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Genebanks provide access to diverse materials for crop improvement. To utilize and evaluate them effectively, core collections, such as the World Rice Core Collection (WRC) in the Genebank at the National Agriculture and Food Research Organization, have been developed. Because the WRC consists of 69 accessions with a high degree of genetic diversity, it has been used for >300 projects. To allow deeper investigation of existing WRC data and to further promote research using Genebank rice accessions, we performed whole-genome resequencing of these 69 accessions, examining their sequence variation by mapping against the Oryza sativa ssp. japonica Nipponbare genome. We obtained a total of 2,805,329 single nucleotide polymorphisms (SNPs) and 357,639 insertion–deletions. Based on the principal component analysis and population structure analysis of these data, the WRC can be classified into three major groups. We applied TASUKE, a multiple genome browser to visualize the different WRC genome sequences, and classified haplotype groups of genes affecting seed characteristics and heading date. TASUKE thus provides access to WRC genotypes as a tool for reverse genetics. We examined the suitability of the compact WRC population for genome-wide association studies (GWASs). Heading date, affected by a large number of quantitative trait loci (QTLs), was not associated with known genes, but several seed-related phenotypes were associated with known genes. Thus, for QTLs of strong effect, the compact WRC performed well in GWAS. This information enables us to understand genetic diversity in 37,000 rice accessions maintained in the Genebank and to find genes associated with different phenotypes.The sequence data have been deposited in DNA Data Bank of Japan Sequence Read Archive (DRA) (Supplementary Table S1).

Burkholderia glumae causes bacterial seedling rot (BSR) and bacterial grain rot (BGR) in rice (Oryza sativa), both of which are important diseases in Japan. We previously evaluated major Japanese cultivars for BGR resistance and selected standard cultivars for resistance assessments. Here, we assessed the BSR occurrence rate in cultivars from the World Rice Collection (WRC) and other sources and found wide variation in resistance. Next, we evaluated major Japanese cultivars for BSR resistance and found that two Japanese landraces, ‘Kujuu’ and ‘Aikoku’, showed “strong” resistance; most others were categorized as “medium” or “medium to weak”. We previously developed a near-isogenic line (RBG1-NIL) by introducing the genomic region containing RBG1, a quantitative trait locus (QTL) for BSR resistance, from ‘Nona Bokra’ (indica) into ‘Koshihikari’ (temperate japonica). The resistance level of RBG1-NIL was “strong”, indicating the effectiveness of RBG1 against BSR. The correlation between BSR and BGR resistance scores was low, indicating that it is necessary to introduce QTLs for resistance from different sources to develop cultivars resistant to both BSR and BGR. On the basis of the screening results, we selected standard cultivars for BSR resistance to cover a range of resistance levels.

Background In temperate rice cultivation regions, japonica rice cultivars are grown preferentially because consumers deem them to have good eating quality, whereas indica rice cultivars have high grain yields and strong heat tolerance but are considered to have poor eating quality. To mitigate the effects of global warming on rice production, it is important to develop novel rice cultivars with both desirable eating quality and resilience to high temperatures. Eating quality and agronomic traits were evaluated in a reciprocal set of chromosome segment substitution lines derived from crosses between a japonica rice cultivar ‘Koshihikari’ and an indica rice cultivar ‘Takanari’. Results We detected 112 QTLs for amylose and protein contents, whiteness, stickiness, hardness and eating quality of cooked rice grains. Almost of ‘Koshihikari’ chromosome segments consistently improved eating quality. Among detected QTLs, six QTLs on chromosomes 1–5 and 11 were detected that increased whiteness and stickiness of cooked grains or decreased their hardness for 3 years. The QTLs on chromosomes 2–4 were not associated with differences in amylose or protein contents. QTLs on chromosomes 1–5 did not coincide with QTLs for agronomic traits such as heading date, culm length, panicle length, spikelet fertility and grain yield. Genetic effects of the detected QTLs were confirmed in substitution lines carrying chromosome segments from five other indica cultivars in the ‘Koshihikari’ genetic background. Conclusion The detected QTLs were associated with differences in eating quality between indica and japonica rice cultivars. These QTLs appear to be widely distributed among indica cultivars and to be novel genetic factors for eating quality traits because their chromosome regions differed from those of the GBSSI (Wx) and SSIIa (Alk) genes. The detected QTLs would be very useful for improvement of eating quality of indica rice cultivars in breeding programs.

