Wild Beet Research Articles

Salinity effect on germination, seedling growth and cotyledon membrane complexes of a Portuguese salt marsh wild beet ecotype

Sugar beet (Beta vulgaris L.) high yields have been achieved through irrigation and this crop is described as coping with mild salinity (40–120 mM NaCl). However, during seed germination, sugar beet is salinity sensitive and soil salinity should not exceed 3 dS/m. Wild beets, ancestors of sugar beet, are naturally able to germinate and grow in saline environments. Salinity tolerance during germination and early seedling development of three Portuguese wild beet ecotypes [Comporta (CMP), Oeiras (OEI), Vaiamonte (VMT)] and one sugar beet cultivar (Isella) was evaluated. Concerning germination, VMT outperformed all the other beets, with 98% (± 2%) of glomerules germinating in 200 mM NaCl after 14 days of scarification. However, in 500 mM NaCl, only CMP was able to initiate and maintain radicle emergence, though in a very small extension (< 3%). On the basis of the relative salinity tolerance index, CMP is the less affected by salinity, despite reduced seedling growth and biomass. Since cotyledons length and membrane proteins abundance were negatively affected by salinity, the hypothesis was raised that membranes functionality, including the photochemical multiprotein complexes, was compromised. To test this hypothesis, a blue-native two-dimensional electrophoresis was applied to CMP seedlings. In the cotyledonary leaves, complexes LC1 and LC4 and several components of LC2, LC5, LC6 complexes were negatively affected by salinity. The components of the complexes of photosystem I and ATP synthase were less abundant what points out to a lower cotyledon capacity for ATP synthesis and ferredoxin reduction. Lower availability of ATP and reduced ferredoxin imply reduced photosynthetic assimilation and, therefore, lower carbon availability for growth.

Registration of Sugar Beet Germplasm Lines CN921‐515 and CN921‐516 with Sugar Beet Cyst Nematode Resistance from Beta vulgaris subsp. maritima

CN921‐515 (Reg. No. GP‐295, PI 669447) and CN921‐516 (Reg. No. GP‐296, PI 669448) sugar beet (Beta vulgaris L.) germplasm lines, with sugar beet cyst nematode (SBCN) resistance derived from Beta vulgaris subsp. maritima L. (Bvm), were developed and released by the USDA‐ARS at Salinas, CA, in cooperation with the Beet Sugar Development Foundation in Denver, CO. CN921‐515 and CN921‐516 are multigerm, green hypocotyl, self‐fertile, partially inbred lines extracted from genetic‐male‐sterile facilitated, random‐mated lines. Greenhouse SBCN screening of individual plants and progeny tests indicate each is likely homozygous resistant. CN921‐306 is the progenitor population and was developed from multiple composite crosses involving &gt;40 Bvm accessions. Therefore, the original wild beet accession that contributed sugar beet cyst nematode resistance to these lines is not known, nor are the allelic relationships with recently released CN12‐446, CN12‐770, and CN72‐652. CN921‐515 and CN921‐516 have a higher frequency of SBCN resistance (higher number of resistant individuals per line) than CN921‐306. CN921‐515 and CN921‐516 also have an increased number of rhizomania‐resistant individuals compared with CN921‐306.

Chloroplastic and nuclear diversity of wild beets at a large geographical scale: Insights into the evolutionary history of the Beta section.

Historical demographic processes and mating systems are believed to be major factors in the shaping of the intraspecies genetic diversity of plants. Among Caryophyllales, the Beta section of the genus Beta, within the Amaranthaceae/Chenopodiaceae alliance, is an interesting study model with species and subspecies (Beta macrocarpa, Beta patula, Beta vulgaris maritima and B.v. adanensis) differing in geographical distribution and mating system. In addition, one of the species, B. macrocarpa, mainly diploid, varies in its level of ploidy with a tetraploid cytotype described in the Canary Islands and in Portugal. In this study, we analyzed the nucleotide diversity of chloroplastic and nuclear sequences on a representative sampling of species and subspecies of the Beta section (except B. patula). Our objectives were (1) to assess their genetic relationships through phylogenetic and multivariate analyses, (2) relate their genetic diversity to their mating system, and (3) reconsider the ploidy status and the origin of the Canarian Beta macrocarpa.

