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Τhe Role of a Gibberellin 20-Oxidase Gene in Fruit Development in Pepper (Capsicum annuum)

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Abstract

Fruit shape is a very important fruit quality character frequently affected by grafting in vegetable plants like pepper. It has already been shown that, similar to tomato, fruit shape in pepper is likely controlled by an Ovate-like gene, CaOvate, the down-regulation of which positively affects fruit elongation. To further understand the molecular mechanisms involved in pepper fruit shape control and the changes imposed by grafting, we have amplified, sequenced, and structurally characterized CaGA20ox1, the target gene of CaOvate, from a long fruit and a round fruit shaped cultivar. The results show that CaGA20ox1 has similar genomic organization to the tomato GA20ox1 and encodes a 375-amino acid polypeptide that shares 89% identity with tomato GA20ox1. We then studied CaGA20ox1 expression in different pepper plant parts and in different developmental stages of flower and fruit development. The expression of the gene was quantified by means of relative quantitative PCR in the developmental stage of 10 days after anthesis fruit of both cultivars. The results showed that there is a significant difference in the expression of the CaGA20ox1 between the two cultivars in this specific stage as well as in the expression of CaGA20ox1 after virus-induced gene silencing (VIGS) of CaOvate. Finally, the 5′ upstream sequences of CaGA20ox1 gene of the two cultivars were examined and compared. These results corroborate our previous findings, where VIGS of CaOvate alters CaGA20ox1 expression, leading to more elongated fruit, and also progress further the understanding of the genes involved in fruit shape control in pepper opening the way for understanding the molecular means of grafting effects.

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Acknowledgments

A. Tsaballa holds a PhD scholarship from the “Alexander S. Onassis” Public Benefit Foundation. This research has been co-financed by the European Union [European Social Fund (ESF)] and Greek national funds through the Operational Program “Education and Lifelong Learning” of the National Strategic Reference Framework (NSRF)—Research Funding Program: Heracletus II. Continuous support for the Institute of Agrobiotechnology/CERTH from the General Secretariat of Research and Technology of Greece is also acknowledged.

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Authors and Affiliations

  1. Department of Genetics and Plant Breeding, School of Agriculture, Aristotle University of Thessaloniki, Thessaloniki, 541 24, Greece

    Aphrodite Tsaballa & Athanasios S. Tsaftaris

  2. Institute of Agrobiotechnology (INA), CERTH, 6th km Charilaou-Thermis Road, Thermi, 570 01, Greece

    Konstantinos Pasentsis & Athanasios S. Tsaftaris

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  1. Aphrodite Tsaballa
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  2. Konstantinos Pasentsis
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  3. Athanasios S. Tsaftaris
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Correspondence to Athanasios S. Tsaftaris.

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Supplementary Fig. 1

Visualization of BLAST output with the Circoletto tool: the GA20ox1 protein sequences from the three Solanaceae plants (Sl Tomato, St potato, Nt tobacco) have been used as queries and the 10 top—according to their BLAST output bit score—pepper ESTs have been used as the database. Ribbons represent the local alignments BLAST has produced, in four colors, blue, green, orange, and red, representing the four quartiles up to the maximum score. The yellow coloring inside the queries’ sequences (colored in gray) represent the Fe(2+) 2-oxoglutarate dioxygenase domain. Each EST bears its original name given in the pepper EST database (http://210.218.199.240/pepper/index.php). EST with the number KS16007H09 bears the GenBank accession number (DOC 246 kb)

Supplementary Table 1

Primers sequences used in the experiments (DOC 29 kb)

Supplementary Table 2

A short depiction of the regulatory elements putative targets of transcription factors to bind to the GA20ox1 5′ upstream region of the two pepper cultivars. The position of each element (in relation to the start codon) is reported as long as whether the element is positioned on the plus (+) or the minus (−) strand and the number of the element (according to the Softberry list http://linux1.softberry.com/berry.phtml?topic=regsitelist). Furthermore, a short characterization and description of the element, the gene where it was first reported and of the corresponding binding factor are included. Finally, selected references for each element are also mentioned (DOC 171 kb)

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Tsaballa, A., Pasentsis, K. & Tsaftaris, A.S. Τhe Role of a Gibberellin 20-Oxidase Gene in Fruit Development in Pepper (Capsicum annuum). Plant Mol Biol Rep 30, 556–565 (2012). https://doi.org/10.1007/s11105-011-0364-8

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