The problems which affect seed germination of D.melanoxylon in the natural environment are also reflected on germination in the Murashige and Skoog medium and in callus induction. Vulnerability to fungal attack is a limitation for successful callus induction and germination in the culture room.

IR64 is one of the world’s most popular rice cultivars. To collect genetic factors involved in controlling its heading date, we developed 70 reciprocal advanced-backcross populations with a total of 6284 individuals at the BC4F2 generation from crosses between Koshihikari and IR64. We detected 29 QTLs associated with heading date on chromosomes 3, 5–8, 10, and 12. Twenty QTLs were located in the same chromosome regions as previously isolated heading date genes (Hd1, Hd6, Hd16, Ghd7, DTH8, Hd17, and Hd18). The rest were located in other chromosome regions. We found more number of QTLs than previous studies using mapping populations of IR64. Fine mapping in additional advanced-backcross populations clearly revealed that QTLs on the long arm of chromosome 7 are overlapping and seem to be a novel genetic factor for heading date because of their different locations from OsPRR37. Our results suggest that the difference in heading date between IR64 and Koshihikari is genetically controlled by many factors, and that a non-functional allele of Hd1 contributes to early heading of IR64 in the genetic background of functional alleles of other heading date QTLs and genes such as Hd6 and Hd16.

Bacterial grain rot (BGR), caused by the bacterial pathogen Burkholderia glumae, is one of the most destructive rice (Oryza sativa) diseases in Japan; however, there are no BGR-resistant cultivars for use in Japan. We previously developed a cut-panicle inoculation method to assess the levels of BGR resistance in the World Rice Collection (WRC). Here, we evaluated major Japanese cultivars for BGR resistance and found that none showed "strong" or "medium to strong" resistance; most were categorized as "medium to weak". On the basis of the screening results, standard cultivars for BGR resistance were selected according to resistance level and relative maturity. Our results indicate that it is necessary to introduce quantitative trait loci (QTLs) from indica or tropical japonica resistant cultivars into Japanese temperate japonica to develop BGR-resistant cultivars for Japan. We previously developed a near-isogenic line (RBG2-NIL) by introducing the genomic region containing RBG2 from 'Kele' (indica) into 'Hitomebore'. In this experiment, we confirmed the resistance level of RBG2-NIL. The resistance score of RBG2-NIL was "medium to strong", indicating its effectiveness against BGR.

The diversification of flowering time in response to natural environments is critical for the spread of crops to diverse geographic regions. In contrast with recent advances in understanding the molecular basis of photoperiodic flowering in rice (Oryza sativa), little is known about how flowering-time diversification is structured within rice subspecies. By analyzing genome sequencing data and a set of 429 chromosome segment substitution lines (CSSLs) originating from 10 diverse rice accessions with wide distributions, we revealed diverse effects of allelic variations for common flowering-time quantitative trait loci in the recipient's background. Although functional variations associated with a few loci corresponded to standing variations among subspecies, the identified functional nucleotide polymorphisms occurred recently after rice subgroup differentiation, indicating that the functional diversity of flowering-time gene sequences was not particularly associated with phylogenetic relationship between rice subspecies. Intensive analysis of the Hd1 genomic region identified the signature of an early introgression of the Hd1 with key mutation(s) in aus and temperate japonica accessions. Our data suggested that, after such key introgressions, new mutations were selected and accelerated the flowering-time diversity within subspecies during the expansion of rice cultivation area. This finding may imply that new genome-wide changes for flowering-time adaptation are one of the critical determinants for establishing genomic architecture of local rice subgroups. In-depth analyses of various rice genomes coupling with the genetically confirmed phenotypic changes in a large set of CSSLs enabled us to demonstrate how rice genome dynamics has coordinated with the adaptation of cultivated rice during the expansion of cultivation area.

A quantitative trait locus (QTL) analysis was conducted on grain appearance in Emi-no-kizuna, a rice cultivar that has superior and stable appearance of the brown rice grain and high tolerance to high temperature stress, by using F3 lines derived from Emi-no-kizuna and Tomohonami. The investigation was performed 2013 and 2015. In summer 2013 the air temperature was higher and the larger differences in grain appearance were observed. In the QTL analysis, a highly contributing locus, qGA8, was detected at the end of the short arm of chromosome 8. Because trends of reduced the occurrence of white immature kernel and increased the percentage of perfect grain were observed in Emi-no-kizuna genotype in both years, qGA8 is likely to be an important QTL that is dominant in the superior grain appearance of Emi-no-kizuna. Also, qGA8 was linked to a QTL associated with days to heading. Another QTL, qGA7, associated with grain appearance was detected on chromosome 7 in 2013. Because no negative correlation was found between the genotype of qGA8 and thousand grain weight, it should be possible to breed cultivars that possess Emi-no-kizuna genotype qGA8 without a decrease in thousand grain weight.