Targeted Next-Generation Sequencing Identification of Mutations in Disease Resistance Gene Analogs (RGAs) in Wild and Cultivated Beets

Resistance gene analogs (RGAs) were searched bioinformatically in the sugar beet (Beta vulgaris L.) genome as potential candidates for improving resistance against different diseases. In the present study, Ion Torrent sequencing technology was used to identify mutations in 21 RGAs. The DNA samples of ninety-six individuals from six sea beets (Beta vulgaris L. subsp. maritima) and six sugar beet pollinators (eight individuals each) were used for the discovery of single-nucleotide polymorphisms (SNPs). Target amplicons of about 200 bp in length were designed with the Ion AmpliSeq Designer system in order to cover the DNA sequences of the RGAs. The number of SNPs ranged from 0 in four individuals to 278 in the pollinator R740 (which is resistant to rhizomania infection). Among different groups of beets, cytoplasmic male sterile lines had the highest number of SNPs (132) whereas the lowest number of SNPs belonged to O-types (95). The principal coordinates analysis (PCoA) showed that the polymorphisms inside the gene Bv8_184910_pkon (including the CCCTCC sequence) can effectively differentiate wild from cultivated beets, pointing at a possible mutation associated to rhizomania resistance that originated directly from cultivated beets. This is unlike other resistance sources that are introgressed from wild beets. This gene belongs to the receptor-like kinase (RLK) class of RGAs, and is associated to a hypothetical protein. In conclusion, this first report of using Ion Torrent sequencing technology in beet germplasm suggests that the identified sequence CCCTCC can be used in marker-assisted programs to differentiate wild from domestic beets and to identify other unknown disease resistance genes in beet.

Crop wild relative populations of Beta vulgaris allow direct mapping of agronomically important genes

Rapid identification of agronomically important genes is of pivotal interest for crop breeding. One source of such genes are crop wild relative (CWR) populations. Here we used a CWR population of <200 wild beets (B. vulgaris ssp. maritima), sampled in their natural habitat, to identify the sugar beet (Beta vulgaris ssp. vulgaris) resistance gene Rz2 with a modified version of mapping-by-sequencing (MBS). For that, we generated a draft genome sequence of the wild beet. Our results show the importance of preserving CWR in situ and demonstrate the great potential of CWR for rapid discovery of causal genes relevant for crop improvement. The candidate gene for Rz2 was identified by MBS and subsequently corroborated via RNA interference (RNAi). Rz2 encodes a CC-NB-LRR protein. Access to the DNA sequence of Rz2 opens the path to improvement of resistance towards rhizomania not only by marker-assisted breeding but also by genome editing.

Deciphering the complex nature of bolting time regulation in Beta vulgaris.

Only few genetic loci are sufficient to increase the variation of bolting time in Beta vulgaris dramatically, regarding vernalization requirement, seasonal bolting time and reproduction type. Beta species show a wide variation of bolting time regarding the year of first reproduction, seasonal bolting time and the number of reproduction cycles. To elucidate the genetics of bolting time control, we used three F3 mapping populations that were produced by crossing a semelparous, annual sugar beet with iteroparous, vernalization-requiring wild beet genotypes. The semelparous plants died after reproduction, whereas iteroparous plants reproduced at least twice. All populations segregated for vernalization requirement, seasonal bolting time and the number of reproduction cycles. We found that vernalization requirement co-segregated with the bolting locus B on chromosome 2 and was inherited independently from semel- or iteroparous reproduction. Furthermore, we found that seasonal bolting time is a highly heritable trait (h 2>0.84), which is primarily controlled by two major QTL located on chromosome 4 and 9. Late bolting alleles of both loci act in a partially recessive manner and were identified in both iteroparous pollinators. We observed an additive interaction of both loci for bolting delay. The QTL region on chromosome 4 encompasses the floral promoter gene BvFT2, whereas the QTL on chromosome 9 co-localizes with the BR 1 locus, which controls post-winter bolting resistance. Our findings are applicable for marker-assisted sugar beet breeding regarding early bolting to accelerate generation cycles and late bolting to develop bolting-resistant spring and winter beets. Unexpectedly, one population segregated also for dwarf growth that was found to be controlled by a single locus on chromosome 9.

