The TFL1 homologue KSN is a regulator of continuous flowering in rose and strawberry

Flowering is a key event in plant life, and is finely tuned by environmental and endogenous signals to adapt to different environments. In horticulture, continuous flowering (CF) is a popular trait introduced in a wide range of cultivated varieties. It played an essential role in the tremendous success of modern roses and woodland strawberries in gardens. CF genotypes flower during all favourable seasons, whereas once-flowering (OF) genotypes only flower in spring. Here we show that in rose and strawberry continuous flowering is controlled by orthologous genes of the TERMINAL FLOWER 1 (TFL1) family. In rose, six independent pairs of CF/OF mutants differ in the presence of a retrotransposon in the second intron of the TFL1 homologue. Because of an insertion of the retrotransposon, transcription of the gene is blocked in CF roses and the absence of the floral repressor provokes continuous blooming. In OF-climbing mutants, the retrotransposon has recombined to give an allele bearing only the long terminal repeat element, thus restoring a functional allele. In OF roses, seasonal regulation of the TFL1 homologue may explain the seasonal flowering, with low expression in spring to allow the first bloom. In woodland strawberry, Fragaria vesca, a 2-bp deletion in the coding region of the TFL1 homologue introduces a frame shift and is responsible for CF behaviour. A diversity analysis has revealed that this deletion is always associated with the CF phenotype. Our results demonstrate a new role of TFL1 in perennial plants in maintaining vegetative growth and modifying flowering seasonality

Identification of superior reference genes for data normalisation of expression studies via quantitative PCR in hybrid roses (Rosa hybrida)

<p>Abstract</p> <p>Background</p> <p>Gene expression studies are a prerequisite for understanding the biological function of genes. Because of its high sensitivity and easy use, quantitative PCR (qPCR) has become the gold standard for gene expression quantification. To normalise qPCR measurements between samples, the most prominent technique is the use of stably expressed endogenous control genes, the so called reference genes. However, recent studies show there is no universal reference gene for all biological questions. Roses are important ornamental plants for which there has been no evaluation of useful reference genes for gene expression studies.</p> <p>Results</p> <p>We used three different algorithms (BestKeeper, geNorm and NormFinder) to validate the expression stability of nine candidate reference genes in different rose tissues from three different genotypes of <it>Rosa hybrida </it>and in leaves treated with various stress factors. The candidate genes comprised the classical "housekeeping genes" (<it>Actin, EF-1α, GAPDH</it>, <it>Tubulin </it>and <it>Ubiquitin</it>), and genes showing stable expression in studies in <it>Arabidopsis </it>(<it>PP2A, SAND, TIP </it>and <it>UBC</it>). The programs identified no single gene that showed stable expression under all of the conditions tested, and the individual rankings of the genes differed between the algorithms. Nevertheless the new candidate genes, specifically, <it>PP2A </it>and <it>UBC</it>, were ranked higher as compared to the other traditional reference genes. In general, <it>Tubulin </it>showed the most variable expression and should be avoided as a reference gene.</p> <p>Conclusions</p> <p>Reference genes evaluated as suitable in experiments with <it>Arabidopsis thaliana </it>were stably expressed in roses under various experimental conditions. In most cases, these genes outperformed conventional reference genes, such as <it>EF1-α </it>and <it>Tubulin</it>. We identified <it>PP2A</it>, <it>SAND </it>and <it>UBC </it>as suitable reference genes, which in different combinations may be used for normalisation in expression analyses via qPCR for different rose tissues and stress treatments. However, the vast genetic variation found within the genus <it>Rosa</it>, including differences in ploidy levels, might also influence expression stability of reference genes, so that future research should also consider different genotypes and ploidy levels.</p

An Autotetraploid Linkage Map of Rose (Rosa hybrida) Validated Using the Strawberry (Fragaria vesca) Genome Sequence

Polyploidy is a pivotal process in plant evolution as it increase gene redundancy and morphological intricacy but due to the complexity of polysomic inheritance we have only few genetic maps of autopolyploid organisms. A robust mapping framework is particularly important in polyploid crop species, rose included (2n = 4x = 28), where the objective is to study multiallelic interactions that control traits of value for plant breeding. From a cross between the garden, peach red and fragrant cultivar Fragrant Cloud (FC) and a cut-rose yellow cultivar Golden Gate (GG), we generated an autotetraploid GGFC mapping population consisting of 132 individuals. For the map we used 128 sequence-based markers, 141 AFLP, 86 SSR and three morphological markers. Seven linkage groups were resolved for FC (Total 632 cM) and GG (616 cM) which were validated by markers that segregated in both parents as well as the diploid integrated consensus map

A survey of flowering genes reveals the role of gibberellins in floral control in rose