To investigate the genetic background responsible for blast resistance in Oryza sativa L. 'Hokuriku 193', QTL analysis was conducted using the F3 lines from the cross [ms-bo] Nekken 2 × Hokuriku 193 that were artificially infected with rice blast fungus (Magnaporthe grisea). QTLs were detected on chromosomes 1, 4, 6 and 12 that correlated with greater blast resistance in the Hokuriku 193-type lines. Notably, the QTL on chromosome 12 had a major effect and localized to the same region where Pi20(t), a broad-spectrum blast resistance gene, is positioned, suggesting strongly that the blast resistance of Hokuriku 193 was controlled by Pi20(t). Also, QTL analysis of the lines found to have no Pi20(t) detected two QTLs on chromosome 4 (qBR4-1 and qBR4-2) and one QTL on chromosome 6 (qBR6), of which qBR4-2 and qBR6 correlated with higher percentages of resistant plants in the Hokuriku 193-type lines. The blast susceptibility of BR_NIL (a NIL of Hokuriku 193 from which Pi20(t) was eliminated) was greater than that of Hokuriku 193, suggesting that elimination of Pi20(t) may markedly increase blast susceptibility. The disease severity of BR_NIL was mild, which might be the effect of qBR4-2 and/or qBR6.

In this study, we investigated the genetic basis of seed shattering and dormancy in Hokuriku 193 and bred an NIL improved these traits. Analysis of an F3 population from Hokuriku 193 × Koshihikari revealed a general correspondence between seed shattering and genotypes at the qSH1 locus, suggesting a strong influence of this locus on the seed shattering in Hokuriku 193. An F2 population from [ms-bo] Nekken 2 × Hokuriku 193 was also analyzed to identify quantitative trait loci (QTLs) for seed dormancy as measured by germination rate in the first December and March after seed harvest. The results revealed a concurrence QTLs of on chromosomes 1, 3, and 6 (qSDo1, qSDo3, qSDo6). In particular, qSDo1 and qSDo6 were considered regions worthy of active modification because they were QTL regions that promoted seed dormancy when carrying Hokuriku 193 genome regions around. SSDo_NIL, a near isogenic line (NIL) derived from Hokuriku 193 by introgressing Nekken 2 alleles only at the qSH1 locus and qSDo1, did not shatter, and its germination rate was significantly higher than that of Hokuriku 193. Yield performance was similar between SSDo_NIL and Hokuriku 193, suggesting that improvement of seed shattering and dormancy does not affect yield.

Morphological traits and two kinds of molecular markers were employed to study the genetic relationships among improved rice (Oryza sativa ) varieties of Indonesia since 1943. Dendrograms based on morphological traits and both molecular markers (simple sequence repeats, SSR and single nucleotide polymorphism, SNP) agreed in separating the varieties into two primary groups. Based on the morphological traits, a larger group (>60 %) contains varieties with smaller sizes compared with those in the smaller group (

Climate resilience of crops is critical for global food security. Understanding the genetic basis of plant responses to ambient environmental changes is key to developing resilient crops. To detect genetic factors that set flowering time according to seasonal temperature conditions, we evaluated differences of flowering time over years by using chromosome segment substitution lines (CSSLs) derived from japonica rice cultivars &ldquo, Koshihikari&rdquo, &times, &ldquo, Khao Nam Jen&rdquo, each with different robustness of flowering time to environmental fluctuations. The difference of flowering times in 9 years&rsquo, field tests was large in &ldquo, (36.7 days) but small in &ldquo, (9.9 days). Part of this difference was explained by two QTLs. A CSSL with a &ldquo, segment on chromosome 11 showed 28.0 days&rsquo, difference, this QTL would encode a novel flowering-time gene. Another CSSL with a segment from &ldquo, in the region around Hd16 on chromosome 3 showed 23.4 days&rdquo, difference. A near-isogenic line (NIL) for Hd16 showed 21.6 days&rsquo, difference, suggesting Hd16 as a candidate for this QTL. RNA-seq analysis showed differential expression of several flowering-time genes between early and late flowering seasons. Low-temperature treatment at panicle initiation stage significantly delayed flowering in the CSSL and NIL compared with &ldquo, Our results unravel the molecular control of flowering time under ambient temperature fluctuations.

