Protein-enriched feedstuff from beet pulp

Of five moulds that were cultivated in a medium containing beet pulp, Trichoderma reesel was superior in protein production (34%) and in conversion of pulp to proteins (18% w/w). Protein ylelds were improved by up to 49% by pre-treatment of the beet pulp, adjustment of the solld/llquid and liquld/solid ratios, the incubation period, nitrogen level and addition of some trace elements as well as with inoculum size. The protein obtained contained all essential amino acids and compared favourably with those of the FAO guldeline and of soy bean oll meal.K.M. Ghanem and M.A. El-Gazaerly are with the Botany Department, Faculty of Science, Alexandria University, Alexandria, Egypt. A.H. El-Refai is with the Microbiological Laboratory, NRC, Cairo, Egypt.     

Effects of organic matter on sulphur oxidation in soil and influence of sulphur oxidation on soil nitrification

Summary The effects of wheat straw and pressed sugar beet pulp on sulphur oxidation were determined in a loam soil amended with 1% (w/w) elemental sulphur. Wheat straw stimulated the oxidation of elemental sulphur over the first 2 to 3 weeks of the incubation period, resulting in an increase in LiCl-extractable sulphate. After 4 to 7 weeks incubation however, the only significant increase in soil sulphate followed the 1% straw addition, while at week 7 sulphate concentrations in the 0.25% and 5.0% straw amended soils were lower than the control. Pressed sugar beet pulp (1% w/w) initially stimulated the oxidation of elemental sulphur in the soil, but by weeks 3 to 7 of the incubation period rates of oxidation in pulp-amended soils were lower than the control. Towards the end of the incubation period however, sulphate concentrations in the amended soils exceeded the control values, significantly so by week 11. The concentration of thiosulphate and tetrathionate also increased in soils receiving sugar beet pulp. Nitrification was inhibited in soils in which sulphur oxidation was actively occurring. Although possible alternatives are mentioned, such inhibition appears to result from a decrease in soil pH brought about by the oxidation of elemental sulphur to sulphuric acid.     

Xylanase production by Trichoderma reesei Rut C-30 grown on L-arabinose-rich plant hydrolysates

The suitability of L-arabinose-rich plant hydrolysates as carbon sources and inducers of xylanase production in Trichoderma reesei Rut C-30 was tested. Significantly higher xylanase activities were obtained in cultures on oat husk and sugar beet pulp hydrolysates than on lactose. In batch culture with oat husk hydrolysate and lactose, the xylanase activity was about 9 times higher ( approximately 510 IU/ml) than in lactose ( approximately 60 IU/ml). Even higher xylanase activity ( approximately 630 IU/ml) was obtained when the batch cultivations were done on sugar beet pulp hydrolysate and lactose. In a fed-batch culture using oat husk hydrolysate-lactose the xylanase activity was as high as 1350 IU/ml in 4 days. The cellulase production clearly decreased when T. reesei was cultured on both hydrolysates compared to the cultivation on lactose. Moreover, the relative amounts of the xylanases I-III were similar regardless the used carbon source.     

Dried sugar beet pulp as a high energy feed for beef cattle

Digestibility trials with 8 lots of pelleted dried sugar beet pulp, carried out with wethers, demonstrated that dried sugar beet pulp is a highly digestible energy source for ruminants. From the digestion coefficients obtained, and a value number of 95, the starch equivalent of the dry matter of dried sugar beet pulp was calculated to be 73.3, while the net energy expressed in EF_r was 619 EF_r per kg dry matter. This is about 90% of the net energy content of barley containing 4% crude fibre in the dry matter.Beef production trials were carried out with 702 young bulls fed on complete dry rations based on dried sugar beet pulp. Two categories of animals were used: 322 baby-beef bulls (intensive system) slaughtered at 13 months of age at an average live-weight of 480 kg; and 380 young bulls coming from pasture (semi-intensive system) at about 250 kg live-weight, and fattened indoors up to at least 550 kg live-weight. With each category, three different rations have been studied. These contained respectively, 50, 60 or 70% pelleted dried sugar beet pulp; the remainder of the rations consisted of respectively 50, 40 or 30% concentrates. The diets were fed ad libitum; straw and water were always available. The three complete dry rations proved to be equally successful for intensive beef production. The carcass quality was good for all animals. The average daily gain obtained with the baby-beef bulls for the three rations respectively was 1 207 g, 1 274 g and 1 172 g; for the second category of bulls the mean growth rates were generally slightly higher: 1 281 g, 1 309 g and 1 357 g.The feed efficiency was higher with the younger animals: the baby-beef bulls (live-weight interval: 150–480 kg) consumed about 2.5 kg protein supplement and 3.5 kg dried sugar beet pulp per kg live-weight gain; while the intake per kg live-weight gain with the bulls of the second category (live-weight interval: 250–560 kg) amounted approximately to 2.75 kg protein supplement and 4 kg dried sugar beet pulp. Within each category of bulls, the feed cost per kg live-weight gain decreased with increasing amounts of dried sugar beet pulp in the rations.     

