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Molecular cloning and characterization of a cytosolic glutamine synthetase gene, a fiber strength-associated gene in cotton

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Abstract

An expressed sequence tag encoding glutamine synthetase (GS) was identified by microarray-based hybridization using fiber mRNAs of allotetraploid Gossypium hirsutum, 7235, a super quality property germplasm line, and TM-1, a genetic standard in G. hirsutum. Northern-blot analysis verified transcript accumulation differences in 8 DPA fibers (including ovules) in the two varieties. The full-length cDNA encoding GS in 7235 was isolated and named GhGS. Sequence analysis revealed that the GhGS was similar to cytosolic GS. Southern-blot analysis showed that tetraploid cotton contained at least one copy of the A sub-genome and the D sub-genome. Genomic GhGS sequences were subsequently isolated from different varieties, TM-1, 7235 and two diploid progenitor cottons, G. herbaceum (A-genome) and G. raimondii (D-genome). Molecular mapping and single-marker analysis revealed that the GhGS was significantly correlated with fiber strength and was mapped to chromosome D7. Additionally, GS activities and total protein of the ovules and the fibers were assayed. The results showed a significantly higher GS activity in 7235 seeds compared to TM-1 seeds at 5 and 8 DPA. Also significant differences were found in total protein content and seed weight at 11 DPA. This suggested that GS promoted the seed-forming process by providing N. On the other hand, in fibers, GS activity and total protein assay indicated a lower total GS activity and longer fiber elongation period in 7235. These results suggest that the respective roles of the GS in ovules and fibers do not completely overlap.

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Abbreviations

GS:

Glutamine synthetase

GS1:

Cytosolic glutamine synthetase

GS2:

Plastidic glutamine synthetase

EST:

Expressed sequence tag

DPA:

Days post-anthesis

References

  • Basra AS, Malik CP (1984) Development of the cotton fiber. Int Rev Cytol 87:65–113

    Article  Google Scholar 

  • Becker TW, Carrayol E, Hirel B (2000) Glutamine synthetase and glutamate dehydrogenase isoforms in maize leaves: localization, relative proportion and their role in ammonium assimilation or nitrogen transport. Planta 211:800–806

    Article  PubMed  CAS  Google Scholar 

  • Brugiere N, Dubois F, Masclaux C, Sangwan RS, Hirel B (2000) Immunolocalization of glutamine synthetase in senescing tobacco (Nicotiana tabacum L.) leaves suggests that ammonia assimilation is progressively shifted to the mesophyll cytosol. Planta 211:519–527

    Article  PubMed  CAS  Google Scholar 

  • Carvalho HG, Lopes-Cardoso IA, Lima LM, Melo PM, Cullimore JV (2003) Nodule-specific modulation of glutamine synthetase in transgenic Medicago truncatula leads to inverse alterations in asparagine synthetase expression. Plant Physiol 133:243–252

    Article  PubMed  CAS  Google Scholar 

  • Cren M, Hirel B (1999) Glulamine synthetase in higher plant: regulation of gene and protein expression from the organ to the cell. Plant Cell Physiol 40:1187–1193

    CAS  Google Scholar 

  • Cui X, Shin H, Song C, Laosinhai W, Amano Y, Brown RM Jr (2001) A putative plant homolog of the yeast beta-1, 3-glucan synthase subunit FKS1 from cotton (Gossypium hirsutum L.) fibers. Planta 213:223–230

    Article  PubMed  CAS  Google Scholar 

  • Dubois F, Brugiere N, Sangwan RS, Hirel B (1996) Localization of tobacco cytosolic glutamine syntheses enzymes and the corresponding transcripts shows organ and cell specific patterns of protein synthesis and gene expression. Plant Mol Biol 31:803–817

    Article  PubMed  CAS  Google Scholar 

  • Edwards JM, Walker EL, Coruzzi GM (1990) Cell-specific expression in transgenic plants reveals nonoverlapping roles for chloroplast and cytosolic glutamine synthetase. Proc Natl Acad Sci USA 87:3459–3463

    Article  PubMed  CAS  Google Scholar 

  • Fuentes SI, Allen DJ, Ortiz-Lopez A, Hernandez G (2001) Overexpression of cytosolic glutamine synthetase increases photosynthesis and growth at low nitrogen concentrations. J Exp Bot 52:1071–1081

    Article  PubMed  CAS  Google Scholar 

  • Fu J, Sampalo R, Gallardo F, Canovas FM, Kirby EG (2003) Assembly of a cytosolic pine glutamine synthetase holoenzyme in leaves of transgenic poplar leads to enhanced vegetative growth in young plants. Plant Cell Environ 26:411–418

    Article  CAS  Google Scholar 

  • Harmer SE, Orford SJ, Timmis JN (2002) Characterisation of six α-expansin genes in Gossypium hirsutum (upland cotton). Mol Genet Genomics 268:1–9

    Article  PubMed  CAS  Google Scholar 

  • Harrison J, Crescenzo MP, Hirel B (2003) Does lowering glutamine synthetase activity in nodules modify nitrogen metabolism and growth of Lotus japonicus L. Plant Physiol 133:253–262

