Overwintering of genetically modified sugar beet, Beta vulgaris L. subsp. vulgaris, as a source for dispersal of transgenic pollen

The potential impact of transgenic crops on community ecology will depend on the distribution and establishment of the new transgenic traits, on the sexual transfer of their new genes to the environment (Bartsch &; Pohl-Orf, 1996) and on the potential ecological impact of the transgenic trait. Flowering and pollen dispersal is important for outcrossing of the genetically engineered trait. For a biennial plant, like the cultivars of Beta vulgaris L., overwintering is normally necessary to become generative and to produce pollen and seeds (Abe et al., 1997), which usually does not happen with sugar beet as a field crop harvested in autumn (Longden 1989). The starting point for the project was a transgenic sugar beet, Beta vulgaris L. subsp. vulgaris (Lange et al., 1998), with rhizomania and herbicide ( Basta®, Liberty®) resistance. Cold tolerance is one of the most important factors for survival of sugar beet in Central- and North-Europe. Among other ways, spreading of transgenic traits into weed beet (Boudry et al., 1993) or wild beet can occur if genetically engineered – biennial – plants survive the winter, flower in spring and spread their pollen. Field experiments were performed with transgenic breeding lines and their hybrids, transgenic and non-transgenic hybrids with Swiss chard and three conventional beet cultivars to evaluate winter survival rates at seven different field sites. We could show that survival of sugar beet – transgenic as well as conventional ones – in Germany and at the Dutch border is possible. Survival rates were well correlated with temperature data and were unexpectedly high. Differences between sugar beet hybrids and breeding lines could be detected but not within different breeding lines or hybrids. There were no differences detectable between transgenic and non-transgenic plants. The data are crucial for the risk assessment of the release of transgenic sugar beet and are the basis for further experiments towards outcrossing and establishment.     

Assessing fitness parameters of hybrids between weed beets and transgenic sugar beets

Herbicide resistance is a desired trait for sugar beet (Beta vulgaris L.) production because it is a low‐competitor crop that requires careful weed management. However, gene flow to weed beet (also B. vulgaris) could jeopardize the weed control strategy by causing the emergence of resistant weed beets; it could also lead to further adaptation of weed beet as a troublesome weed for other crops by selecting more competitive plants. To evaluate the hazard that such a selection process represents, apart the herbicide resistance, we investigated the morphology and reproduction of progeny of weed beets having inherited a herbicide resistance gene. First‐generation resistant weed beet exhibited traits likely counter‐selected. But such crop traits rapidly disappeared with backcrossing to weedy relatives: no biologically relevant difference was noted between resistant and susceptible near‐isogenic siblings in the various experiments. In the absence of resistance selection, our data indicate low chances for weed beet to evolve more competitive forms than present weed beet populations. However, they also suggest that there is no fitness cost limiting transgenes spread.     

Breeding for resistance to rhizomania in sugar beet: A review

Currently rhizomania is the most important disease in sugar beet worldwide, and attack can lead to serious yield losses. The disease is caused by beet necrotic yellow vein virus (BNYVV) that is transmitted by the soil-borne fungus Polymyxa betae. Breeding sugar beet cultivars with resistance to rhizomania is regarded as the most appropriate way to enable continued production of this crop in BNYVV-infested fields and also to slow the spread of the disease. Breeding for resistance started with selection by scoring disease symptoms in field experiments. The development of non-destructive greenhouse tests, with determination of the virus concentration in rootlets using ELISA, has greatly improved the efficiency of selection. In this paper the impact of scientific research on the progress in breeding cultivars with resistance to rhizomania is reviewed. This includes the distribution, composition, and pathogenicity of the virus, the sources of resistance to virus and vector, the genetics of virus resistance, progress with breeding methods, and the use of molecular markers and pathogen-derived resistance. The yields and quality characteristics of recently introduced resistant cultivars now equal those of the commercial susceptible cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Reduced titer of BNYVV in transgenic sugar beets expressing the BNYVV coat protein

Summary Rhizomania is a disease of sugar beet caused by the furovirus beet necrotic yellow vein virus (BNYVV). Coat protein mediated resistance has been reported for a number of viral diseases. This approach to virus resistance was therefore attempted for control of rhizomania. Two constructs of the coat protein gene of BNYVV were introduced into sugar beet by Agrobacterium-mediated transformation. The mRNA level was estimated to be 0.01% of the poly A⁺ RNA. Expression of the coat protein gene was under the detection limit of our western blotting protocol i.e. below 0.01 g/50 g (0.02% of the total soluble protein). One transformation event per construct was tested in a greenhouse assay and in rhizomania infested soil in a field trial. In the greenhouse assay, transgenic plants showed a strong reduction of virus multiplication when compared to non-transgenic plants. This result was confirmed in the field trial, where a significant difference in virus multiplication was shown between plants with and without the coat protein gene.     

