Expression of Mdh1 locus alleles in apozygotic progenies of sugar beet (Beta vulgaris L.)

Expression of Mdh1 alleles has been studied in 60 apozygotic (agamospermic) sugar beet progenies. Seed progenies were obtained by uniparental (pollen less) mode of seed reproduction: selfing of pollen-sterile plants isolated with paper bags. The apozygotic seed progenies demonstrate a disomic gamete autosegregation, i.e., the ratio between genotypes in the progenies correspond to the gamete segregation in a duplex heterozygote of an autotetraploid. It was shown that the ratio between the Mdh1 phenotypes in apozygotic progenies is strongly affected by spontaneous inactivation of one of the alleles. In most progenies, the excess of FF phenotypes and the deficit of SS phenotypes were observed. In our opinion, such deviations in genotype and phenotype frequencies result from conversion of the active Mdh1-S into the inactive Mdh1-S0 allele (epigenetic gene inactivation). The spontaneous inactivation of one allele results in extremely variable frequencies of heterozygous Mdh1-F/Mdh1-S genotypes and phenotypes in the apozygotic seed progenies. The empirical distribution of the frequencies of heterozygous genotypes in the apozygotic seed progenies is given by a negative binomial distribution describing the expected time of occurrence of random events.     

Fruit set variation associated with apozygotic reproduction in sugar beet (Beta vulgaris 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 ofparthenocarpic seeds. Waiter 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 (Beta vulgaris L.)

The ratios of the phenotypic classes of glucosephosphate isomerase (GP12) 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.     

Cytogenetics of autotetraploid sugar beet (Beta vulgaris L.)

Summary Autotetraploid and diploid varieties of sugar beet were investigated for morphology, plant development, root and seed yield. The results obtained from the tetraploid varieties were evaluated according to the number of euploid and aneuploid plants found in each variety. Aneuploid plants often are characterized by delayed growth and poor root or seed yield, which reflects in the average yield of the tetraploid variety. Eutetraploid plants will compete successfully with their diploid counterparts. Until chromosomal stability of euploid plants will be found, aneuploid plants can be eliminated by mechanical selection only, which has to be repeated in each generation. A mechanical selection for euploidy will not only lower the amount of aneuploids in the tetraploid varieties, but also among the triploid hybrid seeds.

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Zusammenfassung Autotetraploide und diploide Zuckerrübensorten wurden auf Morphologie, Wachstum sowie Rüben-und Samenertrag untersucht. Die Ergebnisse der tetraploiden Sorten wurden gemäß den in ihnen gefundenen Anteilen von euploiden und aneuploiden Pflanzen ausgewertet. Aneuploide Pflanzen sind oft durch verlangsamtes Wachstum und schlechten Rüben- oder Samenertrag gekennzeichnet, was sich im Durchschnittsertrag der tetraploiden Sorten widerspiegelt. Eutetraploide Pflanzen können erfolgreich mit den entsprechenden diploiden konkurrieren. Ehe nicht chromosomal stabile eutetraploide Pflanzen gefunden werden, können Aneuploide nur durch mechanische Selektion ausgelesen werden. Diese muß in jeder Generation wiederholt werden. Eine mechanische Selektion auf Aneuploide wird die Anzahl der Aneuploiden nicht nur in den tetraploiden Sorten herabsetzen, sondern auch unter den triploiden Hybridsamen.

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Resumen Se investigó variedades autotetraploides y diploides de remolacha azucarera en relación con la morfología, el desarrollo y el rendimiento en raíz y semilla. Los resultados obtenidos de la variedad tetraploide se evaluaron de acuerdo con el número de plantas euploides y aneuploides halladas. Las plantas aneuploides se caracterizan frecuentemente por un crecimiento retardado y una producción pobre en raíz o semilla, propiedades que se reflejan en el rendimiento medio de las variedades tetraploides. Las plantas eutetraploides pueden competir con éxito con sus correspondientes diploides. Mientras no se alcance la estabilidad cromosómica de las plantas euploides, las aneuploides pueden únicamente eliminarse por medio de una selección mecánica, selección que debe repetirse en cada generación. Una selección mecánica para euploidía no sólo rebajará la proporción de aneuploides en las variedades tetraploides sino tambien dentro de las semillas híbridas triploides.

     

A linkage map of sugar beet (Beta vulgaris L.)

Summary We have established a first linkage map for beets based on RFLP, isozyme and morphological markers. The population studied consisted of 96 F₂ individuals derived from an intraspecific cross. As was expected for outbreeding species, a relatively high degree of polymorphism was found within sugar beet; 47% of the DNA markers were polymorphic for the chosen population. The map consists of 115 independent chromosomal loci designated by 108 genomic DNA probes, 6 isozyme and one morphological marker. The loci cover 789 cM with an average spacing of 6.9 cM. They are dispersed over nine linkage groups corresponding to the haploid chromosome number of Beta species. Eighteen markers (15.4%) showed distorted segregation which, in most instances, can be explained by gametic selection of linked lethal loci. The application of the linkage map in sugar beet breeding is discussed.     

