Piergiorgio Stevanato

The origin of rhizomania resistance in sugar beet

In the last 35 years, breeding has greatly reduced the damages caused by rhizomania in sugar beet crops. After the first encouraging results using the Alba genotypes, the cultivar Rizor represented a substantial step forward and has given good yield improvement in diseased fields in many parts of the world. The original variety and subsequent improved versions continued to offer good performances for about a decade, after which it was surpassed by other hybrids derived in part from the Rizor itself. Further progress in terms of sugar production became possible in 1986, when the Holly monogerm lines were released in USA and Europe. In spite of the incomplete information about the genealogy of the first resistant materials,many evidences and the molecular analyses on the different genotypes suggest a possible common progenitor and lineage. The resistant cultivars have kept the yield at an adequate level, allowing cultivation to continue in countries where the disease has reached epidemic proportions. The case of rhizomania resistance in sugar beet can therefore be considered as one of the most important achievements in plant breeding.

Genetic transformation of the sugar beet plastome

It is very important for the application of chloroplast engineering to extend the range of species in which this technology can be achieved. Here, we describe the development of a chloroplast transformation system for the sugar beet (Beta vulgaris L. ssp. vulgaris, Sugar Beet Group) by biolistic bombardment of leaf petioles. Homoplasmic plastid-transformed plants of breeding line Z025 were obtained. Transformation was achieved using a vector that targets genes to the rrn16/rps12 intergenic region of the sugar beet plastome, employing the aadA gene as a selectable marker against spectinomycin and the gfp gene for visual screening of plastid transformants. gfp gene transcription and protein expression were shown in transplastomic plants. Detection of GFP in Comassie blue-stained gels suggested high GFP levels. Microscopy revealed GFP fluorescence within the chloroplasts. Our results demonstrate the feasibility of engineering the sugar beet chloroplast genome; this technology provides new opportunities for the genetic improvement of this crop and for social acceptance of genetically modified sugar beet plants.

High-throughput RAD-SNP genotyping for characterization of sugar beet genotypes

High-throughput single-nucleotide polymorphism (SNP) genotyping provides a rapid way of developing resourceful sets of markers for delineating genetic structure and for understanding the basis of the taxonomic discrimination. In this paper, we present a panel of 192 SNPs for effective genotyping in sugar beet using a high-throughput marker array technology, QuantStudio 12K Flex system, coupled with Taqman OpenArray technology. The selected SNPs were evaluated for genetic diversity among a set of 150 individuals representing 15 genotypes (10 individuals each) from five cytoplasmic male steriles (CMSs), five pollinators, and five commercial varieties. We demonstrated that the proposed panel of 192 SNPs effectively differentiated the studied genotypes. A higher degree of polymorphism was observed among the CMSs as compared to pollinators and commercial varieties. PCoA and STRUCTURE analysis revealed that CMSs, pollinators, and varieties clustered into three distinct subpopulations. Our results demonstrate the utility of the identified panel of 192 SNPs coupled with TaqMan OpenArray technology as a wide set of markers for high-throughput SNP genotyping in sugar beet.

Genome-enabled predictions for binomial traits in sugar beet populations

BackgroundGenomic information can be used to predict not only continuous but also categorical (e.g. binomial) traits. Several traits of interest in human medicine and agriculture present a discrete distribution of phenotypes (e.g. disease status). Root vigor in sugar beet (B. vulgaris) is an example of binomial trait of agronomic importance. In this paper, a panel of 192 SNPs (single nucleotide polymorphisms) was used to genotype 124 sugar beet individual plants from 18 lines, and to classify them as showing “high” or “low” root vigor.ResultsA threshold model was used to fit the relationship between binomial root vigor and SNP genotypes, through the matrix of genomic relationships between individuals in a genomic BLUP (G-BLUP) approach. From a 5-fold cross-validation scheme, 500 testing subsets were generated. The estimated average cross-validation error rate was 0.000731 (0.073%). Only 9 out of 12326 test observations (500 replicates for an average test set size of 24.65) were misclassified.ConclusionsThe estimated prediction accuracy was quite high. Such accurate predictions may be related to the high estimated heritability for root vigor (0.783) and to the few genes with large effect underlying the trait. Despite the sparse SNP panel, there was sufficient within-scaffold LD where SNPs with large effect on root vigor were located to allow for genome-enabled predictions to work.

