Radish Lines Research Articles

Genome Resources for Identifying SNPs Associated With Eight Horticultural Traits in Commercial Korean Elite Radish (Raphanus sativus) Lines

ABSTRACTRadish (Raphanus sativus), which belongs to the family Brassicaceae, has relatively limited genomic resources, especially for elite lines used in commercial breeding and other agricultural applications. Thus, this study aimed to provide a comprehensive catalogue of genome sequences for 100 elite radish lines used in the Korean industry for commercial breeding purposes. These lines were sequenced and mapped to the elite Bakdal genome. A total of 33,919 high‐quality single nucleotide polymorphisms (SNPs) were identified and were found to be associated with eight distinct phenotypic traits. Five diverse machine learning (ML) models revealed that a subset of 198 SNPs had high predictive potential for the eight horticultural traits. Furthermore, the 100 elite lines were grouped into four clusters based on the eight traits, and their predictive potential was evaluated using the ML models trained using both individual and pooled SNPs. The accuracy ranged from 0.83 to 0.96 for the individually trained models and from 0.84 to 0.95 for the pooled models. This study provides a substantial basis for the advancement of digital/precision radish breeding.

Whole-Genome Sequencing and Analysis of Tumour-Forming Radish (Raphanus sativus L.) Line.

Spontaneous tumour formation in higher plants can occur in the absence of pathogen invasion, depending on the plant genotype. Spontaneous tumour formation on the taproots is consistently observed in certain inbred lines of radish (Raphanus sativus var. radicula Pers.). In this paper, using Oxford Nanopore and Illumina technologies, we have sequenced the genomes of two closely related radish inbred lines that differ in their ability to spontaneously form tumours. We identified a large number of single nucleotide variants (amino acid substitutions, insertions or deletions, SNVs) that are likely to be associated with the spontaneous tumour formation. Among the genes involved in the trait, we have identified those that regulate the cell cycle, meristem activity, gene expression, and metabolism and signalling of phytohormones. After identifying the SNVs, we performed Sanger sequencing of amplicons corresponding to SNV-containing regions to validate our results. We then checked for the presence of SNVs in other tumour lines of the radish genetic collection and found the ERF118 gene, which had the SNVs in the majority of tumour lines. Furthermore, we performed the identification of the CLAVATA3/ESR (CLE) and WUSCHEL (WOX) genes and, as a result, identified two unique radish CLE genes which probably encode proteins with multiple CLE domains. The results obtained provide a basis for investigating the mechanisms of plant tumour formation and also for future genetic and genomic studies of radish.

The homeodomain of the <i>Raphanus sativus</i> WOX4 binds to the promoter of the <i>LOG3</i> cytokinin biosynthesis gene

BACKGROUND: The WOX4 transcription factor plays a crucial role in maintaining the organisation of cambium meristem during secondary growth, but its direct targets are unknown. AIM: The aim of our work were to study the effect of WOX4 overexpression on the root development and gene expression in radish (Raphanus sativus L.), a root crop related to Arabidopsis thaliana, and to search for direct targets of the WOX4 in radish. MATERIALS AND METHODS: Radish line 19 of the St. Petersburg State University radish genetic collection was used. Plants were grown on Murashige–Skoog medium and then in soil at 23оС and 16 h of daylight. Total DNA was extracted from radish seedlings using the CTAB method. The PCR-amplified full-length RsWOX4-2 gene, gene fragments or homeobox sequence were cloned into the vectors for overexpression (pB7WG2D), RNA interference (pH7GWIWG2) and yeast one-hybrid assay (pDEST22), respectively, using the Gateway system. The vectors for overexpression and RNA interference of RsWOX4-2 were transformed into Escherichia coli DH10B and then into Agrobacteium rhizogenes Arqua chemically competent cells. Radish seedlings were transformed with A. rhizogenes containing vectors for overexpression and RNA interference of RsWOX4-2, and GUS-overexpressing A. rhizogenes was used as a control. Total RNA from transgenic radish roots was extracted with Trizol reagent. RNA reverse transcription was performed using dT-18 primers and RevertAid reverse transcriptase. qPCR was performed using the Eva Green reagent kit on a CFX96 thermocycler with fluorescence detection system. Results were processed using the 2–ΔΔCT method. Yeast transformation was performed using the competent Saccharomyces cerevisiae cells of Y2H Gold strain. For the yeast one-hybrid assay, the obtained yeast colonies transformed with plasmids containing TF homeodomain sequence and promoter regions of genes were grown on DDO and TDO selective media with different concentrations of 3-amino-1,2,4-triazole. Statistical processing based on Student’s t-test and graphing were performed using the ggplot2 package for the R programming language (v.4.0.2). RESULTS: Overexpression of the RsWOX4-2 gene affects the structure of the radish root stele and alters the number of vessels and cambium cells. Overexpression and RNA interference of the RsWOX4-2 causes changes in the expression levels of putative target genes with the WOX family transcription factor conserved binding sites in their promoters. Using the yeast one-hybrid assay, we have shown that the DNA-binding homeodomain of RsWOX4-2 interacts with the TAATCC site in the promoter of the RsLOG3 gene, which encodes the enzyme for cytokinin biosynthesis. CONCLUSIONS: We have demonstrated the effect of RsWOX4-2 overexpression on radish root stele and gene expression and identified the RsLOG3 as the putative direct target of the WOX4 transcription factor in radish.

