Many quantitative trait loci (QTLs) for agronomically important traits such as grain yield, disease resistance, and stress tolerance of rice (Oryza sativa L.) have been detected by using segregating populations derived from crosses between indica and japonica subspecies or with wild relatives. However, the QTLs involved in the control of natural variation in agronomic traits among closely related cultivars are still unclear. Decoding the whole genome sequences of Nipponbare and other temperate japonica rice cultivars has accelerated the collection of a huge number of single nucleotide polymorphisms (SNPs). These SNPs are good resource for developing polymorphic DNA markers and for detecting QTLs distributed across all rice chromosomes. The temperate japonica rice cultivar Koshihikari has remained the top cultivar for about 40 years since 1979 in Japan. Unraveling the genetic factors in Koshihikari will provide important insights into improving agronomic traits in temperate japonica rice cultivars. Here we describe recent progress in our studies as an example of genetic analysis in closely related cultivars.
Knowledge of the genetic architecture of flowering and maturity is needed to develop effective breeding strategies in tropical soybean. The aim of this study was to identify haplotypes across multiple environments that contribute to flowering time and maturity, with the purpose of selecting desired alleles, but maintaining a minimal impact on yield-related traits. For this purpose, a genome-wide association study (GWAS) was undertaken to identify genomic regions that control days to flowering (DTF) and maturity (DTM) using a soybean association mapping panel genotyped for single nucleotide polymorphism (SNP) markers. Complementarily, yield-related traits were also assessed to discuss the implications for breeding strategies. To detect either stable or specific associations, the soybean cultivars (N = 141) were field-evaluated across eight tropical environments of Brazil. Seventy-two and forty associations were significant at the genome-wide level relating respectively to DTM and DTF, in two or more environments. Haplotype-based GWAS identified three haplotypes (Gm12_Hap12; Gm19_Hap42 and Gm20_Hap32) significantly co-associated with DTF, DTM and yield-related traits in single and multiple environments. These results indicate that these genomic regions may contain genes that have pleiotropic effects on time to flowering, maturity and yield-related traits, which are tightly linked with multiple other genes with high rates of linkage disequilibrium.
Amylopectin is one of the major determinants of rice (Oryza sativa L.) grain quality, and a large difference in amylopectin is found between two subspecies: japonica and indica. However, the relationship among rice grain quality, indica/japonica genetic background, and amylopectin has not been clearly established. In this study, a series of backcross inbred lines derived from the cross between japonica (cv. Sasanishiki) and indica (cv. Habataki) were used to survey eating and cooking quality (ECQ), rapid visco analyzer (RVA) profiles, and the chain length distribution of amylopectin. The frequency of indica pedigree (Fi) was calculated to analyze the effects of Fi on grain quality and amylopectin. The results showed that the Sasanishiki cultivar was markedly enriched in chain length with DP6-15 and DP34-45 compared to the Habataki. DP34-45 strongly correlated to RVA characteristics, cooking quality, and prolamin content. The Fi also has significant correlations to RVA characteristics and ECQ, but only significantly negative correlation to DP34-45. Seven quantitative trait loci (QTLs) corresponding to amylopectin were mapped, of which three were in agreement with previous findings. The results of this study provide valuable information for amylopectin characteristics in the offspring derived from the subspecies cross, and the novel QTLs may provide new insights to the identification of minor starch synthesis-related genes.
Growing resistant cultivars is the best method of protecting the crops against Potato virus Y (PVY). There are a few sources of PVY resistance/tolerance in tobacco acquired through mass selection, X-ray induced mutagenesis and introgressions from wild Nicotiana species. Here, we compare major sources of PVY resistance/tolerance in inoculation tests using ten PVY isolates collected in Central Europe (Poland and Germany) and differing with their virulence. The diversity of collected isolates was confirmed by DAS-ELISA tests and two PCR assays targeting the most common recombination sites in the PVY genome. We used these isolates in inoculation tests on five resistant cultivars ‘V.SCR’, ‘PBD6’, ‘TN86’, ‘VAM’, ‘Wiślica’, a tolerant breeding line ‘BPA’ and four susceptible cultivars ‘BP-210’, ‘K326’, ‘NC95’, ‘Samsun H’. None of the tested cultivars/breeding lines showed universal resistance against all ten isolates. However, ‘VAM’ and ‘Wiślica’ appeared to be the most effective sources, as they showed no symptoms and gave negative DAS-ELISA tests for four out of ten tested PVY isolates. In contrast, tolerance of the breeding line ‘BPA’ was effective against all tested isolates, because inoculation did not lead to development of full disease symptoms in that breeding line.
