Isolation and characterisation of cytosolic glutamine synthetase (GSe) genes and association with grain protein content in durum wheat
Agata Gadaleta A B , Domenica Nigro A , Ilaria Marcotuli A , Angelica Giancaspro A , Stefania L. Giove A and Antonio Blanco A
+ Author Affiliations
- Author Affiliations
A Department of Soil, Plant and Food Sciences, Section of Genetics and Plant Breeding, University of Bari ‘Aldo Moro’, Via G. Amendola 165/A – 70126 Bari, Italy.
B Corresponding author. Email: agata.gadaleta@agr.uniba.it
Crop and Pasture Science 65(1) 38-45 https://doi.org/10.1071/CP13140
Submitted: 24 April 2013 Accepted: 23 August 2013 Published: 17 December 2013
Abstract
Glutamine synthetase (GS) enzyme (EC 6.3.1.2) plays a central role in assimilating ammonia produced in the leaf from metabolic processes, spanning from assimilation to transamination reactions and catabolic processes. GS is located in both cytoplasm (GS1, GSe and GSr) and plastids (GS2) of plant cells. Glutamine and glutamate, produced by the concerted action of GS and glutamate synthase, are then transported from the leaf to the developing sinks or grain in wheat. The goal of the present study was to characterise GSe genes and to assess the linkage with grain protein content, an important quantitative trait controlled by multiple genes. Here, we report the isolation of the complete cytosolic GS gene sequences of the durum wheat cvv. ‘Ciccio’ and ‘Svevo’ (characterised by low and high protein content, respectively). GSe-A4 located on 4A chromosome comprises 12 exons separated by 11 introns, while the GSe-B4 gene on 4B chromosome comprises 11 exons separated by 10 introns. Quantitative real-time PCR indicated different expression levels of GSe-A4 and GSe-B4 genes in the two wheat cvv. ‘Ciccio’ and ‘Svevo’. The two GSe genes were significantly associated to quantitative trait loci for grain protein content.
Additional keywords: candidate gene, nitrogen metabolism, sequencing, wheat.
References
Bernard S, Habash DZ (2009) The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling. New Phytologist 182, 608–620.| The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXmtVWqtr4%3D&md5=cfc44568680d09ab517a8180cf45a5d8CAS | 19422547PubMed |
Bernard SM, Blom Møller AL, Dionisio G, Kichey T, Jahn TP, Dubois F, Baudo M, Lopes MS, Terce’-Laforgue T, Foyer CH, Parry MAJ, Forde BG, Araus JL, Hirel B, Schjoerring JK, Habash DZ (2008) Gene expression, cellular localisation and function of glutamine synthetase isozymes in wheat (Triticum aestivum L.). Plant Molecular Biology 67, 89–105.
| Gene expression, cellular localisation and function of glutamine synthetase isozymes in wheat (Triticum aestivum L.).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXks1Krt7Y%3D&md5=c150d820af2724b5060325492decafedCAS | 18288574PubMed |
Blanco A, Mangini G, Giancaspro A, Giove S, Colasuonno P, Simeone R, Signorile A, De Vita P, Mastrangelo AM, Cattivelli L, Gadaleta A (2012) Relationships between grain protein content and grain yield components through quantitative trait locus analyses in a recombinant inbred line population derived from two elite durum wheat cultivars. Molecular Breeding 30, 79–92.
| Relationships between grain protein content and grain yield components through quantitative trait locus analyses in a recombinant inbred line population derived from two elite durum wheat cultivars.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XnvFOlurk%3D&md5=ef139faa2e30dc99926e83a1cebddb2bCAS |
Canovas F, Avila C, Canton FR, Canas R, de la Torre F (2007) Ammonium assimilation and amino acid metabolism in conifers. Journal of Experimental Botany 58, 2307–2318.
| Ammonium assimilation and amino acid metabolism in conifers.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXos1yktrc%3D&md5=7c0595871c34cce090c5cd9b5e648d2cCAS | 17490997PubMed |
Cavanagh CR, Chao S, Wang S, Huang BE, Stephen S, Kiani S, et al (2013) Genome-wide comparative diversity uncovers multiple targets of selection for improvement in hexaploid wheat landraces and cultivars. Proceedings of the National Academy of Sciences of the United States of America 110, 8057–8062.
