Bulk RNA-Seq analysis to dissect the regulation of stigma position in tomato.

KEY MESSAGE: Transcriptomic analysis of tomato genotypes contrasting for stigma position suggests that stigma insertion occurred by the disruption of a process that finds a parallel in Arabidopsis gynoecium development. Domestication of cultivated tomato (Solanum lycopersicum L.) included the transition from allogamy to autogamy that occurred through the loss of self-incompatibilty and the retraction of the stigma within the antheridial cone. Although the inserted stigma is an established phenotype in modern tomatoes, an exserted stigma is still present in several landraces or vintage varieties. Moreover, exsertion of the stigma is a frequent response to high temperature stress and, being a cause of reduced fertility, a trait of increasing importance. Few QTLs for stigma position have been described and only one of the underlying genes identified. To gain insights on genes involved in stigma position in tomato, a bulk RNA sequencing (RNA-Seq) approach was adopted, using two groups of contrasting genotypes. Phenotypic analysis confirmed the extent and the stability of stigma position in the selected genotypes, whereas they were highly heterogeneous for other reproductive and productive traits. The RNA-Seq analysis yielded 801 differentially expressed genes (DEGs), 566 up-regulated and 235 down-regulated in the genotypes with exserted stigma. Validation by quantitative PCR indicated a high reliability of the RNA-Seq data. Up-regulated DEGs were enriched for genes involved in the cell wall metabolism, lipid transport, auxin response and flavonoid biosynthesis. Down-regulated DEGs were enriched for genes involved in translation. Validation of selected genes on pistil tissue of the 26 single genotypes revealed that differences between bulks could both be due to a general trend of the bulk or to the behaviour of single genotypes. Novel candidate genes potentially involved in the control of stigma position in tomato are discussed.

Fine mapping of the parthenocarpic fruit ( pat) mutation in tomato.

The parthenocarpic fruit ( pat) gene of tomato is a recessive mutation conferring parthenocarpy, which is the capability of a plant to set seedless fruits in the absence of pollination and fertilization. Parthenocarpic mutants offer a useful method to regulate fruit production and a suitable experimental system to study ovary and fruit development. In order to map the Pat locus, two populations segregating from the interspecific cross Lycopersicon esculentum x Lycopersicon pennellii were grown, and progeny plants were classified as parthenocarpic or wild-type by taking into account some characteristic aberrations affecting mutant anthers and ovules. Through bulk segregant analysis, we searched for both random and mapped AFLPs linked to the target gene. In this way, the Pat locus was assigned to the long arm of chromosome 3, as also confirmed by the analysis of a set of L. pennellii substitution and introgression lines. Afterwards, the Pat position was refined by using simple sequence repeats (SSRs) and conserved ortholog set (COS) markers mapping in the target region. The tightest COSs were converted into CAPS or SCAR markers. At present, two co-dominant SCAR markers encompassing a genetic window of 1.2 cM flank the Pat locus. Considering that these markers are orthologous to Arabidopsis genes, a positional cloning exploiting the tomato- Arabidopsis microsynteny seems to be a short-term objective.