Molecular genetic investigation of sugar beet (Beta vulgaris 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.     

Thidiazuron-induced organogenesis and somatic embryogenesis in sugar beet (Beta vulgaris L.)

Summary Improved in vitro tissue culture systems are needed to facilitate the application of recombinant DNA technology to the improvement of sugar beet germplasm. The effects of N ⁶-benzyladenine (BA) and thidiazuron (TDZ) pretreatment on adventitious shoot and somatic embryogenesis regeneration were evaluated in a range of sugar beet breeding lines and commercial varieties. Petiole explants showed higher frequencies of direct adventitious shoot formation and produced more shoots per explant than leaf lamina explants. TDZ was more effective than BA for the promotion of shoot formation. The optimal TDZ concentrations were 2.3–4.6 μM for the induction of adventitious shoot regeneration. Direct somatic embryogenesis from intact seedlings could be induced by either BA or TDZ. TDZ-induced somatic embryogenesis occurred on the lower surface of cotyledons at concentrations of 0.5–2μM and was less genotype-dependent than with Ba. A high frequency of callus induction could be obtained from seedlings and leaf explants, but only a few of the calluses derived from leaf explants could regenerate to plants via indirect somatic embryogenesis. These results demonstrated that TDZ could prove to be a more effective cytokinin for in vitro culture of sugar beet than BA. Rapid and efficient regeneration of plants using TDZ may provide a route for the production of transgenic sugar beet following Agrobacterium-mediated transformation.     

Variability of cathodic peroxidases in sugar beet (Beta vulgaris L.) cultivars

Eighteen varieties of sugar beet (Beta vulgaris L.), originating from various European countries, were compared in respect of peroxidase variability level. They were cultivated in the same experimental plot. The cultivars differed in ploidy level: one variety was tetraploid, three were diploid and 14 varieties were triploid. The cathodic peroxidase system is controlled by four independent genes, of which only one is polymorphic. Consequently, the varieties were characterised by frequencies of 3 allozymes belonging to one locus. Only one variety proved to be fully monomorphic. Genetic similarities between the cultivars were illustrated by a dendrogram (UPGMA) and show different groups of varieties not related to their ploidy level.     

Biolistic transformation of highly regenerative sugar beet (Beta vulgaris L.) leaves

Leaves of greenhouse-grown sugar beet (Beta vulgaris L.) plants that were first screened for high regeneration potential were transformed via particle bombardment with the uidA gene fused to the osmotin or proteinase inhibitor II gene promoter. Stably transformed calli were recovered as early as 7 weeks after bombardment and GUS-positive shoots regenerated 3 months after bombardment. The efficiency of transformation ranged from 0.9% to 3.7%, and stable integration of the uidA gene into the genome was confirmed by Southern blot analysis. The main advantages of direct bombardment of leaves to regenerate transformed sugar beet include (1) a readily available source of highly regenerative target tissue, (2) minimal tissue culture manipulation before and after bombardment, and (3) the overall rapid regeneration of transgenic shoots.     

Flavin excretion from roots of iron-deficient sugar beet (Beta vulgaris L.)

The characteristics of flavin excretion from iron-deficient sugar-beet roots have been studied. Roots from iron-deficient sugar beet excreted flavins when plants were allowed to decrease the pH of the nutrient solution, but not when plants were grown in nutrient solutions buffered at high pH. As shown by reversed-phase high-performance liquid chromatography, the two major flavins whose excretion was induced by iron deficiency were different from riboflavin, FMN and FAD. These flavins have been identified as riboflavin 3′-sulfate and riboflavin 5′-sulfate by electrospray-mass spectrometry, inductively coupled plasma emission spectroscopy, infrared spectrometry and¹H-nuclear magnetic resonance. We have characterized the time courses of accumulation of the different flavins in the nutrient solution and considered several possible roles for flavin excretion in iron acquisition.     

Restriction fragment map of sugar beet (Beta vulgaris L.) chloroplast DNA

Summary A restriction endonuclease fragment map of sugar beet chloroplast DNA (ctDNA) has been constructed with the enzymes SmaI, PstI and PvuII. The ctDNA was found to be contained in a circular molecule of 148.5 kbp. In common with many other higher plant ctDNAs, sugar beet ctDNA consists of two inverted repeat sequences of about 20.5 kbp separated by two single-copy regions of different sizes (about 23.2 and 84.3 kbp). Southern hybridization analyses indicated that the genes for rRNAs (23S+16S) and the large subunit of ribulose 1,5-bisphosphate carboxylase were located in the inverted repeats and the large single-copy regions, respectively.     

Ultrastructural expression of cytoplasmic male sterility in sugar beet (Beta vulgaris L.)

