BREEDING SCIENCE

Phenotyping and a genome-wide association study of elite lines of pearl millet
Kambara K, Gupta SK, Takano T and Tsugama D
Pearl millet ( (L.) R. BR.) is a cereal crop mainly grown in India and sub-Saharan Africa. In pearl millet, genes and genomic regions associated with traits are largely unknown. Pearl millet parental lines bred at the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) are useful for the production and breeding of pearl millet. However, the phenotypic diversity of these lines has not been fully evaluated. In this study, 16 traits of 107 of those parental lines were assessed with field trials in Japan, and a genome-wide association study (GWAS) was performed using these phenotypic data to identify the genomic regions and genes associated with those traits. The GWAS revealed genomic regions associated with culm height and pigmentation of the shoot basal part (PS). The genomic region associated with PS contained a homolog of (), a gene involved in anthocyanin accumulation in . The homolog can be a candidate for a gene involved in regulating PS in pearl millet. These results provide a better understanding of the phenotypic diversity of pearl millet and its genetic background.
Genetic mapping and quantitative trait loci analysis for pistillate flowers per node and multi-pistillate flower traits in the F cucumber population
Anankul N, Sattayachiti W, Onmanee N, Chanmoe S, Bundithya W and Kumchai J
This study focused on cucumbers' multi-pistillate flower (MPF) trait, which is essential for high yields. A genetic linkage map was constructed using a population of 219 F plants to analyze quantitative trait loci (QTL) associated with MPF traits. Crossbreeding of EWSCU-809 (MPF) with EWSCU-989 (single pistillate flower: SPF) generated an F hybrid that self-pollinated to form an F population. Based on 244 single nucleotide polymorphic markers across seven cucumber chromosomes, a linkage map facilitated QTL analysis considering average pistillate flowers (PFs) per node and nodes with MPF traits. The results indicated a 9:6:1 epistatic ratio in the F populations, revealing recessive allele control of the MPF trait in gynoecious plants. Three QTLs (, , ) on chromosomes 2, 3, and 7 were associated with average PFs per node, explaining 5.6 to 10.3% of phenotypic variation. Four QTLs (, , , ) on chromosomes 2, 3, 4, and 7 were linked to the presence of nodes with MPF traits, explaining 5.8 to 10.6% of phenotypic variance. Notably, QTL regions overlapped between the two datasets, suggesting pleiotropic effects, particularly on chromosomes 3 and 7. These reliable QTLs have the potential to improve breeding programs, enhance PF development, and increase cucumber yields.
Development of SNP genotyping assays for heading date in rice
Kitazawa N, Shomura A, Mizubayashi T, Ando T, Hayashi N, Yabe S, Matsubara K, Ebana K, Yamanouchi U and Fukuoka S
Heading date (HD) is a crucial agronomic trait, controlled by multiple loci, that conditions a range of geographical and seasonal adaptations in rice ( L.). Therefore, information on the HD genotypes of cross parents is essential in marker-assisted breeding programs. Here, we used the Fluidigm 96-plex SNP genotyping platform to develop genotyping assays to determine alleles at 41 HD loci (29 previously characterized genes and 12 quantitative trait loci [QTLs], including a newly detected QTL). The genotyping assays discriminated a total of 144 alleles (defined on the basis of the literature and publicly available databases) and QTLs. Genotyping of 377 cultivars revealed 3.5 alleles per locus on average, a higher diversity of , , , and than that of the other loci, and the predominance of the reference ('Nipponbare') genotype at 30 of the 41 loci. HD prediction models using the data from 200 cultivars showed good correlation ( > 0.69,  < 0.001) when tested with 22 cultivars not included in the prediction models. Thus, the developed assays provide genotype information on HD and will enable cost-effective breeding.