Sugar beets homozygous for resistance to beet cyst nematode (Heterodera schachtii Schm.), developed from monosomic additions of Beta procumbens to B. vulgaris

Summary Monosomic additions of Beta vulgaris x B. procumbens with resistance to beet cyst nematode (Heterodera schachtii Schm.) were used for the production of resistant diploids through incorporation of the B. procumbens chromosome fragment bearing the resistance gene(s) into one of the sugar beet chromosomes. The heterozygotes obtained accordingly were selfed for producing homozygotes. These homozygotes differed morphologically from commercial sugar beet varieties, but produced reasonable amounts of pollen. Female transmission of resistance was 100%, whereas male transmission, apart from some exceptions, was more than 90%. The number of hypersensitivity reactions to penetrated larvae was related to the degree of susceptibility. Larval development was severely retarded in the resistant plants, preventing most of them to produce cysts. If cysts were formed, their content was considerably less as compared to those in the susceptible plants.     

Multiplicative models for cultivar by location interaction in testing sugar beets for resistance to beet necrotic yellow vein virus

Summary Sugar beet cultivars were evaluated for resistance to beet necrotic yellow vein virus (BNYVV) on various locations in two consecutive years. Resistance levels of cultivars were measured by virus assays of plants from the field and the greenhouse. Infection levels in the fields were characterised by sampling plants of a susceptible indicator cultivar. For each year, statistical analyses were performed on two-way tables of cultivar by location for yield and quality parameters. In analysis of variance (ANOVA) significant main effects and significant cultivar by location interaction were found for all parameters (P<0.05). Interactions were further investigated by multiplicative models. In the Additive Main effects and Multiplicative Interaction effects (AMMI) model, interaction was written as the product of a cultivar score and a location score. Cultivar interaction scores were highly correlated to virus concentrations of the cultivars, and location interaction scores to virus concentrations of the susceptible indicator cultivar. Main effects of cultivars and locations were less clearly related to virus concentrations than interaction effects. In general, virus concentrations of plants from a greenhouse test gave higher correlations than virus concentrations of plants from the field. In the factorial regression model, virus concentrations were incorporated in the model. The model can be understood as a two-way ANOVA, with greenhouse virus concentrations and virus concentration of the indicator cultivar as concomitant variables on the cultivar and location factor. Results of analyses with both multiplicative interaction models showed that interactions of all yield and quality parameters can be described in terms of virus concentrations. Therefore, the relative performance of susceptible and partially resistant cultivars in infested fields can be estimated by means of three independent parameters, (i) the level of resistance determined in a greenhouse experiment, (ii) the yield and quality in non-infested fields, and (iii) the level of infection in the field.Abbreviations AMMI model Additive Main effects and Multiplicative Interaction effects model - ANOVA analysis of variance - BNYVV beet necrotic yellow vein virus - ELISA enzyme-linked immunosorbent assay - -amino N -amino nitrogen - K Kalium (potassium) - Na Natrium (sodium)     

Breeding sugar beets with globe-shaped roots: selection and agronomical performance

Losses and costs during and after harvesting of sugar beets can be reduced by modifying the traditional shape of the roots. Crosses have been made between sugar beets and globe-shaped red table beets, and selection for root shape and sugar content was carried out in the progenies. The root yield of the selected material is equal to that of commercial varieties. The sugar content is still too low and the contents of juice impurities too high, but further selection can improve the material in these respects. The amount of dirt tare can be 40–60% less than that of the control variety.     

The inheritance of resistance to beet necrotic yellow vein virus (BNYVV) in B. vulgaris subsp. maritima, accession WB42: Statistical comparisons with Holly-1-4

In this study, the inheritance of resistance to Beet necrotic yellow vein virus (BNYVV) in accessions Holly-1-4and WB42 was investigated. Crosses between both resistant sources and susceptible parents were carried out and F₁F₂ and BC₁ populations were obtained. Virus concentrations in WB42and its F₁ populations were lower than in Holly-1-4. Observed ratios of susceptible and resistant plants in segregating populations of Holly-1-4 as well as WB42 were in agreement with hypothesis of one dominant major gene. Segregation of plants in F₂ populations obtained from crosses betweenHolly-1-4 and WB42 revealed that the resistance genes in Holly-1-4 and WB42 were nonallelic and linked loci. This revised version was published online in July 2006 with corrections to the Cover Date.     