| Genome-wide comparative diversity uncovers multiple targets of selection for improvement in hexaploid wheat landraces and cultivars.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhtV2ksL%2FJ&md5=2c5723a0e4d21c78550e63c9066fe8f5CAS | 23630259PubMed |
Chen Q, Silflow CD (1996) Isolation and characterization of glutamine synthetase genes in Chlamydomonas reinhardtii. Plant Physiology 112, 987–996.
| Isolation and characterization of glutamine synthetase genes in Chlamydomonas reinhardtii.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XntVWrtrk%3D&md5=e03a55ca6421ca557505f5bb4561f748CAS | 8938407PubMed |
Endo TR, Gill BS (1996) The deletion stocks of common wheat. The Journal of Heredity 87, 295–307.
| The deletion stocks of common wheat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28Xls1Clt7s%3D&md5=a317b3864c3e5b59ef21a3ae57466056CAS |
Fontaine J, Ravel C, Pageau K, Heumez E, Dubois F, Hirel B, Le Gouis J (2009) A quantitative genetic study for elucidating the contribution of glutamine synthetase, glutamate dehydrogenase and other nitrogen-related physiological traits to the agronomic performance of common wheat. Theoretical and Applied Genetics 119, 645–662.
| A quantitative genetic study for elucidating the contribution of glutamine synthetase, glutamate dehydrogenase and other nitrogen-related physiological traits to the agronomic performance of common wheat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXpsVeisr8%3D&md5=521c3e3cebfcee5ca222f9e162780a19CAS | 19513687PubMed |
Gadaleta A, Giancaspro A, Giove SL, Zacheo S, Mangini G, Simeone R, Signorile A, Blanco A (2009) Genetic and physical mapping of new EST-derived SSRs on the A and B genome chromosomes of wheat. Theoretical and Applied Genetics 118, 1015–1025.
| Genetic and physical mapping of new EST-derived SSRs on the A and B genome chromosomes of wheat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXis12gsLw%3D&md5=e4907beae4d7d456356800113604eefdCAS | 19183861PubMed |
Gadaleta A, Nigro D, Giancaspro A, Blanco A (2011) The glutamine synthetase (GS2) genes in relation to grain protein content of durum wheat. Functional & Integrative Genomics 11, 665–670.
| The glutamine synthetase (GS2) genes in relation to grain protein content of durum wheat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhsV2lsrjP&md5=93ad737903347d32033d4cc51fff6af7CAS |
Gallais A, Hirel B (2004) An approach to the genetics of nitrogen use efficiency in maize. Journal of Experimental Botany 55, 295–306.
| An approach to the genetics of nitrogen use efficiency in maize.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXms1egug%3D%3D&md5=269a729d5d2ec90dd8f3ead30e7d214fCAS | 14739258PubMed |
Gebhardt CG, Li L, Pajerowska-Mukthar K, Achenbach U, Sattarzadeh A, Bormann C, Ilarionova E, Ballvora A (2007) Candidate gene approach to identify genes underlying quantitative traits and develop diagnostic markers in potato. Crop Science 47, S106–S111.
| Candidate gene approach to identify genes underlying quantitative traits and develop diagnostic markers in potato.Crossref | GoogleScholarGoogle Scholar |
Habash DZ, Bernard S, Schondelmaier J, Weyen J, Quarrie SA (2007) The genetics of nitrogen use in hexaploid wheat: N utilisation, development and yield. Theoretical and Applied Genetics 114, 403–419.
| The genetics of nitrogen use in hexaploid wheat: N utilisation, development and yield.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXosFKrtA%3D%3D&md5=b0e87053c559dba7d25bd661ae76198dCAS | 17180378PubMed |
Hirel B, Lea PJ (2001) Ammonia assimilation. In ‘Plant nitrogen’. (Eds PJ Lea, J-F Morot-Gaudry) pp. 79–99. (Springer-Verlag: Berlin)
Hirel B, Le Gouis J, Ney B, Gallais A (2007) The challenge of improving nitrogen use efficiency in crop plants: towards a more central role for genetic variability and quantitative genetics within integrated approaches. Journal of Experimental Botany 58, 2369–2387.