Suitability of commercial beet molasses fractions as substrates fro polyhydroxyalkanoate production byAzotobacter vinelandii UWD

Summary Beet molasses that had been fractionated commercially by ion exclusion resulted in two waste-streams: extract molasses (EM) and concentrated separator by-product (CSB). Only EM at 4–5% w/v contained sufficient sugar to promote polyhydroxybutyrate (PHB) formation byAzotobacter vinelandii UWD, but the yield of PHB/protein was less than that obtained in unfractionated beet molasses. EM and especially CSB added at 0.5–2.0% w/v to media containing a variety of sugar sources promoted an increased yield of PHB/protein. The best use of these beet molasses fractions was, therefore, as a minor addition to media containing sugars to increase PHB yield, but not as a primary substrate for PHB production.     

Chelating agents improve enzymatic solubilization of pectinaceous co-processing streams

This study investigates the hypothesis that loosening of the egg-box structure by presence of divalent ion chelating agents during enzymatic degradation of homogalacturonan (HG) can improve enzymatic polysaccharide solubilization on pectinaceous, agro-industrial co-processing streams. The influence of different levels of ethylene-diaminetetraacetic acid (EDTA), citric acid, oxalic acid, and phosphate was assessed in relation to enzymatic solubilization of isopropanol precipitatable oligo- and polysaccharides from sugar beet pulp, citrus peel, and two types of potato pulp. The two types of potato pulp were FiberBind 400, a dried commercial potato pulp product, and PUF, a dried calcium reduced product, respectively. The enzymatic treatment consisted of 1% (w/w) of substrate treated with pectin lyase from Aspergillus nidulans and polygalacturonase from A. aculeatus [each dosed at 1.0% (w/w) enzyme/substrate] at 60 °C, pH 6.0 for 1 min. Characterization of the released fractions demonstrated a significantly improved effect of chelating agents for polysaccharide solubilization from FiberBind 400, PUF, and citrus peel, whereas only low amounts of polysaccharides were solubilized from the sugar beet pulp. The results substantiated the importance of chelating agents during enzymatic extraction of pectinaceous polysaccharides. Lower levels of chelating agents were required for the calcium-reduced potato pulp substrate (PUF) indicating the significance of calcium cross-linking in HG in relation to the enzymatic solubilization yields. The effect of the chelating agents correlated to their dissociation constants (pK_a values) and calcium binding constants and citric acid and EDTA exerted highest effects. Maximum polysaccharide yield was obtained for FiberBind 400 where the enzymatic treatment in presence of citric acid yielded 22.5% (w/w) polysaccharides of the initial substrate dry matter.     

Influence of different fibre sources on digestibility and nitrogen and energy balances in growing pigs

Abstract

The present study was undertaken to investigate how three different fibre sources, sugar beet pulp, soya bean hulls and pectin residue, constituting 15% of diets for growing pigs, influenced daily body gain, feed conversion, apparent faecal digestibility and nitrogen and energy balances. Eight castrated crossbreed pigs (30 – 80 kg live weight) were used in a replicated 4 × 4 Latin-square design with one control diet and three fibre containing diets. Daily body weight gain and feed conversion were not affected by the dietary treatments. The apparent faecal digestibility of organic matter (OM) and energy were significantly lower for the fibre diets (OM: 0.81 – 0.85; energy: 0.78 – 0.83) compared to the control diet (OM: 0.88; energy: 0.86). The apparent faecal digestibility of crude protein (CP) was lower for the fibre diets (0.71 – 0.78) compared to the control diet (0.83), although it was only significantly lower for the sugar beet pulp and pectin residue diets. The pectin residue diet, which contained the highest amount of dietary fibre, lignin and insoluble non-starch polysaccharides, had the lowest digestibility of OM, CP and energy. There was a tendency (p = 0.07) for a diet effect on retained nitrogen in proportion to digested nitrogen, where the sugar beet pulp and pectin residue diets had numerically the highest values. Heat production and retained energy in proportion to metabolizable energy intake were not affected by fibre inclusion. It was concluded that the inclusion of sugar beet pulp, soya bean hulls and pectin residue in diets for growing pigs decreased the apparent faecal digestibility and in the diets with sugar beet pulp and pectin residue higher utilization of digested nitrogen for retention compensated for the lower amount of digested nitrogen.     

