Isolation, characterization, and chromosomal location of a gene encoding the Δ1‐pyrroline‐5‐carboxylate synthetase in Arabidopsis thaliana

Savouré, Arnould; Jaoua, Samir; Hua, Xuejun; Ardiles, Wilson; Montagu, Marc Van; Verbruggen, Nathalie

doi:10.1016/0014-5793(95)00935-3

Cited by 183 publications

(142 citation statements)

References 36 publications

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“…The hybridization and washing were performed as described for the Southern analysis. The same experiment was performed with the P5CS cDNA (Savoure et al, 1995) used as a probe. The membrane was stained with methylene blue to verify the equal amount, loading, and transfer of RNA.…”

Section: Total Rna Extraction and Northern Hybridizationmentioning

confidence: 99%

“…Regulation of the glutamate route is relatively well documented: moth bean (Vigna aconitifolia) P5CS enzyme synthesized in Escherichia coli is allosterically inhibited by Pro (Hu et al, 1992). The stimulation of Pro biosynthesis under salt and water stress is also correlated with an increase in the levels of P5CR and P5CS mRNA (Hu et al, 1992;Verbruggen et al, 1993;Savoure et al, 1995;Yoshiba et al, 1995;Strizhof et al, 1997).…”

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confidence: 99%

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Isolation of the Ornithine-δ-Aminotransferase cDNA and Effect of Salt Stress on Its Expression inArabidopsis thaliana1

et al. 1998

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To evaluate the relative importance of ornithine (Orn) as a precursor in proline (Pro) synthesis, we isolated and sequenced a cDNA encoding the Orn-␦-aminotransferase (␦-OAT) from Arabidopsis thaliana. The deduced amino acid sequence showed high homology with bacterial, yeast, mammalian, and plant sequences, and the N-terminal residues exhibited several common features with a mitochondrial transit peptide. Our results show that under both salt stress and normal conditions, ␦-OAT activity and mRNA in young plantlets are slightly higher than in older plants. This appears to be related to the necessity to dispose of an easy recycling product, glutamate. Analysis of the expression of the gene revealed a close association with salt stress and Pro production. In young plantlets, free Pro content, ⌬ 1 -pyrroline-5-carboxylate synthase mRNA, ␦-OAT activity, and ␦-OAT mRNA were all increased by salt-stress treatment. These results suggest that for A. thaliana, the Orn pathway, together with the glutamate pathway, plays an important role in Pro accumulation during osmotic stress. Conversely, in 4-week-old A. thaliana plants, although free Pro level also increased under salt-stress conditions, the ␦-OAT activity appeared to be unchanged and ␦-OAT mRNA was not detectable. ⌬ 1 -pyrroline-5-carboxylate synthase mRNA was still induced at a similar level. Therefore, for the adult plants the free Pro increase seemed to be due to the activity of the enzymes of the glutamate pathway.

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Section: Total Rna Extraction and Northern Hybridizationmentioning

confidence: 99%

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confidence: 99%

Isolation of the Ornithine-δ-Aminotransferase cDNA and Effect of Salt Stress on Its Expression inArabidopsis thaliana1

et al. 1998

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“…We have demonstrated that the Glu pathway is predominant under the conditions of osmotic stress . In Vigna aconitifolia and Arabidopsis, the first two steps of the Pro biosynthesis from Glu are catalyzed by ⌬ 1 -pyrroline-5-carboxylate synthetase (P5CS), a bifunctional enzyme with activities of ␥-glutamyl kinase (␥-GK) and Glu-5-semialdehyde (GSA) dehydrogensae (or ␥-glutamyl phosphate reductase; Hu et al, 1992;Savoure et al, 1995;Yoshiba et al, 1995). In tomato, it has been reported that there are two Pro loci in the nuclear genome: one specifies a bifunctional P5CS (tomPro2) and the other one (tomPro1) apparently encodes a polycistronic mRNA that directs the synthesis of ␥-GK and GSA dehydrogenase as two separate peptides (GarciaRios et al, 1997).…”

