Study of genetic diversity in potato cultivars using PCR analysis of organelle DNA

The genetic diversity of 98 potato cultivars of Russian and foreign breeding was studied using of PCR with organelle-specific primers. The polymorphism of both plastid (atpE, trnG/trnR) and mitochondrial (rps10, atp6) loci was revealed. Eight different haplotypes were detected in the sample of cultivars studied. Comparatively low polymorphism of organelle DNA in the potato cultivars was demonstrated: most cultivars (91 or 92.9%) possessed only two haplotypes (I and II); 62 cultivars of them had the same “cultural” cytoplasmic type (haplotype I). The breeding cultivars of the Russian and foreign origin did not differ from each other in frequency of basic haplotypes.     

Polymorphism of chloroplast microsatellite DNA loci in Russian potato cultivars

The cpSSR method was for the first time used to analyze the plastome in 29 Russian potato Solanum tuberosum cultivars. The informativeness coefficient H and the allele variability of the NTCP6, NTCP8, and NTCP9 loci were determined. In total, 14 allelic variants of the microsatellite cpDNA loci were identified. The NTCP8 and NTCP9 cpSSR displayed the highest allele polymorphism in the cultivars examined. A cultivar-specific haplotypes of the chloroplast genome were observed for 20 out of 29 cultivars.     

An extreme cytoplasmic bottleneck in the modern European cultivated potato (Solanum tuberosum) is not reflected in decreased levels of nuclear diversity

We have used the polymorphic chloroplast (cp) and nuclear simple sequence repeats (SSRs) to analyse levels of cytoplasmic and nuclear diversity in the gene pool of the European cultivated potato (Solanum tuberosum ssp. tuberosum). Primers designed from the complete chloroplast sequence of tobacco (Nicotiana tabacum) were used to amplify polymorphic products in a range of potato cultivars. Combining the data from seven polymorphic cpSSR loci gave 26 haplotypes, one of which (haplotype A) accounted for 151 out of the 178 individuals studied and corresponded to the T-type cytoplasm previously identified in cultivated potatoes using chloroplast restriction fragment length polymorphism analysis. Phylogenetic and diversity analyses of the relationships between cpSSR haplotypes confirmed much higher levels of cytoplasmic diversity outwith the T-type group. Diversity levels at eight nuclear SSR loci, however, were not significantly different between cytoplasmic groups, suggesting a severe maternal bottleneck in the evolution of the modern cultivated potato. These results highlight the importance in quantifying levels of cytoplasmic as well as nuclear diversity and confirm the need for a change in breeding practices to increase levels of non-T-type cytoplasm in the cultivated gene pool, thus helping reduce problems associated with pollen sterility. This may be facilitated by germplasm analysis using cpSSRs, which will allow efficient selection of diverse cytoplasm donors.     

Intraspecific polymorphism of the sucrose synthase genes in Russian and Kazakh potato cultivars

In 12 different Russian and Kazakh potato cultivars, the polymorphism of the glucosyltransferase domain of the sucrose synthase gene was first examined, as well as the polymorphism of the sucrose synthase domain fragment of the same gene in the potato cultivars of Kazakh breed. It was demonstrated that the examined sequences contained point mutations, as well as insertions and deletions, including those not described earlier. Amino acid substitutions specific to heat- and drought-tolerant varieties were also identified and could be associated with the development of abiotic stress resistance.     

Trends in genetic diversity change of spring bread wheat cultivars released in Russia in 1929-2003

Using genealogy analysis, we studied genetic diversity of 340 cultivars of spring bread wheat that were released on the territory of Russia in 1929-2003. Trends in the temporal change of genetic diversity were inferred from analysis of a set of n x m matrices, where n is the number of the released cultivars and m is the number of original ancestors. The pool of original ancestors of the spring bread wheat cultivars for the total period of study included 255 landraces, of which 88 were from the former USSR and modern Russia. The original ancestors showed great differences in their presence in the cultivar sets examined and, consequently, in their importance for the gene pool of Russian spring wheats. The distributions of contributions of dominant original ancestors to cultivar diversity were significantly different in different regions, indicating that the ancestors were specific for the cultivation conditions. During the last 75 years, the genetic diversity of the spring bread wheat cultivars has been increasing owing to the wide use of foreign material in Russian breeding programs. However, our analysis showed that about 60 landraces, including the Russian ones, were lost during the studied time period. The lost part makes up 35% of the gene pool of the Russian original ancestors. It is reasonable to assume that the lost landraces carried a gene complex f or adaptation to specific Russian environments. Specificity of the contributions of the original ancestors in the sets of cultivars produced in different breeding centers was established. A comparative analysis of genetic similarity of cultivars was carried out using coefficients of parentage. Significant differences in this parameter between breeding institutes and regions of cultivation were revealed.     