Screening corn hybrids for early-stage drought stress tolerance using SPAR phenotyping platform
Lone AA, Hussan SU, Jumaa SH, Dar ZA and Reddy KR
An experiment was conducted comprising of six corn hybrids that were subjected to drought and irrigated environment in separate columns in soil-plant-atmosphere-research (SPAR) cubes. The treatments and hybrids in SPAR cubes were replicated four times and a two factorial randomized complete block design (RCBD) was used to analyze the effect of drought on hybrids and their effects on traits. Significant drought × hybrid interactions were observed for most of the parameters. All the traits observed under this study were affected by drought conditions. Root volume (RV) and root shoot ratio (RSR) increased, and number of root tips (NRT), number of root forks (NRF), and number of root crossings (NRC) were drastically reduced under drought conditions. The photosynthetic rate (Phot) declined by 57.96% and electron transport rate (ETR) by 54.60% and was negatively correlated with plant height (PH) and root number (RN) during drought stress. Chlorophyll content (SPAD) showed a non-significant correlation with all the traits. As per results, there were significant differences among corn hybrids for different traits studied under the SPAR setup, which indicates that this setup successfully creates differences in treatments. A cumulative drought stress response index (CDSRI) was worked out. DKC-6581 and N61X-3110 were found to be highly drought tolerant as per our findings.
THB1, a putative transmembrane protein that causes hybrid breakdown in rice
Wakabayashi T and Kato K
Hybrid breakdown is a post-zygotic reproductive isolation that hinders genetic exchange between species or populations in both animals and plants. Two complementary recessive genes, () and , cause hybrid breakdown in rice (). The present study delimited the locus to a 9.1-kb sequence, containing a single gene encoding a putative transmembrane protein with unknown functions. Haplotype analysis of in the two core collections of 119 accessions revealed that these accessions were divided into 22 haplotypes. A test cross with carrier showed that haplotype2 (H2) was assigned to and was restricted to . A nonsynonymous nucleotide polymorphism (SNP) specific to H2 was identified as a causal mutation in . A test cross with carrier indicated that six accessions, including , , and , carried . These results suggest that has recently evolved in , whereas arose in an ancient and introgressed into the present three subgroups. Furthermore, we developed a derived cleaved amplified polymorphic sequence (dCAPS) marker to detect causal SNP in . Our findings provide new insights into reproductive isolation and may benefit rice breeding.
Substitution mapping and characterization of brown planthopper resistance genes from traditional rice cultivar 'Rathu Heenati' ( L.)
Shar SBD, Nguyen CD, Sanada-Morimura S, Zheng SH and Fujita D
The brown planthopper (BPH; Stål) is a devastating pest that causes severe rice yield losses in Asia. Introducing multiple BPH resistance genes into rice cultivars is an effective and sustainable way to mitigate yield losses. A traditional rice cultivar, 'Rathu Heenati', has durable BPH resistance due to multiple resistance genes (including and ) and quantitative trait loci (QTLs). However, these genes have not been used in Japanese rice breeding owing to limited genetic information. To identify markers tightly linked to and introgressed into the 'Sagabiyori' (susceptible) genetic background, we performed substitution mapping. was delimited between RM3132 and RM589 on chromosome 6, and between RM16493 and RM16531 on chromosome 4. We also performed QTL analysis to identify additional BPH resistance genes from 'Rathu Heenati' and detected a QTL, denoted as , on chromosome 3. The effect of pyramiding and was significantly greater against virulent BPH populations than that of either gene alone. The combination of , and from 'Rathu Heenati' might be facilitated to improve commercial Japanese cultivars with more robust BPH resistance.
Development of a laboratory test using stem cuttings to measure resistance to foot rot disease caused by in sweetpotato
Tabuchi H, Kobayashi A, Kawata Y, Okada Y and Ohdaira Kobayashi Y
Over the last several years, foot rot caused by has become the most destructive sweetpotato disease in the southernmost region of Japan. Breeding of cultivars resistant to foot rot is required for effective and low-cost management. Field tests are often used to evaluate resistance of cultivars, but this approach has several limitations, including a long test period of several months and the requirement of field isolation and labor-intensive procedures. To minimize these issues, we have developed an easier and faster laboratory method using stem cuttings for the resistance test by optimizing four parameters: the number of unfolded leaves per cuttings, the positions of stems from which a cutting was prepared, the adequate number of culture days after inoculation, and the density of conidia of at inoculation. Significant correlation was detected between the resistance indices of the laboratory test and the field test, namely, the length of the rotted part of a stem and the proportion of the plants rotted at the basal part of a stem, respectively. These results indicated that the laboratory test could indirectly evaluate the foot rot resistance of sweetpotato stems in the field and will be helpful to breed resistant cultivars.