Development of S Haplotype-Specific Markers to Identify Genotypes of Self-Incompatibility in Radish (Raphanus sativus L.).

Radish (Raphanus sativus L.), a root vegetable belonging to the Brassicaceae family, is considered one of the representative crops displaying sporophytic self-incompatibility (SSI). The utilization of a self-incompatibility system in F1 breeding can improve the efficiency of cross-combinations, leading to a reduction in breeding time and aiding in the development of novel F1 varieties. The successful implementation of this system necessitates the rapid and accurate identification of S haplotypes in parental lines. In this study, we identified a total of nine S haplotypes among 22 elite radish lines through Sanger sequencing. Subsequently, we obtained sequences for showing a 95% similarity to nine S haplotypes, along with sequences identified by other researchers using BLAST. Following this, multiple sequence alignment (MSA) was conducted to identify SRK and SLG sequence similarities, as well as polymorphisms within the class I and II groups. Subsequently, S haplotype-specific marker sets were developed, targeting polymorphic regions of SRK and SLG alleles. These markers successfully amplified each of the nine S haplotypes. These markers will play a crucial role in the rapid and precise identification of parental S haplotypes in the radish F1 breeding process, proving instrumental in the radish F1 purity test.

Basic research in the developmental genetics on the model of tumor growth in higher plants

Today, thanks to use the modern methods of biotechnology, the molecular mechanism underlying different aspects of plant development are started to open up. Along with the traditional methods of genetic analysis, plant developmental genetics actively uses the technics of genetic engineering and “omics” technologies. One of the problems of plant developmental genetics is the study of tumor growth in plants as a model for revealing the mechanisms of systemic control of cell division. Tumor is a pathological structure emerging as a result of uncontrolled proliferation of a group of cells leaving the systemic control of growth rate, cell differentiation and proliferation. Therefore, the elucidation of the mechanisms of tumor formation may help to identify the key regulators of systemic mechanisms controlling cell proliferation and differentiation. Tumor-like structures are found in almost all multicellular organisms, including higher plants. Pathogen-induced tumors, which make up the majority of neoplasms in higher plants, develop under the influence of infectious agents (bacteria, viruses, fungi, nematodes, insects, etc.) which create a niche for their own habitation in the host plant’s organism mostly by shifting the phytohormonal balance and sometimes activation of the meristematic competence of plant cells or modulation of plant cell cycle. At the same time, much rarer spontaneous tumors of higher plants are formed in plants with certain genotypes (mutants, interspecific hybrids, inbred lines) in the absence of any pathogen, which makes them closer to animal tumors. In particular, in the genetic collection of radish (Raphanus sativusL.), the inbred lines that form spontaneous tumors on the taproot during the flowering period were obtained many years ago. The connection between the spontaneous tumor formation in these lines and the altered balance of the main phytohormones, as well as ectopic expression of meristem-specific genes, was previously demonstrated. We have analyzed the differential gene expression in the spontaneous tumors of radish versus the lateral roots using the RNA-seq method. Data were obtained indicating the increased expression of genes associated with cell division and growth (especially genes that regulate G2-M transition and cytokinesis) in the spontaneous tumor. Among genes downregulated in tumor tissue, genes participating in the response to stress and wounding, as well as in the biosynthesis of glucosinolates, were enriched. Subsequently, we also performed whole genome sequencing of two closely related radish lines, contrasting in their ability to spontaneous tumor formation. In the coding genes of tumorous line, we have identified numerous SNPs and InDel which lead to frameshift and are probably associated with the tumor formation trait. Thus, spontaneous tumor formation in inbred lines of radish is probably under complex polygenic control. Testing of the relationship of these polymorphisms with tumor formation has begun. Our data will help to elucidate the mechanisms of spontaneous tumor development in higher plants.
 The conference was held with support of the Ministry of Science and Higher Education of the Russian Federation in accordance with agreement No. 075-15-2022-322 date 22.04.2022 on providing a grant in the form of subsidies from the Federal budget of Russian Federation. The grant was provided for state support for the creation and development of a World-class Scientific Center “Agrotechnologies for the Future”.