Summary The development of microspore mother cells (MMC) and tapetum in male-fertile and male-sterile anthers of Beta vulgaris L. was compared at the electron microscope level. These studies were complemented by morphometric analyses of mitochondria in both tissues through successive stages of microsporogenesis. The earliest irregularities in the ultrastructure of male-sterile anthers were noted within the tapetum at the tetrad stage. These disturbances were initially expressed by a slight reduction in mitochondrial size and the appearance of concentric configurations of endoplasmic reticulum. As development proceeded, a further decrease in mitochondrial size become more conspicuous and was accompanied by a reduction in ribosome population and a failure of the tapetum to produce Ubisch bodies. This failure to produce Ubisch bodies is reflected in the underdevelopment of sterile microspore exine.     

Genetic analysis of bolting after winter in sugar beet (Beta vulgaris L.)

Key message

This study reveals for the first time a major QTL for post-winter bolting resistance in sugar beet ( Beta vulgaris L.). The knowledge of this QTL is a major contribution towards the development of a winter sugar beet with controlled bolting behavior.

Abstract

In cool temperate climates, sugar beets are currently grown as a spring crop. They are sown in spring and harvested in autumn. Growing sugar beet as a winter crop with an extended vegetation period fails due to bolting after winter. Bolting after winter might be controlled by accumulating genes for post-winter bolting resistance. Previously, we had observed in field experiments a low post-winter bolting rate of 0.5 for sugar beet accession BETA 1773. This accession was crossed with a biennial sugar beet with regular bolting behavior to develop a F₃ mapping population. The population was grown in the greenhouse, exposed to artificial cold treatment for 16 weeks and transplanted to the field. Bolting was recorded twice a week from May until October. Post-winter bolting behavior was assessed by two different factors, bolting delay (determined as days to bolt after cold treatment) and post-winter bolting resistance (bolting rate after winter). For days to bolt, means of F₃ families ranged from 25 to 164 days while for bolting rate F₃ families ranged from 0 to 1. For each factor one QTL explaining about 65 % of the phenotypic variation was mapped to the same region on linkage group 9 with a partially recessive allele increasing bolting delay and post-winter bolting resistance. The results are discussed in relation to the potential use of marker-assisted breeding of winter sugar beets with controlled bolting.     

The "crystals" in the sugar beet (Beta vulgaris L.) leaves

Crystal containing cells widely distributed in plant tissues, though the origin of the crystals and their functions are still opened to question. Membrane vesicles in beet leaves are visible in electronic microscope. They originate in cytoplasm and penetrate into vacuole by pinocytosis with participation of tonoplast. In light microscope, vesicles are luminous likewise crystals in crystal cells. Such vesicles-"crystals" fulfill crystal cells also. The content of vesicles-"crystals" are electronic transparent at every path of leaf development. It was proposed that distinct vesicles-"crystals" in cytoplasm and vacuole and mass of them in crystal cells, vein bundles, and epidermal cells--all of them are lytic compartments. Later, obviously, true crystals are formed.     

Multiple-line cross quantitative trait locus mapping in sugar beet (Beta vulgaris L.)

Linkage mapping based on multiple-line crosses is a promising strategy for mapping quantitative trait loci (QTL) underlying important agronomic traits. The main goal of this survey was to study the advantages of QTL mapping across versus within biparental populations using experimental data from three connected sugar beet (Beta vulgaris L.) populations evaluated for beet yield and potassium and sodium content. For the combined analysis across populations, we used two approaches for cofactor selection. In Model A, we assumed identical cofactors for every segregating population. In contrast, in Model B we selected cofactors specific for every segregating population. Model A performed better than Model B with respect to the number of QTL detected and the total proportion of phenotypic variance explained. The QTL analyses across populations revealed a substantially higher number of QTL compared to the analyses of single biparental populations. This clearly emphasizes the potential to increase QTL detection power with a joint analysis across biparental populations.     

A refined method for ovule culture in sugar beet (Beta vulgaris L.)

Induction of gynogenesis through ovule culture is a valuable tool to produce haploid and doubled haploid plants in sugar beet (Beta vulgaris L.). However, there is still large room for refining the method. In this study we investigated the gynogenic response of cultured ovules of three sugar beet genotypes, the effect of the application to inflorescences of different pretreatments with mannitol at 4ºC and with 5-azacytidine and 2,4-D, and the effect of the use of different basal culture media and sucrose concentrations. The response was evaluated in terms of percentages of induction of gynogenesis, embryogenesis and callogenesis, as well as of regenerated plants. We showed that a pretreatment with 0.5 M mannitol at 4 °C for 4 days, and with 50 µM 5-AzaC for 1 h, notably improved the percentage of embryogenesis and plant regeneration. Besides, the use of MS basal medium and 60 g/L sucrose was also found beneficial. This study provides new ways to improve the efficiency of haploid induction and plant regeneration through ovule culture in sugar beet, and is potentially applicable to ovule culture in other crops.