Genetic Diversity and Physiological Performance of Portuguese Wild Beet (Beta vulgaris spp. maritima) from Three Contrasting Habitats.

The establishment of stress resilient sugar beets (Beta vulgaris spp. vulgaris) is an important breeding goal since this cash crop is susceptible to drought and salinity. The genetic diversity in cultivated sugar beets is low and the beet wild relatives are useful genetic resources for tolerance traits. Three wild beet populations (Beta vulgaris spp. maritima) from contrasting environments, Vaiamonte (VMT, dry inland hill), Comporta (CMP, marsh) and Oeiras (OEI, coastland), and one commercial sugar beet (Isella variety, SB), are compared. At the genetic level, the use of six microsatellite allowed to detect a total of seventy six alleles. It was observed that CMP population has the highest value concerning the effective number of alleles and of expected heterozygosity. By contrast, sugar beet has the lowest values for all the parameters considered. Loci analysis with STRUCTURE allows defining three genetic clusters, the sea beet (OEI and CMP), the inland ruderal beet (VMT) and the sugar beet (SB). A screening test for progressive drought and salinity effects demonstrated that: all populations were able to recover from severe stress; drought impact was higher than that from salinity; the impact on biomass (total, shoot, root) was population specific. The distinct strategies were also visible at physiological level. We evaluated the physiological responses of the populations under drought and salt stress, namely at initial stress stages, late stress stages, and early stress recovery. Multivariate analysis showed that the physiological performance can be used to discriminate between genotypes, with a strong contribution of leaf temperature and leaf osmotic adjustment. However, the separation achieved and the groups formed are dependent on the stress type, stress intensity and duration. Each of the wild beet populations evaluated is very rich in genetic terms (allelic richness) and exhibited physiological plasticity, i.e., the capacity to physiologically adjust to changing environments. These characteristics emphasize the importance of the wild beet ecotypes for beet improvement programs. Two striking ecotypes are VMT, which is the best to cope with drought and salinity, and CMP which has the highest root to shoot ratio. These genotypes can supply breeding programs with distinct goals.

Evolutionary and Biogeographic Insights on the Macaronesian Beta-Patellifolia Species (Amaranthaceae) from a Time-Scaled Molecular Phylogeny.

The Western Mediterranean Region and Macaronesian Islands are one of the top biodiversity hotspots of Europe, containing a significant native genetic diversity of global value among the Crop Wild Relatives (CWR). Sugar beet is the primary crop of the genus Beta (subfamily Betoideae, Amaranthaceae) and despite the great economic importance of this genus, and of the close relative Patellifolia species, a reconstruction of their evolutionary history is still lacking. We analyzed nrDNA (ITS) and cpDNA gene (matK, trnH-psbA, trnL intron, rbcL) sequences to: (i) investigate the phylogenetic relationships within the Betoideae subfamily, and (ii) elucidate the historical biogeography of wild beet species in the Western Mediterranean Region, including the Macaronesian Islands. The results support the Betoideae as a monophyletic group (excluding the Acroglochin genus) and provide a detailed inference of relationships within this subfamily, revealing: (i) a deep genetic differentiation between Beta and Patellifolia species, which may have occurred in Late Oligocene; and (ii) the occurrence of a West-East genetic divergence within Beta, indicating that the Mediterranean species probably differentiated by the end of the Miocene. This was interpreted as a signature of species radiation induced by dramatic habitat changes during the Messinian Salinity Crisis (MSC, 5.96–5.33 Mya). Moreover, colonization events during the Pleistocene also played a role in shaping the current diversity patterns among and within the Macaronesian Islands. The origin and number of these events could not be revealed due to insufficient phylogenetic resolution, suggesting that the diversification was quite recent in these archipelagos, and unravelling potential complex biogeographic patterns with hybridization and gene flow playing an important role. Finally, three evolutionary lineages were identified corresponding to major gene pools of sugar beet wild relatives, which provide useful information for establishing in situ and ex situ conservation priorities in the hotspot area of the Macaronesian Islands.

