the control of 35S promoter into 84K poplar(Populus alba×***).PtPEPCK1 gene is well-known for its role in ***,our data confi rmed that it has signifi cant eff ects on amino acid biosynthesis and nitrogen *** and fl uorescence microscopy indicate that PtPEPCK1 is specifi cally expressed in the cytoplasm of the spongy and palisade *** of PtPEPCK1 was characterized through transcriptomics and *** metabolites concentration of the ornithine cycle and its precursors also increased,of which N-acetylornithine was up-regulated almost 50-fold and ornithine *** were accompanied by a massive increase in levels of several amino ***,overexpression of PtPEPCK1 increases amino acid levels with urea cycle disorder.

关键词： Amino acid metabolism Metabolome Nitrogen metabolism Phosphoenolpyruvate carboxylase kinase Transcriptome Urea cycle

Physiological characteristics of the primitive CO_(2) concentrating mechanism in PEPC transgenic rice

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Science China(Life Sciences) 2003年第4期46卷 438-446页

作者：焦德茂匡廷云李霞戈巧英黄雪清郝乃斌白克智 Institute of Agrobiological Geneticsand Physiology Jiangsu Academyof Agriculturalsciences Institute of Botany Chinese Academy of Sciences

The relationship between carbon assimilation and high-level expression of the maize PEPC in PEPC transgenic rice was studied by comparison to that in the untransformed rice, japonica kitaake. Stomatal conductance and photosynthetic rates in PEPC transgenic rice were higher than those of untransformed rice, but the increase of stomatal conductance had no statistical correlation with that of photosynthetic rate. Under high levels of light intensity, the protein contents of PEPC and CA were increased significantly. Therefore the photosynthetic capacity was increased greatly (50%) with atmospheric CO2 supply. While CO2 release in leaf was reduced and the compensation point was lowered correspondingly under CO2 free conditions. Treatment of the rice with the PEPC-specific inhibitor DCDP showed that overexpression of PEPC and enhancement of carbon assimilation were related to the stability of Fv/Fm. Labeling with 14CO2 for 20 s showed more 14C was distributed to C4 primary photosynthate asperate in PEPC transgenic rice, suggesting that there exists a limiting C4 photosynthetic mechanism in leaves. These results suggest that the primitive CO2 concentrating mechanism found in rice could be reproduced through metabolic engineering, and shed light on the physiological basis for transgenic breeding with high photosynthetic efficiency.

关键词： PEPC transgenic rice, phosphoenopyruvate carboxylase (PEPC), carbonic anhydrase (CA), stomatal conductance, CO2 concentrating mechanism.

The characteristics of CO_2 assimilation of photosynthesis and chlorophyll fluorescence in transgenic PEPC rice

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Chinese Science Bulletin 2001年第13期46卷 1080-1084页

作者： JIAO Demao, LI Xia, HUANG Xueqing, CHI Wei, KUANG Tingyun & Ku SB Maurice1. Institute of Agrobiological Genetics and Physiology, Jiangsu Academy of Agricultural Sciences, Nangjing 210014, China 2. Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, China 3. Botany Department, Washington State University, Pullman, WA 99164-4236, USA Institute of Agrobiological Genetics and Physiology Jiangsu Academy of Agricultural Sciences Nangjing China Institute of Botany The Chinese Academy of Sciences Beijing China Botany Department Washington State University Pullman USA

With PEPC, PPDK, NADP-ME and PEPC+ PPDK transgenic and untransformed rice (Orysa sativa L.), the activities of related C4 photosynthesis enzymes, the chlorophyll fluorescence parameters, CO2 exchange and other physiological indexes were compared, in which the physiological characteristics of PEPC transgenic rice were mainly studied. The results were as follows: (i) The activities of PEPC in PEPC transgenic rice were 20-fold higher than those in untransformed rice; the light-saturation photosynthetic rates and the carboxylation efficiency of PEPC transgenic rice were increased by 55% and 50% more than those of untransformed rice, respectively, while the CO2 compensation point decreased by 27%. (ii) The PS II photochemical efficiency (Fv/Fm) and photochemical quenching (qp) of transgenic PEPC rice decreased less in comparison with those of untransformed rice after the treatment with high light intensity (3 h) or methyl viologen (MV), a photooxidative reagent, which demonstrated that the tolerance of PEPC

关键词： transgenic rice photosynthetic characteristics photoin-hibition of photosynthesis chlorophyll fluorescence phosphoenopy-ruvate carboxylase (PEPC).