Analysis of microsatellite locus polymorphism in potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis>) cultivars of Russian breeding

Polymorphism of microsatellite loci of the nuclear genome was examined in 29 cultivars and accessions of wild potato (S. tuberosum, S. stoloniferum, S. demissum, and S. phureja). Nine SSR markers, most informative (PIC = 0.61–0.92) for genotyping of the cultivars of Russian breeding were selected. Polymorphism of the selected SSR loci was characterized, and prevailing, as well as unique SSR allele phenotypes were described. A total of 87 allele phenotypes were identified. The highest number of allele phenotypes was detected for the SSR1 (17), ST83/84 (12), and STRBCS1b (12) loci. The least numbers of allele phenotypes were typical of the ST47/48 (5) and STWIN12G (6) loci. Based on the microsatellite loci analyzed, for each of the cultivars examined, its allele formula was established. The latter can be uses as the cultivar molecular genetic passport. Diagnostic sets of most informative loci (SSR markers), enabling identification of the genotypes of all potato cultivars of Russian breeding examined, were determined     

Application of high-resolution DNA melting for genotyping and variant scanning of diploid and autotetraploid potato

The ideal marker system for tetraploid potato would be dosage-sensitive and have the ability to distinguish heterozygous genotypes with multiple haplotypes within the genomic region targeted by the marker. The objective of this study was to evaluate the utility of high-resolution DNA melting (HRM) for genotyping and polymorphism detection in diploid and tetraploid potato. Amplicon scanning, unlabelled probe, and short amplicon assays were developed for four candidate genes affecting tuber skin and flesh colour, and starch, and a marker linked to nematode resistance. Genotyping a set of 95 potato clones revealed several examples of clones with three distinct haplotypes. Combined probe and amplicon analysis identified between 29 and 44 unique genotypes for the same assays. Assays developed for four of the five target genes are suitable for marker-assisted selection in potato breeding programs. This study illustrates the use of HRM in potato genetics. Further advances in the technology and associated data analysis should make HRM a useful tool for basic and applied studies of potato.     

Trends in Genetic Diversity Change of Spring Bread Wheat Cultivars Released in Russia in 1929–2003

Using genealogy analysis, we studied genetic diversity of 340 cultivars of spring bread wheat that were released on the territory of Russia in 1929–2003. Trends in the temporal change of genetic diversity were inferred from analysis of a set of n × m matrices, where n is the number of the released cultivars and m is the number of original ancestors. The pool of original ancestors of the spring bread wheat cultivars for the total period of study included 255 landraces, of which 88 were from the former USSR and modern Russia. The original ancestors showed great differences in their presence in the cultivar sets examined and, consequently, in their importance for the gene pool of Russian spring wheats. The distributions of contributions of dominant original ancestors to cultivar diversity were significantly different in different regions, indicating that the ancestors were specific for the cultivation conditions. During the last 75 years, the genetic diversity of the spring bread wheat cultivars has been increasing owing to the wide use of foreign material in Russian breeding programs. However, our analysis showed that about 60 landraces, including the Russian ones, were lost during the studied time period. The lost part makes up 35% of the gene pool of the Russian original ancestors. It is reasonable to assume that the lost landraces carried a gene complex f or adaptation to specific Russian environments. Specificity of the contributions of the original ancestors in the sets of cultivars produced in different breeding centers was established. A comparative analysis of genetic similarity of cultivars was carried out using coefficients of parentage. Significant differences in this parameter between breeding institutes and regions of cultivation were revealed.     

Polymorphism of chloroplast microsatellite DNA loci in Russian potato cultivars

The cpSSR method was for the first time used to analyze the plastome in 29 Russian potato Solanum tuberosum cultivars. The informativeness coefficient H and the alleles of the NTCP6, NTCP8, and NTCP9 loci were determined. In total, 14 allelic variants of the microsatellite cpDNA loci were identified. The NTCP8 and NTCP9 cpSSR displayed the greatest allele polymorphism in the cultivars examined. A cultivar-specific haplotype of the chloroplast genome was observed for 20 out of 29 cultivars.     

