QTL analysis of Cercospora leaf spot resistance in sugar beet

The inheritance of Cercospora leaf spot resistance in sugar beet was investigated by means of quantitative trait loci (QTL) analysis of a segregating population of 193 individuals, using 110 AFLP and 35 restriction fragment length polymorphism (RFLP) markers. Five QTL were found through composite interval mapping on linkage groups 1, 2, 3, and 9, respectively, two of which were linked on linkage group 3. The significance of these QTL was tested by permutation analysis The QTL had mostly additive, but also certain negative dominance effects; all the resistance alleles came from the Cercospora-resistant parent. Each quantitative trait locus accounted for 7-18% of the phenotypic variation, leaving 37% of the variation unexplained. The results are discussed in relation to the potential use of marker-assisted breeding for Cercospora leaf spot resistance in sugar beet.     

Marker analysis for quantitative traits in sugar beet

Many economically important traits are inherited quantitatively and are analysed by breeders in replicated field trials. If dense maps are available, chromosomal regions containing quantitative trait loci (QTL) can be identified and this opens up the possibility of preselecting for quantitative traits in the laboratory. In this study, QTL analysis for yield and yield components in sugar beet is used in two different populations tested in several environments in both populations, QTL were detected for all traits investigated, and their predictive value in breeding schemes was analysed by correlating predicted with observed values. Tolerance to Rhizomania, caused by a gene on chromosome 3, was the main source of genotype‐environment interaction in one population, allowing selection on a QTL basis within macro‐environments with or without Rhizomania infestation, respectively. No clear results were found for the second population tested in environments with and with‐out Cercospora infestation. Consequences for breeding strategies are discussed.     

甜菜无融合生殖单体附加系M14大孢子发生期间细胞壁胼胝质的变化

应用脱色苯胺蓝诱导荧光法观察了甜菜单体附加系M14（Beta vulgaris L., VV+1C、2n=18+1）正常有性生殖与二倍体孢子生殖时，大孢子发生期间细胞壁内胼胝质的变化，结果如下。韭型（Allium odorum-type）胚囊大孢子发生时，自大孢子母细胞的珠孔端细胞壁内出现胼胝质荧光，并逐渐扩展到整个细胞壁，中期Ⅰ至末期Ⅰ细胞壁呈现胼胝质荧光。二分体时，珠孔端大孢子细胞壁内胼胝质荧光消失，二分体之间的横壁以及合点端功能大孢子的侧壁上荧光明显。二倍体功能大孢子的合点端细胞壁内的胼胝质荧光消失。单核胚囊形成后，其细胞壁内无胼胝质荧光，而退化的大孢子细胞壁胼胝质荧光显著。蝶须型（Antennaria-type）胚囊大孢子发生时，大孢子母细胞、二倍体功能大孢子的细胞壁均无胼胝质荧光。蓼型（Polygonum-type）胚囊大孢子母细胞减数分裂时，其珠孔端细胞壁出现胼胝质荧光，并逐渐扩展到整个细胞壁。二分体、三分体、四分体时期，胼胝质荧光主要存在于大孢子之间的横壁上，侧壁内胼胝质荧光较弱。退化的大孢子细胞壁胼胝质荧光明显，功能大孢子细胞壁上缺少胼胝质荧光。此外，本文还讨论了大孢子母细胞减数分裂与细胞壁内沉积胼胝质之间的相关性。     

Chromosome localisation of genes for resistance to Heterodera schachtii, Cercospora beticola and Polymyxa betae using sets of Beta procumbens and B. patellaris derived monosomic additions in B. vulgaris

Beet cyst nematodes (BCN, Heterodera schachtii), Cercospora beticola, and rhizomania, caused by the beet necrotic yellow vein virus (BNYVV) and vectored by the soil-borne fungus Polymyxa betae, are the most serious diseases of sugar beet ( Beta vulgaris subsp. vulgaris). The wild Beta species of section Procumbentes are known to be completely resistant to H. schachtii, C. beticola and P. betae. Alien monosomic additions (2n=19), plants of cultivated beet (2n=18) carrying different individual chromosomes of B. procumbens (2n=18) or B. patellaris (2n=36), were tested in greenhouse experiments for resistance to these pathogens. Gene(s) conferring full resistance to the beet cyst nematode in B. patellaris are located on chromosome 1.1, and the other tested chromosomes of B. patellaris are not involved in the expression of resistance. Artificial inoculation under greenhouse conditions, with in vitro produced inoculum of C. beticola and spot-percentage rating of the disease intensity, showed that the high level of resistance that was observed in the wild species B. procumbens and B. patellaris was not found in any of the monosomic additions tested. It was suggested that genes on various chromosomes of the wild species are needed to express full resistance, and that the chromosomes of group 7 of B. patellaris and chromosome 7 of B. procumbens have the largest effect. The greenhouse tests for resistance to P. betae in B. patellaris derived monosomic additions showed that the addition families of group 4.1 have a strong partial resistance, while the addition families of group 8.1 appeared to be completely resistant to the pathogen. Resistance to P. betae in the two wild species as well as in the two resistant addition types did not exclude infection with BNYVV, but resulted in a considerable reduction of the virus concentration. It was concluded that resistance to the vector would complement virus resistance, and may provide a more effective and durable control of rhizomania. This revised version was published online in July 2006 with corrections to the Cover Date.     