Differential occurrence of beet cryptic viruses—A new tool for germplasm characterization and evolutionary studies in beets?

Summary Beet cryptic viruses occur differentially in cultivars of chard (only BCV 1) and red beet (only BCV 2). As landraces of these crops and wild beets can contain both of the viruses, the differential occurrence in cultivars should indicate founder effects in the course of evolution of chard and red beet. Consequently, the genetic base of those crops should be considered as narrow. On the basis of the presence or absence of the viruses in beet plants within the populations conclusions can be drawn concerning the uniformity of cultivars maintained in the collections, the characterization of chard and beet landraces and some evolutionary tendencies within the variable Beta vulgaris-complex.     

Partial resistance of sugarbeet to beet cyst eelworm (Heterodera schachtii Schm.)

Summary Within Beta vulgaris and B. maritima origins some partial resistance to beet cyst eelworm was found which could be raised to a very limited extent by selection. However after the second backcross to commercial sugar beet varieties and successive selection of the inbreds this resistance was lost. It was demonstrated that in the rootsystem of resistant plants as much nematodes penetrate and develop as in susceptible ones but the ratio between males and females is different. It was therefore quite probable that this resistance is polyfactorial and merely recessive.     

Inheritance of resistance to pendimethalin herbicide induced stem damage in soybean

Selective herbicides are valuable weed control tools; however, selectivity is not always complete, resulting in crop damage. Stem breakage, lodging, and enlarged hypocotyls (brittle bean syndrome) limit yields of soybean [Glycine max (L.) Merr.] genotypes treated with pendimethalin [N-(1-ethylpropyl)-3,4-dimethyl-2,6-dinitrobenzenamine]. Developing genotypes with resistance to pendimethalin injury would eliminate or reduce this problem. Genetic studies were conducted to determine the inheritance of resistance to pendimethalin induced stem damage. The F_2:3 progeny of crosses involving resistant ('Asgrow A4715' and 'Flyer') and susceptible (`Essex' and K87-7-95) genotypes were screened in a greenhouse. Each genotype was treated with 1.68 kg ha^-1 pendimethalin preemergence and irrigated as needed. Plants were scored at V4 for stem breakage. Progeny distributions indicated that resistance to brittle bean syndrome damage behaved as a quantitative trait. Dominance for stem breakage was expressed in the population of A4715 × Essex. Flyer was more sensitive to herbicide damage than Asgrow A4715 because it has fewer genes for resistance or different alleles. The F_2:3 variance component heritability estimates ranged from 0.19 to 0.52. Gain from selection for resistance to pendimethalin injury is possible, and resistant progeny can be recovered from segregating populations. This revised version was published online in August 2006 with corrections to the Cover Date.     

Natural variation within the genus Beta and its possible use for breeding sugar beet: A review

Summary Sugar beet is a relatively young crop, which supposedly has a narrow genetic base. Natural variation occurring in primitive beet varieties and in wild Beta species has been used for breeding sugar beet. This paper reviews information on desirable characteristics in Beta germplasm and the attempts made for the introgression of such characters into commercial breeding material. After an introduction on the availability of germplasm and the possibilities of hybridisation, attention is focussed on the mating system (especially male sterility), on morphological and physiological characteristics, including yield and sugar content, and on resistances to diseases and pests.     

Peroxidase activity as a possible test for distinguishing s-plasm from n-plasm in sugar beet (Beta vulgaris L.)

Summary The O-type lines (N-plasm) of sugar beet (Beta vulgaris L.) proved to have a consistently lower peroxidase activity (49.2–62.7%) than their male sterile (MS) equivalents (S-plasm).Similar differences in peroxidase activity (45.4–56.3%) were found when O-type lines were compared with unrelated MS lines. They were also observed in different parts of the plant (cotyledons, hypocotyl + radicle, true leaves) and were reproducible.The fertile lines could be grouped by their level of peroxidase activity in categories of either S-plasm or N-plasm.The peroxidase activity test could be used for selection of N-plasm lines from fertile sugar beet populations and for selection of MS and O-type lines of higher stability.     

Induction of cytoplasmic male sterility and fertility reversion by mutagenesis; negative evidence from a study on sugar beet

Summary Attempts have been made to induce cytoplasmic male sterility (cms) and cytoplasmically inherited fertility reversion in sugar beet using various mutagenic agents by repeating techniques previously described. Despite the previous report of successful induction of cms and reversion at very high frequency it was not possible to repeat this success. Analysis of M₂ plants at the DNA level demonstrated no major rearrangements in chloroplast or mitochondrial DNA.     