Genome-wide association analysis was used to discover genes related to soybean grain weight per plant and 100-grain weight
Sun T, Zhang Q, Liu L, Tang Y, Wang J, Wang K, Yuan B and Wang P
As an essential grain, oil, and feed crop worldwide, soybean plays a crucial role. Developing high-yielding and high-quality soybean varieties is a critical goal for breeders. The grain weight per plant and 100-grain weight directly impact the soybean yield. This study combined genotypic data from the population with phenotypic data. Based on genome-wide association analysis (GWAS), GLM and MLM analysis models were used to locate the Gm04_21489088, Gm04_15703616, and Gm04_46466250 are loci related to soybean grain weight per plant, and find the Gm09_20334173, Gm04_39518612 and Gm04_39518624 are loci related to 100-seed weight. After performing a reference comparison, we conducted gene annotation and identified candidate genes and , potentially associated with grain weight per plant in soybeans. These genes are primarily involved in protein synthesis and cell differentiation processes. The candidate gene , associated with the 100-grain weight trait, was successfully annotated. The analysis revealed that the gene primarily involves enzyme activity, suggesting its potential role in regulating grain weight. These findings offer valuable insights into the mechanism of soybean yield and serve as a critical theoretical foundation and genetic resource for cultivating new soybean germplasm with high yield. These findings are of immense significance for future research endeavors to achieve high-yielding soybean varieties.
Identification of as a positive awn regulator from two wild rice species
Mizutani M, Murase R, Aoki SI, Sato Y, Yamagata Y, Yasui H, Yoshimura A, Ashikari M and Bessho-Uehara K
The awn is a bristle-like appendage that protrudes from the seed tip and plays a critical role in preventing feed damage and spreading habitats in many grass species, including rice. While all wild species in the genus have awns, this trait has been eliminated in domesticated species due to its obstructive nature to agricultural processes. To date, several genes involved in awn development have been identified in wild rice, and which are ancestral species of cultivated rice in Asia and Africa, respectively. However, the responsible genes for awn development have not been identified in other wild rice species even though multiple QTLs have been reported previously. In this study, we identified gene responsible for awn development in two wild rice species, and . encodes a cytochrome P450 enzyme and is homologous to , a known brassinosteroid biosynthesis enzyme in rice. The identification of provides insight into a distinct molecular mechanism underlying awn development that occurs in geographically separated environments.
Identification of a major QTL conferring resistance to derived from the winter wheat 'Hokkai 240' on chromosome 2AS
Kawaguchi K, Ohki T, Ishikawa G, Sayama M, Terasawa Y, Oda S, Fujita M, Ito M and Hatta K
Wheat yellow mosaic disease is a soilborne disease caused by (WYMV). Symptoms include yellow mosaic coloring of leaves, stunting, and growth inhibition. Here we conducted a detailed analysis of resistance to this virus in winter wheat 'Hokkai 240' by carrying out inoculation tests of WYMV and conducting field tests. The resistance level observed in 'Hokkai 240' was compared with those in varieties harboring known resistance genes. In the inoculation tests, 'Hokkai 240' showed resistance to WYMV Pathotypes I and II and partial resistance to Pathotype III. This result was contrary to the sensitive responses to the three pathotypes exhibited by the variety harboring resistance gene on chromosome 2DL. In fields infected with WYMV Pathotypes II and III, 'Hokkai 240' plants exhibited few disease symptoms and little proliferation of the virus. By analyzing the quantitative trait loci (QTLs) in recombinant inbred lines from a cross between 'Hokkai 240' and 'Nanbukomugi', a single major QTL, , from 'Hokkai 240', which had significant effects on Pathotypes II and III of the virus, was detected in the proximity of and mapped on chromosome 2AS. These results indicate that may be useful for developing broad resistance to WYMV in wheat breeding programs.
Genomic diversity of the Japanese wheat core collection and selection of alleles for agronomic traits in the breeding process
Mizuno N, Kobayashi F, Morita T and Handa H
Combining high-throughput genotyping data with the latest wheat genomic information provided more detailed information on the genetic diversity of the Japanese wheat core collection (JWC). Analysis of genomic population structure divided the JWC accessions into three populations: northeast Japan accessions, native and southwest Japan accessions, and modern accessions showing mixed breeding patterns. This indicates that Japanese wheat varieties have a background of native genomes from southwest Japan incorporating valuable genes from various exotic lines, which is supported by the history of Japanese wheat breeding. Association analyses of several agronomic traits have revealed how genes or alleles have been selected in Japanese wheat breeding and how they differ from those in other regions of the world. This analysis of the JWC collection is expected to contribute not only to the elucidation of genetic diversity in Japanese wheat accessions but also to future wheat breeding by providing a new genetic resource.
