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Nodule-enhanced expression of a sucrose phosphate synthase gene member (MsSPSA) has a role in carbon and nitrogen metabolism in the nodules of alfalfa (Medicago sativa L.).

ABSTRACT: Sucrose phosphate synthase (SPS) catalyzes the first step in the synthesis of sucrose in photosynthetic tissues. We characterized the expression of three different isoforms of SPS belonging to two different SPS gene families in alfalfa (Medicago sativa L.), a previously identified SPS (MsSPSA) and two novel isoforms belonging to class B (MsSPSB and MsSPSB3). While MsSPSA showed nodule-enhanced expression, both MsSPSB genes exhibited leaf-enhanced expression. Alfalfa leaf and nodule SPS enzymes showed differences in chromatographic and electrophoretic migration and differences in V (max) and allosteric regulation. The root nodules in legume plants are a strong sink for photosynthates with its need for ATP, reducing power and carbon skeletons for dinitrogen fixation and ammonia assimilation. The expression of genes encoding SPS and other key enzymes in sucrose metabolism, sucrose phosphate phosphatase and sucrose synthase, was analyzed in the leaves and nodules of plants inoculated with Sinorhizobium meliloti. Based on the expression pattern of these genes, the properties of the SPS isoforms and the concentration of starch and soluble sugars in nodules induced by a wild type and a nitrogen fixation deficient strain, we propose that SPS has an important role in the control of carbon flux into different metabolic pathways in the symbiotic nodules.

SUBMITTER: Aleman L

PROVIDER: S-EPMC3881968 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature

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Nodule-enhanced expression of a sucrose phosphate synthase gene member (MsSPSA) has a role in carbon and nitrogen metabolism in the nodules of alfalfa (Medicago sativa L.).

Aleman Lorenzo L Ortega Jose Luis JL Martinez-Grimes Martha M Seger Mark M Holguin Francisco Omar FO Uribe Diana J DJ Garcia-Ibilcieta David D Sengupta-Gopalan Champa C

Planta 20091108 2

Sucrose phosphate synthase (SPS) catalyzes the first step in the synthesis of sucrose in photosynthetic tissues. We characterized the expression of three different isoforms of SPS belonging to two different SPS gene families in alfalfa (Medicago sativa L.), a previously identified SPS (MsSPSA) and two novel isoforms belonging to class B (MsSPSB and MsSPSB3). While MsSPSA showed nodule-enhanced expression, both MsSPSB genes exhibited leaf-enhanced expression. Alfalfa leaf and nodule SPS enzymes s ...[more]

PMID: 19898977

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Project description:Screening high nitrogen (N) efficiency crops is crucial to utilize resources rationally and reduce N losses. In this research, the biomass, morphological and N-related parameters of 28 alfalfa (Medicago sativa L.) cultivars were assessed at seedling stage. Then, we selected representative materials to compare the changes in stem-leaf dry weight (SDW), total root length (RL) and plant N accumulation (PNA) during whole period. Lastly, we analyzed the expressions of NRT2 and AMT1 genes of alfalfa cultivars. The correlation coefficients between SDW, PDW, RL, RV, SNA, RNA, and PNA were all in the range of 0.522∼0.996. The coefficient of variations of SDW, PDW, RL, RV, SNA and PNA were all more than 20% under low and medium N levels. Though the comprehensive evaluation and cluster analysis, the comprehensive value of LW6010, Gannong NO.5, Longmu 806, Giant 2, Giant 601, Zhaodong, Crown were greater than 0.5 under low and medium N levels; the comprehensive value of Gannong NO.3, Gannong NO.4, Xinjiangdaye, Xinmu NO.1 were less than 0.5 under low N level, but were greater than 0.5 under medium N level. The comprehensive value of Gannong NO.7 Gannong NO.9, Longmu 801, Gongnong NO.3, Elite, Sadie 10, Giant 551 were greater than 0.5 under low N level, but were lesser than 0.5 under medium N level; and those of Longdong, Gannong NO.8, Gongnong NO.1, Reindee, Goldqueen, Weston, Tourists, Giant 6, Algonquin, Sadie 7 were lesser than 0.5 under low and medium N levels. Four N efficiency types of alfalfa cultivars were classified: (1) Very efficient; (2) Efficient; (3) Anti-efficient; and (4) Inefficient.The SDW, RL and PNA of LW6010 were higher than Longdong in each growth period. The expressions of NRT2 and AMT1 genes were highest for LW6010, and lowest for Longdong. So, N efficiency parameters assessed at seedling stage include: SDW, PDW, RL, RV, SNA and PNA. We developed new classification system of N efficiency types of alfalfa cultivars. It proved its effectiveness on 28 alfalfa in China.

Project description:BackgroundAlfalfa (Medicago sativa L.) is a widely adapted perennial forage crop that has high biomass production potential. Enhanced cellulose content in alfalfa stems would increase the value of the crop as a bioenergy feedstock. We examined if increased expression of sucrose synthase (SUS; EC 2.4.1.13) would increase cellulose in stem cell walls.ResultsAlfalfa plants were transformed with a truncated alfalfa phosphoenolpyruvate carboxylase gene promoter (PEPC7-P4) fused to an alfalfa nodule-enhanced SUS cDNA (MsSUS1) or the β-glucuronidase (GUS) gene. Strong GUS expression was detected in xylem and phloem indicating that the PEPC7-P4 promoter was active in stem vascular tissue. In contrast to expectations, MsSUS1 transcript accumulation was reduced 75-90 % in alfalfa plants containing the PEPC7-P4::MsSUS1 transgene compared to controls. Enzyme assays indicated that SUS activity in stems of selected down-regulated transformants was reduced by greater than 95 % compared to the controls. Although SUS activity was detected in xylem and phloem of control plants by in situ enzyme assays, plants with the PEPC7-P4::MsSUS1 transgene lacked detectable SUS activity in post-elongation stem (PES) internodes and had very low SUS activity in elongating stem (ES) internodes. Loss of SUS protein in PES internodes of down-regulated lines was confirmed by immunoblots. Down-regulation of SUS expression and activity in stem tissue resulted in no obvious phenotype or significant change in cell wall sugar composition. However, alkaline/neutral (A/N) invertase activity increased in SUS down-regulated lines and high levels of acid invertase activity were observed. In situ enzyme assays of stem tissue showed localization of neutral invertase in vascular tissues of ES and PES internodes.ConclusionsThese results suggest that invertases play a primary role in providing glucose for cellulose biosynthesis or compensate for the loss of SUS1 activity in stem vascular tissue.

Dataset Information

Nodule-enhanced expression of a sucrose phosphate synthase gene member (MsSPSA) has a role in carbon and nitrogen metabolism in the nodules of alfalfa (Medicago sativa L.).

Publications

Nodule-enhanced expression of a sucrose phosphate synthase gene member (MsSPSA) has a role in carbon and nitrogen metabolism in the nodules of alfalfa (Medicago sativa L.).

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