Molecular Adaptation of Peanut Metabolic Pathways to Wide Variations of Mineral Ion Composition and Concentration

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American Journal of Plant Sciences 2012年第1期3卷 33-50页

作者： Godson O. Osuji Tassine K. Brown Sanique M. South Justin C. Duncan Dwiesha Johnson Shanique Hyllam 不详

Plant evolution, nutritional genomics, and mineral nutrition have been well documented but no studies have focused on the molecular adaptation of crop metabolism to wide variations of mineral ion composition and concentration. Diversification of peanut species from primary centers of domestication in South America depended on metabolic adaptation to the mineral ion conditions of the newer habitats. Understanding the diversification molecular biology of peanut metabolic pathways will permit the synthesis of the best mineral ion combinations for doubling CO2 assimilation. Valencia and Virginia cultivars belong to different subspecies of the tetraploid Arachis hypogaea. They were planted in the absence and presence of up to 99 mM (equivalent to 166 moles per hectare) of different mineral ions. Molecular properties of the primary metabolic pathways were studied by Northern analyses using Valencia GDH-synthesized RNAs as probes for Virginia mRNA and GDH-synthesized RNAs. Messenger RNAs are silenced by homologous RNAs synthesized by GDH. Peanut cellulose was analyzed by gravimetry;and fatty acids by HPLC. Complementary DNA probes made from Valencia GDH-synthesized RNAs hybridized perfectly to Virginia mRNAs and GDH-synthesized RNAs. Wide variations in mineral ion compositions and concentrations induced the GDHs of Valencia and Virginia to synthesize RNAs that differentially down-regulated the mRNAs encoding phosphate translocator, granule-bound starch synthase, phosphoglucomutase, glucosyltransferase, acetyl CoA carboxylase, nitrate reductase, and NADH-glutamate synthase so that the percent weights of oil (41.53 ± 8.75) and cellulose (30.29 ± 3.12) were similar in the control and mineral-treated peanuts. Therefore, RNA sequences that defined the molecular adaptation of mRNAs encoding the enzymes of primary metabolism were the same in the varietal types of A. hypogaea, in agreement with genetic data suggesting that tetraploid Arachis evolved relatively recently from the wi

关键词： Glutamate Dehydrogenase-Synthesized RNA mRNA Silencing Phosphate Translocator Acetyl CoA carboxylase Cellulosic Biomass Northern Analysis HPLC Fatty Acids

Anaplerosis in cancer: Another step beyond the warburg effect

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American Journal of Molecular Biology 2012年第4期2卷 291-303页

作者： Estefania Ochoa-Ruiz Rodrigo Diaz-Ruiz Facultad de Medicina Programa de Posgrado en Ciencias Médicas Odontológicas y de la Salud Universidad Nacional Autónoma de México Mexico City Mexico Unidad de Genética de la Nutrición Instituto de Investigaciones Biomédicas Universidad Nacional Autónoma de México Mexico City Mexico

Biosynthesis is up-regulated in tumors and thus the demand for anabolic intermediates is increased. The metabolic routes providing the building blocks for macromolecules are thus a very attractive target as they are not normally up-regulated in a normal quiescent cell. Some routes for glycolysis-derived intermediates production have been identified, but these do not constitute the whole pool of biosynthetic molecules in the cell, as many of these derive from mitochondria in the Krebs cycle. Indeed, this metabolic pathway is considered a “biosynthetic hub” from which anabolism is fed. If a metabolite efflux is indeed occurring, anaplerotic reactions must keep a steady supply of substrates. In spite of this obvious relevance of anaplerosis, it has been poorly characterized in the malignant cell context. Glutaminolysis and and pyruvate carboxylation are two pathways that function in an anaplerotic fashion. In spite of the increasing evidence implicating these two processes in cancer metabolism their role as intermediate providers is overlooked. In this review we analyze the implications of an active anaplerosis in cancer and we discuss experimental evidence showing the relevance of these metabolic routes in tumor physiology.

关键词： Anaplerosis Biosynthesis Cancer Glutaminase Krebs Cycle Metabolism Mitochondria Pyruvate carboxylase Warburg Effect

Atypical presentations of propionic acidemia

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Health 2012年第9期4卷 629-633页

作者： Ali M. Al-Asmari AbdulKarim S. Al-Makadma Department of Pediatrics Children’s Hospital King Fahad Medical City KSA

One of the most common recessively inherited organic acidemias is the Propionic Acidosis (PA) which results from Propionyle-CoA carboxylase (PCC) enzyme deficiency that is necessary for the catabolism of the branched chain Amino Acids and other metabolites. Classically this disease presented with high anion gap metabolic acidosis with its clinical consequences. We report 4 patients who presented to our facility with sepsis like picture and no metabolic acidosis. All of them were found to have high ammonia level. Diagnosis was confirmed by tandem MS/MS and urine Gas Chromatography/ Mass Spectrometry (GC/MS). All of them were treated supportively and by supplementation of adequate calories and PA formula. The different presentations may be very well attributed to the PCC molecular defects heterogeneity. Mutations in both genes PCCA and PCCB can cause PA with more frequent heterogeneity of PCCA gene. In spite of the fact that PCCB gene is responsible for the most of the oriental cases, our first patient condition was attributed to PCCA gene with a rare mutation which was not described in the literatures.

关键词： Acidemia Atypical Presentation Hyperammonemia Organic Propionic Propionyle-CoA carboxylase