Single candidate gene for salt tolerance of (Ohwi) Ohwi & Ohashi identified by QTL mapping, whole genome sequencing and triplicated RNA-seq analyses
Ito M, Ohashi H, Takemoto M, Muto C, Seiko T, Noda Y, Ogiso-Tanaka E, Nagano AJ, Takahashi Y, Furukawa J, Monden Y and Naito K
Salt tolerance has been an important issue as a solution for soil salinization and groundwater depletion. To challenge this issue, genetic diversity of wild plants must be harnessed. Here we report a discovery of a candidate gene for salt tolerance in , one of the coastal species in the genus . Using intraspecific variation, we performed a forward genetic analysis and identified a strong QTL region harboring ~200 genes. To further narrow down the candidate genes, we performed a comparative transcriptome analysis, using the genome sequence of azuki bean () as a reference. However the detected differentially-expressed genes (DEGs) did not include those related to salt tolerance. As we suspected that the target gene in is missing in , we sequenced the whole genome sequence of with long-reads. By re-analyzing the transcriptome data with the new reference genome, we successfully identified as a candidate gene, which was missing not only in but also in the salt-sensitive accession of . Further comparative analysis revealed that the tolerant genotypes conserved the ancestral form of the locus, while the sensitive genotypes did not. We also emphasize the pitfalls in our study, such as position effect in a growth chamber, missing important genes in the reference genome, and limited reproducibility of RNA-seq experiments.
Development of genomic and genetic resources facilitating molecular genetic studies on untapped Myanmar rice germplasms
Furuta T, Saw OM, Moe S, Win KT, Hlaing MM, Hlaing ALL, Thein MS, Yasui H, Ashikari M, Yoshimura A and Yamagata Y
To counteract the growing population and climate changes, resilient varieties adapted to regional environmental changes are required. Landraces are valuable genetic resources for achieving this goal. Recent advances in sequencing technology have enabled national seed/gene banks to share genomic and genetic information from their collections including landraces, promoting the more efficient utilization of germplasms. In this study, we developed genomic and genetic resources for Myanmar rice germplasms. First, we assembled a diversity panel consisting of 250 accessions representing the genetic diversity of Myanmar varieties, including an elite lowland variety, Inn Ma Yebaw (IMY). Our population genetic analyses illustrated that the diversity panel represented Myanmar varieties well without any apparent population structure. Second, de novo genome assembly of IMY was conducted. The IMY assembly was constructed by anchoring 2888 contigs, which were assembled from 30× coverage of long reads, into 12 chromosomes. Although many gaps existed in the IMY genome assembly, our quality assessments indicated high completeness in the gene-coding regions, identical to other near-gap-free assemblies. Together with dense variant information, the diversity panel and IMY genome assembly will facilitate deeper genetic research and breeding projects that utilize the untapped Myanmar rice germplasms.
Detection and validation of QTLs for green stem disorder of soybean ( (L.) Merr.)
Ogata D, Taguchi-Shiobara F, Uchikawa O, Miyazaki M and Ishibashi Y
In mechanically harvested soybean, green stem disorder (GSD) is an undesirable trait that causes green-stained seeds, which are graded lower in Japan. To obtain DNA markers for reduced GSD, we conducted a quantitative trait locus (QTL) analysis for 2 years using F and F lines from a cross between 'Suzuotome' (less GSD) and 'Fukuyutaka' (more GSD). We validated the effect of a detected QTL for GSD by first identifying F or F plants in which one or more markers in the QTL region were heterozygous. The F or F progeny of each plant was used to form a pair consisting of two groups in which the QTL region was homozygous for either the 'Suzuotome' or 'Fukuyutaka' allele in a similar genetic background, and the two groups within each pair were compared for GSD. Over 3 years of testing, the 'Suzuotome' allele of a QTL on chromosome 6 was found to reduce the level of GSD. This novel QTL was mapped to the region around DNA marker W06_0130, and was not closely linked to QTLs for important agronomic traits including yield components. Using this marker, the low level of GSD from 'Suzuotome' could be conferred to 'Fukuyutaka' or other high-GSD cultivars.