Histone acetylation influences the transcriptional activation of POX in Beta vulgaris L. and Beta maritima L. under salt stress

Acetylation of histone proteins is a type of chromatin modification which facilitates the activation of genes. Recent studies brought up the importance of this reversible and rapid process for the regulation of gene expression especially in plant defense against a variety of environmental stresses. Deciphering the exact mechanisms of chromatin modifications under abiotic stress conditions is important for improving crop plants’ performance and yield. In a previous study we compared the salt stress responses of Beta vulgaris (sugar beet) and Beta maritima (wild beet). In accordance with those results we suggested that chromatin remodeling can be an active process in the regulation of genes related to salt stress tolerance of these plants. Therefore we performed ChIP assay in control and salt stressed (250 and 500 mM NaCl) plants and compared the enrichment of acetylation in the associated chromatin sites. We found that the transcriptional activation of one peroxidase (POX) encoding gene was associated with the elevated levels of acetylation in H3K9 and H3K27 sites. The acetylation patterns were remarkably different between two species in which the highest acetylation levels were found at H3K9 and H3K27 in wild beet and sugar beet respectively.

Evaluation of rhizomania-resistance segregating sequences and overall genetic diversity pattern among selected accessions of Beta and Patellifolia. Potential implications of breeding for genetic bottlenecks in terms of rhizomania resistance

AbstractSugar beet is hypothesized to have a narrowed genetic base due to its origin as White Silesian Beet and from numerous breeding selections and practices. High sugar quality, yield of recoverable sugar, cytoplasmic-male sterility system, monogermity, pests and disease resistance and bolting resistance constitute some of the adaptations that significantly influenced the existing genetic background of the crop. In this study we aimed to evaluate the extent of genetic diversity existing in wild beet representatives of Beta and Patellifolia and sugar beet cultivars, with a special focus on the complex Beta vulgaris. Another purpose was to determine the potential usefulness and conformity of selected molecular markers in different groups of materials in the context of rhizomania resistance. To reach these goals, molecular RAPD, ISSR techniques, literature-selected rhizomania resistance-segregating sequences as well as mitochondrial markers were used. The comparison of genetic diversity in wild and cultivated Beta forms shows that the population differentiation values and distance values are relatively high in cultivars. Moreover, the diversity component seemed to be compromised rather on the level of population (Hs) than in total (Ht) in cultivars. Our results shed a new light on the expected genetic bottlenecks existing in cultivars and revealed features specific for individual taxa (Patellifolia, Corollinae). Some degree of distinctiveness was suggested between genetic determinants of rhizomania resistance in modern cultivars in comparison with wild resistance sources. In addition, we document here an internal heterogeneity existing in selected wild/weedy accessions at the level of crucial sequences using high resolution melting.

Transcriptome Analysis of Beta macrocarpa and Identification of Differentially Expressed Transcripts in Response to Beet Necrotic Yellow Vein Virus Infection.

