Plants with a winter growth habit flower earlier when exposed for several weeks to cold temperatures, a process called vernalization. We report here the positional cloning of the wheat vernalization gene VRN2, a dominant repressor of flowering that is downregulated by vernalization. Loss of function of VRN2, whether by natural mutations or deletions, resulted in spring lines, which do not require vernalization to flower. Reduction of the RNA level of VRN2 by RNA interference accelerated flowering time of transgenic winter wheat plants more than a month.Common wheat (Triticum aestivum L.) is one of the primary grains consumed by humans and is grown in very different environments. This wide adaptability has been favored by the existence of wheat varieties with different growth habits. Winter wheats require a long exposure to low temperatures to flower (vernalization) and are sown in the fall, whereas spring wheats do not have a vernalization requirement and can be planted in spring or fall. The genes from the vernalization pathway prevent flower development during the winter, providing protection for the temperature-sensitive floral organs against the cold.VRN1 and VRN2 are the central genes in the vernalization pathway in wheat, barley, and other temperate cereals. These two genes have strong epistatic interactions and are likely part of the same regulatory pathway (1, 2). In both diploid wheat (Triticum monococcum L.)

Apomixis is a route of asexual reproduction through seeds, that progresses in the absence of meiosis and fertilization to generate maternal clonal progenies. Gametophytic apomicts are usually polyploid and probably arose from sexual ancestors through a limited number of mutations in the female reproductive pathway. A differential display analysis was carried out on immature inflorescences of sexual and apomictic tetraploid genotypes of Paspalum notatum, in order to identify genes associated with the emergence of apospory. Analysis of approximately 10,000 transcripts led to the identification of 94 high-quality differentially expressed sequences. Assembling analysis, plus validation, rendered 65 candidate unigenes, organized as 14 contigs and 51 singletons. Thirty-four unigenes were isolated from apomictic plants and 31 from sexual ones. A total of 45 (69.2%) unigenes were functionally categorized. While several of the differentially expressed sequences appeared to be components of an extracellular receptor kinase (ERK) signal transduction cascade, others seemed to participate in a variety of central cellular processes like cell-cycle control, protein turnover, intercellular signalling, transposon activity, transcriptional regulation and endoplasmic reticulum-mediated biosynthesis. In silico mapping revealed that a particular group of five genes silenced in apomictic plants clustered in a rice genomic area syntenic with the region governing apospory in Paspalum notatum and Brachiaria brizantha. Two of these genes mapped within the set of apo-homologues in P. notatum. Four genes previously reported to be controlled by ploidy were identified among those expressed differentially between apomictic and sexual plants. In situ hybridization experiments were performed for selected clones.

Grain protein content (GPC) is an important factor in pasta and breadmaking quality, and in human nutrition. It is also an important trait for wheat growers because premium prices are frequently paid for wheat with high GPC. A promising source for alleles to increase GPC was detected on chromosome 6B of Triticum turgidum var. dicoccoides accession FA-15-3 (DIC). Two previous quantitative trait locus (QTL) studies found that the positive effect of DIC-6B was associated to a single locus located between the centromere and the Nor-B2 locus on the short arm of chromosome 6B. Microsatellite markers Xgwm508 and Xgwm193 flanking the QTL region were used in this study to develop 20 new homozygous recombinant substitution lines (RSLs) with crossovers between these markers. These 20 RSLs, plus nine RSLs developed in previous studies were characterized with four new RFLP markers located within this chromosome segment. Grain protein content was determined in three field experiments organized as randomized complete block designs with ten replications each. The QTL peaks for protein content were located in the central region of a 2.7-cM interval between RFLP markers Xcdo365 and Xucw67 in the three experiments. Statistical analyses showed that almost all lines could be classified unequivocally within low-and high-protein groups, facilitating the mapping of this trait as a single Mendelian locus designated Gpc-6B1. The Gpc-6B1 locus was mapped 1.5-cM proximal to Xcdo365 and 1.2-cM distal to Xucw67. These new markers can be used to reduce the size of the DIC chromosome segment selected in markerassisted selection programs. Markers Nor-B2 and Xucw66 flanking the previous two markers can be used to select against the DIC segment and reduce the linkage drag during the transfer of Gpc-6B1 into commercial bread and pasta wheat varieties. The precise mapping of the high GPC gene, the high frequency of recombinants recovered in the targeted region, and the recent development of a tetraploid BAC library including the Gpc-6B1 DIC allele are the first steps towards the map-based cloning of this gene.

The concentration of yellow carotenoid pigments in durum wheat grain is an important quality criterion and is determined both by their accumulation and by their degradation by lipoxygenase enzymes (Lpx loci). The existence of a duplication at the Lpx-B1 locus and the allelic variation for a deletion of the Lpx-B1.1 copy is reported. This deletion was associated with a 4.5-fold reduction in lipoxygenase activity and improved pasta color (Po0.0001) but not semolina color, suggesting reduced pigment degradation during pasta processing. A molecular marker for the deletion was mapped on chromosome 4B in a population obtained from the cross between durum line UC1113 and variety Kofa. A second lipoxygenase locus, designated Lpx-A3, was mapped on the homoeologous region on chromosome 4A and was associated with semolina and pasta color (Po0.01) but not with lipoxygenase activity in the mature grain. Selection for both the UC1113 allele for Lpx-A3 and the Kofa Lpx-B1.1 deletion resulted in a 10% increase in yellow scores for dry pasta relative to the opposite allele combination. This result indicates that the markers and the new allelic variants reported here will be useful tools to manipulate the wheat Lpx loci and to improve pasta color. r