Evaluation of genetic diversity and root traits of sea beet accessions of the Adriatic Sea coast

Thirty-nine sea beet [Beta vulgaris L. ssp. maritima (L.) Arcang.] accessions of the Adriatic coast were screened genetically and for their adaptive morpho-functional root traits in order to identify new sources of abiotic resistances for sugar beet breeding programs. Genetic diversity was evaluated with 21 microsatellites markers that identified 44 polymorphic alleles. Sea beets grouped into two main clusters: the West and the East Adriatic coast groups, with the latter showing higher genetic diversity. Among sea beet accessions with desirable root traits, four accessions have proved to be interesting for sugar beet [B. vulgaris (L.) ssp. vulgaris] breeding aimed to improve tolerance to nutritional stresses. Lastovo (ID 29) and Zut (ID 34) accessions were characterized by the highest values of RER, TRL, FRL and RSA still maintaining a high value of RTD, while Grado (ID 21) an Portic (ID 23) accessions were characterized by the highest RTD, but with low values of RER, TRL, FRL and RSA parameters.

Beneficial Bacteria Isolated from Grapevine Inner Tissues Shape Arabidopsis thaliana Roots.

We investigated the potential plant growth-promoting traits of 377 culturable endophytic bacteria, isolated from Vitis vinifera cv. Glera, as good biofertilizer candidates in vineyard management. Endophyte ability in promoting plant growth was assessed in vitro by testing ammonia production, phosphate solubilization, indole-3-acetic acid (IAA) and IAA-like molecule biosynthesis, siderophore and lytic enzyme secretion. Many of the isolates were able to mobilize phosphate (33%), release ammonium (39%), secrete siderophores (38%) and a limited part of them synthetized IAA and IAA-like molecules (5%). Effects of each of the 377 grapevine beneficial bacteria on Arabidopsis thaliana root development were also analyzed to discern plant growth-promoting abilities (PGP) of the different strains, that often exhibit more than one PGP trait. A supervised model-based clustering analysis highlighted six different classes of PGP effects on root architecture. A. thaliana DR5::GUS plantlets, inoculated with IAA-producing endophytes, resulted in altered root growth and enhanced auxin response. Overall, the results indicate that the Glera PGP endospheric culturable microbiome could contribute, by structural root changes, to obtain water and nutrients increasing plant adaptation and survival. From the complete cultivable collection, twelve promising endophytes mainly belonging to the Bacillus but also to Micrococcus and Pantoea genera, were selected for further investigations in the grapevine host plants towards future application in sustainable management of vineyards.

Identification and Validation of a SNP Marker Linked to the Gene HsBvm-1 for Nematode Resistance in Sugar Beet

The beet-cyst nematode (Heterodera schachtii Schmidt) is one of the major pests of sugar beet. The identification of molecular markers associated with nematode tolerance would be helpful for developing tolerant varieties. The aim of this study was to identify single nucleotide polymorphism (SNP) markers linked to nematode tolerance from the Beta vulgaris ssp. maritima source WB242. A WB242-derived F2 population was phenotyped for host-plant nematode reaction revealing a 3:1 segregation ratio of the tolerant and susceptible phenotypes and suggesting the action of a gene designated as HsBvm-1. Bulked segregant analysis (BSA) was used. The most tolerant and susceptible individuals were pooled and subjected to restriction site associated DNA sequencing (RAD-Seq) analysis, which identified 7,241 SNPs. A subset of 384 candidate SNPs segregating between bulks were genotyped on the 20 most-tolerant and most-susceptible individuals, identifying a single marker (SNP192) showing complete association with nematode tolerance. Segregation of SNP192 confirmed the inheritance of tolerance by a single gene. This association was further validated on a set of 26 commercial tolerant and susceptible varieties, showing the presence of the SNP192 WB242-type allele only in the tolerant varieties. We identified and mapped on chromosome 5 the first nematode tolerance gene (HsBvm-1) from Beta vulgaris ssp. maritima and released information on SNP192, a linked marker valuable for high-throughput, marker-assisted breeding of nematode tolerance in sugar beet.