BackgroundRhizomania is one of the most devastating diseases of sugar beet. It is caused by Beet necrotic yellow vein virus (BNYVV) transmitted by the obligate root-infecting parasite Polymyxa betae. Beta macrocarpa, a wild beet species widely used as a systemic host in the laboratory, can be rub-inoculated with BNYVV to avoid variation associated with the presence of the vector P. betae. To better understand disease and resistance between beets and BNYVV, we characterized the transcriptome of B. macrocarpa and analyzed global gene expression of B. macrocarpa in response to BNYVV infection using the Illumina sequencing platform.ResultsThe overall de novo assembly of cDNA sequence data generated 75,917 unigenes, with an average length of 1054 bp. Based on a BLASTX search (E-value ≤ 10−5) against the non-redundant (NR, NCBI) protein, Swiss-Prot, the Gene Ontology (GO), Clusters of Orthologous Groups of proteins (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, there were 39,372 unigenes annotated. In addition, 4,834 simple sequence repeats (SSRs) were also predicted, which could serve as a foundation for various applications in beet breeding. Furthermore, comparative analysis of the two transcriptomes revealed that 261 genes were differentially expressed in infected compared to control plants, including 128 up- and 133 down-regulated genes. GO analysis showed that the changes in the differently expressed genes were mainly enrichment in response to biotic stimulus and primary metabolic process.ConclusionOur results not only provide a rich genomic resource for beets, but also benefit research into the molecular mechanisms of beet- BNYV Vinteraction.

Two male sterility-inducing cytoplasms of beet (Beta vulgaris) are genetically distinct but have closely related mitochondrial genomes: implication of a substoichiometric mitochondrial DNA molecule in their evolution

I-12CMS(2) and I-12CMS(3) are sugar beet lines with different sources of cytoplasmic male sterility (CMS) derived from wild beets in Turkey and Pakistan, respectively. We established that I-12CMS(2) has a genetically distinct cytoplasm, but its mitochondrial genome is very similar to I-12CMS(3). Male fertility was assessed in F1 hybrids produced with a common pollen parent. Fertility in the F1's carrying the I-12CMS(3) cytoplasm exceeded that of the F1's with the I-12CMS(2) cytoplasm. Organization of the I-12CMS(2) and I-12CMS(3) mitochondrial genomes were compared based on their physical maps. Mitochondrial genomes of the two strains were largely collinear, except for a large deletion in the noncoding region of I-12CMS(2). Because a mitochondrial orf129 in the I-12CMS(3) cytoplasm is associated with a male sterility phenotype and preservation of orf129 was evident in I-12CMS(2), I-12CMS(2) orf129 was investigated in detail. I-12CMS(2) plants contained three to five times more ORF129 protein than did I-12CMS(3) plants. A single nucleotide substitution, present in the putative promoter region of orf129, appeared to be responsible for the differential accumulation of orf129 transcript. A long N-terminal extension of atp6 is a common feature of some beet CMSs and is found in I-12CMS(2), but the amino acid sequence is unique. I-12CMS(3) mitochondria, but not I-12CMS(2) mitochondria, were found to be heteroplasmic. This heteroplasmy is characterized by a substoichiometric DNA molecule(s) that has at least two I-12CMS(2)-type mitochondrial loci, suggesting the possibility that the I-12CMS(2) mitochondrial genome might have evolved from such a substoichiometric DNA molecule in I-12CMS(3) mitochondria.

Characterization of wild Beta populations in and adjacent to sugar beet fields in the Imperial Valley, California

