Crystals in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.) leaves

Crystal-containing cells (C-cells) are widely spread in plant tissues; however, the origin of the crystals and their functions remain a subject of discussion. In sugar beet leaves, the membrane vesicles seen in an electron microscope appear in the cytoplasm and penetrate the vacuole by pinocytosis with the participation of tonoplast. In a light microscope, the vesicles fluoresce like crystals in C-cells. These crystal vesicles also fill the C-cells. The content of crystal vesicles is electron-transparent at all stages of leaf development. It is suggested that both individual crystal vesicles in the cytoplasm and in vacuoles and their agglomerations in C cells, vascular bundles, and epidermal cells are lytic compartments. Later, true crystals seem to be formed.     

Molecular genetic investigation of sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

Molecular genetic studies of sugar beet (Beta vulgaris L.) are reviewed as a basis for the development of genomics of this species. The methods used to study structural and functional genomics are considered. The results and their application to increase the efficiency of sugar beet breeding are discussed.     

Multi-trait association mapping in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

Association mapping promises to overcome the limitations of linkage mapping methods. The main objective of this study was to examine the applicability of multivariate association mapping with an empirical data set of sugar beet. A total of 111 diploid sugar beet inbreds was selected from the seed parent heterotic pool to represent a broad diversity with respect to sugar content (SC). The inbreds were genotyped with 26 simple sequence repeat markers chosen according to their map positions in proximity to previously identified quantitative trait loci for SC. For SC and beet yield (BY), the genotypic variances were highly significant (P < 0.01). Based on the global test of the bivariate mixed-model approach, four markers were significantly associated with SC, BY, or both at a false discovery rate of 0.025. All four markers were significantly (P < 0.05) associated with BY but only two with SC. The identification of markers associated with SC, BY, or both indicated that association mapping can be successfully applied in a sugar beet breeding context for detection of marker-phenotype associations. Furthermore, based on our results multivariate association mapping can be recommended as a promising tool to discriminate with a high mapping resolution between pleiotropy and linkage as reasons for co-localization of marker-phenotype associations for different traits.     

Leaf area prediction model for sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.) cultivars

In two successive years (2003 and 2004), a set of 16 commercial sugar beet cultivars was established in Randomized Complete Block experiments at two sites in central Greece. Cultivar combination was different between years, but not between sites. Leaf sampling took place once during the growing season and leaf area, LA [cm²], leaf midvein length, L [cm] and maximum leaf width, W [cm] were determined using an image analysis system. Leaf parameters were mainly affected by cultivars. Leaf dimensions and their squares (L², W²) did not provide an accurate model for LA predictions. Using L×W as an independent variable, a quadratic model (y = 0.003 x² − 1.3027 x + 296.84, r ² = 0.970, p<0.001, n = 32) provided the most accurate estimation of LA. With compromises in accuracy, the linear relationship between L×W and LA (y = 0.5083 x + 31.928, r ² = 0.948, p<0.001, n = 32) could be used as a prediction model thanks to its simplicity.     

Efficient somatic embryogenesis in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.) breeding lines

Efficient regeneration via somatic embryogenesis (SE) would be a valuable system for the micropropagation and genetic transformation of sugar beet. This study evaluated the effects of basic culture media (MS and PGo), plant growth regulators, sugars and the starting plant material on somatic embryogenesis in nine sugar beet breeding lines. Somatic embryos were induced from seedlings of several genotypes via an intervening callus phase on PGo medium containing N⁶-benzylaminopurine (BAP). Calli were mainly induced from cotyledons. Maltose was more effective for the induction of somatic embryogenesis than was sucrose. There were significant differences between genotypes. HB 526 and SDM 3, which produced embryogenic calli at frequencies of 25–50%, performed better than SDM 2, 8, 9 and 11. The embryogenic calli and embryos produced by this method were multiplied by repeated subculture. Histological analysis of embryogenic callus cultures indicated that somatic embryos were derived from single- or a small number of cells. 2,4-dichlorophenoxyacetic acid (2,4-D) was ineffective for the induction of somatic embryogenesis from seedlings but induced direct somatic embryogenesis from immature zygotic embryos (IEs). Somatic embryos were mainly initiated from hypocotyls derived from the cultured IEs in line HB 526. Rapid and efficient regeneration of plants via somatic embryogenesis may provide a system for studying the molecular mechanism of SE and a route for the genetic transformation of sugar beet.     