Preharvest sprouting (PHS) of durum wheat (Triticum turgidum ssp. durum (Desf.) Husn.) is an important problem in Japan, where the rainy season overlaps with the harvest season. Since there are few PHS-resistant genetic resources in durum wheat, we introduced an R-gene for red seeds, the MFT gene, and the QPhs-5AL QTL, all of which are associated with PHS resistance, into durum wheat from a PHS-resistant bread wheat (T. aestivum L.) cultivar, ‘Zenkoujikomugi’ (Zen), by backcross breeding. Developed near isogenic lines (NILs) with red seeds had a lower percentage germination (PG) and germination index (GI) than the recurrent parent, and seed color had the greatest effect. A NIL combining all three sequences had the lowest GI and PG, with a similar GI to that of ‘Shiroganekomugi’ bread wheat. Among NILs with white seeds, a NIL combining MFT and QPhs-5AL had the lowest GI and PG. As the combination of all three sequences from Zen conferred PHS resistance on durum wheat, PHS-resistant genetic resources in bread wheat can be used in breeding durum wheat.
Grain size and weight are important determinants of rice yield. The identification of beneficial genes from wild rice that have been lost or weakened in cultivated rice has become increasingly important for modern breeding strategies. In this study, we constructed a set of chromosome segment substitution lines (CSSLs) of wild rice, Oryza rufipogon with the indica cultivar 9311 genetic background. Four grain-related traits, i.e., grain length (GL), grain width (GW), length-width ratio (LWR), and thousand grain weight (TGW), were screened across six environments. A total of 37 quantitative trait loci (QTLs) were identified in these environments and mapped to 12 chromosomes. Sixteen QTLs were detected in at least two environments, and two QTL clusters were observed on Chr. 4 and Chr. 8. Based on a comparative analysis with QTLs identified in previous studies, the CSSLs between Oryza rufipogon accessions and 9311 had high genetic diversity. Among the sixteen stable QTLs, seven for TGW, LWR, GL, and GW were not previously identified, indicating potentially novel alleles from wild rice. These CSSLs provide powerful tools for functional studies and the cloning of essential genes in rice; furthermore, we identified elite germplasm for rice variety improvement.
In wheat (Triticum aestivum L.) high temperature (≥30°C) during grain filling leads to considerable reduction in grain yield. We studied 400 multiple synthetic derivatives (MSD) lines to examine the genetic variability of heat stress–adaptive traits and to identify new sources of heat tolerance to be used in wheat breeding programs. The experiment was arranged in an augmented randomized complete block design in four environments in Sudan. A wide range of genetic variability was found in most of the traits in all environments. For all traits examined, we found MSD lines that showed better performance than their parent ‘Norin 61’ and two adapted Sudanese cultivars. Using the heat tolerance efficiency, we identified 13 highly heat-tolerant lines and several lines with intermediate heat tolerance and good yield potential. We also identified lines with alleles that can be used to increase wheat yield potential. Our study revealed that the use of the MSD population is an efficient way to explore the genetic variation in Ae. tauschii for wheat breeding and improvement.
Genetic variation in blast resistance was clarified in 334 Bangladesh rice accessions from 4 major ecotypes (Aus, Aman, Boro and Jhum). Cluster analysis of polymorphism data of 74 SSR markers separated these accessions into cluster I (corresponding to the Japonica Group) and cluster II (corresponding to the Indica Group). Cluster II accessions were represented with high frequency in all ecotypes. Cluster II was further subdivided into subclusters IIa and IIb. Subcluster IIa accessions were represented with high frequency in only Aus and Jhum ecotypes. Cluster I accessions were more frequent in the Aman ecotype than in other ecotypes. Distinct variations in resistance were found, and accessions were classified into 4 groups (A1, A2, B1 and B2) based on their reactions to standard differential blast isolates. The most susceptible group was A2 (which included susceptible variety Lijiangxintuanheigu, most of the differential varieties, and a few Bangladesh accessions), followed in order by A1, B2 and B1 (the most resistant). Accessions from 4 ecotypes fell with different frequencies into each of these resistance groups. These results demonstrated that Japonica Group accessions were found mainly in Aman, and Indica Group accessions were distributed across all ecotypes. Susceptible accessions were limited in Aus and Aman.
