Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses

Mary S. Kalamaki; Dimitris Alexandrou; Diamanto Lazari; Georgios Merkouropoulos; Vasileios Fotopoulos; Irini Pateraki; Alexandros Aggelis; Armando Carrillo-López; Maria J. Rubio-Cabetas; Angelos K. Kanellis

doi:10.1093/jxb/erp072

Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses

Mary S. Kalamaki, Dimitris Alexandrou, Diamanto Lazari, Georgios Merkouropoulos, Vasileios Fotopoulos, Irini Pateraki, Alexandros Aggelis, Armando Carrillo-López, Maria J. Rubio-Cabetas, Angelos K. Kanellis

56 Citations (Scopus)

Abstract

A single copy of the N-acetyl-L-glutamate synthase gene (SlNAGS1) has been isolated from tomato. The deduced amino acid sequence consists of 604 amino acids and shows a high level of similarity to the predicted Arabidopsis NAGS1 and NAGS2 proteins. Furthermore, the N-terminus ArgB domain and the C-terminus ArgA domain found in SlNAGS1 are similar to the structural arrangements that have been reported for other predicted NAGS proteins. SlNAGS1 was expressed at high levels in all aerial organs, and at basic levels in seeds, whereas it was not detected at all in roots. SlNAGS1 transcript accumulation was noticed transiently in tomato fruit at the red-fruit stage. In addition, an increase of SlNAGS1 transcripts was detected in mature green tomato fruit within the first hour of exposure to low oxygen concentrations. Transgenic Arabidopsis plants have been generated expressing the SlNAGS1 gene under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Three homozygous transgenic lines expressing the transgene (lines 1-7, 3-8, and 6-5) were evaluated further. All three transgenic lines showed a significant accumulation of ornithine in the leaves with line 3-8 exhibiting the highest concentration. The same lines demonstrated higher germination ability compared to wild-type (WT) plants when subjected to 250 mM NaCl. Similarly, mature plants of all three transgenic lines displayed a higher tolerance to salt and drought stress compared to WT plants. Under most experimental conditions, transgenic line 3-8 performed best, while the responses obtained from lines 1-7 and 6-5 depended on the applied stimulus. To our knowledge, this is the first plant NAGS gene to be isolated, characterized, and genetically modified.

Original language	English
Journal	Journal of Experimental Botany
Volume	60
Issue number	6
Pages (from-to)	1859-1871
Number of pages	13
ISSN	0022-0957
DOIs	https://doi.org/10.1093/jxb/erp072
Publication status	Published - 2009
Externally published	Yes

Keywords

Arabidopsis thaliana
Arginine biosynthesis
Citrulline
Drought tolerance
N-acetyl-L-glutamate synthase
Ornithine
Salt stress
Seed germination
Transgenic plants

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10.1093/jxb/erp072

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Kalamaki, M. S., Alexandrou, D., Lazari, D., Merkouropoulos, G., Fotopoulos, V., Pateraki, I., Aggelis, A., Carrillo-López, A., Rubio-Cabetas, M. J., & Kanellis, A. K. (2009). Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses. Journal of Experimental Botany, 60(6), 1859-1871. https://doi.org/10.1093/jxb/erp072

Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses. / Kalamaki, Mary S.; Alexandrou, Dimitris; Lazari, Diamanto et al.

In: Journal of Experimental Botany, Vol. 60, No. 6, 2009, p. 1859-1871.

Research output: Contribution to journal › Journal article › Research › peer-review

Kalamaki, MS, Alexandrou, D, Lazari, D, Merkouropoulos, G, Fotopoulos, V, Pateraki, I, Aggelis, A, Carrillo-López, A, Rubio-Cabetas, MJ & Kanellis, AK 2009, 'Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses', Journal of Experimental Botany, vol. 60, no. 6, pp. 1859-1871. https://doi.org/10.1093/jxb/erp072

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title = "Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses",

abstract = "A single copy of the N-acetyl-L-glutamate synthase gene (SlNAGS1) has been isolated from tomato. The deduced amino acid sequence consists of 604 amino acids and shows a high level of similarity to the predicted Arabidopsis NAGS1 and NAGS2 proteins. Furthermore, the N-terminus ArgB domain and the C-terminus ArgA domain found in SlNAGS1 are similar to the structural arrangements that have been reported for other predicted NAGS proteins. SlNAGS1 was expressed at high levels in all aerial organs, and at basic levels in seeds, whereas it was not detected at all in roots. SlNAGS1 transcript accumulation was noticed transiently in tomato fruit at the red-fruit stage. In addition, an increase of SlNAGS1 transcripts was detected in mature green tomato fruit within the first hour of exposure to low oxygen concentrations. Transgenic Arabidopsis plants have been generated expressing the SlNAGS1 gene under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Three homozygous transgenic lines expressing the transgene (lines 1-7, 3-8, and 6-5) were evaluated further. All three transgenic lines showed a significant accumulation of ornithine in the leaves with line 3-8 exhibiting the highest concentration. The same lines demonstrated higher germination ability compared to wild-type (WT) plants when subjected to 250 mM NaCl. Similarly, mature plants of all three transgenic lines displayed a higher tolerance to salt and drought stress compared to WT plants. Under most experimental conditions, transgenic line 3-8 performed best, while the responses obtained from lines 1-7 and 6-5 depended on the applied stimulus. To our knowledge, this is the first plant NAGS gene to be isolated, characterized, and genetically modified.",