Utilization of agro-industrial orange peel and sugar beet pulp wastes for fungal endo- polygalacturonase production

The pectinase enzymes are involved in several industrial applications, and industrial waste is one of the largest environmental pollutants, so this study aims to Endo-polygalacturonase (endo-PG) producing using Aspergillus niger AUMC 4156, Penicillium oxalicum AUMC 4153 and P. variotii AUMC 4149 by using some agro-industrial wastes (dried orange peel and sugar beet pulp) as a sole raw carbon source for degradation these waste in the process of urban wastes disposal. The fermentation process was carried out as a submerged culture technique under both shaken and static culture conditions. A. niger AUMC 4156 was the most promising producer of endo-PG under static conditions while P. oxalicum AUMC 4153 was the highest producer of endo-PG under shaken conditions. Sugar beet pulp proved to be the most preferable to orange peel as the only source of carbon in both shaken and static cultures. The medium that encompassing orange peel as a single carbon source afforded the highest protein content with all tested fungal strains in stirred and static cultures in comparison with sugar beet pulp. The highest activity of endo-polygalacuronase that produced using A. niger AUMC 4156 and P. oxalicum AUMC 4153 was achieved by using sugar beet pulp at 3% concentration under static cultures, meanwhile maximal enzyme activity produced by both fungal strains required 2% sugar beet pulp under shaken cultures. Sugar beet pulp showed promised potential as a good inducer for endo-polygalacturoase production, and enzymes production depended on fungal strains, culture medium, and submerged fermentation conditions.     

Influence of different fibre sources on digestibility and nitrogen and energy balances in growing pigs

The present study was undertaken to investigate how three different fibre sources, sugar beet pulp, soya bean hulls and pectin residue, constituting 15% of diets for growing pigs, influenced daily body gain, feed conversion, apparent faecal digestibility and nitrogen and energy balances. Eight castrated crossbreed pigs (30-80 kg live weight) were used in a replicated 4 x 4 Latin-square design with one control diet and three fibre containing diets. Daily body weight gain and feed conversion were not affected by the dietary treatments. The apparent faecal digestibility of organic matter (OM) and energy were significantly lower for the fibre diets (OM: 0.81-0.85; energy: 0.78-0.83) compared to the control diet (OM: 0.88; energy: 0.86). The apparent faecal digestibility of crude protein (CP) was lower for the fibre diets (0.71-0.78) compared to the control diet (0.83), although it was only significantly lower for the sugar beet pulp and pectin residue diets. The pectin residue diet, which contained the highest amount of dietary fibre, lignin and insoluble non-starch polysaccharides, had the lowest digestibility of OM, CP and energy. There was a tendency (p = 0.07) for a diet effect on retained nitrogen in proportion to digested nitrogen, where the sugar beet pulp and pectin residue diets had numerically the highest values. Heat production and retained energy in proportion to metabolizable energy intake were not affected by fibre inclusion. It was concluded that the inclusion of sugar beet pulp, soya bean hulls and pectin residue in diets for growing pigs decreased the apparent faecal digestibility and in the diets with sugar beet pulp and pectin residue higher utilization of digested nitrogen for retention compensated for the lower amount of digested nitrogen.     

Effects of pressed beet pulp silage inclusion in maize-based rations on performance of high-yielding dairy cows and parameters of rumen fermentation

Beet pulp contains high amounts of pectins that can reduce the risk of rumen disorders compared to using feedstuffs high in starch. The objective was to study the effects of inclusion of ensiled pressed beet pulp in total mixed rations (TMR) for high-yielding dairy cows. Two TMR containing no or about 20% (on dry matter (DM) basis) beet pulp silage were used. The beet pulp silage mainly replaced maize silage and corn cob silage. The TMR were intentionally equal in the concentrations of energy and utilisable crude protein (CP) at the duodenum. TMR were fed to 39 and 40 dairy cows, respectively, for 118 days. The average daily milk yield was about 43 kg/day. No significant differences in milk yield and milk fat or milk protein content were detected. DM intake of cows was significantly reduced by the inclusion of beet pulp silage (23.0 v. 24.5 kg/day). However, a digestibility study, separately conducted with sheep, showed a significantly higher organic matter digestibility and metabolisable energy concentration for the TMR that contained beet pulp silage. In vitro gas production kinetics indicated that the intensity of fermentation was lower in the TMR that contained beet pulp silage. In vitro production of short-chain fatty acids, studied using a Rusitec, did not differ between the TMR. However, the inclusion of beet pulp silage in the ration caused a significant reduction in the efficiency of microbial CP synthesis in vitro. The amino acid profile of microbial protein remained unchanged. It was concluded that beet pulp silage has specific effects on ruminal fermentation that may depress feed intake of cows but improve digestibility. An inclusion of beet pulp silage of up to 20% of DM in rations for high-yielding dairy cows is possible without significant effects on milk yield and milk protein or milk fat.     