| The challenge of improving nitrogen use efficiency in crop plants: towards a more central role for genetic variability and quantitative genetics within integrated approaches.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXos1ykt74%3D&md5=385094725eb34c1069aad972dab64ce3CAS | 17556767PubMed |
Hu J, Vick BA (2003) Target region amplification polymorphism: a novel marker technique for plant genotyping. Plant Molecular Biology Reporter 21, 289–294.
| Target region amplification polymorphism: a novel marker technique for plant genotyping.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXhtVylu7s%3D&md5=2c2f59f22927388e3f15523c3a34d944CAS |
Marone D, Laidò G, Gadaleta A, Colasuonno P, Ficco DBM, Giancaspro A, Giove S, Panio G, Russo MA, De Vita P, Cattivelli L, Papa R, Blanco A, Mastrangelo AM (2012) A high-density consensus map of A and B wheat genomes. Theoretical and Applied Genetics 125, 1619–1638.
| A high-density consensus map of A and B wheat genomes.Crossref | GoogleScholarGoogle Scholar | 22872151PubMed |
McNally S, Hirel B, Gadal P, Mann AF, Stewart GR (1983) Glutamine synthetases of higher plants – evidence for specific isoform content related to their possible physiological role and their compartmentation within the leaf. Plant Physiology 72, 22–25.
| Glutamine synthetases of higher plants – evidence for specific isoform content related to their possible physiological role and their compartmentation within the leaf.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL3sXktF2ksLw%3D&md5=728d2a98fafa7178668bab11c93e3953CAS | 16662965PubMed |
Menzo V, Giancaspro A, Giove SL, Nigro D, Zacheo S, Colasuonno P, Marcotuli I, Incerti O, Blanco A, Gadaleta A (2013) TRAP molecular markers as a system for saturation of the genetic map of durum wheat. Euphytica
| TRAP molecular markers as a system for saturation of the genetic map of durum wheat.Crossref | GoogleScholarGoogle Scholar |
Miflin BJ, Habash DZ (2002) The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops. Journal of Experimental Botany 53, 979–987.
| The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XivFSnur8%3D&md5=3c53ceabf55dbe27b1b1b2d8afd6fc1aCAS | 11912240PubMed |
Obara M, Sato T, Sasaki S, Kashiba K, Nagano A, Nakamura I, Ebitani T, Yano M, Yamaya T (2004) Identification and characterization of a QTL on chromosome 2 for cytosolic glutamine synthetase content and panicle number in rice. Theoretical and Applied Genetics 110, 1–11.
| Identification and characterization of a QTL on chromosome 2 for cytosolic glutamine synthetase content and panicle number in rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXotVGrtA%3D%3D&md5=677862ecdd9a5bed3b005ed4ae7acf81CAS | 15549232PubMed |
Pajerowska KM, Parker JE, Gebhardt C (2005) Potato homologs of Arabidopsis thaliana genes functional in defence signalling – identification, genetic mapping, and molecular cloning. Molecular Plant-Microbe Interactions 18, 1107–1119.
| Potato homologs of Arabidopsis thaliana genes functional in defence signalling – identification, genetic mapping, and molecular cloning.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXhtVCgtr7P&md5=b20ba7c4a8b5b63179da23c53f8e3d92CAS | 16255250PubMed |
Paolacci A, Tanzarella O, Porceddu E, Ciaffi M (2009) Identification and validation of reference genes for quantitative RT-PCR normalization in wheat. BMC Molecular Biology 10, –11.
| Identification and validation of reference genes for quantitative RT-PCR normalization in wheat.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXjtFSjurg%3D&md5=f24d58c1bce1ddff0fd6edfa067d2efeCAS | 19232096PubMed |
Pereira S, Pissarra J, Sunkel C, Salema R (1996) Tissue specific distribution of glutamine synthetase in potato tubers. Annals of Botany 77, 429–432.
| Tissue specific distribution of glutamine synthetase in potato tubers.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XjvFOktLc%3D&md5=ac9760d2f499de430b15c9949157ba84CAS |
Sakurai N, Hayakawa T, Nakamura T, Yamaya T (1996) Changes in the cellular localization of cytosolic glutamine synthetase protein in vascular bundles of rice leaves at various stages of development. Planta 200, 306–311.
| Changes in the cellular localization of cytosolic glutamine synthetase protein in vascular bundles of rice leaves at various stages of development.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28Xms1Cns74%3D&md5=77f369d53302380c98828836af170872CAS |
Sears ER (1954) The aneuploids of common wheat. College of Agriculture, Agricultural Experiment Station Bulletin 572, University of Missouri Columbia, MO.