keywords = "Arabidopsis thaliana, Arginine biosynthesis, Citrulline, Drought tolerance, N-acetyl-L-glutamate synthase, Ornithine, Salt stress, Seed germination, Transgenic plants",

author = "Kalamaki, {Mary S.} and Dimitris Alexandrou and Diamanto Lazari and Georgios Merkouropoulos and Vasileios Fotopoulos and Irini Pateraki and Alexandros Aggelis and Armando Carrillo-L{\'o}pez and Rubio-Cabetas, {Maria J.} and Kanellis, {Angelos K.}",

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doi = "10.1093/jxb/erp072",

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pages = "1859--1871",

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T1 - Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses

AU - Kalamaki, Mary S.

AU - Alexandrou, Dimitris

AU - Lazari, Diamanto

AU - Merkouropoulos, Georgios

AU - Fotopoulos, Vasileios

AU - Pateraki, Irini

AU - Aggelis, Alexandros

AU - Carrillo-López, Armando

AU - Rubio-Cabetas, Maria J.

AU - Kanellis, Angelos K.

PY - 2009

Y1 - 2009

N2 - A single copy of the N-acetyl-L-glutamate synthase gene (SlNAGS1) has been isolated from tomato. The deduced amino acid sequence consists of 604 amino acids and shows a high level of similarity to the predicted Arabidopsis NAGS1 and NAGS2 proteins. Furthermore, the N-terminus ArgB domain and the C-terminus ArgA domain found in SlNAGS1 are similar to the structural arrangements that have been reported for other predicted NAGS proteins. SlNAGS1 was expressed at high levels in all aerial organs, and at basic levels in seeds, whereas it was not detected at all in roots. SlNAGS1 transcript accumulation was noticed transiently in tomato fruit at the red-fruit stage. In addition, an increase of SlNAGS1 transcripts was detected in mature green tomato fruit within the first hour of exposure to low oxygen concentrations. Transgenic Arabidopsis plants have been generated expressing the SlNAGS1 gene under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Three homozygous transgenic lines expressing the transgene (lines 1-7, 3-8, and 6-5) were evaluated further. All three transgenic lines showed a significant accumulation of ornithine in the leaves with line 3-8 exhibiting the highest concentration. The same lines demonstrated higher germination ability compared to wild-type (WT) plants when subjected to 250 mM NaCl. Similarly, mature plants of all three transgenic lines displayed a higher tolerance to salt and drought stress compared to WT plants. Under most experimental conditions, transgenic line 3-8 performed best, while the responses obtained from lines 1-7 and 6-5 depended on the applied stimulus. To our knowledge, this is the first plant NAGS gene to be isolated, characterized, and genetically modified.

AB - A single copy of the N-acetyl-L-glutamate synthase gene (SlNAGS1) has been isolated from tomato. The deduced amino acid sequence consists of 604 amino acids and shows a high level of similarity to the predicted Arabidopsis NAGS1 and NAGS2 proteins. Furthermore, the N-terminus ArgB domain and the C-terminus ArgA domain found in SlNAGS1 are similar to the structural arrangements that have been reported for other predicted NAGS proteins. SlNAGS1 was expressed at high levels in all aerial organs, and at basic levels in seeds, whereas it was not detected at all in roots. SlNAGS1 transcript accumulation was noticed transiently in tomato fruit at the red-fruit stage. In addition, an increase of SlNAGS1 transcripts was detected in mature green tomato fruit within the first hour of exposure to low oxygen concentrations. Transgenic Arabidopsis plants have been generated expressing the SlNAGS1 gene under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Three homozygous transgenic lines expressing the transgene (lines 1-7, 3-8, and 6-5) were evaluated further. All three transgenic lines showed a significant accumulation of ornithine in the leaves with line 3-8 exhibiting the highest concentration. The same lines demonstrated higher germination ability compared to wild-type (WT) plants when subjected to 250 mM NaCl. Similarly, mature plants of all three transgenic lines displayed a higher tolerance to salt and drought stress compared to WT plants. Under most experimental conditions, transgenic line 3-8 performed best, while the responses obtained from lines 1-7 and 6-5 depended on the applied stimulus. To our knowledge, this is the first plant NAGS gene to be isolated, characterized, and genetically modified.

KW - Arabidopsis thaliana

KW - Arginine biosynthesis

KW - Citrulline

KW - Drought tolerance

KW - N-acetyl-L-glutamate synthase

KW - Ornithine

KW - Salt stress

KW - Seed germination

KW - Transgenic plants

U2 - 10.1093/jxb/erp072

DO - 10.1093/jxb/erp072

M3 - Journal article

C2 - 19357433

AN - SCOPUS:65349085058

SN - 0022-0957

VL - 60

SP - 1859

EP - 1871

JO - Journal of Experimental Botany

JF - Journal of Experimental Botany

IS - 6

ER -

Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SlNAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses

Abstract

Keywords

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