10个牛品种线粒体12S rRNA基因多态性分析

以鲁西牛、渤海黑牛、大别山牛、南阳牛、晋南牛、秦川牛、中国西门塔尔牛、日本和牛、海福特牛和安格斯牛10个牛品种共计353头牛为研究对象,利用单链构象多态性（SSCP）分子标记方法对其线粒体DNA 12S rRNA基因进行了多态性分析,结果发现该基因出现分布频率不同的3种单倍型（Ⅰ、Ⅱ、Ⅲ）,其中单倍型Ⅰ出现的频率最低,仅在晋南牛1个个体中发现;单倍型Ⅱ在中国本地黄牛中都存在,在中国西门塔尔牛中的分布频率很低,但在3个国外品种海福特牛、安格斯牛和日本和牛中不存在;单倍型Ⅲ在这10个牛品种中均有分布,但以国外品种最丰富。计算平均多态信息含量,发现我国地方品种在0.232～0.423之间,基本属于中度多态,说明我国地方黄牛品种在12S rRNA上多态性比较丰富,国外品种较为贫乏;但是单倍型Ⅰ频率很低有丢失的趋势,需要对其进行保护。     

SSR polymorphism of modern cultivars and autochthonous forms of the pear tree from North Caucasus

Genetic similarity and relatedness within the set of pear genotypes including autochthonous Circassian cultivars from North Caucasus, European cultivars, accessions of Pyrus caucasica Fed., and modern Russian cultivars were estimated on the basis of analysis of SSR loci. The level of polymorphism for the studied loci varied from 11 to 15 alleles per locus in the set of 29 samples of pears. A higher level of allelic polymorphism of SSR loci was revealed for a set of P. caucasica samples in comparison with modern cultivated cultivars: from 9 to 12 alleles for P. caucasica and from 6 to 8 alleles for modern cultivars. Specific alleles for the mentioned groups of pears were identified. UPGMA clustering revealed two distinct groups: one includes P. caucasica accessions and autochthonous Caucasian cultivars and the other group includes all cultivated European and Russian pear cultivar. The results support the hypothesis of an isolated gene pool formation of autochthonous pear cultivars of the North Caucasus and their probable origin from the wild P. caucasica.     

Identification of SNPs in Closely Related Temperate Japonica Rice Cultivars Using Restriction Enzyme-Phased Sequencing

Very low polymorphism in the germplasm typically used by breeding programs poses a significant bottleneck with regards to molecular breeding and the exploitation of breeding materials for quantitative trait analyses. California rice cultivars, derived from a very small base of temperate japonica germplasm and having a relatively brief breeding history, are a good example. In this study, we employed a reduced representation sequencing approach called Restriction Enzyme Site Comparative Analysis (RESCAN) to simultaneously identify and genotype single nucleotide polymorphisms (SNPs) in forty-five rice cultivars representing the majority of the 100 year-old breeding history in California. Over 20,000 putative SNPs were detected relative to the Nipponbare reference genome which enabled the identification and analysis of inheritance of pedigree haplotypes. Haplotype blocks distinguishing modern California cultivars from each other and from the ancestral short grain temperate japonica cultivars were easily identified. Reduced representation sequencing methods such as RESCAN are a valuable alternative to SNP chip genotyping and low coverage whole genome sequencing.     

Y-chromosome DNA haplotypes in north African populations

The frequency distribution of Y-chromosome haplotypes at DNA polymorphism p49/TaqI was studied in a sample of 505 North Africans from Mauritania, Morocco, Algeria, Tunisia, Libya, and Egypt. A particularly high frequency (55.0%) of Y-haplotype 5 (A2, C0, D0, F1, I1) was observed in these populations, with a relative predominance in those of Berber origin. Examination of the relative frequencies of other haplotypes in these populations, mainly haplotype 4 (the "African" haplotype), haplotype 15 (the "European" haplotype), and haplotypes 7 and 8 (the "Near-East" haplotypes), permit useful comparisons with neighboring peoples living in sub-Saharan Africa, Europe, and the Near East.     

Analysis of Genetic Diversity of Spring Durum Wheat (<Emphasis Type="Italic">Triticum durum</Emphasis> Desf.) Cultivars Released in Russia in 1929–2004