The genetic variation in resistance to blast (Pyricularia oryzae Cavara) in 195 rice accessions comprising 3 species of the AA genome complex (Asian rice [Oryza sativa L.], African rice [Oryza glaberrima Steud.] and wild rice [Oryza barthii]) was investigated based on their patterns of reaction to standard differential blast isolates (SDBIs) and SSR marker polymorphism data. Cluster analysis of the polymorphism data of 61 SSR markers identified 3 major clusters: cluster A (mainly Japonica Group or upland accessions), cluster B (mainly Indica Group or lowland accessions) and cluster C (O. glaberrima and O. barthii). The accessions were classified again into 3 resistance groups based on reactions to SDBIs: group Ia (susceptible), group Ib (middle resistance) and group II (high resistance). Group Ia included only a few differential varieties, susceptible controls and the Japonica Group cultivar Nipponbare. Accessions in clusters A and B included all 3 resistance groups and showed a wide variation in blast resistance, but cluster C contained only group Ib. These results demonstrated that variations in Asian rice (O. sativa) accessions in West Africa were skewed toward high resistance and that variations in O. glaberrima and O. barthii were limited and lower than the Asian rice accessions.
Wheat flours are used to produce bread, pasta, breakfast cereals, and biscuits; the various properties of these end-products are attributed to the gluten content, produced as seed storage proteins in the wheat endosperm. Thus, genes encoding gluten protein are major targets of wheat breeders aiming to improve the various properties of wheat flour. Here, we describe a novel compensating wheat–Thinopyrum elongatum Robertsonian translocation (T1AS.1EL) line involving the short arm of wheat chromosome 1A (1AS) and the long arm of Th. elongatum chromosome 1E (1EL); we developed this line through centric breakage-fusion. Compared to the common wheat cultivars Chinese Spring and Norin 61, we detected two additional 1EL-derived high-molecular-weight glutenin subunits (HMW-GSs) in the T1AS.1EL plants. Based on the results of an SDS-sedimentation volume to estimate the gluten strength of T1AS.1EL-derived flour, we predict that T1AS.1EL-derived flour is better suited to bread-making than Chinese Spring- and Norin 61-derived flour and that this is because of its greater gluten diversity. Also, we were able to assign 33 of 121 wheat PCR-based Landmark Unique Gene markers to chromosome 1E of Th. elongatum. These markers can now be used for further chromosome engineering of the Th. elongatum segment of T1AS.1EL.
Heading date is an important event to ensure successful seed production. Although foxtail millet (Setaria italica (L.) P.Beauv.) is an important foodstuff in semiarid regions around the world, the genetic basis determining heading date is unclear. To identify genomic regions regulating days to heading (DTH), we conducted a QTL-seq analysis based on combining whole-genome re-sequencing and bulked-segregant analysis of an F2 population derived from crosses between the middle-heading cultivar Shinanotsubuhime and the early-heading cultivar Yuikogane. Under field conditions, transgressive segregation of DTH toward late heading was observed in the F2 population. We made three types of bulk samples: Y-bulk (early-heading), S-bulk (late-heading) and L-bulk (extremely late-heading). By genome-wide comparison of SNPs in the Y-bulk vs. the S-bulk and the Y-bulk vs. the L-bulk, we identified two QTLs associated with DTH. The first QTL, qDTH2, was detected on chromosome 2 from the Y-bulk and S-bulk comparison. The second QTL, qDTH7, was detected on chromosome 7 from the Y-bulk and L-bulk comparison. The Shinanotsubuhime allele for qDTH2 caused late heading in the F2 population, whereas the Yuikogane allele for qDTH7 led to extremely late heading. These results suggest that allelic differences in both qDTH2 and qDTH7 determine regional adaptability in S. italica.