Rice blast caused by Pyricularia oryzae (syn. Magnaporthe oryzae) is a destructive disease of rice worldwide that poses a serious threat to rice production. Strengthening blast resistance is an important objective in rice breeding programs. Race-specific resistance genes (R-genes) confer complete resistance to blast, but new races of blast pathogen can overcome it. After the first report of breakdown of resistance conferred by the R-gene Pik in 1963, this type of resistance has frequently been broken 1–7 years after the release of resistant varieties to farmers in Japan and other countries. To overcome this genetic vulnerability, Japanese rice breeders have focused on the use of race-nonspecific resistance in Japanese upland rice varieties whose resistance has been maintained for a long time. However, linkage drag between genes controlling this type of blast resistance and undesired traits has hindered its use. Therefore, researchers genetically dissected race-nonspecific resistance to rice blast. Among detected QTLs, a single recessive resistance gene, pi21, was identified by map-based cloning. The use of pi21 has improved durable resistance in rice breeding programs.

IR64 is one of the world’s most popular rice cultivars. To collect genetic factors involved in controlling its heading date, we developed 70 reciprocal advanced-backcross populations with a total of 6284 individuals at the BC4F2 generation from crosses between Koshihikari and IR64. We detected 29 QTLs associated with heading date on chromosomes 3, 5–8, 10, and 12. Twenty QTLs were located in the same chromosome regions as previously isolated heading date genes (Hd1, Hd6, Hd16, Ghd7, DTH8, Hd17, and Hd18). The rest were located in other chromosome regions. We found more number of QTLs than previous studies using mapping populations of IR64. Fine mapping in additional advanced-backcross populations clearly revealed that QTLs on the long arm of chromosome 7 are overlapping and seem to be a novel genetic factor for heading date because of their different locations from OsPRR37. Our results suggest that the difference in heading date between IR64 and Koshihikari is genetically controlled by many factors, and that a non-functional allele of Hd1 contributes to early heading of IR64 in the genetic background of functional alleles of other heading date QTLs and genes such as Hd6 and Hd16.

The fungal pathogen Pyricularia oryzae causes blast, a severe disease of rice (Oryza sativa L.). Improving blast resistance is important in rice breeding programs. Inoculation tests have been used to select for resistance genotypes, with DNA marker-based selection becoming an efficient alternative. No comprehensive DNA marker system for race-specific resistance alleles in the Japanese rice breeding program has been developed because some loci contain multiple resistance alleles. Here, we used the Fluidigm SNP genotyping platform to determine a set of 96 single nucleotide polymorphism (SNP) markers for 10 loci with race-specific resistance. The markers were then used to evaluate the presence or absence of 24 resistance alleles in 369 cultivars; results were 93.5% consistent with reported inoculation test-based genotypes in japonica varieties. The evaluation system was successfully applied to high-yield varieties with indica genetic backgrounds. The system includes polymorphisms that distinguish the resistant alleles at the tightly linked Pita and Pita-2 loci, thereby confirming that all the tested cultivars with Pita-2 allele carry Pita allele. We also developed and validated insertion/deletion (InDel) markers for ten resistance loci. Combining SNP and InDel markers is an accurate and efficient strategy for selection for race-specific resistance to blast in breeding programs.

Analysis of the genetic structure of Indonesian Oryza sativa and O. rufipogon using neighbour-joining trees based on single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers revealed that O. sativa in Indonesia is separated from O. rufipogon. Accessions of O. sativa in this study were differentiated into two major groups, indica and tropical japonica, excluding some varieties. SSR and SNP markers revealed the high value of differentiation (F ST) and genetic distance (D) between indica and tropical japonica and we discovered four loci by SNP markers and one locus by SSR markers that play a role in differentiation between indica and tropical japonica. Interestingly, genetic diversity (H) in O. rufipogon was lower than that in O. sativa, however H in O. rufipogon was the highest and H in tropical japonica was the lowest when O. sativa was divided into two groups. Inbreeding coefficient (Fst) showed evidences that gene flow (Nm) between species and within species might be one of the mechanisms related to the diversification and differentiation of Indonesian rice germplasm by asymmetric pattern between species and within O. sativa as revealed by SSR and SNP markers. In addition, we found evidences on stabilizing selection in Indonesian rice germplasm and they might be the reasons why Indonesian rice germplasm did not differentiate due to source location of landrace. However, we found a weak relation between SSR and SNP markers probably due to highly polymorphic in SSR and the different properties of both markers.