Association between resistance to Cercospora and yield in commercial sugarbeet hybrids

This report documents the difficulty breeders have experienced in combining resistance to Cercospora leaf spot (causal agent Cercospora beticola Sacc.) with high yield in sugarbeet (Beta vulgaris L.). Forty commercial hybrids, all recommended for Cercospora-threat areas, were grown in a Cercospora-free and a diseased (inoculated) environment in 1991 and 1992. A 2.9 Mg/ha decrease in root yield associated with each increment increase in susceptibility confirmed that under a severe epiphytotic (1991) Cercospora resistance provided substantial protection. Under less-severe disease conditions (1992) there was no apparent relationship between yield and resistance, suggesting that the benefits of resistance were similar to the yield potential sacrificed to obtain the resistance. In the absence of the disease, root yields increased 2.7 Mg/ha for each increment of increased susceptibility. There was no evidence of association between sucrose concentration and resistance in the Cercospora-free environment. Despite the limited efforts and/or success in developing resistant commercial hybrids, the demonstrated ability of Cercospora to produce fungicide-resistant strains and the possibility that effective fungicides will not be available provide incentives to seek genetic resistance through breeding efforts.     

A marker-assisted analysis of bolting tendency in sugar beet (Beta vulgaris L.)

Bolting tendency in sugar beet varies among breeding lines and cultivars. Four crosses were made between breeding lines susceptible and resistant to bolting in order to study the genetic basis of bolting tendency. Bolting percentage in F₂, after 8 weeks of low temperature treatment, varied among the crosses, suggesting a complicated genetic control of bolting tendency. Different segregation ratios were observed, in particular, between families derived from the bolting F₁ plants and those from their non-bolting siblings, the former families showing a significantly higher bolting percentage than the latter. A marker-assisted analysis with seven isozyme loci, Ak1, Gdh2, Idh1, Lap, Mdh1, Pgi2 and Pgm1, revealed that a locus with marked effect on bolting tendency was located near Idh1. Because of a close linkage of Idh1 with B for annuality, the gene tagged by Idh1 appeared to be equivalent or similar to B′ for easy bolting allelic to B as reported by Owen et al. (1940). The results obtained suggest that the B locus may not only have an important role in determining growth habits but also control various degrees of bolting tendency in individual sugar beet plants. A linked pair, Ak1-Lap, and Pgi2 also were found to affect bolting tendency, although their effects varied depending on the crosses and families tested. This revised version was published online in July 2006 with corrections to the Cover Date.     

Combined linkage maps and QTLs in sugar beet (Beta vulgaris L.) from different populations

The construction of genetic maps is an expensive and time-consuming process. The breeder is therefore interested in using maps developed from other mapping populations but this is only possible if the genetic structure is similar for the chromosomal regions of interest. In this paper, maps of three populations of sugar beet (Beta vulgaris L.) with common polymorphic marker loci are compared. Maps were constructed with MAPMAKER 3.0 and JOINMAP 2.0. Both mapping programs gave, in general, the same order for common markers. However, the number of common markers was too low to construct a combined map for all chromosomes. For one population, in contrast to the other two, the map constructed with MAPMAKER 3.0 was much longer than that constructed with JOINMAP 2.0. For two of these populations yield traits were also available from different environments. For quantitative trait loci (QTL) analysis of the yield data, the packages MAPMAKER/QTL 1.1 and PLABQTL were used. No QTL common for the two populations could be detected. The program and the version used strongly influenced the estimated positions of QTLs. There was also a strong interaction with environments.     