Sugar beet resistance to rhizomania: state of the art and perspectives.

The productivity of sugar beet is strongly limited by several biotic stresses, among them rhizomania (induced by Beet necrotic yellow vein virus, BNYVV) which causes yield losses of 20–50% or more. The only way to control this disease is the use of resistant varieties. Sources of resistances have been found in the ancestor of the cultivated beets Beta vulgaris L. ssp. maritima (L.) Arcang. Rz1 is the major resistance gene present within commercial sugar beet varieties. This resistance was recognized as monogenic and dominant. Experimental evidence highlighted that Rz1 originated from sea beets belonging to Munerati’s genetic pool. The development of molecular markers linked to Rz1 allows the use of marker assisted selection (MAS) to introgress this gene into pollinator lines, to assess the trueness of hybridity and to remove off-type individuals. MAS speeds variety development and reduces production costs guaranteeing at the same time a high qualitative standard in variety development. Recent studies have shown an emergence of new BNYVV strains with increased virulence that could overcome Rz1 resistance. Therefore, exploitation of new genetic sources of resistance and the pyramiding of several resistance genes into new breeding lines is becoming a main priority for sugar beet breeding companies to maintain an adequate resistance level to rhizomania.

The Sea Beet (Beta vulgaris L. ssp. maritima) of the Adriatic Coast as Source of Resistance for Sugar Beet

ConclusionsThe sea beet is considered the ancestral species from which the different types of cultivated beet originate. Its hybridation is easy because of its evolutionary proximity, unlike with other species of the GenusBeta. These traits have long made the sea beet the subject of research aimed at targeting and isolating its useful characters which could then be transferred to cultivated beet.The sea beet collected at the mouth of the Po di Levante river in the early 1900s has given more appreciable results than any of the other biotypes from different origins. Indeed, it was from this biotype that today’s most developed genetic resistance to serious diseases such as cercospora and rhizomania was isolated.

Transcriptome and Cell Physiological Analyses in Different Rice Cultivars Provide New Insights Into Adaptive and Salinity Stress Responses

Salinity tolerance has been extensively investigated in recent years due to its agricultural importance. Several features, such as the regulation of ionic transporters and metabolic adjustments, have been identified as salt tolerance hallmarks. Nevertheless, due to the complexity of the trait, the results achieved to date have met with limited success in improving the salt tolerance of rice plants when tested in the field, thus suggesting that a better understanding of the tolerance mechanisms is still required. In this work, differences between two varieties of rice with contrasting salt sensitivities were revealed by the imaging of photosynthetic parameters, ion content analysis and a transcriptomic approach. The transcriptomic analysis conducted on tolerant plants supported the setting up of an adaptive program consisting of sodium distribution preferentially limited to the roots and older leaves, and in the activation of regulatory mechanisms of photosynthesis in the new leaves. As a result, plants resumed grow even under prolonged saline stress. In contrast, in the sensitive variety, RNA-seq analysis revealed a misleading response, ending in senescence and cell death. The physiological response at the cellular level was investigated by measuring the intracellular profile of H2O2 in the roots, using a fluorescent probe. In the roots of tolerant plants, a quick response was observed with an increase in H2O2 production within 5 min after salt treatment. The expression analysis of some of the genes involved in perception, signal transduction and salt stress response confirmed their early induction in the roots of tolerant plants compared to sensitive ones. By inhibiting the synthesis of apoplastic H2O2, a reduction in the expression of these genes was detected. Our results indicate that quick H2O2 signaling in the roots is part of a coordinated response that leads to adaptation instead of senescence in salt-treated rice plants.