Exhaustive studies on flowering control in annual plants have provided a framework for exploring this process in other plant species, especially in perennials for which little molecular data are currently available. Rose is a woody perennial plant with a particular flowering strategy—recurrent blooming, which is controlled by a recessive locus (RB). Gibberellins (GA) inhibit flowering only in non-recurrent roses. Moreover, the GA content varies during the flowering process and between recurrent and non-recurrent rose. Only a few rose genes potentially involved in flowering have been described, i.e. homologues of ABC model genes and floral genes from EST screening. In this study, we gained new information on the molecular basis of rose flowering: date of flowering and recurrent blooming. Based on a candidate gene strategy, we isolated genes that have similarities with genes known to be involved in floral control in Arabidopsis (GA pathway, floral repressors and integrators). Candidate genes were mapped on a segregating population, gene expression was studied in different organs and transcript abundance was monitored in growing shoot apices. Twenty-five genes were studied. RoFT, RoAP1 and RoLFY are proposed to be good floral markers. RoSPY and RB co-localized in our segregating population. GA metabolism genes were found to be regulated during floral transition. Furthermore, GA signalling genes were differentially regulated between a non-recurrent rose and its recurrent mutant. We propose that flowering gene networks are conserved between Arabidopsis and rose. The GA pathway appears to be a key regulator of flowering in rose. We postulate that GA metabolism is involved in floral initiation and GA signalling might be responsible for the recurrent flowering character

Development and validation of an SNP-based new set of markers useful for early selection for Sharka disease in apricot ( P. armeniaca )

Sharka disease, due to the Plum pox virus, is a major threat to apricot trees. Most of the accessions world-wide and particularly those cultivated in Western Europe and the Mediterranean area are susceptible to the disease. A few resistance sources have been identified to date in cultivars developed in the US from a basis of Central Asian germplasm, all of these sharing the same genomic region associated with resistance to sharka in chromosome 1. Based on this, Simple sequence repeats (SSR) markers framing the MATH genes possibly responsible for the resistance have been developed in this region and a set of 2 SSRs (PGS1.21 and PGS1.24) and an indel (ZP002) was selected for identifying and discarding susceptible individuals at the plantlet stage of the apricot breeding process. However SSR markers could be difficult to implement by small teams due to the investment in hands-on time and equipment. As a consequence we turned to single nucleotide polymorphisms (SNP) to develop an alternative set of markers that could be cheaper and more easily implemented. Taking advantage of the numerous apricot genomic-sequences available at the UGAFL, we identified resistant/susceptible SNP/indel haplotypes and developed a set of three primer triplets (SP353, SP870, and SP871) based on the competitive allele-specific PCR (KASP™) technique by LGC genomics. This set was validated using a wide range of apricot accessions from different geographic origins and genetic backgrounds, concurrently with the original set of three SSRs. The new set gave similar results as the SSR-based set, was more convenient and, as a result, is now routinely used for early selection of apricot seedlings in our breeding programs

Genetic structure of a worldwide germplasm collection of Prunus armeniaca L

Analyses of genetic diversity and phylogenetic relationships illuminate the origin and domestication of crop species and have important implications for plant breeding programs and the conservation of genetic resources. Apricot (Prunus armeniaca L.) genetic resources in collections that are widely dispersed around the world were assessed using 25 simple sequence repeat (SSR) markers. Analysis of allelic diversity among SSR data revealed at least a double pattern of diffusion from Central Asia to eastern and western countries with a clear east-west loss of genetic diversity related to the genetic bottleneck during apricot domestication. Structure and phylogenetic analysis indicated that accessions from Central Asia and China were genetically most diverse, suggesting that this large region constitutes the apricot center of origin

Genetic structure of a worldwide germplasm collection of <em>Prunus armeniaca</em> L. species

International audienceAnalyses of genetic diversity and phylogenetic relationships illuminate the origin and domestication of crop species and have important implications for plant breeding programs and the conservation of genetic resources. Apricot ( Prunus armeniaca L.) genetic resources in collections that are widely dispersed around the world were assessed using 25 simple sequence repeat ( SSR) markers. Analysis of allelic diversity among SSR data revealed at least a double pattern of diffusion from Central Asia to eastern and western countries with a clear east-west loss of genetic diversity related to the genetic bottleneck during apricot domestication. Structure and phylogenetic analysis indicated that accessions from Central Asia and China were genetically most diverse, suggesting that this large region constitutes the apricot center of origin

Genomic approach to study floral development genes in Rosa sp.

International audienceCultivated for centuries, the varieties of rose have been selected based on a number of flower traits. Understanding the genetic and molecular basis that contributes to these traits will impact on future improvements for this economically important ornamental plant. In this study, we used scanning electron microscopy and sections of meristems and flowers to establish a precise morphological calendar from early rose flower development stages to senescing flowers. Global gene expression was investigated from floral meristem initiation up to flower senescence in three rose genotypes exhibiting contrasted floral traits including continuous versus once flowering and simple versus double flower architecture, using a newly developed Affymetrix microarray (Rosa1_Affyarray) tool containing sequences representing 4765 unigenes expressed during flower development. Data analyses permitted the identification of genes associated with floral transition, floral organs initiation up to flower senescence. Quantitative real time PCR analyses validated the mRNA accumulation changes observed in microarray hybridizations for a selection of 24 genes expressed at either high or low levels. Our data describe the early flower development stages in Rosa sp, the production of a rose microarray and demonstrate its usefulness and reliability to study gene expression during extensive development phases, from the vegetative meristem to the senescent flower