Decolorization of sugar syrups using commercial and sugar beet pulp based activated carbons

Sugar syrup decolorization was studied using two commercial and eight beet pulp based activated carbons. In an attempt to relate decolorizing performances to other characteristics, surface areas, pore volumes, bulk densities and ash contents of the carbons in the powdered form; pH and electrical conductivities of their suspensions and their color adsorption properties from iodine and molasses solution were determined. The color removal capabilities of all carbons were measured at 1/100 (w/w) dosage, and isotherms were determined on better samples. The two commercial activated carbons showed different decolorization efficiencies; which could be related to their physical and chemical properties. The decolorization efficiency of beet pulp carbon prepared at 750 degrees C and activated for 5h using CO2 was much better than the others and close to the better one of the commercial activated carbons used. It is evident that beet pulp is an inexpensive potential precursor for activated carbons for use in sugar refining.     

Stilbella aciculosa: A Potential Biocontrol Fungus against Rhizoctonia solani

A semi‐solid fermentation product of the potential biocontrol fungus Stilbella aciculosa was formulated on wheat bran: water (1:1, w/w) and incubated 5, 10 and 15 days before addition to soil infested with the pathogen Rhizoctonia solani. Generally, preparations did not reduce survival of the pathogen in infested beet seed but they did prevent saprophytic growth of the pathogen from beet seed into soil. The magnitude of reduction by the 15‐day‐old inoculum was greater than that by the 5‐day‐old inoculum. Ten‐day‐old bran preparations of S. aciculosa at rates of 0.5 and 1.0% (w/w) in soil prevented post‐emergence damping‐off of cotton, radish and sugar beet in the glasshouse and a rate of 1.0% gave stands similar to those in the non‐infested control soil. The antagonist, grown on perlite formulated with molasses, cornmeal, alfalfa tissue or corn stover, prevented damping‐off of cotton in a naturally infested soil. However, the stands were not as great as that in soil planted with pentachloronitrobenzene (PCNB)‐treated seed. Toxic metabolites, produced by S. aciculosa developing on various substrates, slightly inhibited the growth of R. solani in culture and induced cytoplasmic leakage of the pathogen mycelium.     

Release of ferulic acid from agroindustrial by-products by the cell wall-degrading enzymes produced by Aspergillus niger I-1472

Aspergillus niger I-1472 was grown on sugar beet pulp to produce cell wall polysaccharide-degrading enzymes, including feruloyl esterases. Compared to enzymatic activities measured in commercially available mixtures previously used for the release of ferulic acid, the A. niger enzymes were more various. These enzymes were tested to release ferulic acid from sugar beet pulp, maize bran, or autoclaved maize bran. They were as efficient as the commercial mixture to release ferulic acid from sugar beet pulp. On the other hand, they were much more efficient to release ferulic acid from maize bran after autoclaving pretreatment, as 95% of ferulic acid ester were solubilized. Thus, A. niger enzymes exhibited a high interest in the release of ferulic acid from various agro-industrial by-products.     

Solid state fermentation for the production of α-amylase from Penicillium chrysogenum using mixed agricultural by-products as substrate

Production of α-amylase from local isolate, Penicillium chrysogenum, under solid-state fermentation (SSF) was carried out in this study. Different agricultural by-products, such as wheat bran (WB), sunflower oil meal (SOM), and sugar beet oil cake (SBOC), were used as individual substrate for the enzyme production. WB showed the highest enzyme activity (750 U/gds). Combination of WB, SOM, and SBOC (1:3:1 w/w/w) resulted in a higher enzyme yield (845 U/gds) in comparison with the use of the individual substrate. This combination was used as mixed solid substrate for the production of α-amylase from P. chrysogenum by SSF. Fermentation conditions were optimized. Maximum enzyme yield (891 U/gds) was obtained when SSF was carried out using WB + SOM + SBOC (1:3:1 w/w/w), having initial moisture of 75%, inoculum level of 20%, incubation period of 7 days at 30°C. Galactose (1% w/w), urea and peptone (1% w/w), as additives, caused increase in the enzyme activity.     