Molecular markers linked to rhizomania resistance in sugar beet, Beta vulgaris, from two different sources map to the same linkage group

Rhizomania, one of the most important diseases of sugar beet, is caused by beet necrotic yellow vein virus, a Furovirus vectored by the fungus Polymyxa betae Keskin. Reduction of the production losses caused by this disease can only be achieved by using tolerant cultivars. The objective of this study was the identification and mapping of random amplified polymorphic DNA (RAPD) markers linked to a rhizomania resistance gene. The RAPD markers were identified using bulked segregant analysis in a segregating population of 62 individuals derived by intercrossing plants of the resistant commercial hybrid GOLF, and the resistance locus was positioned in a molecular marker linkage map made with a different population of 50 GOLF plants. The resistance locus, Rr1, was mapped to linkage group III of our map of Beta vulgaris L. ssp. vulgaris, which consisted of 76 RAPDs, 20 restriction fragment length polymorphisms (RFLPs), three sequence characterized amplified regions (SCARs) and one sequence tagged site (STS). In total, 101 molecular markers were mapped over 14 linkage groups which spanned 688.4 cM with an average interval length of 8.0 cM. In the combined map, Rr1 proved to be flanked by the RAPD loci RA411₁₈₀₀ and AS7₁₁₀₀ at 9.5 and 18.5cM, respectively. Moreover, in our I₂ population, we found that a set of markers shown by Barzen et al. (1997) to be linked to the ‘Holly’ type resistance gene was also linked to the ‘GOLF’-type resistance gene. These results appeared to indicate that the rhizomania resistance gene present in the GOLF hybrid could be the same gene underlying resistance in ‘Holly’-based resistant genotypes. Two other explanations could be applied: first, that two different alleles at the same locus could have been selected; second, that two different genes at two different but clustered loci underwent the selection process.     

The efficiency of between and within full-sib family selection in a recurrent selection programme in sugar beet (Beta vulgaris L.)

The efficiency of between and within family selection in a full-sib family recurrent selection programme was investigated using data from three cycles of recurrent selection. Correlations between mid-parent values and offspring were high for sugar content and juice purity characters, but low for root weight and sugar yield. This suggests that single root selection within full-sib families is effective in the improvement of sugar content and juice purity, but ineffective for root weight and sugar yield. Root weight and sugar yield will respond better to between full-sib family selection. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and mapping of random amplified polymorphic DNA markers linked to a rhizomania resistance gene in sugar beet (Beta vulgaris L.) by bulked segregant analysis

Bulked segregant analysis was employed to identify random amplified polymorphic DNA (RAPD) markers linked to a gene that confers rhizomania resistance to a sugar beet line created from a Holly Sugar Company breeding population (USA). Polymorphism revealed with 160 arbitrary 10-mer oligonucleotide primers was screened in two bulks produced by separately pooling the individual DNAs from the six most resistant and the six most susceptible plants of an F₂ population segregating for rhizomania resistance. A study of the F₂ individuals showed that 19 primers generated 44 polymorphic markers which were then grouped into nine linkage groups. By analysis of variance, 12 were shown to have a significant effect upon the level of resistance and were mapped on a segment 22.3 cM long. A quantitative trait locus (QTL) of resistance was identified and located in a 4.6cM interval between two markers. It accounted for 67.4% of the observed variation and almost all the genetic variation. These results suggest that the identified QTL corresponds to a unique major gene conditioning the Holly resistance studied, which we have named Rz-l.     

甜菜无融合生殖单体附加系M14大孢子发生的超微结构

应用透射电镜技术探求甜菜单体附加系M14 (Beta vulgaris L.,VV+1C, 2n=18+1)有性生殖大孢子与二倍体孢子生殖大孢子发生时的超微结构特征,了解有性生殖和无融合生殖起始细胞的超微结构差别。甜菜单体附加系M14为兼性无融合生殖体,大孢子发生有韭型、蝶须型和蓼型三种类型。韭型和蓼型大孢子发生时均起源于大孢子母细胞,二分体之前从形态结构上难以区分;减数分裂前期I,细胞核中出现核液泡,形成联会复合体,细胞质改组,细胞壁上沉积胼胝质。韭型大孢子发生时,只进行减数第一次分裂,不发生减数第二次分裂,形成二分体,珠孔端二分体细胞退化,合点端二分体细胞发育为二倍体功能大孢子。蓼型二分体的珠孔端细胞在减数第二次分裂前或分裂过程中退化,合点端细胞经减数第二次分裂形成两个细胞,构成三分体,最终合点端大孢子发育为单倍体功能大孢子。蝶须型大孢子发生是M14中唯一的二倍体孢子无融合生殖方式,其大孢子发生时大孢子母细胞不发生减数分裂,不出现核液泡,未形成联会复合体,无细胞质改组,细胞壁上缺乏胼胝质的沉积和缺乏胞间连丝,这些可作为二倍体孢子无融合生殖的鉴定指标。     

Correlation between polyovules and number of pistil lobes (stigmatic rays) in sugar beet (Beta vulgaris L.)