Identification of SNPs and InDels probably associated with the development of spontaneous tumors in radish (<i>Raphanus sativus</i> L.)

Radish (Raphanus sativus L.) is an agronomically important root crop belonging to the Brassicaceae family. A genetic collection of inbred radish lines, which was based on contrasting genetically determined traits including trait ability to develop tumors, was created at St. Petersburg State University in the middle of the 20th century [1]. Full genetics network controlling this trait still remains unclear [2], and elucidation of its mechanisms can help reveal the key regulators of systemic mechanisms that control cell proliferation and differentiation.
 The purpose of this work is to identify single nucleotide polymorphisms (SNPs) and InDels in genes which are candidates for participation in the tumor development process within in the tumor-forming radish line compared to the non-tumor radish line.
 We have assembled the genomes of two lines contrasting in the ability to tumorigenesis, annotated them, aligned sequences per assembly, identified candidate genes and differences in the structure of these genes in contrasting radish lines using bioinformatics tools. Bioinformatics data were confirmed by the sequencing by Sanger method.
 As a result, in the tumor-forming radish line we identified 151 genes with InDels in their coding regions, which led to various variants of frameshift. Moreover, we detected 39 genes with single nucleotide substitutions (SNPs). According to the gene pathway enrichment analysis, the corresponding genes were classified into the several groups.
 Thus, the data obtained will allow us to clarify in more detail the genetic control mechanisms of the tumor development in radish.
 The work was supported by the Ministry of Science and Higher Education of the Russian Federation in accordance with the agreement No. 075-15-2022-322.

Development of Molecular Markers for Predicting Radish (Raphanus sativus) Flesh Color Based on Polymorphisms in the RsTT8 Gene.

Red radish (Raphanus sativus L.) cultivars are a rich source of health-promoting anthocyanins and are considered a potential source of natural colorants used in the cosmetic industry. However, the development of red radish cultivars via conventional breeding is very difficult, given the unusual inheritance of the anthocyanin accumulation trait in radishes. Therefore, molecular markers linked with radish color are needed to facilitate radish breeding. Here, we characterized the RsTT8 gene isolated from four radish genotypes with different skin and flesh colors. Sequence analysis of RsTT8 revealed a large number of polymorphisms, including insertion/deletions (InDels), single nucleotide polymorphisms (SNPs), and simple sequence repeats (SSRs), between the red-fleshed and white-fleshed radish cultivars. To develop molecular markers on the basis of these polymorphisms for discriminating between radish genotypes with different colored flesh tissues, we designed four primer sets specific to the RsTT8 promoter, InDel, SSR, and WD40/acidic domain (WD/AD), and tested these primers on a diverse collection of radish lines. Except for the SSR-specific primer set, all primer sets successfully discriminated between red-fleshed and white-fleshed radish lines. Thus, we developed three molecular markers that can be efficiently used for breeding red-fleshed radish cultivars.

Transcriptomic Analysis of Radish (Raphanus sativus L.) Spontaneous Tumor.

Spontaneous tumors can develop in different organs of various plant species without any pathogen infection and, as a rule, appear in plants with a certain genotype: Mutants, interspecific hybrids, etc. In particular, among the inbred lines of radish (Raphanus sativus L.), lines that form spontaneous tumors on the taproot during the flowering period were obtained many years ago. In this work, we analyzed the differential gene expression in the spontaneous tumors of radish versus the lateral roots using the RNA-seq method. Data were obtained indicating the increased expression of genes associated with cell division and growth (especially genes that regulate G2-M transition and cytokinesis) in the spontaneous tumor. Among genes downregulated in the tumor tissue, genes participating in the response to stress and wounding, mainly involved in the biosynthesis of jasmonic acid and glucosinolates, were enriched. Our data will help elucidate the mechanisms of spontaneous tumor development in higher plants.