In vitro fermentability and physicochemical properties of fibre substrates and their effect on bacteriological and morphological characteristics of the gastrointestinal tract of newly weaned piglets

Abstract

Fermentability of fibre has a great impact on the bacterial flora along the gastrointestinal tract of newly weaned piglets. Therefore, this parameter was determined by incubating in vitro different fibre substrates (chicory roots, sugar beet pulp, wheat bran and corn cobs) with contents of jejunum or caecum sampled from slaughtered pigs. Incubating with small intestinal contents, lactic acid was the only fermentation product. Fermentability was highest for chicory roots, followed by wheat bran and sugar beet pulp, while corn cobs were not fermented. Based on SCFA formed in the incubations with caecal contents, ranking of the fermentability of the fibre substrates was in the same order. The effect of adding different fibre substrates to diets of newly weaned piglets on bacteriological and morphological aspects of the gastrointestinal tract was also investigated. In Experiment 1 three groups of five piglets, weaned at four weeks of age, received a control feed (C), C supplemented with corn cobs (50 g/kg) or with chicory roots (20 g/kg). In Experiment 2, diet C was supplemented with sugar beet pulp (120 g/kg) or with wheat bran (75 g/kg). After three weeks animals were euthanized and digesta were sampled from stomach, proximal and distal jejunum, caecum and colon. Furthermore, mucosal scrapings were prepared and tissue samples were taken from jejunum, caecum and colon. Viscosity was determined for jejunal, caecal and colon contents. Corn cobs in the feed increased the number of total bacteria, lactobacilli and bifidobacteria in the stomach and proximal duodenum, while a decreased count of streptococci in distal jejunum contents was noted. Chicory roots increased the counts of Escherichia coli in the distal jejunum and on the mucosa, while sugar beet pulp decreased the number of lactobacilli on the mucosa only. Wheat bran seemed to increase the count of E. coli in jejunal digesta and on the mucosa, and also the number of lactobacilli in the stomach and jejunum. Bifidobacterial numbers were increased but only in the proximal part of the jejunum. Fibre substrates affected the concentration of lactate and SCFA in different parts of the intestinal tract. Feeding corn cobs increased villus length in the proximal jejunum by 13%. The number of intra-epithelial lymphocytes in the villous epithelium of proximal and distal jejunum was decreased by corn cobs and chicory roots supplementation while beet pulp and wheat bran had the opposite effect. In Experiment 1, apoptotic index of the mucosa of the distal jejunum was very low and decreased when corn cobs were fed. Mitotic index in the crypts was only affected by the wheat bran diet and a small decrease was noted. It was concluded that the fermentability of fibre was not an ideal criterion for predicting its effects on the flora. The effect of fibres on viscosity of digesta was negligible probably explaining the lack of clear and consistent influences on the intestinal mucosa.     

Use of the GFP reporter as a vital marker for Agrobacterium-mediated transformation of sugar beet (Beta vulgaris L.)

Molecular approaches to sugar beet improvement will benefit from an efficient transformation procedure that does not rely upon exploitation of selectable marker genes such as those which confer antibiotic or herbicide resistance upon the transgenic plants. The expression of the green fluorescent protein (GFP) signal has been investigated during a program of research that was designed to address the need to increase the speed and efficiency of selection of sugar beet transformants. It was envisaged that the GFP reporter could be used initially as a supplement to current selection regimes in order to help eliminate “escapes” and perhaps eventually as a replacement marker in order to avoid the public disquiet associated with antibiotic/herbicide-resistance genes in field-released crops. The sgfp-S65T gene has been modified to have a plant-compatible codon usage, and a serine to threonine mutation at position 65 for enhanced fluorescence under blue light. This gene, under the control of the CaMV 35S promoter, was introduced into sugar beet via Agrobacterium-mediated transformation. Early gene expression in cocultivated sugar beet cultures was signified by green fluorescence several days after cocultivation. Stably transformed calli, which showed green fluorescence at a range of densities, were obtained at frequencies of 3–11% after transferring the inoculated cultures to selection media. Cocultivated shoot explants or embryogenic calli were regularly monitored under the microscope with blue light when they were transferred to media without selective agents. Green fluorescent shoots were obtained at frequencies of 2–5%. It was concluded that the sgfp-S65T gene can be used as a vital marker for noninvasive screening of cells and shoots for transformation, and that it has potential for the development of selectable marker-free transgenic sugar beet.     