Populations of wild Beta L. species exist as weeds in commercial sugar beet (Beta vulgaris L. subspecies vulgaris) fields in the Imperial Valley, California. Significant losses to sugar yield and quality result if these wild plants are not removed. In cases of extreme infestation, fields are abandoned without harvest. No selective chemicals are available to differentiate conventional sugar beet from wild relatives and hand removal is labor intensive and expensive. Planting sugar beet varieties with tolerance to glyphosate is a potential solution for infested fields, but risk of gene flow to adjacent wild relatives must be determined. Previous research identified these populations as either Beta vulgaris L. subspecies maritima (L.) Arcang. or Beta macrocarpa Guss. This distinction is critical because B. v. subsp. maritima will readily cross hybridize with cultivated sugar beet while B. macrocarpa rarely will. In April 2011, we collected herbarium samples, mature seed, and leaf tissue from wild Beta populations in 25 infested sugar beet fields throughout the Imperial Valley. Bolting cultivated beets were identified at two locations. Taxonomy of whole plant herbarium samples was unclear due to wild beet stem elongation when under competition with sugar beet plants for canopy light. Morphology of plants from collected seed grown in non-competitive conditions assigned taxonomy of these populations to B. macrocarpa. We used molecular tools to determine the genetic structure of wild Beta populations throughout the Imperial Valley. Extracted DNA was genotyped with 22 simple sequence repeat molecular markers and evaluated for population structure. The bolting beet samples were clearly separated from the majority of B. macrocarpa samples, except for two. The remaining wild populations were further divided into two subgroups suggesting exchange of genetic information or a common ancestor.

Differences between the rhizosphere microbiome of Beta vulgaris ssp. maritima-ancestor of all beet crops-and modern sugar beets.

The structure and function of the plant microbiome is driven by plant species and prevailing environmental conditions. Effectuated by breeding efforts, modern crops diverge genetically and phenotypically from their wild relatives but little is known about consequences for the associated microbiota. Therefore, we studied bacterial rhizosphere communities associated with the wild beet B. vulgaris ssp. maritima grown in their natural habitat soil from coastal drift lines (CS) and modern sugar beets (Beta vulgaris ssp. vulgaris) cultivated in CS and potting soil (PS) under greenhouse conditions. Analysis of 16S rRNA gene fingerprints and pyrosequencing-based amplicon libraries revealed plant genotype- and soil-specific microbiomes. Wild beet plants harbor distinct operational taxonomic units (OTUs) and a more diverse bacterial community than the domesticated sugar beet plants. Although the rhizospheres of both plant genotypes were dominated by Proteobacteria and Planctomycetes, 37.5% of dominant OTUs were additionally detected in the wild beet rhizosphere. Analysis of the cultivable fraction confirmed these plant genotype-specific differences at functional level. The proportion of isolates displayed in vitro activity against phytopathogens was lower for wild beet (≤45.8%) than for sugar beet (≤57.5%). Conversely, active isolates from the wild beet exhibited stronger ability to cope with abiotic stresses. From all samples, active isolates of Stenotrophomonas rhizophila were frequently identified. In addition, soil type-specific impacts on the composition of bacterial communities were found: Acidobacteria, Chloroflexi, and Planctomycetes were only detected in plants cultivated in CS; whereas Bacteroidetes and Proteobacteria dominated in PS. Overall, in comparison to modern sugar beets, wild beets were associated with taxonomically and functionally distinct microbiomes.

Cytosine Methylation of an Ancient Satellite Family in the Wild Beet Beta procumbens

DNA methylation is an essential epigenetic feature for the regulation and maintenance of heterochromatin. Satellite DNA is a repetitive sequence component that often occurs in large arrays in heterochromatin of subtelomeric, intercalary and centromeric regions. Knowledge about the methylation status of satellite DNA is important for understanding the role of repetitive DNA in heterochromatization. In this study, we investigated the cytosine methylation of the ancient satellite family pEV in the wild beet Beta procumbens. The pEV satellite is widespread in species-specific pEV subfamilies in the genus Beta and most likely originated before the radiation of the Betoideae and Chenopodioideae. In B. procumbens, the pEV subfamily occurs abundantly and spans intercalary and centromeric regions. To uncover its cytosine methylation, we performed chromosome-wide immunostaining and bisulfite sequencing of pEV satellite repeats. We found that CG and CHG sites are highly methylated while CHH sites show only low levels of methylation. As a consequence of the low frequency of CG and CHG sites and the preferential occurrence of most cytosines in the CHH motif in pEV monomers, this satellite family displays only low levels of total cytosine methylation.