Transcriptome analysis of crown gall in radish (Raphanus sativus L.) inbred lines

AbstractStudying aberrations of normal developmental processes like plant tumour initiation provides a great opportunity for understanding of systemic control of plant cells differentiation. Most well‐studied example of plant tumour is the crown gall caused by Agrobacterium tumefaciens; much less studied are spontaneous tumours developing in plants with certain genotype, regardless of agrobacterial infection. In the present study, we used RNA‐seq to analyse the changes in gene expression at the early stages of development of A. tumefaciens‐induced tumours in radish seedlings. In our studies we used two closely related radish inbred lines contrasting in their ability to form spontaneous tumours. RNA‐seq and qPCR analysis showed significant changes in the expression levels for numerous genes involved in different metabolic and developmental processes, and Gene Ontology analysis indicated many biological processes probably involved in the crown gall development in radish. For example, a unique result of our research is upregulation of genes related to photomorphogenesis and downregulation of senescence‐associated genes in the young crown gall tumours. Differences in gene expression between the analysed radish lines included upregulation of pathways related to chromatin remodelling and protein targeting to chloroplast in the tumorous line along with downregulation of pathways of chaperone‐mediated complex assembly and anthocyanin biosynthesis.

Genome-wide identification and expression profiling of MYB transcription factor genes in radish (Raphanus sativus L.)

Radish (Raphanus sativus L.), an important root vegetable crop of the Brassicaceae family, has a high level of anthocyanin accumulation in its pigment root tissues. It was reported that MYB transcription factors (TFs) play vital roles in plant development and anthocyanin metabolism, and the PAP1/2 could promote expression of anthocyanin biosynthesis genes. In this study, a total of 187 radish MYB genes (RsMYBs) were identified in the radish genome and clustered into 32 subfamilies. Among them, 159 RsMYBs were localized on nine radish chromosomes. Interestingly, 14 RsMYBs exhibited differential expression profiles in different taproot developmental stages among four differently colored radish lines. A number of RsMYBs were highly expressed in the pigmented root tissues at the maturity stage, several genes including RsMYB41, RsMYB117, and RsMYB132 being homologous to PAP1/2, showed high expression levels in the red skin of NAU-YH (red skin-white flesh) taproot, while RsMYB65 and RsMYB159 were highly expressed in the purple root skin of NAU-YZH (purple skin-red flesh), indicating that these RsMYBs might positively regulate the process of anthocyanin accumulation in radish taproot. These results would provide valuable information for further functional characterization of RsMYBs, and facilitate clarifying the molecular mechanism underlying anthocyanin biosynthesis in radish.

Genotype × Environment interaction: AMMI analysis of radish (Raphanus sativus) genotypes in different environments of Punjab

The study entailed the determination of Genotype × Environment (G × E) interaction through Additive Main effects and Multiplicative Interaction (AMMI) analysis to identify radish (Raphanus sativus L.) genotypes suitable for cultivation in the province of Punjab, India. Twelve newly developed advanced breeding lines of radish were sown on six different dates for two years, i.e. 2014-15 and 2015-2016, in a randomized complete block design, replicated thrice. The explicit analysis of the AMMI revealed pronounced performance differences among the genotypes due to environments, explained by high percentage of GEI through IPCA1 and IPCA2 (65.48 %-82.79%). The AMMI analysis clearly delineated the mega environments and provided information on the cultivars exhibiting enhanced performance in those mega-environments on the basis of stability, performance and genetic difference between genotypes and environments. Temperature and photoperiod were the major contributing factors in G × E interactions. RL-30 excelled across the environments in terms of root length, root weight, plant weight and total marketable yield whilst, RB-21 performed best under hostile environments (high – ve score on IPCA axis).

Gibberellin Promotes Bolting and Flowering via the Floral Integrators RsFT and RsSOC1-1 under Marginal Vernalization in Radish.

