Identification of a stable epi-allele associated with flower development and low bunch compactness in a somatic variant of Tempranillo Tinto

Abstract

Grapevine cultivars are vegetatively propagated to preserve their varietal characteristics. However, spontaneous somatic variations that occur and are maintained during cycles of vegetative growth offer opportunities for the natural improvement of traditional grape cultivars. One advantageous trait for winegrowing is reduced bunch compactness, which decreases the susceptibility to pests and fungal diseases and favor an even berry ripening. In this study, we examined a loose bunch somatic variant of Tempranillo Tinto cultivar (TT) to elucidate the molecular basis underlying this variation. The variant displayed a masculinized flower phenotype, characterized by an underdeveloped yet functional gynoecium, with reduced development of the style, stigma and septum, as compared to the complete hermaphroditic flowers typically observed in TT. Genetic analysis of its self-progeny demonstrated the co-segregation of the masculinized flower phenotype with the hermaphrodite allele of the grape sex locus (SDR)^[1]. While genome re-sequencing did not identify any genetic variation within the SDR locus, an RNA-seq analysis identified one SDR-located gene over-expressed in the somatic variant compared to a control TT clone. The hermaphrodite allele was specifically over-expressed in the variant. Both Illumina WGBS and Nanopore epigenomic analyses identified a hermaphrodite allele-specific hyper-methylated region upstream of the upregulated gene. Analysis of chromatin conformation capture (3C) revealed a three-dimensional rearrangement of the locus, including the presence of a novel 36 kb chromatin loop delimiting the hyper-methylated region, which could be responsible for the over-expression and the phenotype. These findings indicate that somatic epi-alleles in the SDR locus can determine variation in gynoecium development leading to decreased fruit set and looser bunches in grapevine.

Acknowledgements: This study is part of the projects BIO-2017-86375-R and PID2020-120183RB-I00, and PRE-2018-086017 grant funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”. This study is also part of the vWISE project.

1)  Massonnet, M., Cochetel, N., Minio, A., Vondras, A. M., Lin, J., Muyle, A., Garcia, J. F., Zhou, Y., Delledonne, M., Riaz, S., Figueroa-Balderas, R., Gaut, B. S., & Cantu, D. (2020). The genetic basis of sex determination in grapes. Nature Communications, 11(1), 1–12. DOI: 10.1038/s41467-020-16700-z