Association mapping in multiple segregating populations of sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

Association mapping in multiple segregating populations (AMMSP) combines high power to detect QTL in genome-wide approaches of linkage mapping with high mapping resolution of association mapping. The main objectives of this study were to (1) examine the applicability of AMMSP in a plant breeding context based on segregating populations of various size of sugar beet (Beta vulgaris L.), (2) compare different biometric approaches for AMMSP, and (3) detect markers with significant main effect across locations for nine traits in sugar beet. We used 768 F _n (n = 2, 3, 4) sugar beet genotypes which were randomly derived from 19 crosses among diploid elite sugar beet clones. For all nine traits, the genotypic and genotype × location interaction variances were highly significant (P < 0.01). Using a one-step AMMSP approach, the total number of significant (P < 0.05) marker-phenotype associations was 44. The identification of genome regions associated with the traits under consideration indicated that not only segregating populations derived from crosses of parental genotypes in a systematic manner could be used for AMMSP but also populations routinely derived in plant breeding programs from multiple, related crosses. Furthermore, our results suggest that data sets, whose size does not permit analysis by the one-step AMMSP approach, might be analyzed using the two-step approach based on adjusted entry means for each location without losing too much power for detection of marker-phenotype associations.     

Fruit set variation associated with apozygotic reproduction in sugar beet <Emphasis Type=Italic>Beta vulgaris</Emphasis> L.

Fruit set upon apozygotic reproduction was studied for four years in the msSOAN-5 pollen-sterile inbred sugar beet line. The progenies obtained from pollen-sterile plants by apozygotic reproduction had both fruits with normal seeds and parthenocarpic fruits without seeds, which was not an occasional event. Growth conditions proved to strongly affect the fruit set and seed quality. For instance, water deficiency during early plant development increased the proportion of parthenocarpic seeds. Water deficiency combined with a lower temperature during flowering additionally caused a substantial decrease in the total number of fruits. Under the same growth conditions, related accessions did not differ in seed productivity, but varied in the proportion of normal (with seeds) and parthenocarpic (without seeds) fruits.     

Autosegregation of enzyme loci in agamospermous progenies of triploid plants of sugar beet (<Emphasis Type=Italic>Beta vulgaris</Emphasis> L.)

The ratios of the phenotypic classes of glucosephosphate isomerase (GPI2) and malate dehydrogenase (MDH1 and MDH2) were studied in agamospermous progenies of triploid sugar beet plants. The ratio of the phenotypic classes of these enzymes corresponds to the calculations based on the assumption of polyteny of chromosomes carrying alleles of the enzyme loci accompanied by the loss of extra copies of the alleles in the first division of a cell entering embryogenesis. An increase in the gene dosage due to polyteny leads to the appearance in the progeny with a definite frequency of alleles that were absent in the original parental plant. The notions of “meiotic autosegregation” and “mitotic autosegregation” characteristic of meiotic and mitotic agamospermy are introduced, as well as the term locus polygenotype characterizing not only the allelic composition and the number of chromosomes, but also the number of chromatids carrying alleles of the marker locus in the cell before its entry into embryogenesis.     

Using the method of natural samples for studying the variability in sugar beat (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.) agamospermous seed progeny

The existence of natural genetic and natural mixed samples of seeds was theoretically grounded and experimentally demonstrated. A natural genetic sample is detected when analyzing the seed agamospermous progeny in the sequence coinciding with the sequence of seed setting, i.e., in the order they are located on a branch. The method of natural genetic samples has demonstrated the presence of individual regions with nonrandom distribution of phenotypic classes of agamospermous seeds on the branches of sugar beet plants. This phenotype distribution reflects the state of somatic cells on extended branch regions and the manifestation of this state in seed-bud cells. In turn, the genotype of each seed reflects the state of the seed-bud cell (apozygote) that gave rise to embryo development. We also demonstrated heterogeneity in the ratios of enzyme phenotypes of the seeds set on individual branches, regarding it as a result of the changes arising in somatic tissues at the moment of branching. Analysis of the ratios of seed phenotypes on individual branches of the same plant was named the method of natural mixed samples. The combination of natural genetic and natural mixed samples provides for identifying and analyzing various modes of seed setting (gamospermy and agamospermy) and studying the factors that influence the variability in these processes. We postulated that the nonrandom distribution and ratio of the seed phenotypes on plant branches resulted from a nonrandom endoreduplication of the homologous regions in homologous chromosomes carrying the marker locus alleles.     