Gibberellic acid (GA) is one of the factors that promotes flowering in radish (Raphanus Sativus L.), although the mechanism mediating GA activation of flowering has not been determined. To identify this mechanism in radish, we compared the effects of GA treatment on late-flowering (NH-JS1) and early-flowering (NH-JS2) radish lines. GA treatment promoted flowering in both lines, but not without vernalization. NH-JS2 plants displayed greater bolting and flowering pathway responses to GA treatment than NH-JS1. This variation was not due to differences in GA sensitivity in the two lines. We performed RNA-seq analysis to investigate GA-mediated changes in gene expression profiles in the two radish lines. We identified 313 upregulated, differentially expressed genes (DEGs) and 207 downregulated DEGs in NH-JS2 relative to NH-JS1 in response to GA. Of these, 21 and 8 genes were identified as flowering time and GA-responsive genes, respectively. The results of RNA-seq and quantitative PCR (qPCR) analyses indicated that RsFT and RsSOC1-1 expression levels increased after GA treatment in NH-JS2 plants but not in NH-JS1. These results identified the molecular mechanism underlying differences in the flowering-time genes of NH-JS1 and NH-JS2 after GA treatment under insufficient vernalization conditions.

Transcriptome sequencing assisted discovery and computational analysis of novel SNPs associated with flowering in Raphanus sativus in-bred lines for marker-assisted backcross breeding

The sequencing of radish genome aids in the better understanding and tailoring of traits associated with economic importance. In order to accelerate the genomics assisted breeding and genetic selection, transcriptomes of 33 radish inbred lines with diverse traits were sequenced for the development of single nucleotide polymorphic (SNP) markers. The sequence reads ranged from 2,560,543,741 bp to 20,039,688,139 bp with the GC (%) of 47.80–49.34 and phred quality score (Q30) of 96.47–97.54%. A total of 4951 polymorphic SNPs were identified among the accessions after stringent filtering and 298 SNPs with efficient marker assisted backcross breeding (MAB) markers were generated from the polymorphic SNPs. Further, functional annotations of SNPs revealed the effects and importance of the SNPs identified in the flowering process. The SNPs were predominantly associated with the four major flowering related transcription factors such as MYB, MADS box (AG), AP2/EREB, and bHLH. In addition, SNPs in the vital flowering integrator gene (FT) and floral repressors (EMBRYONIC FLOWER 1, 2, and FRIGIDA) were identified among the radish inbred lines. Further, 50 SNPs were randomly selected from 298 SNPs and validated using Kompetitive Allele Specific PCR genotyping system (KASP) in 102 radish inbred lines. The homozygosity of the inbred lines varied from 56 to 96% and the phylogenetic analysis resulted in the clustering of inbred lines into three subgroups. Taken together, the SNP markers identified in the present study can be utilized for the discrimination, seed purity test, and adjusting parental combinations for breeding in radish.

Results and prospects of European radish breeding

Relevance Currently, the first place goes to the creation of new high-yielding varieties and hybrids F1 crops that meet the requirements of modern industrial technologies. Heterosis hybrids have high yield, marketability, equalization, friendly yield and resistance to disease. Methods When developing new varieties and hybrids of radish suitable for mechanized harvesting, attention should be paid to the strength of the attachment of the leaf outlet and its upright position, the uniformity of immersion in the soil and the easy pull-out of root crops. Studies were performed in a protected (heated film greenhouse) and open ground in All-Russian Scientific Research Institute of Vegetable Growing – Branch of the FSBSI Federal Scientific Vegetable Center. Results The results of breeding work with European radish are presented. According to the results of expert evaluation, the state register of breeding achievements included two varieties of European summer radish – Bianca, Sirius and one hybrid of European winter radish – Tsyganskiy Baron F1. The new varieties are distinguished by uniformity in shape, length, diameter of the root crop, compact leaf rosette, form a root crop in a short and long daylight, recommended for cultivation under film shelters and in the open ground (early spring and summer sowing time). As a result of repeated inbreeding, inbred lines of European winter radish (brought to the seventh generation) were created, characterized by high equalization of the root crop, with strict self-compatibility. These lines were included in the process of creating heterosis hybrids. Evaluation of the obtained hybrids was carried out in the open ground. A hybrid of radish European winter Tsyganskiy Baron F1 – medium-ripe, flat-rounded root, black, weighing 220-260 g. Perspective hybrid in relation to the standard has a more aligned form of the root, with a rich black color and smooth surface. Tsyganskiy Baron F1 on the above indicators exceeds the standard night, resistant to stressful conditions.