    Article  PubMed  CAS  Google Scholar 

  • Hirel B, Phillipson B, Murchie E, Suzuki A, Kunz C, Ferrario S, Limami A, Chaillou S, Deleens E, Brugiere N, Chaumont-Bonnet M, Foyer C, Morot-Gaudry JF (1997) Manipulating the pathway of ammonia assimilation in transgenic non-legumes and legumes. Z Pflanzeneraehr Bodenkd 160:283–290

    Article  CAS  Google Scholar 

  • Hirel B, Bertin P, Quillere I, Bourdoncle W, Attagnant C, Dellay C, Gouy A, Cadiou S, Retailliau C, Falque M, Gallais A (2001) Towards a better understanding of the genetic and physiological basis for nitrogen use efficiency in maize. Plant Physiol 125:1258–1270

    Article  PubMed  CAS  Google Scholar 

  • Ireland RJ, Lea PJ (1999) The enzymes of glutamine, glutamate, asparagine, and aspartate metabolism. In: Singh BK (ed) Plant amino acids: biochemistry and biotechnology. Marcel Dekker, New York, pp 49–109

    Google Scholar 

  • Ishiyama K, Inoue E, Watanabe-Takahashi A, Obara M, Yamaya T, Takahashi H (2004) Kinetic properties and ammonium-dependent regulation of cytosolic isoenzymes of glutamine synthetase in Arabidopsis. J Biol Chem 279:16598–16605

    Article  PubMed  CAS  Google Scholar 

  • Jiang JX, Zhang TZ (2003) Extraction of total RNA in cotton tissues with CTAB-acidic phenolic method. Cotton Sci 15:166–167

    Google Scholar 

  • Jing ZP, Gallardo F, Pascual MB, Sampalo R, Romero J, Torres de Navarra A, Canovas FM (2004) Improved growth in a field trial of transgenic hybrid poplar overexpressing glutamine synthetase. New Phytol 164:137–145

    Article  CAS  Google Scholar 

  • Kim HJ, Triplett BA (2004) Cotton fiber germin-like protein I. Molecular cloning and gene expression. Planta 218:516–524

    Article  PubMed  CAS  Google Scholar 

  • Kohel RJ, Richmond TR, Lewis CF (1970) Texas marker-1 Description of genetic standard for Gossypium hirsutum L. Crop Sci 10:670–671

    Google Scholar 

  • Kozaki A, Takeba G (1996) Photorespiration protects C3 plants from photooxidation. Nature 384:557–560

    Article  CAS  Google Scholar 

  • Laosinchai W, Cui X, Brown RM Jr (2000) A full cDNA of cotton cellulose synthase has high homology with the Arabidopsis RSW1 gene and cotton CelA1 (Accession No. AF 200453) (PGR 00–002). Plant Physiol 122:291

    Article  Google Scholar 

  • Lea PJ, Robinson SA, Stewart GR (1990) The enzymology and metabolism of glutamine, glutamate and asparagine. In: Miflin BJ, Lea PJ (eds) The biochemistry of plants: intermediary nitrogen metabolism, vol 16. Academic, San Diego, pp 121–157

    Google Scholar 

  • Li XB, Fan XP, Wang XL, Cai L, Yang WC (2005) The cotton ACTIN1 gene is functionally expressed in fibers and participates in fiber elongation. Plant Cell 17:859–875

    Article  PubMed  CAS  Google Scholar 

  • Martin A, Lee J, Kichey T, Gerentes D, Zivy M, Tatout C, Dubois F, Balliau T, Valot B, Davanture M, Terce-Laforgue T, Quillere I, Coque M, Gallais A, Gonzalez-Moro MB, Bethencourt L, Habash DZ, Lea PJ, Charcosset A, Perez P, Murigneux A, Sakakibara H, Edwards KJ, Hirel B (2006) Two cytosolic glutamine synthetase isoforms of maize are specifically involved in the control of grain production. Plant Cell 18:3252–3274

    Article  PubMed  CAS  Google Scholar 

  • Masclaux C, Valadier MH, Brugiere N, Morot-Gaudry JF, Hirel B (2000) Characterization of the sink/source transition in tobacco (Nicotiana tabacum L.) shoots in relation to nitrogen management and leaf senescence. Planta 211:510–518

    Article  PubMed  CAS  Google Scholar 

  • Nicholas KB, Nicholas HB Jr (1997) GeneDoc: a tool for editing and annotating multiple sequence alignments. Distributed by the author (available at http://www.psc.edu/biomed/ genedoc)

  • Ochs G, Schoth G, Trischler M, Kosemund K, Wild A (1999) Complexity and expression of the glutamine synthetase multi-gene family in the amphidiploid crop Brassica napus. Plant Mol Biol 39:395–405

    Article  PubMed  CAS  Google Scholar 

  • Oliveira IC, Brears T, Knight TJ, Clark A, Coruzzi GM (2002) Overexpression of cytosolic glutamine synthetase. Relation to nitrogen, light, and photorespiration. Plant Physiol 129:1170–1180