Sears ER (1966) Nullisomic-tetrasomic combinations in hexaploid wheat. In ‘Chromosome manipulations and plant genetics’. (Eds R Riley, KR Lewis) pp. 20–45. (Oliver and Boyd: Edinburgh)
Sears ER, Sears LMS (1978) The telocentric chromosomes of common wheat. In ‘Proceedings of the 5th International Wheat Genetics Symposium’. (Ed. S Ramanujam) pp. 389–407. (Indian Society of Genetics and Plant Breeding: New Delhi)
Sharp PJ, Kreis M, Shewry PR, Gale MD (1988) Location of β-amylase sequences in wheat and its relatives. Theoretical and Applied Genetics 75, 286–290.
| Location of β-amylase sequences in wheat and its relatives.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1cXhs1amsLY%3D&md5=0e7051161df45cd7e82357578665d632CAS |
Somers JD, Isaac P, Edwards K (2004) A high-density microsatellite consensus map for bread wheat (Triticum aestivum L.). Theoretical and Applied Genetics 109, 1105–1114.
| A high-density microsatellite consensus map for bread wheat (Triticum aestivum L.).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXotlKqtbs%3D&md5=c7f6d3f4e640f5312875128b2ad7ffd3CAS |
Tabuchi M, Sugiyama K, Ishiyama K, Inoue E, Sato T, Takahashi H, Yamaya T (2005) Severe reduction in growth rate and grain filling of rice mutants lacking OsGS1;1, a cytosolic glutamine synthetase. The Plant Journal 42, 641–651.
| Severe reduction in growth rate and grain filling of rice mutants lacking OsGS1;1, a cytosolic glutamine synthetase.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXlt1ynsrY%3D&md5=03e95f964077ab469285e754c156fb6bCAS | 15918879PubMed |
Tobin AK, Yamaya T (2001) Cellular compartmentation of ammonium assimilation in rice and barley. Journal of Experimental Botany 52, 591–604.
| Cellular compartmentation of ammonium assimilation in rice and barley.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXksFKht7k%3D&md5=3fe7bab2b696312a671adc7732229157CAS | 11373307PubMed |
Van Ooijen J (2006) ‘Joinmap 4, Software for the calculation of genetic linkage maps in experimental populations.’ (Kyazma B.V.: Wageningen, The Netherlands)
Wallsgrove RM, Turner JC, Hall NP, Kendall AC, Bright SWJ (1987) Barley mutants lacking chloroplast glutamine synthetase biochemical and genetic analysis. Plant Physiology 83, 155–158.
| Barley mutants lacking chloroplast glutamine synthetase biochemical and genetic analysis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2sXhsFeitLw%3D&md5=1f1f09e128df05c479476af8892e67f9CAS | 16665193PubMed |
Wang S, Wong D, Forrest K, Allen A, Chao S, et al (2013) Characterization of polyploid wheat genomic diversity using the high-density 90 000 SNP array. Plant Biotechnology Journal, in press.
Weeden NF (1981) Genetic and biochemical implications of the endosymbiotic origin of the chloroplast. Journal of Molecular Evolution 17, 133–139.
| Genetic and biochemical implications of the endosymbiotic origin of the chloroplast.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL3MXktl2jsLY%3D&md5=a79c20add8436217a3a1c2dbfb36e540CAS | 7265265PubMed |
Yamaya T, Obara M, Nakajima H, Sasaki S, Hayakawa T, Sato T (2002) Genetic manipulation and quantitative-trait loci mapping for nitrogen recycling in rice. Journal of Experimental Botany 53, 917–925.
| Genetic manipulation and quantitative-trait loci mapping for nitrogen recycling in rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XivFSntbs%3D&md5=c55b67cd14d3fb95e3873c63d56bbcb3CAS | 11912234PubMed |