Micro-morphology of common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) nodules undergoing senescence

We studied morphological changes over time by nodules formed on the root system of the common bean (Phaseolus vulgaris L.). Two cultivars, Bayomex and Cacahuate 72 with growth habit Type I and the Rhizobium etli strain CE-3 were used. The results showed the collapse of the infected zone, degradation of the cell walls and membranes, changes in the number and distribution of the starch granules, appearance of protein granules, and disintegration of the central tissue of the nodule with ageing. Additionally, we describe the influence of time on the progress of the nodular senescence.     

Growth and photosynthetic efficiency promotion of sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.) by endophytic bacteria

Very little is known about the physiological interactions between plants and endophytic bacteria. We investigated the impact of three endophytic bacteria, Bacillus pumilus 2-1, Chryseobacterium indologene 2-2, and Acinetobacter johnsonii 3-1, on the photosynthetic capacity and growth of sugar beet. Endophyte-free plants were obtained first and infected with the bacteria. Measurements of total chlorophyll content revealed very significant differences between endophyte-free beet plants and some infected by endophytic bacteria. The maximum photochemical yield (Fv/Fm) was used to determine any photosynthetic effect on plants caused by biotic or abiotic factors. After 30 days of growth, there was significantly higher Fv/Fm for endophyte-infected than endophyte-free plants. The light response curves of beet showed that photosynthetic capacity was significantly increased in endophyte-infected plants. Photosynthesis of endophyte-free plants was saturated at 1,300 μmol m⁻² s⁻¹, whereas endophyte-infected plants were not saturated at the irradiance used. The effect seemed to be due to promotion of electron transport in the thylakoid membranes. Promotion of photosynthetic capacity in sugar beet was due to increased chlorophyll content, leading to a consequent increased carbohydrate synthesis. It is possible that the increased maximum yield of photosynthesis in sugar beet was promoted by phytohormones and produced by the bacteria.     

<Emphasis Type="Italic">Bacillus anthracis</Emphasis>, <Emphasis Type="Italic">Francisella tularensis</Emphasis> and <Emphasis Type="Italic">Yersinia pestis</Emphasis>. The most important bacterial warfare agents — <Emphasis Type="Italic">review</Emphasis>

There are three most important bacterial causative agents of serious infections that could be misused for warfare purposes: Bacillus anthracis (the causative agent of anthrax) is the most frequently mentioned one; however, Fracisella tularensis (causing tularemia) and Yersinia pestis (the causative agent of plague) are further bacterial agents enlisted by Centers for Disease Control and Prevention into the category A of potential biological weapons. This review intends to summarize basic information about these bacterial agents. Military aspects of their pathogenesis and the detection techniques suitable for field use are discussed.     

<Emphasis Type="Italic">Pseudocercospora griseola</Emphasis> Causing Angular Leaf Spot on <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> Produces 1,8-Dihydroxynaphthalene-Melanin

Pseudocercospora griseola is the causal agent of angular leaf spot of common bean (ALS). It has undergone parallel coevolution with its host and two major groups have been defined, “Andean” (P. griseola f. griseola) and “Mesoamerican” (P. griseola f. mesoamericana). The aim of this study was to analyze the nature and the level of the dark pigment synthesized by the representatives of each group. After 21 days of incubation on potato dextrose agar medium, P. griseola f. griseola isolate S3b developed colonies with diameters of 17.5 ± 1.3 mm and concentric rings of pigmentation. Isolate T4 of P. griseola f. mesoamericana presented smaller colonies (9.9 ± 0.3 mm) with a uniform dark-gray color. Both isolates, S3b and T4, produced the same pigment, a 1,8-dihydroxynaphthalene-melanin, although different in quantity and structural features as suggested by the IR spectrum. The P. griseola f. griseola isolate S3b had a higher growth rate and melanin content as well as smaller sensitivity to melanin synthesis inhibitors compared to the isolate T4 of P. griseola f. mesoamericana. These results suggest a possible link between melanin and growth in P. griseola.     