The polyovules phenomenon in sugar beet has the same importance for breeding as multigermity, because only mono-ovulic monogerm plants can be used for mechanized cultivation. The polyovules trait was investigated in two Japanese lines, mono-ovulic line NK-219-O and polyovulic line NK-183-cms. Plants of the former had three pistil lobes (stigmatic rays) in each flower, a single ovule per ovary, a single seed in the mature fruit and a single seedling germinated in subsequent tests. Plants of the latter had 5–6 pistil lobes in each flower, 2–3 ovules per ovary, double-seeds and double-seedlings. The correlations between a Pistil Lobes Index (PLI) and thenumber of ovules per ovary, percentage of double-seeds inan X-ray test, and percentage of double-seedlings in germination tests were 0.885, 0.838, and 0.749, respectivelyindicating close associations. These results are important both in sugar beet breeding for indirect selection to eliminate the undesirable polyovules trait and for botanicalinvestigations of the relationship among different parts of the gynoecium.     

The use of bulked segregant analysis to accumulate RAPD markers near a locus for beet cyst nematode resistance in Beta vulgaris

Bulked segregant analysis (BSA) was used to accumulate RAPD markers near the beet cyst nematode resistance locus Hsl^pro-1 of sugar beet (Beta vulgaris L.). Graphical genotypes constructed from RFLP data were utilized to select F₂ individuals in (1) the construction of pools of plants used in the initial screening for polymorphisms, and (2) the selection of individual plants used to confirm the potential linkage. The pooled DNA samples were screened for polymorphisms using 668 RAPD primers. Forty-four candidate markers potentially linked to the region were analysed further using 14 segregating individuals. Close linkage was confirmed for 17 of the markers. Four of the RAPD markers were assigned map coordinates within the RFLP map. Three of these markers extended the RFLP map by 3cM. Altogether, the 8cM target interval contains 10 RFLP and 17 RAPD markers, corresponding to an average marker density of 0.3cM in the Hsl_pro-1 region.     

Mapping of rhizoctonia root rot resistance genes in sugar beet using pathogen response‐related sequences as molecular markers

Rhizoctonia root and crown rot caused by the fungus Rhizoctonia solani is a serious disease of sugar beet. An F_2:3 population from a cross between a resistant and a susceptible parent has been tested for R. solani resistance and a genetic map has been constructed from the corresponding F₂ parents. The map encompasses 38 expressed sequence tags (ESTs) with high similarity to genes which are involved in resistance reactions of plants (R‐ESTs) and 25 bacterial artificial chromosomes (BACs) containing nucleotide binding site (NBS)‐motifs typical for disease resistance genes. Three quantitative trait loci (QTL) for R. solani resistance were found on chromosomes 4, 5 and 7 collectively explaining 71% of the total phenotypic variation. A number of R‐ESTs were mapped in close distance to the R. solani resistance QTL. In contrast, the NBS‐BACs mapped to chromosomes 1, 3, 7 and 9 with two major clusters of NBS‐BACs on chromosome 3. No linkage between NBS‐BACs and R. solani resistance QTL was found. The data are discussed with regard to using R‐ESTs and NBS markers for mapping quantitative disease resistances.     

Early-bolting trait and RAPD markers in the specific monosomic addition line of radish carrying the e-chromosome of Brassica oleracea

The specific monosomic addition line of radish, Raphanus sativus, carrying the e chromosome of Brassica oleracea (2n = 19, e‐type MAL) with the genetic background of the late‐bolting cv.‘Tokinashi’ was produced by successive backcrossing of the original e‐type MAL of radish that showed early bolting in the genetic background of the cv. ‘Shogoin’. The early‐bolting trait specific to the e‐type MAL was constantly expressed in the backcrossed progenies (BC₂, BC₃ and BC₄), whereas the reverted radish‐like plants (2n =18) were gradually converted to bolting as late as ‘Tokinashi’. The added e‐chromosome expressed an epistatic effect against the genome of Japanese radish. Its early‐bolting trait was dominant to the late‐bolting trait of ‘Tokinashi’ which may be under the control of a few genes. Moreover, e‐type specific RAPD markers detected in eight primers were invariably transmitted in the backcrossed progenies by ‘Tokinashi’. From the analysis of the characteristics to the e‐type MAL and e‐type specific RAPD markers, it is suggested that the e‐added chromosome of kale (B. oleracea) was transmitted from generation to generation without any recombination with the radish chromosome. The gene(s) for the early‐bolting trait detected in this study may be useful for breeding work in radish, especially in the tropical areas.     

