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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.)

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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.

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Acknowledgments

This work was supported by the National Institutes of Health (Grant numbers GMO-8136, GMO-61222, GMO-7667), National Science Foundation (Grant numbers NSF-DBI0619747, NSF-0331446), and by the Agricultural Experiment Station at New Mexico State University.

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Authors and Affiliations

  1. Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, NM, 88003, USA

    Jose Luis Ortega, Francisco Omar Holguin, Diana J. Uribe & Champa Sengupta-Gopalan

  2. Graduate Program in Molecular Biology, New Mexico State University, Las Cruces, NM, 88003, USA

    Lorenzo Aleman, Martha Martinez-Grimes, Mark Seger, David Garcia-Ibilcieta & Champa Sengupta-Gopalan

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  1. Lorenzo Aleman
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  2. Jose Luis Ortega
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  8. Champa Sengupta-Gopalan
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Correspondence to Champa Sengupta-Gopalan.

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J. L. Ortega is the joint first author.

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425_2009_1043_MOESM1_ESM.eps

Fig. S1 Phenotype of alfalfa plants grown in the greenhouse in Magenta boxes 28d after inoculation with either Sinorhizobium meliloti 2011 (WT) or S. meliloti 1491 (Fix−) strains. Five boxes of plants inoculated with the WT strain and 9 boxes of plants inoculated with the Fix- strain are shown. Supplementary material 1 (EPS 7945 kb)

425_2009_1043_MOESM2_ESM.eps

Fig. S2 Analysis of SPS protein in the leaves and nodules of different legumes. Protein (50 μg) from leaf and nodule extracts from alfalfa (Ms), Phaseolus vulgaris (Pv), Pisum sativum (Ps) and Lotus japonicus (Lj) was separated by SDS PAGE followed by Western blot analysis using anti-SPS antibodies. The SPS antibody was immunoselected against SPS immobilized to a nylon membrane. The migration of the molecular weight standards is as shown. Supplementary material 2 (EPS 2300 kb)

425_2009_1043_MOESM3_ESM.eps

Fig. S3 Phylogenetic analysis of the SPS predicted protein sequences from alfalfa, Medicago truncatula, Arabidopsis and tobacco. Protein sequences from alfalfa MsSPSA, MsSPSB, MsSPSB3 (NCBI accession numbers AAR31210, ABW89596 and ACN89831), and M. truncatula SPSA and SPSB forms (M. truncatula sequencing resources gene numbers AC144657_7, AC157648_2_3, and CU424494_10), were clustered together with the A, B and C isoforms from Arabidopsis and tobacco (AtSPSA1, AtSPSA2, AtSPSB, AtSPSC, NtSPSA, NtSPSB and NtSPSC; NCBI accession numbers AAK09427, ABW89596, NP_197528, NP_196672, NP_171984, NP_192750, AAF06792, ABA64521 and ABA64520, respectively). The tree was generated using Geneious Pro 4.7 (Biomatters) under default parameters. The numbers on each node are the bootstrap proportion values for 1000 replicates. Bar represents the Jukes-Cantor genetic distance. Supplementary material 3 (EPS 397 kb)

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Aleman, L., Ortega, J.L., Martinez-Grimes, M. et al. 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.). Planta 231, 233–244 (2010). https://doi.org/10.1007/s00425-009-1043-y

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