Cloning of Tomato (Lycopersicon esculentum Mill.) Arginine Decarboxylase Gene and Its Expression during Fruit Ripening

Abstract

Arginine decarboxylase (ADC) is the first enzyme in one of the two pathways of putrescine biosynthesis in plants. The genes encoding ADC have previously been cloned from oat and Fscherichia coli. Degenerate oligonucleotides corresponding to two conserved regions of ADC were used as primers in polymerase chain reaction amplification of tomato (Lycopersicon esculentum Mill.) genomic DNA, and a 1.05-kb fragment was obtained. This genomic DNA fragment encodes an open reading frame of 350 amino acids showing about 50% identity with the oat ADC protein. Using this fragment as a probe, we isolated severa1 partial ADC cDNA clones from a tomato pericarp cDNA library. The 5' end of the coding region was subsequently obtained from a genomic clone containing the entire ADC gene. The tomato ADC gene contains an open reading frame encoding a polypeptide of 502 amino acids and a predicted molecular mass of about 55 kD. The predicted amino acid sequence exhibits 47 and 38% identity with oat and E. coli ADCs, respectively. Cel blot hybridization experiments show that, in tomato, ADC is encoded by a single gene and is expressed as a transcript of approximately 2.2 kb in the fruit pericarp and leaf tissues. During fruit ripening the amount of ADC transcript appeared to peak at the breaker stage. No significant differences were seen when steady-state ADC mRNA levels were compared between normal versus long-keeping Alcobaca (alc) fruit, although alc fruit contain elevated putrescine levels and ADC activity at the ripe stage. The lack of correlation between ADC activity and steady-state mRNA levels in alc fruit suggests a translational and/ or posttranslational regulation of ADC gene expression during tomato fruit ripening.

Molecular cloning and functional identification of a plant ornithine decarboxylase cDNA

“…Fainter bands could also be detected after high-stringency washes that may represent partial digests or related sequences. The Southern blot data are consistent with the possibility of there being more than one ODC gene, in contrast with the situation reported for ADC in tomato, which may be present in only one copy [36].…”

“…The predicted amino acid sequence of the plant ODC was aligned with ADC of oat [21], ADC of tomato [36], ADC of E. coli [38], ODC of yeast [51], ODC of Neurospora crassa [52], ODC of human [53], ODC of mouse [54], ODC of Xenopus [55], ODC of Trypanosoma brucei [56], ODC of Drosophila [57], ODC of Leishmania donovanii [58], DapDC of Pseudomonas aeruginosa [59] and the product of the tabA gene of Pseudomonas syringae cv. tabaci [41].…”

Regulation of Arabidopsis thaliana (L.) Heynh Arginine Decarboxylase by Potassium Deficiency Stress

“…The level of ARGdc message did not change, although there was a 10-fold increase in ARGdc enzyme activity. Rastogi et al (1993) similarly showed that there was no change in ARGdc mRNA levels during tomato fruit ripening, although there was an increase in ARGdc specific activity. Perez-Amador et al (1995) reported that ARGdc enzyme activity and mRNA levels accumulated in developing parthenocarpic www.plantphysiol.org on May 10, 2018 -Published by Downloaded from Copyright © 1996 American Society of Plant Biologists.…”

The Cloning of Two Tomato Lipoxygenase Genes and Their Differential Expression during Fruit Ripening

“…RNA for blot analysis of fruit stages was extracted from pericarp tissue as outlined by Rastogi et al (1993). The method of Chang et al (1993) was used to extract RNA from other tissues.…”