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Research ArticleCELL BIOLOGY AND SIGNAL TRANSDUCTION
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LeCTR1, a Tomato CTR1-Like Gene, Demonstrates Ethylene Signaling Ability in Arabidopsis and Novel Expression Patterns in Tomato

Julie Leclercq, Lori C. Adams-Phillips, Hicham Zegzouti, Brian Jones, Alain Latché, James J. Giovannoni, Jean-Claude Pech, Mondher Bouzayen
Julie Leclercq
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Lori C. Adams-Phillips
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Hicham Zegzouti
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Brian Jones
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Alain Latché
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James J. Giovannoni
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Jean-Claude Pech
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Mondher Bouzayen
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Published November 2002. DOI: https://doi.org/10.1104/pp.009415

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    Fig. 1.

    Sequence comparison between tomato LeCTR1 (AAl87456) and LeCTR2 (AJ005077), AtCTR1 (CAB82938), AtEDR1 (AAG31143), and R. norvegicus C-Raf (P11345) proteins. Identities between proteins are indicated by shaded squares. The kinase catalytic domain is located in the C-terminal side, including the 11 subdomains (roman numerals). The sequence consensus for the ATP-binding site and Ser/Thr protein kinase are also indicated (… … and - - - - - -, respectively). Arabic numbers indicate the starting of each exon for LeCTR1 and AtCTR1. (Figure continues on facing page.)

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    Fig. 2.

    Comparison of the genomic structure of the tomatoLeCTR1 (AY079048) and the Arabidopsis AtCTR1 gene (L08790). Black portions represent the introns, white portions represent the exons, and gray portions represent the untranslated region. Arrows indicate that each exon of LeCTR1 gene correspond to its homolog in the AtCTR1 gene.

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    Fig. 3.

    Phenotypes of the transgenicLeCTR1-overexpressing lines (27, 17, and 104) compared with that of Arabidopsis wild type and the ctr1-1 mutant. A, Adult plants at the rosette stage grown in the greenhouse. B, Adult plants at the flowering stage grown in the greenhouse. C, Four-day-old seedlings grown in the light. D, Four-day-old etiolated seedlings.

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    Fig. 4.

    Ethylene response of the transgenicLeCTR1-overexpressing lines (27, 17, 104) compared with that of wild type and the ctr1-1 mutant. Etiolated seedlings untreated or treated with increasing ethylene concentration (0.01, 0.1, 1, and 10 μL L−1) were grown for 3 d before monitoring hypocotyl length. Each histogram represents the mean of 30 measurements and the vertical bars indicate the confidence interval.

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    Fig. 5.

    Effect of ethylene on the apical hook curvature of the transgenic LeCTR1-complemented lines (27, 17, and 104) compared with that of wild type and the ctr1-1 mutant. ○, Wild type; +, ctr1-1 mutant; Δ, line 27; ⋄, line 17; ■, line 104. The level of apical curvature was estimated visually for 30 seedlings using a scale ranging from 0 to 4 (0, no apical hook; 1, 90° curvature; 2, 180° curvature; 3, beginning of hook formation; and 4, full hook). The experiment was repeated twice.

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    Fig. 6.

    Effect of Glc on the development of the transgenicLeCTR1-complemented lines (27, 17, and 104) compared with wild type and the ctr1-1 mutant. For each line, 50 seedlings are grown on Murashige and Skoog medium containing 6% (w/v) Glc during 10 d in the light. Wild type, ctr1-1 mutant, or LeCTR1-complemented lines grown in a sealed box with air (A) or in the presence of 10 μL L−1 ethylene (B). The experiment was repeated three times.

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    Fig. 7.

    Molecular analysis of the transgenicLeCTR1-complemented lines (27, 17, and 104) compared with wild type and the ctr1-1 mutant. A, Southern analysis of the transgenic lines with a NPTII probe. Numbers indicate the fragment size in kilobase pairs. Line 27 contained two insertions, whereas line 17 and 104 contained only one copy of the transgene. B, Northern-blot analysis of LeCTR1 and basic chitinase transcript accumulation. Equal loading of the gel with the RNA samples is checked by ethidium bromide staining (bottom).

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    Fig. 8.

    Ethylene-dependent and tissue-specific expression of LeCTR1 in tomato. The levels of LeCTR1transcripts were assessed by real-time quantitative PCR. The experiments were carried out in triplicate. A, LeCTR1 mRNA accumulation was monitored in the root (R), hypocotyl (H), cotyledon (C), unopened buds (B), flower at anthesis (FA), senescent flowers (FS), young fruit 7 DPA (IG), mature green fruit (MG), breaker fruit (Br), breaker + 3 (Br+3), breaker + 7 (Br+7), abscission zone (Ab), callus (Cal), and leaf (L). ΔΔCt on the y axis refers to the fold difference in LeCTR1 expression relative to the leaf. B, Ethylene responsiveness of LeCTR1 in mature green fruit treated with 20 μL L−1 ethylene. ΔΔCt on the y axis refers to the fold difference inLeCTR1 expression relative to the control. C,LeCTR1 ethylene regulation in root and leaves. Ethylene treatment was performed as in B. ΔΔCt on the y axis refers to the fold difference in LeCTR1 expression relative to air-treated root and leaf, respectively.

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LeCTR1, a Tomato CTR1-Like Gene, Demonstrates Ethylene Signaling Ability in Arabidopsis and Novel Expression Patterns in Tomato
Julie Leclercq, Lori C. Adams-Phillips, Hicham Zegzouti, Brian Jones, Alain Latché, James J. Giovannoni, Jean-Claude Pech, Mondher Bouzayen
Plant Physiology Nov 2002, 130 (3) 1132-1142; DOI: 10.1104/pp.009415

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LeCTR1, a Tomato CTR1-Like Gene, Demonstrates Ethylene Signaling Ability in Arabidopsis and Novel Expression Patterns in Tomato
Julie Leclercq, Lori C. Adams-Phillips, Hicham Zegzouti, Brian Jones, Alain Latché, James J. Giovannoni, Jean-Claude Pech, Mondher Bouzayen
Plant Physiology Nov 2002, 130 (3) 1132-1142; DOI: 10.1104/pp.009415
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Plant Physiology: 130 (3)
Plant Physiology
Vol. 130, Issue 3
Nov 2002
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