Based on genealogical analysis, the genetic diversity of 78 spring durum wheat cultivars released in Russia in 1929–2004 have been examined. The temporal trends of change in diversity were studied using series of n × m matrices (where n is the number of the cultivars and m is the number of original ancestors) and calculating coefficients of parentage in sets of cultivars released in particular years. The pool of original ancestors of spring durum wheat cultivars includes 90 landraces and old varieties, more than a half (57%) of which originate from European countries, including Russia and Ukraine (45%). The original ancestors strongly differ in the frequency of presence in the cultivar pedigrees. Landraces Beloturka, Sivouska, Kubanka (T. durum Desf.), Transbaikalian emmer, Yaroslav emmer (T. dicoccum Schuebl.), Poltavka (T. aestivum L.), and the original ancestors of cultivars Kharkov 46, Narodnaya, and Melanopus 1932 enter in the pedigrees of more than half of cultivars created within the framework of various breeding programs. At that, their distribution by cultivars from different breeding centers strongly varies. Analysis of temporal dynamics of genetic diversity, based on genetic profiles and coefficients of parentage, has shown that the genetic diversity of Russian durum wheats increased during the period examined. Nevertheless, genetic erosion of the local material—a loss of approximately 20% of the pool of Russian original ancestors—has been found. The contribution of the original ancestors to the pedigrees of different cultivars, constructed in different breeding centers and recommended for cultivation in different regions, has been estimated. The variation of the released cultivars was highest in the Lower Volga region and lowest in the Ural region. In all, the lower threshold of genetic diversity in all regions does not reach the critical level, corresponding to the similarity of half-sibs. The set of modern cultivars included in the Russian Official List 2004 has a cluster structure.     

High resolution DNA melting markers for identification of <Emphasis Type="Italic">H1</Emphasis>-linked resistance to potato cyst nematode

Although Sequence-Characterized Amplified Region (SCAR) markers linked to the potato H1 locus, which confers resistance to pathotypes Ro1 and Ro4 of the potato cyst nematode (PCN) Globodera rostochiensis, have been reported, robust markers that enable estimation of allele dosage would improve the quality of information obtained from genotyping parental accessions (cultivars/breeding lines) and progeny populations within breeding programmes. With this in mind, we have developed single nucleotide polymorphism (SNP)-based molecular markers flanking the H1 resistance gene, using genomic re-sequence data from five elite tetraploid accessions. The published TG689 and 57R primer sequences were used in a Basic Local Alignment Search Tool (BLAST) examination of the reference potato genome, and SNPs within the vicinity of these primer regions were identified and targeted for designing probe-based High Resolution Melting (HRM) SNP assays. Evaluation of the subsequently developed HRM markers, TG689_1P and 57R_1P, against the publicly available SCAR markers, TG689 and 57R, indicated that the HRM markers enabled more reliable marker-trait association than the SCARs. Additionally, allelic dosage estimates for the H1 locus were also derived using the TG689_1P marker, providing a tool to optimise parental and progeny selections in PCN resistance breeding.     

Analysis of diversity of Russian and Ukrainian bread wheat (Triticum aestivum L.) cultivars for high-molecular-weight glutenin subunits

The allelic diversity of high-moleculat-weght glutenin subunits (H WIGS) in Russian and Ukrainian bread wheat cultivars was analyzed. The diversity of spring wheat cultivars for alleles of the Glu-1 loci is characterized by medium values of the polymorphism index (polymorphism information content, PlC), and in winter wheats it varies from high at the Glu-A1 locus to low at the Glu-D1 locus. The spring and winter cultivars differ significantly in the frequencies of alleles of the glutenin loci. The combination of the Glu-A1b, Glu-B1c, and Glu-D1a alleles prevails among the spring cultivars, and the combination of the Glu-A1a, Glu-B1c, and Glu-D1d alleles prevails among the winter cultivars. The distribution of the Glu-1 alleles significantly depends on the moisture and heat supply in the region of origin of the cultivars. Drought resistance is associated with the Glu-D1a allele in the spring wheat and with the Glu-B1b allele in the winter wheat. The sources of the Glu-1 alleles were identified in the spring and wheat cultivars. The analysis of independence of the distribution of the spring and winter cultivars by the market classes and by the alleles of the HMWGS loci showed a highly significant association of the alleles of three Glu-1 loci with the market classes in foreign cultivars and independence or a weak association in the Russian and Ukrainian cultivars. This seems to be due to the absence of a statistically substantiated system of classification of the domestic cultivars on the basis of their quality.     