Bright yellow color, Wrmness and low cooking loss are important factors for the production of good-quality pasta products. However, the genetic factors underlying those traits are still poorly understood. To Wll this gap we developed a population of 93 recombinant inbred lines (RIL) from the cross between experimental line UC1113 (intermediate pasta quality) with the cultivar Kofa (excellent pasta quality). A total of 269 markers, including 23 SNP markers, were arranged on 14 linkage groups covering a total length of 2,140 cM. Samples from each RIL from Wve diVerent environments were used for complete pasta quality testing and the results from each year were used for QTL analyses. The combined eVect of diVerent loci, environment and their interactions were analyzed using factorial ANOVAs for each trait. We identiWed major QTLs for pasta color on chromosomes 1B, 4B, 6A, 7A and 7B. The 4B QTL was linked to a polymorphic deletion in the Lpx-B1.1 lipoxygenase locus, suggesting that it was associated with pigment degradation during pasta processing. The 7B QTL for pasta color was linked to the Phytoene synthase 1 (Psy-B1) locus suggesting diVerence in pigment biosynthesis. QTLs aVecting pasta Wrmness and cooking loss were detected on chromosomes 5A and 7B, and in both cases they were overlapping with QTL for grain protein content and wet gluten content. These last two parameters were highly correlated with pasta Wrmness (R > 0.71) and inversely correlated to cooking loss (R < ¡0.37). The location and eVect of other QTLs aVecting grain size and weight, gluten strength, mixing properties, and ash content are also discussed.

To overcome environmental stress, plants develop physiological responses that are triggered by genetic or epigenetic changes, some of which involve DNA methylation. It has been proposed that apomixis, the formation of asexual seeds without meiosis, occurs through the temporal or spatial deregulation of the sexual process mediated by genetic and epigenetic factors influenced by the environment. Here, we explored whether there was a link between the occurrence of apomixis and various factors that generate stress, including drought stress, in vitro culture, and intraspecific hybridization. For this purpose, we monitored the embryo sacs of different weeping lovegrass (Eragrostis curvula [Schrad.] Nees) genotypes after the plants were subjected to these stress conditions. Progeny tests based on molecular markers and genome methylation status were analyzed following the stress treatment. When grown in the greenhouse, the cultivar Tanganyika INTA generated less than 2% of its progeny by sexual reproduction. Plants of this cultivar subjected to different stresses showed an increase of sexual embryo sacs, demonstrating an increased expression of sexuality compared to control plants. Plants of the cv. Tanganyika USDA did not demonstrate the ability to generate sexual embryo sacs under any conditions and is therefore classified as a fully apomictic cultivar. We found that this change in the prevalence of sexuality was correlated with genetic and epigenetic changes analyzed by MSAP and AFLPs profiles. Our results demonstrate that different stress conditions can alter the expression of sexual reproduction in facultative tetraploid apomictic cultivars and when the stress stops the reproductive mode shift back to the apomixis original level. These data together with previous observations allow us to generate a hypothetical model of the regulation of apomixis in weeping lovegrass in which the genetic/s region/s that condition apomixis, is/are affected by ploidy, and is/are subjected to epigenetic control.

The aim of this work was to map quantitative trait loci (QTLs) associated with flour yellow color (Fb*) and yellow pigment content (YPC) in durum wheat (Triticum turgidum L. var. durum). Additionally, QTLs affecting flour redness (Fa*) and brightness (FL*) color parameters were investigated. A population of 93 RILs (UC1113 9 Kofa) was evaluated in three locations of Argentina over 2 years. High heritability values ([94%) were obtained for Fb* and YPC, whereas FL* and Fa* showed intermediate to high values. The main QTLs affecting Fb* and YPC overlapped on chromosome arms 4AL (4AL.2), 6AL (6AL.2), 7AS, 7AL, 7BS (7BS.2) and 7BL (7BL.2).The 7BL.1 QTL included the Psy-B1 locus, but one additional linked QTL was detected. A novel minor QTL located on 7AS affected Fb*, with an epistatic effect on YPC. An epistatic interaction occurred between the 7AL and 7BL.2 QTLs. The 4AL.2 QTL showed a strong effect on Fb* and was involved in two digenic epistatic interactions. The 6AL.2 QTL explained most of the variation for Fb* and YPC. The main QTLs affecting FL* and Fa* were located on 2BS and 7BL, respectively. These results confirm the complex inheritance of flour color traits and open the possibility of developing perfect markers to improve pasta quality in Argentinean breeding programs.Electronic supplementary material The online version of this article (

A long-standing goal in plant breeding has been the ability to confer apomixis to agriculturally relevant species, which would require a deeper comprehension of the molecular basis of apomictic regulatory mechanisms. Eragrostis curvula (Schrad.) Nees is a perennial grass that includes both sexual and apomictic cytotypes. The availability of a reference transcriptome for this species would constitute a very important tool toward the identification of genes controlling key steps of the apomictic pathway. Here, we used Roche/454 sequencing technologies to generate reads from inflorescences of E. curvula apomictic and sexual genotypes that were de novo assembled into a reference transcriptome. Near 90% of the 49568 assembled isotigs showed sequence similarity to sequences deposited in the public databases. A gene ontology analysis categorized 27448 isotigs into at least one of the three main GO categories. We identified 11475 SSRs, and several of them were assayed in E curvula germoplasm using SSR-based primers, providing a valuable set of molecular markers that could allow direct allele selection. The differential contribution to each library of the spliced forms of several transcripts revealed the existence of several isotigs produced via alternative splicing of single genes. The reference transcriptome presented and validated in this work will be useful for the identification of a wide range of gene(s) related to agronomic traits of E. curvula, including those controlling key steps of the apomictic pathway in this species, allowing the extrapolation of the findings to other plant species.