Major genes for resistance to beet necrotic yellow vein virus (BNYVV) in Beta vulgaris

Summary Inheritance of resistance to beet necrotic yellow vein virus (BNYVV) was studied in segregating F2 and backcross families obtained from crosses between resistant plants of the sugar beet selection Holly-1-4 or the wild beet accession Beta vulgaris subsp. maritima WB42 and susceptible parents. Greenhouse tests were carried out, in which seedlings were grown in a mixture of sand and infested soil. Virus concentrations of BNYVV in the rootlets were estimated by ELISA. To discriminate resistant and susceptible plants, mixtures of normal distributions were fitted to log₁₀ virus concentrations, estimated for segregating F1, F2 and BC populations of both accessions. The hypothesis that Holly-1-4 contained one single dominant major gene was accepted. For WB42, results fitted with the hypotheses that resistance was based on either one (or more) dominant major gene(s) showing distorted segregation, or two complementary dominant genes, which are both required for resistance. Resistance from WB42 appeared to be more effective against BNYVV than resistance from Holly-1-4.This research was carried out as part of a PhD study at the Graduate School Experimental Plant Sciences (EPS), Department of Virology, Wageningen, The Netherlands     

Sources of resistance to diseases of sugar beet in related Beta germplasm: II. Soil-borne diseases

Between 580 and 700 accessions of related cultivated and wild species of the genus Beta were assessed for resistance to four soil-borne diseases of sugar beet: two seedling damping-off diseases caused by the fungi Aphanomyces cochlioides and Pythium ultimum and two diseases of more mature plants, Rhizoctonia root and crown rot, caused by the fungus R. solani, and Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV), a furovirus transmitted by the plasmodiophorid Polymyxa betae. Analysis of resistance data (assessed on an international standardised 1–9 scale of Resistance Scores) indicated that the highest levels of resistance ({RS} 2) to A. cochlioides and P. ultimum were to be found amongst accessions of the more distantly related sections Corollinae (93% of accessions tested) and Procumbentes (10%), respectively; although useful levels could also be found in the more closely related, and sexually compatible, section Beta (1–6%). Resistance to Rhizoctonia was also found in section Beta (5–7%), depending on whether field or glasshouse tests were used, but there was little evidence of generally high levels of resistance to Rhizomania among accessions of this section. None of the accessions of sections Corollinae and Procumbentes exhibited any notable resistance to Rhizoctonia. However, all sections Procumbentes and some sections Corollinae (4%) accessions were highly resistant to Rhizomania. Individuals with high levels of resistance to Rhizomania were identified from within some section Beta and Corollinae accessions, in which there was evidence of segregation.     

An evaluation of single plant randomised field trials of sugar beet (Beta vulgaris L.)

Summary Diploid, monogerm families of sugar beet were grown in transplanted field trials randomised on a single-plant basis. Individual roots were weighed and a range of chemical characters were assessed. There was no significant difference in performance between plants that had members of their own family as nearest neighbours within the row and plants that had randomly assigned neighbours. Good correlations were found between the performance of families in single-plant trials and their performance in conventional plot trials. We conclude that single-plant randomisation could prove useful for genetical studies and for breeding trials particularly when seed or facilities are limited.     

Evaluation of resistance to verticillium wilt in diploid,wild potato interspecific hybrids

Summary Verticillium wilt (V. albo-atrum Reinke & Berthold or V. dahliae Kleb) threatens potato (Solanum tuberosum L.) production in most growing areas of the world. Genetic resistance offers the most cost-effective and environmentally-sound control measure. However, there is a dearth of genetic and breeding information on resistance to verticillium wilt in potato, because of obscure parentage of some standard cultivars and the complex segregation at the tetraploid level. The wide range of genetic variability in wild relatives of potatoes can be potentially useful as a source of disease resistance, such as verticillium wilt resistance. Six diploid, wild, interspecific Solanum hybrids involving resistant x resistant and susceptible x resistant crosses, were assayed for verticillium wilt resistance under greenhouse conditions to evaluate their potential as sources of verticillium wilt resistance. The cross between S. gourlayi Oka. and S. chacoense Bitt. and its reciprocal had the most resistant progenies based on mean colony counts. No simple mode of inheritance can be proposed based on the observed segregation ratios. However, the continuous distributions observed on verticillium wilt disease response among hybrid families indicate that inheritance of resistance may be polygenic and complex. In addition, skewness of colony count distributions toward the resistance parents were characteristic of all resistant x susceptible crosses suggesting that resistance may be dominant. By contrast, the susceptible x susceptible cross showed a more normal distribution. Overall, the cross between S. gourlayi and S. chacoense showed the most potential as a source of verticillium wilt resistance.