Spatial distribution and basic ecology of Heterodera schachtii and H. betae wild populations developing on sea beet, Beta vulgaris ssp. maritima

Most populations of crop pathogens have wild relative populations from which they can originate but for which basic knowledge of their ecological requirementsin naturais scarce. However, the study of spatial distribution and ecology of wild pathogen populations may help control them in crops through a better understanding of the environmental factors driving population dynamics. Here, we focused onHeterodera schachtiiandH. betae, two cyst nematodes that cause severe damage to sugar beet (Beta vulgarisssp.vulgaris) crops and can develop on a wild beet relative, the sea beet (B. vulgarisssp.maritima). We investigated the occurrence of both nematode species in the wild and explored some environmental factors that may influence their geographical distribution. To do so, we sampled the wild hostB. v.ssp.maritimaalong the European Atlantic and North Sea coastlines. Results showed thatH. schachtiimainly occurred in the colder environments of northern Europe, whereasH. betaewas preferentially distributed in the warm environments of southern Europe. It was previously established thatH. betaeonly recently appeared in The Netherlands, which are in the north of Europe. Thus, our results do not support this hypothesis. Overall, this study accurately documents the geographical occurrence of two nematode crop pest species in the wild and helps identify the main environmental factors affecting their distribution range.

Genetic diversity and linkage disequilibrium analysis in elite sugar beet breeding lines and wild beet accessions

Linkage disequilibrium decay in sugar beet is strongly affected by the breeding history, and varies extensively between and along chromosomes, allowing identification of known and unknown signatures of selection. Genetic diversity and linkage disequilibrium (LD) patterns were investigated in 233 elite sugar beet breeding lines and 91 wild beet accessions, using 454 single nucleotide polymorphisms (SNPs) and 418 SNPs, respectively. Principal coordinate analysis suggested the existence of three groups of germplasm, corresponding to the wild beets, the seed parent and the pollen parent breeding pool. LD was investigated in each of these groups, with and without correction for genetic relatedness. Without correction for genetic relatedness, in the pollen as well as the seed parent pool, LD persisted beyond 50 centiMorgan (cM) on four (2, 3, 4 and 5) and three chromosomes (2, 4 and 6), respectively; after correction for genetic relatedness, LD decayed after <6 cM on all chromosomes in both pools. In the wild beet accessions, there was a strong LD decay: on average LD disappeared after 1 cM when LD was calculated with a correction for genetic relatedness. Persistence of LD was not only observed between distant SNPs on the same chromosome, but also between SNPs on different chromosomes. Regions on chromosomes 3 and 4 that harbor disease resistance and monogermy loci showed strong genetic differentiation between the pollen and seed parent pools. Other regions, on chromosomes 8 and 9, for which no a priori information was available with respect to their contribution to the phenotype, still contributed to clustering of lines in the elite breeding material.

The Spontaneous Plants’ Recipes Project for enhancing Mediterranean Diet profile change: A sustainable strategy for obesity prevention in childhood, and for contributing to biodiversity maintenance. An intervention of the Italian Food and Nutrition Public Provincial Service

