Abstract
Key message
Seedlessness, one of the most desired traits in fleshy fruits, can be obtained altering solely AGL11 gene, a D -class MADS-box. Opposite to overlap** functions described for ovule identity.
Abstract
AGAMOUS like-11 (AGL11) is a D-class MADS-box gene that determines ovule identity in model species. In grapevine, VviAGL11 has been proposed as the main candidate gene responsible for seedlessness because ovules develop into seeds after fertilization. Here, we demonstrate that AGL11 has a direct role in the determination of the seedless phenotype. In grapevine, broad expression analysis revealed very low expression levels of the seedless allele compared to the seeded allele at the pea-size berry stage. Heterozygous genotypes have lower transcript accumulation than expected considering the diploid nature of grapevine, thereby revealing that the dominant phenotype previously described for seedlessness is based on its expression level. In a seeded somatic variant of Sultanina (Thompson Seedless) that has well-developed seeds, Sultanine Monococco, structural differences were identified in the regulatory region of VviAGL11. These differences affect transcript accumulation levels and explain the phenotypic differences between the two varieties. Functional experiments in tomato demonstrated that SlyAGL11 gene silencing produces seedless fruits and that the degree of seed development is proportional to transcript accumulation levels. Furthermore, the genes involved in seed coat development, SlyVPE1 and SlyVPE2 in tomato and VviVPE in grapevine, that are putatively controlled by SlyAGL11 and VviAGL11, respectively, are expressed at lower levels in silenced tomato lines and in seedless grapevine genotypes. In conclusion, this work provides evidence that the D-class MADS-box AGL11 plays a major and direct role in seed development in fleshy fruits, providing a valuable tool for further analysis of fruit development.
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Acknowledgments
This research was supported by FONDECYT grant 1120532, FONDEF grant G09i1007 and the BIOFRUTALES consortium. We are very grateful to Loïc Le Cunff and colleagues from the grapevine team INRA Montpellier for providing assistance and precious samples.
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Communicated by P. Puigdomenech.
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Supplementary material 1: Primers used in this study. Target name, target accession number, primers name, primers sequence and use are listed (PDF 84 kb)
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Supplementary material 2: The somatic variant of Sultanina, Sultanine Monococco, is a chimera with mutations derived from mitotic recombination events in the regulatory region of the VviAGL11 MADS-Box gene. Multiple alignment of eight alleles isolated from genomic DNA from Sultanine Monococco, Sultanina and reference Pinot Noir (PN44024, (Jaillon et al. 2007)). Sultanina alleles are indicated as seeded (188) and seedless (198) alleles based on genotype by marker p3_VvAGL11 (Mejia et al. 2011). Four recombinant alleles (SuMo 1 2, 3, and 5) and two original alleles (SuMo 4 and 6) are represented by green or orange fragments to denote seedless and seeded alleles, respectively (JPEG 1244 kb)
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Supplementary material 3: Spatio-temporal expression of SlyAGL11 analyzed by histochemical GUS assay. Transgenic lines expressing GUS reporter gene under the control of SlyAGL11 promoter (pSlyAGL11 of 1568 bp) were used to analyze patterns of expression between pre and post anthesis stages: Stage nine (petals approaching top of sepals and develo** ovules) according to Brukhin et al. (2003), anthesis, eight DPA (where a peak of expression was detected by RT-qPCR), 18 and 30 DPA (JPEG 14532 kb)
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Ocarez, N., Mejía, N. Suppression of the D-class MADS-box AGL11 gene triggers seedlessness in fleshy fruits. Plant Cell Rep 35, 239–254 (2016). https://doi.org/10.1007/s00299-015-1882-x
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DOI: https://doi.org/10.1007/s00299-015-1882-x