Polyploid QTL-seq revealed multiple QTLs controlling steamed tuber texture and starch gelatinization temperature in sweetpotato
Yamakawa H, Mizubayashi T and Tanaka M
Sweetpotato () includes diverse cultivars with flesh textures ranging from dry to moist. Moist-fleshed cultivars often contain starch with a lower gelatinization temperature (GT). To elucidate the genetic determinants of flesh texture and starch GT, we conducted a QTL analysis using F progenies obtained from a cross between dry-fleshed and moist-fleshed cultivars, 'Benikomachi' (BK) and 'Amahazuki' (AH), by using an updated polyploid QTL-seq pipeline. Flesh texture was assessed based on the wet area ratio (WAR) observed on the cut surface of steamed tubers, as progenies with dry and moist flesh exhibited low and high WAR values, respectively, demonstrating a strong correlation. Three QTLs were found to regulate the WAR. Notably, two AH-derived alleles at 4.30 Mb on Itr_chr05 and 21.01 Mb on Itr_chr07, along with a BK-derived allele at 2.89 Mb on Itr_chr15, were associated with increased WAR. Starch GT, which displayed no correlation with either flesh texture or WAR, was distinctly influenced by two QTLs: a GT-increasing BK-derived allele at 1.74 Mb on Itr_chr05 and a GT-decreasing AH-derived allele at 30.16 Mb on Itr_chr12. Consequently, we developed DNA markers linked to WAR, offering a promising avenue for the targeted breeding of sweetpotato with the desired flesh textures.
Diversity of salt tolerance in , wild related species of the azuki bean ()
Ogiso-Tanaka E, Chankaew S, Isemura T, Marubodee R, Kongjaimun A, Baba-Kasai A, Okuno K, Ehara H and Tomooka N
is a wild species closely related to the azuki bean (), with salt-tolerance abilities. The present study aimed to explore the genetic and salt tolerance diversity within the species, by evaluating the phylogenetic relationships of 55 accessions of including 25 newly collected from the Gotō Islands and Iki in Nagasaki Prefecture, Japan. We conducted salt-tolerance analysis for 48 of the accessions, including 18 of the newly collected accessions. Phylogenetic analysis based on single-nucleotide polymorphisms obtained from MIGseq and RADseq analyses revealed the genetic diversity of to reflect the geographic arrangement of the habitat islands. Korean accessions formed one clade, while Japanese accessions predominantly grouped into Uku Island and Fukue Island subclades. Within this population, we identified "G4-2" (JP248291) as the most salt tolerant, surpassing even the previously reported "Ukushima" accession. Both accessions collected from Uku Island, with accessions belonging to the Uku Island subclade exhibiting a strong trend of salt tolerance. Our results strongly suggest the occurrence of genetic mutations conferring enhanced salt tolerance in specific clade and region. This study highlights the potential of genetic analyses for identifying regions suitable for collecting valuable genetic resources for stress tolerance.
DNAMarkMaker: streamlining ARMS and CAPS marker development from resequencing data with NGS short reads
Segawa T, Saiga S, Takata M, Kumazawa R, Hara M, Yamakawa H and Takagi H
DNA markers serve as essential tools in breeding selection and genetic analysis. However, developing DNA markers can be time-consuming and labor-intensive due to the need to identify polymorphisms between cultivars/lines and to design suitable primers. To address these challenges, we have developed DNAMarkMaker, a tool designed to automate the process of primer design for Amplification Refractory Mutation System (ARMS) and Cleaved Amplified Polymorphic Sequences (CAPS) markers, utilizing resequencing data. One key feature of DNAMarkMaker is its user-friendly graphical user interface (GUI), ensuring its accessibility and ease of use, even for researchers not well-versed in bioinformatics. We confirmed DNAMarkMaker's applicability by developing DNA markers for rice, potato, and turnip-each representing distinct genome structures: homozygous diploid, heterozygous autotetraploid, and heterozygous diploid, respectively. DNAMarkMaker will contribute to the rapid and efficient development of DNA markers, accelerating breeding and genetic analysis in various crops.