Since 2009 the SIAN is monitoring BMI and dietary habits of 5.220 children, (48.6% males) aged 3–14 years, providing a multi‐sectoral community‐oriented intervention. It was articulated with a core focus on Mediterranean Diet, a proxy of healthy nutrition recommendation. Traditional food and cookery recipes were collected and processed by children. Different types of cultivated and spontaneous plants are still a niche component of Mediterranean dietary profile and wild beet, chard (beta vulgaris), borage, chicory, fennel, cabbages and cardoons are still used in Mediterranean cookery: addressing children to recognizing, collecting, cooking and eating spontaneous plants is a strategic tool supported by the collaboration of teachers, relatives (grandmothers), parents and volunteers. Preliminary results. Initial prevalence of Obesity was 27.6%. At the second year of intervention, a significant drop to 21.27% was already reported with a concurrent increase of Mediterranean Diet Adherence. No such effect was observed by the current educational programs provided nationwide (Italy prevalence: 34.0%). Within the Spontaneous Plants’ Recipes Project at the third year of intervention there is a satisfactory adherence.ConclusionThis exercise allows a closer approach to natural environment, traditional food, green plants, fresh and cooked, and to a more active daily life, moving children in the countryside.

A sugar beet (Beta vulgaris L.) reference FISH karyotype for chromosome and chromosome‐arm identification, integration of genetic linkage groups and analysis of major repeat family distribution

We developed a reference karyotype for B. vulgaris which is applicable to all beet cultivars and provides a consistent numbering of chromosomes and genetic linkage groups. Linkage groups of sugar beet were assigned to physical chromosome arms by FISH (fluorescent in situ hybridization) using a set of 18 genetically anchored BAC (bacterial artificial chromosome) markers. Genetic maps of sugar beet were correlated to chromosome arms, and North-South orientation of linkage groups was established. The FISH karyotype provides a technical platform for genome studies and can be applied for numbering and identification of chromosomes in related wild beet species. The discrimination of all nine chromosomes by BAC probes enabled the study of chromosome-specific distribution of the major repetitive components of sugar beet genome comprising pericentromeric, intercalary and subtelomeric satellites and 18S-5.8S-25S and 5S rRNA gene arrays. We developed a multicolor FISH procedure allowing the identification of all nine sugar beet chromosome pairs in a single hybridization using a pool of satellite DNA probes. Fiber-FISH was applied to analyse five chromosome arms in which the furthermost genetic marker of the linkage group was mapped adjacently to terminal repetitive sequences on pachytene chromosomes. Only on two arms telomere arrays and the markers are physically linked, hence these linkage groups can be considered as terminally closed making the further identification of distal informative markers difficult. The results support genetic mapping by marker localization, the anchoring of contigs and scaffolds for the annotation of the sugar beet genome sequence and the analysis of the chromosomal distribution patterns of major families of repetitive DNA.

Contribution of groundkeepers vs. weed beet to gene escape from sugar beet (Beta vulgaris spp.). Consequences for growing genetically-modified sugar beet – A modelling approach

Weed beet cannot be controlled by herbicides in sugar beet (except via height-selective applicators) as it is a crop relative, descending from accidentally flowering sugar beet (Beta vulgaris) crop plants either because of vernalization during cold springs, or presence of a dominant bolting allele in sugar beet seed lots due to cross-pollination by annual wild beet (B. vulgaris ssp. maritima) in seed production areas. A second, minor source of weed beet are crop roots lost during harvest. These roots (“groundkeepers”) can reproduce in the year after sugar beet and potentially contribute to weed beet dynamics and gene flow. Bolting, flowering and seed production timing and potential of groundkeepers were measured in field experiments. Bolting and flowering were faster in groundkeepers vs. weed beet; flower and seed production was lower in groundkeepers but the latter were less sensitive to competitive crops. The measured parameters were used to introduce a ground-keeper life-cycle into the GeneSys-Beet model which quantifies the effects of cropping systems on weed beet in landscapes. Simulations over several years showed weed beet dynamics to be more sensitive to groundkeeper parameter values than to root loss at sugar beet harvest. Groundkeepers were identified as a key source of weed beet populations and of gene escape from novel sugar beet varieties (e.g. genetically-modified herbicide-tolerant varieties) in the absence of crop bolters. The control of the latter, either by manual weeding or by genetic improvement of sugar beet varieties, was shown to be essential for controlling weed beet populations and avoid the advent of herbicide-tolerant weed beet.