Applications of biotechnology and genomics in potato improvement

Potato is the third most important global food crop and the most widely grown noncereal crop. As a species highly amenable to cell culture, it has a long history of biotechnology applications for crop improvement. This review begins with a historical perspective on potato improvement using biotechnology encompassing pathogen elimination, wide hybridization, ploidy manipulation and applications of cell culture. We describe the past developments and new approaches for gene transfer to potato. Transformation is highly effective for adding single genes to existing elite potato clones with no, or minimal, disturbances to their genetic background and represents the only effective way to produce isogenic lines of specific genotypes/cultivars. This is virtually impossible via traditional breeding as, due to the high heterozygosity in the tetraploid potato genome, the genetic integrity of potato clones is lost upon sexual reproduction as a result of allele segregation. These genetic attributes have also provided challenges for the development of genetic maps and applications of molecular markers and genomics in potato breeding. Various molecular approaches used to characterize loci, (candidate) genes and alleles in potato, and associating phenotype with genotype are also described. The recent determination of the potato genome sequence has presented new opportunities for genomewide assays to provide tools for gene discovery and enabling the development of robustly unique marker haplotypes spanning QTL regions. The latter will be useful in introgression breeding and whole‐genome approaches such as genomic selection to improve the efficiency of selecting elite clones and enhancing genetic gain over time.     

Identification of agronomically important QTL in tetraploid potato cultivars using a marker–trait association analysis

Key message

Nineteen tuber quality traits in potato were phenotyped in 205 cultivars and 299 breeder clones. Association analysis using 3364 AFLP loci and 653 SSR-alleles identified QTL for these traits.

Abstract

Two association mapping panels were analysed for marker–trait associations to identify quantitative trait loci (QTL). The first panel comprised 205 historical and contemporary tetraploid potato cultivars that were phenotyped in field trials at two locations with two replicates (the academic panel). The second panel consisted of 299 potato cultivars and included recent breeds obtained from five Dutch potato breeding companies and reference cultivars (the industrial panel). Phenotypic data for the second panel were collected during subsequent clonal selection generations at the individual breeding companies. QTL were identified for 19 agro-morphological and quality traits. Two association mapping models were used: a baseline model without, and a more advanced model with correction for population structure and genetic relatedness. Correction for population structure and genetic relatedness was performed with a kinship matrix estimated from marker information. The detected QTL partly not only confirmed previous studies, e.g. for tuber shape and frying colour, but also new QTL were found like for after baking darkening and enzymatic browning. Pleiotropic effects could be discerned for several QTL.     

Analysis of genetic diversity of spring durum wheat (Triticum durum Desf.) cultivars released in Russia in 1929-2004

Based on genealogical analysis, the genetic diversity of 78 spring durum wheat cultivars released in Russia in 1929-2004 have been examined. The temporal trends of change in diversity were studied using series of n x m matrices (where n is the number of the cultivars and m is the number of original ancestors) and calculating coefficients of parentage in sets of cultivars released in particular years. The pool of original ancestors of spring durum wheat cultivars includes 90 landraces and old varieties, more than a half (57%) of which originate from European countries, including Russia and Ukraine (45%). The original ancestors strongly differ in the frequency of presence in the cultivar pedigrees. Landraces Beloturka, Sivouska, Kubanka (T. durum Desf.), Transbaikalian emmer, Yaroslav emmer (T. dicoccum Schuebl.), Poltavka (T. aestivum L.), and the original ancestors of cultivars Kharkov 46, Narodnaya, and Melanopus 1932 enter in the pedigrees of more than half of cultivars created within the framework of various breeding programs. At that, their distribution by cultivars from different breeding centers strongly varies. Analysis of temporal dynamics of genetic diversity, based on genetic profiles and coefficients of parentage, has shown that the genetic diversity of Russian durum wheats increased during the period examined. Nevertheless, genetic erosion of the local material-a loss of approximately 20% of the pool of Russian original ancestors-has been found. The contribution of the original ancestors to the pedigrees of different cultivars, constructed in different breeding centers and recommended for cultivation in different regions, has been estimated. The variation of the released cultivars was highest in the Lower Volga region and lowest in the Ural region. In all, the lower threshold of genetic diversity in all regions does not reach the critical level, corresponding to the similarity of half-sibs. The set of modern cultivars included in the Russian Official List 2004 has a cluster structure.     

Polymorphism of a short fragment of the mitochondrial genome control region (D-loop) in the Siberian roe deer <Emphasis Type="Italic">Capreolus pygargus</Emphasis> Pallas, 1771 (Artiodactyla,Cervidae) from the Russian Far East

Genetic diversity of Siberian roe deer Capreolus pygargus, 1771 from the Russian Far East was studied based on polymorphism analysis of a mtDNA control region fragment (390 bp). Three phylogenetic lines were found in the animals examined. The trend for change of haplotypes of different phylogroups was shown to occur from north to south in Sikhote-Alin. The haplotype distribution of Siberian roe deer in the Russian Far East correlated with data on morphologiocal variability.