Simultaneous saccharification and protein enrichment fermentation of sugar beet pulp

Summary A product with 40 % protein content was obtained from sugar beet pulp (1.25–2.0 mm) in 48 h one stage (simultaneous) saccharification/fermentation process under optimized conditions using a specific enzyme mixture andCandida tropicalis strain, also saving about 40 % enzymes in comparison to a 2-stage process.     

Newly Isolated Bacillus gibsonii S-2 Capable of using Sugar Beet Pulp for Alkaline Pectinase Production

Summary A Bacillus strain capable of growing under highly alkaline conditions was isolated from a sample of alkaline soil. The isolate was a Gram-positive, spore-forming, aerobic, alkaliphilic bacterium and was designated as strain S-2. Growth of the strain was observed in the pH range of 7–12 and temperature range of 4–40 °C. Sequence analysis of the 16S rDNA of the strain revealed more than 99% homology with the strains of Bacillus gibsonii. The S-2 strain was confirmed as B. gibsonii by comparing its physiological and biochemical characteristics with the B. gibsonii DSM 8722 strain. The S-2 strain could use sugar beet pulp as the carbon source as well as the pectinase inducer to produce extracellular alkaline pectinase by solid-state fermentation. The maximum polygalacturonase yield of 3600 U/g dry sugar beet pulp was obtained at 35 °C after 48 h of incubation.     

Azetidine-2-carboxylic acid in the food chain

Azetidine-2-carboxylic acid (Aze) 1 is a non-protein amino acid present in sugar beets and in table beets (Beta vulgaris). It is readily misincorporated into proteins in place of proline 2 in many species, including humans, and causes numerous toxic effects as well as congenital malformations. Its role in the pathogenesis of disease in humans has remained unexplored. Sugar beet agriculture, especially in the Northern Hemisphere, has become widespread during the past 150 years, and now accounts for nearly 30% of the world’s supply of sucrose. Sugar beet byproducts are also used as a dietary supplement for livestock. Therefore, this study was undertaken as an initial survey to identify Aze-containing links in the food chain. Herein, we report the presence of Aze 1 in three sugar beet byproducts that are fed to farm animals: sugar beet molasses, shredded sugar beet pulp, and pelleted sugar beet pulp.     

Cytoplasmic male sterility in hybrids of sterile wild beet (Beta vulgaris ssp. maritima) and O-type fertile sugar beet (Beta vulgaris L.): molecular analysis of mitochondrial and nuclear genomes

The organization of the mitochondrial genome of B3, B4 and B5generations of hybrids created by backcrossing sterile wild beet Betamaritima with a fertile O-type sugar beet line was studied usingrestriction fragment length polymorphism (RFLP) analysis. Random amplifiedpolymorphic DNA (RAPD) analysis was used to study restoration of the fertile(O-type) sugar beet genotype in hybrids after multiple backcrossings.Restriction of mtDNAs from the cytoplasm of B. maritimaandhybrids revealed BamHI, EcoRI andXhoI restriction patterns different from those for sterileand fertile sugar beet lines. The most conspicuous feature of our accession ofsterile wild beet mtDNA was the absence of the 10.7-kbEcoRI fragment detected in the cytoplasm of S-type sterileB. maritima and sugar beet. The hybridization of digestedmtDNAs with coxII, atpA andatp6 homologous probes revealed alterations within thesegene loci that distinguished wild beet and hybrids from sugar beets.Characteristic hybridization profiles for the wild beet and B3, B4 and B5hybrids were observed for all probes regardless of the restrictase used todigest mtDNA. Notable changes in atpA andatp6 genes resulted when probes that comprised the5flanking sequences of these genes and a small part of the coding sequences wereused. RFLP analysis of the sterile B. maritimamitochondrial genome further supported the unique character of this source ofwild beet sterility. The genotypic differences between hybrids and parentalaccessions were determined by scoring PCR-RAPD reaction products for nineselected primers. The diversity of the B. maritimagenotyperesulted in a lower genetic similarity index in comparison with hybrids,sterileand fertile lines of sugar beet. The dendrogram obtained after cluster analysisdistinguished hybrids as a group that differed from wild beet and themaintainersugar beet line used for backcrossing. These results may indicate incompleterestoration of the fertile sugar beet genotype in hybrids.