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.     

Ecological aspects of transgenic sugar beet: transfer and expression of herbicide resistance in hybrids with wild beets

Summary An increasing number of genetically engineered cultivars of several crops is being experimentally released into the environment. In future, crops with new transgenic traits will probably play an important role in agricultural practice. The long-term effect of transgenes on community ecology will depend on the distribution and establishment of transgenic plants in the wild, on the sexual transfer of their new genes to the environment and on the potential ecological impact of the transgenic trait. The starting point was the use of transgenic sugar beet lines, Beta vulgaris subspec. vulgaris var. altissima DÖLL (Helm 1957), with transgenes coding for rhizomania and herbicide (BASTA^®) resistance. The first two questions to answer were: Can the transgenes be transferred via pollen to wild beets, Beta vulgaris subspec. maritima (L.) ARCANG. or cultivated relatives such as red beet or spinach beet and are they expressed in the hybrids? Can transgenes be monitored in young Beta vulgaris-hybrids? The experimental transfer of transgenes was conducted in 1993 at a field location in northern Germany. The beets were hand-pollinated with transgenic pollen. In a non destructive biotest, the hybrid seedlings were tested for herbicide resistance. Transgenic plants showed no noxious phenotypic effects whereas control plants developed leaf necroses. All herbicide resistant hybrids within the biotest were assumed to be transgenic.     

中国甜菜主要种质资源抗病性鉴定与评价

旨在鉴定与评价不同生态区甜菜主要种质资源的抗病性,为种质资源在抗病育种上科学有效利用提供依据。以甜菜3个生态区的313份主要种质资源为试材,在自然发病条件下,按照甜菜的规范标准,调查种质的发病级数,计算病情指数,划分抗性级别。鉴定出高抗甜菜褐斑病种质资源85份、抗病89份,抗甜菜根腐病种质资源11份,抗甜菜丛根病种质资源3份,抗甜菜白粉病种质资源28份。同时鉴定出兼抗2种病害的种质资源,高抗褐斑病兼抗根腐病种质资源5份、抗褐斑病兼抗根腐病种质资源5份、抗褐斑病兼抗丛根病种质资源2份,高抗褐斑病兼抗白粉病种质资源20份。结果表明,中国甜菜种质资源比较匮乏,除抗褐斑病种质资源外,抗根腐病、丛根病和白粉病的资源不但数量少,而且抗性级别低。     

Identification of wheat–Thinopyrum intermedium 2Ai-2 ditelosomic addition and substitution lines with resistance to barley yellow dwarf virus

Among the regenerated plants derived from immature hybrid embryos of wheat–Thinopyrum intermedium disomic addition line Z6 × common wheat variety ‘Zhong8601’, a plant with a telocentric chromosome and barley yellow dwarf virus (BYDV) resistance was obtained. The telocentric chromosome paired with an entire Thinopyrum chromosome to form a heteromorphic bivalent at meiotic metaphase I. Genomic in situ hybridization showed that the telosome originated from Th. intermedium. Two ditelosomic additions and one disomic substitution were identified among the offspring of the plant. Two random amplified polymorphic DNA molecular markers were identified among 150 random primers used to detect the different arms of the alien chromosome. These might be useful for developing translocation lines with BYDV resistance.     

Expression in vitro of resistance to Heterodera schachtii in hairy roots of an alien monotelosomic addition plant of Beta vulgaris,transformed by Agrobacterium rhizogenes

Summary Hairy roots, induced by Agrobacterium rhizogenes, were obtained of nematode susceptible beet plants (Beta vulgaris) and of the nematode resistant alien monotelosomic addition AN5, carrying a telosome from B. patellaris. The additional telosome was found to be stably present in vitro in the roots of AN5. The hairy root cultures were inoculated with larvae of the beet cyst nematode Heterodera schachtii. On the root culture of AN5 significantly less cysts developed than on the other root cultures. These results indicate that the resistance to the beet cyst nematode is expressed in the roots after transformation and can be monitored under in vitro conditions.