Interaction between genetic regions responsible for the starch properties in non-glutinous rice varieties in Hokkaido, Japan
Ikegaya T
Starch properties are the major determinants of grain quality and food characteristics in rice ( L.). Understanding the interactions between genetic regions responsible for starch properties will lead to the development of rice cultivars with desirable characteristics. This study investigated the genetic effect and interaction between , a low-amylose quantitative trait locus (QTL), and the genetic region around (). Both these factors are responsible for the starch properties of the Hokkaido breeding population. The amylose content, pasting temperature, and amylopectin chain-length distribution were compared using F lines derived from the cross between the lower amylose content and lower pasting temperature strain 'Hokkai332 (, )' and the higher amylose content and higher pasting temperature variety 'Kitagenki (-, )'. The genotype exhibited low amylose content and reduced the hardness of boiled rice but increased the ratio of amylopectin long chains and did not alter the pasting temperature. In contrast, the genotype was associated with pasting temperature but did not affect the amylose content and hardness of boiled rice. It was suggested that appropriately selecting genotypes of these genetic regions and QTL would allow the fine-tuning of starch properties of cooked rice suitable for future demand.
Seed abortion caused by the combination of two duplicate genes in the progeny from the cross between and
Toyomoto D, Shibata Y, Uemura M, Taura S, Sato T, Henry R, Ishikawa R and Ichitani K
Seed development is an essential phenomenon for all sexual propagative plant species. The functional allele at () or () loci is essential for seed development for and . In the present study, we performed fine mapping of , narrowing down the area of interest to 333kb on chromosome 6. Haplotype analysis around the locus of accessions indicated that they shared the DNA polymorphism, suggesting that they have a common abortive allele at the locus. Linkage analysis of the candidate gene showed that it was located on chromosome 4. The candidate was confirmed using a population in which both the and genes were segregating. The chromosomal region covering the gene was predicted to contain 30 protein-coding genes in . Five of these genes have conserved DNA sequences in the chromosomal region of the gene on chromosome 4, and not on chromosome 6, of . These results suggest that these five genes could be candidates for , and that their orthologous genes located on chromosome 4 of could be candidates for .
Narrowing down a major QTL region reveals () as the candidate gene controlling flowering time in mungbean ()
Amkul K, Laosatit K, Lin Y, Yimram T, Chen J, Yuan X, Chen X and Somta P
Flowering time is an important agronomic trait that is highly correlated with plant height, maturity time and yield in mungbean. Up to present, however, molecular basis of flowering time in mungbean is poorly understood. Previous studies demonstrated that flowering time in mungbean is largely controlled by a major QTL on linkage group 2 (LG2). In this study, the QTL on the LG2 in mungbean was investigated using F and F populations derived from a cross between mungbean cultivar Kamphaeng Saen 2 (KPS2) and wild mungbean accession ACC41. The QTL was narrowed down to a genome region of 164.87 Kb containing a phytochrome gene, designated , encoding phytochrome E (phyE), a known photoreceptor modulating flowering time. Compared to of the wild ACC41, of KPS2 contained several single nucleotide polymorphisms (SNPs) causing amino acid changes. Those SNPs were also found in other mungbean cultivars. Some amino acid changes were predicted to occur in the regulatory region of phytochromes. Gene expression analysis revealed that in KPS2 was expressed significantly higher than that in ACC41. These results showed that is the candidate gene controlling flowering time in the mungbean.
NARO historical phenotype dataset from rice breeding
Matsushita K, Onogi A and Yonemaru JI
Data from breeding, including phenotypic information, may improve the efficiency of breeding. Historical data from breeding trials accumulated over a long time are also useful. Here, by organizing data accumulated in the National Agriculture and Food Research Organization (NARO) rice breeding program, we developed a historical phenotype dataset, which includes 6052 records obtained for 667 varieties in yield trials in 1991-2018 at six NARO research stations. The best linear unbiased predictions (BLUPs) and principal component analysis (PCA) were used to determine the relationships with various factors, including the year of cultivar release, for 15 traits, including yield. Yield-related traits such as the number of grains per panicle, plant weight, grain yield, and thousand-grain weight increased significantly with time, whereas the number of panicles decreased significantly. Ripening time significantly increased, whereas the lodging degree and protein content of brown rice significantly decreased. These results suggest that panicle-weight-type high-yielding varieties with excellent lodging resistance have been selected. These trends differed slightly among breeding locations, indicating that the main breeding objectives may differ among them. PCA revealed a higher diversity of traits in newer varieties.