    Article  PubMed  CAS  Google Scholar 

  • Ortega JL, Temple SJ, Bagga S, Ghoshroy S, Sengupta-Gopalan C (2004) Biochemical and molecular characterization of transgenic Lotus japonicus plants constitutively over-expressing a cytosolic glutamine synthetase gene. Planta 219:807–818

    Article  PubMed  CAS  Google Scholar 

  • Ruan YL, Llewellyn DJ, Furbank RT (2003) Suppression of sucrose synthase gene expression represses cotton fiber cell initiation, elongation, and seed development. Plant Cell 15:952–964

    Article  PubMed  CAS  Google Scholar 

  • Shen XL, Guo WZ, Lu QX, Zhu XF, Yuan YL, Zhang TZ (2007) Genetic mapping of quantitative trait loci for fiber quality and yield trait by RIL approach in Upland cotton. Euphytica 155:371–380

    Article  CAS  Google Scholar 

  • Song XL, Wang K, Guo WZ, Zhang J, Zhang TZ (2005) A comparison of genetic maps constructed from haploid and BC1 mapping populations from the same crossing between Gossypium hirsutum L. × G. barbadense L. Genome 48:378–390

    Article  PubMed  CAS  Google Scholar 

  • Tabuchi M, Sugiyama T, Ishiyama K, Inoue E, Sato T, Takahashi H, Yamaya T (2005) Severe reduction in growth and grain filling of rice mutants lacking OsGS1:1, a cytosolic glutamine synthetase 1:1. Plant J 42:641–655

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–4882

    Article  Google Scholar 

  • Van Ooijen JW, Voorrips RE (2001) JoinMapR Version 3.0: software for the calculation of genetic linkage maps. CPRO-DLO, Wageningen

  • Vincent R, Fraiser V, Chaillou S, Limami MA, Deleens E, Phillipson B, Douat C, Boutin J-P, Hirel B (1997) Overexpression of a soybean gene encoding cytosolic glutamine synthetase in shoots of transgenic Lotus corniculatus L. plants triggers changes in ammonium assimilation and plant development. Planta 201:424–433

    Article  PubMed  CAS  Google Scholar 

  • Wallsgrove RM, Turner JC, Hall NP, Kendall AC, Bright SJW (1987) Barley mutants lacking chloroplast glutamine synthetase—biochemical and genetic analysis. Plant Physiol 83:155–158

    PubMed  CAS  Google Scholar 

  • Wang HY, Yu Y, Chen ZL, Xia GX (2005) Functional characterization of Gossypium hirsutum profilin 1 gene (GhPFN1) in tobacco suspension cells. Planta 222:594–603

    Article  PubMed  CAS  Google Scholar 

  • Wang K, Song XL, Han ZG, Guo WZ, Yu JZ, Sun J, Pan JJ, Kohel RJ, Zhang TZ (2006) Complete assignment of the chromosomes of Gossypium hirsutum L. by translocation and fluorescence in situ hybridization mapping. Theor Appl Genet 113:73–80

    Article  PubMed  CAS  Google Scholar 

  • Wang S, Wang JW, Yu N, Li CH, Luo B, Gou JY, Wang LJ, Chen XY (2004) Control of plant trichome development by a cotton fiber MYB gene. Plant Cell 16:2323–2334

    Article  PubMed  CAS  Google Scholar 

  • Wilkins TA, Jernstedt JA (1999) Molecular genetics of developing cotton fibers. In: Basra AS (ed) Cotton fibers: developmental biology, quality improvement, and textile processing. Binghamton, New York, pp 231–269

    Google Scholar 

  • Wu Y, Machado AC, White RG, Llewellyn DJ, Dennis ES (2006) Expression profiling identifies genes expressed early during lint fibre initiation in cotton. Plant Cell Physiol 47:107–127

    Article  PubMed  CAS  Google Scholar 

  • Zhang TZ, Yuan YL, Guo WZ, Yu J, Kohel RJ (2001) SSR molecular tagging of QTLs for fiber strength in upland cotton. Sci Agric Sin 34:363–366

    CAS  Google Scholar 

  • Zhu YQ, Xu KX, Luo B, Wang JW, Chen XY (2003) An ATP-binding cassette transporter GhWBC1 from elongating cotton fibers. Plant Physiol 133:580–588

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This program was financially supported in part by the State Key Basic Research and Development Plant of China (2002CB111301), the Key Project of Chinese Ministry of Education (10418), and the 111 project (B08025).

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Research Institute, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, China

    He Yajun, Guo Wangzhen, Shen Xinlian & Zhang Tianzhen

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  1. He Yajun
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  2. Guo Wangzhen
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  3. Shen Xinlian
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  4. Zhang Tianzhen
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Correspondence to Zhang Tianzhen.

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Yajun, H., Wangzhen, G., Xinlian, S. et al. Molecular cloning and characterization of a cytosolic glutamine synthetase gene, a fiber strength-associated gene in cotton. Planta 228, 473–483 (2008). https://doi.org/10.1007/s00425-008-0751-z

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