Plant breeding programs in local regions may have genetic and phenotypic variations that are desirable and shape adaptability during the establishment of local populations. Despite the characterization of genetic population structures in various kinds of populations, the effects of variations in phenotype on agro-economical traits currently remain unclear. In the present study, we evaluated phenotypic changes in 26 agro-economical traits among the local population during rice breeding programs in Hokkaido. Wide variations were observed in all 26 agro-economical traits with continuous distributions. In order to elucidate improvements in these agro-economic traits during rice breeding programs in Hokkaido, values were compared between genetic population structures. Traits were classified into four patterns based on the timing of significant differences. Patterns A and B showed significant differences once and twice, respectively. Pattern C gradually showed significant differences. Pattern D showed no significant differences for the desired directions. Based on the changes in phenotype observed in the present study and the genetic population structure for the local population in Hokkaido, a model of the artificial selection for phenotypes in genetic diversity among the local population during plant breeding programs has been proposed.
The fertility conversion of photoperiod thermo-sensitive genic male sterility (P/TGMS) lines in rice is mostly regulated by the P/TGMS genes in different environmental conditions. A point mutation with G-C on the pms3 (p/tms12-1) gene regulates the pollen fertility of Nongken58S and a large amount of Nongken58S-derived lines. In this study, we developed and designed a functional co-dominant marker according to the SNP loci for the pms3 (p/tms12-1) gene. We can differentiate the SNP loci in pms3 (p/tms12-1) gene from another TGMS lines and inbred cultivars using the dpms3-54 marker. The results showed that 376 bp band was detected in the homozygous genotype for pms3 (p/tms12-1), while 359 bp band was detected in the homozygous genotype for non-pms3 (p/tms12-1), two bands with 376 and 359 bp were detected in the heterozygous genotype. The dpms3-54 marker can be used to test the purity of two-line hybrid rice seeds and to divide each of F1 plant into homozygous and heterozygous genotypes at the seedling stage. Thus, this study provide a useful functional marker to detect pms3 (p/tms12-1) gene in different genetic resources and populations, which can be applied to the development and breeding of P/TGMS lines in two-line hybrid rice system with marker-assisted selection.
Rice brown spot (BS), caused by Bipolaris oryzae, is one of the major diseases of rice in Japan. Quantitative resistance has been observed in local cultivars (e.g., CH45), but no economically useful resistant variety has been bred. Using simple sequence repeat (SSR) polymorphic markers, we conducted quantitative trait locus (QTL) analysis of BS resistance in backcross inbred lines (BILs) from a cross between indica CH45 (resistant) and japonica Koshihikari (susceptible). On the basis of field disease evaluations in 2015 and 2016, four QTLs contributing to BS resistance were identified on chromosomes 2 (qBSR2-kc), 7 (qBSR7-kc), 9 (qBSR9-kc), and 11 (qBSR11-kc). The ‘CH45’ alleles at qBSR2-kc, qBSR7-kc, and qBSR11-kc and the ‘Koshihikari’ allele at qBSR9-kc increased resistance. The major QTL qBSR11-kc explained 23.0%–25.9% of the total phenotypic variation. Two QTLs (qBSR9-kc and qBSR11-kc) were detected in both years, whereas the other two were detected only in 2016. Genetic markers flanking these four QTLs will be powerful tools for marker-assisted selection to improve BS resistance.
Simple sequence repeat (SSR) is a popular tool for individual fingerprinting. The long-core motif (e.g. tetra-, penta-, and hexa-nucleotide) simple sequence repeats (SSRs) are preferred because they make it easier to separate and distinguish neighbor alleles. In the present study, a new set of 8 tetra-nucleotide SSRs in potato (Solanum tuberosum) is reported. By using these 8 markers, 72 out of 76 cultivars obtained from Japan and the United States were clearly discriminated, while two pairs, both of which arose from natural variation, showed identical profiles. The combined probability of identity between two random cultivars for the set of 8 SSR markers was estimated to be 1.10 × 10−8, confirming the usefulness of the proposed SSR markers for fingerprinting analyses of potato.