Mitochondrial orf463 causing male sterility in radish is possessed by cultivars belonging to the ‘Niger’ group

In addition to Ogura cytoplasmic male sterility (CMS), which is used worldwide in radish and Brassica crops, another type of CMS was found and named DCGMS. A mitochondrial candidate gene (orf463) of DCGMS was identified, but the distribution of this gene in radish species has not been clarified to date. We found that orf463 is distributed specifically in black radish cultivars belonging to the ‘Niger’ group and a line of wild species, Raphanus raphanistrum. The progeny tests using the cultivars having orf463 as a female parent demonstrated that the cytoplasm induces male sterility in radishes. Although the DNA sequences of orf463 possessed by the ‘Niger’ group cultivars are identical to those of DCGMS, the orf463 sequence of the wild radish line contained nine nucleotide substitutions, seven of which were nonsynonymous. All radishes with orf463 had both DCGMS-type and ‘Black radish’- type mitochondrial structures according to multiplex PCR. Based on these results, the mechanism of rearrangement between the two mitochondrial types was studied.

A new male sterile line of “Duanye-13” radish (Raphanus sativus L.) produced by ethyl methanesulfonate mutagenesis

Mutations in radish can be induced by exposure to ethyl methanesulfonate (EMS), and this method is suitable for obtaining target traits in new radish materials. We investigated the optimal concentration of EMS for mutagenesis on the radish variety Duan-Ye No. 13 (DY13). Treating DY13 radish seeds with 0.5% EMS led to the development of sterile mutants. During 3 years of continuous hybridization and tracing, male fertility remained stable at 100% sterility. Morphological observations showed that the anthers of these male sterile were wrinkled, curled, and browned; disintegration of the wall structure at the pollen tetrad stage caused eventual pollen abortion. Detection of the molecular marker ORF138-F/R confirmed that the target band could be amplified in this radish line; however, the band was not amplified in the corresponding maintainer line. Together, these results show that male sterile lines were successfully selected from DY13 radishes, demonstrating a new and effective method for breeding male sterile radish lines.

Genetic control of regeneration processes of radish plants in vitro: from phenotype to genotype

This review highlights the years of research on the genetics of in vitro regeneration in higher plants conducted at the Department of Genetics and Biotechnology of Saint Petersburg State University. The genetic collection of radish (Raphanus sativus) created at the department by selfing of individual plants from three cultivars was used as a model in these studies. Some radish inbred lines from the genetic collection form spontaneous tumors in the roots and are also used to study mechanisms of tumor growth in higher plants. It was revealed that radish lines that differed in the ability to form tumors also contrastingly differed in the reaction of their explants to auxin and cytokinin in vitro, which reflects a difference in the levels of these hormones in the tissues of related tumorous and non-tumorous radish lines. Moreover, high concentrations of cytokinins in cultural medium induced tumor formation in the regenerated plants of tumorous radish lines. The presence of meristematic zones in spontaneous tumors in radish lines, as well as in crown gall tumors induced by Agrobacterium tumefaciens and cytokinin-induced tumors made it possible to reveal the role of the main meristem regulators, such as KNOX and WOX family transcription factors and the CLAVATA system, in both the process of tumor growth and regeneration in plants. Analysis of the expression of meristem-specific genes during the development of spontaneous and induced tumors in radish as well as in regenerated radish plants confirmed this assumption.

IIdentification and Variation of Major Aliphatic Glucosinolates in Doubled Haploid Lines of Radish (Raphanus sativus L.)

IIdentification and Variation of Major Aliphatic Glucosinolates in Doubled Haploid Lines of Radish (Raphanus sativus L.)

Sequence Variation Analysis of Flowering-time Genes in Two Radish Lines with Different Flowering Time

Sequence Variation Analysis of Flowering-time Genes in Two Radish Lines with Different Flowering Time

THE STAGES OF HETEROTIC HYBRIDS F1 DEVELOPMENT IN EUROPEAN RADISH

The scheme of development of two-line of hybrids F 1 in European radish based on self-incompatibility includes five stages, as follows: 1 – selection of self-incompatible lines, common and specified combining ability estimation; 2 – inbreeding and selection to make the lines homozygous for morphological traits, common and specified combining ability estimation; maintenance and reproduction of self-incompatible lines; 4 – production of hybrid seeds. The research work on assessment of hybrid F 1 that were obtained from cross of eight self-incompatible lines of European winter radish by the Griffing’s method was carried out in 2016. The assessment of length, diameter and yield of radish root was performed. According to the root shape the heterotic hybrids F 1 were divided into three groups: rounded-flat, 48.2%; round, 50.0%; and flatten-round, 1.8%. The level of root marketability of hybrids F 1 reached 100%. As a result of the work the promising hybrid combination distinguished by high uniformity, marketability and high yield were selected out.