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Removal of Feedback Inhibition of Δ1-Pyrroline-5-Carboxylate Synthetase Results in Increased Proline Accumulation and Protection of Plants from Osmotic Stress

et al. 2000

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The ⌬ 1 -pyrroline-5-carboxylate synthetase (P5CS; EC not assigned) is the rate-limiting enzyme in proline (Pro) biosynthesis in plants and is subject to feedback inhibition by Pro. It has been suggested that the feedback regulation of P5CS is lost in plants under stress conditions. We compared Pro levels in transgenic tobacco (Nicotiana tabacum) plants expressing a wild-type form of Vigna aconitifolia P5CS and a mutated form of the enzyme (P5CSF129A) whose feedback inhibition by Pro was removed by site-directed mutagenesis. Transgenic plants expressing P5CSF129A accumulated about 2-fold more Pro than the plants expressing V. aconitifolia wild-type P5CS. This difference was further increased in plants treated with 200 mM NaCl. These results demonstrated that the feedback regulation of P5CS plays a role in controlling the level of Pro in plants under both normal and stress conditions. The elevated Pro also reduced free radical levels in response to osmotic stress, as measured by malondialdehyde production, and significantly improved the ability of the transgenic seedlings to grow in medium containing up to 200 mM NaCl. These findings shed new light on the regulation of Pro biosynthesis in plants and the role of Pro in reducing oxidative stress induced by osmotic stress, in addition to its accepted role as an osmolyte.

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“…Genes specifying GK have been cloned from moth bean (Vigna aconitifolia), Arabidopsis, rice (Oryza sativa), and tomato (Hu et al, 1992;Savouré et al, 1995;Yoshiba et al, 1995;Maggio et al, 1996;García-Ríos et al, 1997;Igarashi et al, 1997;Strizhov et al, 1997). The genes that were cloned from moth bean, Arabidopsis, and rice encode a P5CS made up of a hybrid GK and GPR.…”

mentioning

confidence: 99%

“…High salinity or dehydration results in increased accumulation of Pro in Arabidopsis, rice, and moth bean roots, and has been shown to be accompanied by an increase in the P5CS message level (Hu et al, 1992;Savouré et al, 1995;Yoshiba et al, 1995;Igarashi et al, 1997). …”

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confidence: 99%

Comparative Analysis of the Regulation of Expression and Structures of Two Evolutionarily Divergent Genes for Δ1-Pyrroline-5-Carboxylate Synthetase from Tomato

Fujita

Maggio

Garcia-Rios³

et al. 1998

Plant Physiology

112

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We isolated two tomato (Lycopersicon esculentum) cDNA clones, tomPRO1 and tomPRO2, specifying ⌬ 1 -pyrroline-5-carboxylate synthetase (P5CS), the first enzyme of proline (Pro) biosynthesis. tomPRO1 is unusual because it resembles prokaryotic polycistronic operons (M.G. García-Ríos, T. Fujita, P.C. LaRosa, R.D. Locy, J.M. Clithero, R.A. Bressan, L.N. Csonka [1997] Proc Natl Acad Sci USA 94: 8249-8254), whereas tomPRO2 encodes a fulllength P5CS. We analyzed the accumulation of Pro and the tomPRO1 and tomPRO2 messages in response to NaCl stress and developmental signals. Treatment with 200 mM NaCl resulted in a >60-fold increase in Pro levels in roots and leaves. However, there was a <3-fold increase in the accumulation of the tomPRO2 message and no detectable induction in the level of the tomPRO1 message in response to NaCl stress. Although pollen contained approximately 100-fold higher levels of Pro than other plant tissues, there was no detectable increase in the level of either message in pollen. We conclude that transcriptional regulation of these genes for P5CS is probably not important for the osmotic or pollenspecific regulation of Pro synthesis in tomato. Using restriction fragment-length polymorphism mapping, we determined the locations of tomPRO1 and tomPRO2 loci in the tomato nuclear genome. Sequence comparison suggested that tomPRO1 is similar to prokaryotic P5CS loci, whereas tomPRO2 is closely related to other eukaryotic P5CS genes.

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Isolation, characterization, and chromosomal location of a gene encoding the Δ¹‐pyrroline‐5‐carboxylate synthetase in Arabidopsis thaliana

Cited by 183 publications

References 36 publications

Isolation of the Ornithine-δ-Aminotransferase cDNA and Effect of Salt Stress on Its Expression inArabidopsis thaliana1

Isolation of the Ornithine-δ-Aminotransferase cDNA and Effect of Salt Stress on Its Expression inArabidopsis thaliana1

Removal of Feedback Inhibition of Δ1-Pyrroline-5-Carboxylate Synthetase Results in Increased Proline Accumulation and Protection of Plants from Osmotic Stress

Comparative Analysis of the Regulation of Expression and Structures of Two Evolutionarily Divergent Genes for Δ1-Pyrroline-5-Carboxylate Synthetase from Tomato

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