A two-stage pretreatment of seedlings improves adventitious shoot regeneration in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

The effects of a two-stage pretreatment of seedlings on the subsequent shoot regeneration capacity were investigated. Pretreated seedlings were obtained by germinating seeds on three different germination media and then further culturing on six different growth media. Lamina and petiole explants of two sugar beet (Beta vulgaris L.) breeding lines were then excised from the pretreated seedlings and cultured on five different shoot regeneration media. In both breeding lines, petiole explants produced significantly more shoots than lamina explants with higher frequencies of organogenic capacities; petiole explants of the lines M1195 and ELK345 produced a mean of 2.1 and 2.7 shoots per explant while their lamina explants produced 1.5 and 2.2 shoots per explant, respectively. A genotypic variation was evident as the line ELK345 was more productive for shoot development from both types of explants. In overall comparisons of different germination, growth and regeneration media, germination medium was most effective when supplemented with 0.5 mg/l 6-benzyladenine (BA) while both growth and regeneration media were most productive when contained a combination of 0.25 mg/l BA and 0.10 mg/l indole-3-butyric acid (IBA). Of all the treatments tested, the highest mean number of shoots per explant (8.3 shoots) and frequency of organogenic explants (75.6%) were obtained on regeneration medium supplemented with 0.25 mg/l BA and 0.10 mg/l IBA when petiole explants of the line ELK345 were excised from the seedlings that had been germinated on medium containing 0.5 mg/l BA followed by further growth on medium containing 0.25 mg/l BA and 0.10 mg/l IBA.     

Comparative proteomic analysis of apomictic monosomic addition line of <Emphasis Type="Italic">Beta corolliflora</Emphasis> and <Emphasis Type="Italic">Beta vulgaris</Emphasis> L. in sugar beet

Apomixis refers to a process in which plants produce seed without fertilization through female syngamy that produces embryos genetically identical to the maternal parent. In sugar beet, interspecific hybrids between diploid Beta vulgaris and tetraploid Beta corolliflora were established and monosomic addition line M14 was selected because of the apomictic phenotype. By using two-dimensional electrophoresis gels we identified the proteins which were differently expressed between the M14 and B. vulgaris. A total of 27 protein spots which varied expressed between lines were isolated and successfully identified with MALDI-TOF MS. Among them five protein spots were found to be only presented in M14 and two protein spots only expressed in Beta. According to their functional annotations described in Swissprot database, these proteins were, respectively, involved in important biological pathways, such as cell division, functionally classified using the KEGG functional classification system. The result may be useful for us to better understand the genetic mechanism of apomixes.     

Frequencies of the <Emphasis Type="Italic">DRB1</Emphasis>, <Emphasis Type="Italic">DQA1</Emphasis>, <Emphasis Type="Italic">DQB1</Emphasis> and <Emphasis Type="Italic">TNFA</Emphasis> alleles in immigrant population of west Siberia

Based on population analysis of the DRB1, DQA1, DQB1 and TNFA allele frequency distribution patterns, regional features of immunogenetic structure of the population of West Siberia were investigated. Statistically significant linkage disequilibrium within the HLA class II region, as well as between the TNFA and DRB1, DQA1, and DQB1 was demonstrated. Population frequency distribution patterns of two- and multilocus haplotypes were examined.     

Isolation of a <Emphasis Type="Italic">Ve</Emphasis> homolog, <Emphasis Type="Italic">mVe1</Emphasis>, and its relationship to verticillium wilt resistance in <Emphasis Type="Italic">Mentha longifolia</Emphasis> (L.) Huds

As a step toward greater understanding of the genetics of verticillium wilt resistance in plants, we report the sequencing of a candidate wilt resistance gene, mVe1, from the mint diploid model species, Mentha longifolia (Lamiaceae). mVe1 is a putative homolog of tomato (Solanum lycopersicum L.) verticillium wilt (Ve) resistance genes. The mVe1 gene has a coding region of 3,051 bp. The predicted mVe1 protein contains a leucine-rich repeat domain, a common feature of plant disease resistance proteins. We compared 13 mVe1 alleles from three mint species. These alleles shared 96.2–99.6% nucleotide identity. We analyzed four M. longifolia populations segregating with respect to mVe1 alleles and wilt resistance versus susceptibility and found one association between mVe1 genotype and wilt phenotype. We conclude that mVe1 may play a role in mint verticillium wilt resistance, but variation for resistance in our segregating progenies is likely polygenic. Therefore, further investigations of mVe1 and identification of additional candidate genes are both warranted.