We investigated the natural variations in the flag leaf morphology of rice. We conducted a principal component analysis based on nine flag leaf morphology traits using 103 accessions from the National Institute of Agrobiological Sciences Core Collection. The first component explained 39% of total variance, and the variable with highest loading was the width of the flag leaf (WFL). A genome-wide association analysis of 102 diverse Japanese accessions revealed that marker RM6992 on chromosome 4 was highly associated with WFL. In analyses of progenies derived from a cross between Takanari and Akenohoshi, the most significant quantitative trait locus (QTL) for WFL was in a 10.3-kb region containing the NARROW LEAF 1 (NAL1) gene, located 0.4 Mb downstream of RM6992. Analyses of chromosomal segment substitution lines indicated that a mutation (G1509A single-nucleotide mutation, causing an R233H amino acid substitution in NAL1) was present at the QTL. This explained 13 and 20% of total variability in WFL and the distance between small vascular bundles, respectively. The mutation apparently occurred during rice domestication and spread into japonica, tropical japonica, and indica subgroups. Notably, one accession, Phulba, had a NAL1 allele encoding only the N-terminal, or one-fourth, of the wild-type peptide. Given that the Phulba allele and the histidine-type allele showed essentially the same phenotype, the histidine-type allele was regarded as malfunctional. The phenotypes of transgenic plants varied depending on the ratio of histidine-type alleles to arginine-type alleles, raising the possibility that H233-type products function differently from and compete with R233-type products.

Insertion-deletion (indel) polymorphisms, such as simple sequence repeats, have been widely used as DNA markers to identify QTLs and genes and to facilitate rice breeding. Recently, next-generation sequencing has produced deep sequences that allow genome-wide detection of indels. These polymorphisms can potentially be used to develop high-accuracy polymerase chain reaction (PCR)-based markers. Here, re-sequencing of 5 indica, 2 aus, and 3 tropical japonica cultivars and Japanese elite cultivar 'Koshihikari' was performed to extract regions containing large indels (10-51 bp) shared by diverse cultivars. To design indel markers for the discrimination of genomic regions between 'Koshihikari' and other diverse cultivars, we subtracted the indel regions detected in 'Koshihikari' from those shared in other cultivars. Two sets of indel markers, KNJ8-indel (shared in eight or more cultivars, including 'Khao Nam Jen' as a representative tropical japonica cultivar) and C5-indel (shared in five to eight cultivars), were established, with 915 and 9,899 indel regions, respectively. Validation of the two marker sets by using 23 diverse cultivars showed a high PCR success rate (≥95%) for 83.3% of the KNJ8-indel markers and 73.9% of the C5-indel markers. The marker sets will therefore be useful for the effective breeding of Japanese rice cultivars.

Lodging has been a major roadblock to attaining increased crop productivity. In an attempt to understand the mechanism for culm strength in rice, we isolated an effective quantitative trait locus (QTL), STRONG CULM3 (SCM3), the causal gene of which is identical to rice TEOSINTE BRANCHED1 (OsTB1), a gene previously reported to positively control strigolactone (SL) signaling. A near-isogenic line (NIL) carrying SCM3 showed enhanced culm strength and increased spikelet number despite the expected decrease in tiller number, indicating that SL also has a positive role in enhancing culm strength and spikelet number. We produced a pyramiding line carrying SCM3 and SCM2, another QTL encoding APO1 involved in panicle development. The NIL-SCM2+SCM3 showed a much stronger culm than NIL-SCM2 and NIL-SCM3 and an increased spikelet number caused by the additive effect of these QTLs. We discuss the importance of utilizing suitable alleles of these STRONG CULM QTLs without inducing detrimental traits for breeding.

Effective control of blast, a devastating fungal disease of rice caused by Pyricularia oryzae, is an important breeding objective to increase and stabilize worldwide rice production. Use of quantitative resistance to blast is considered a promising strategy because such resistance is more durable than race-specific qualitative resistance. Quantitative resistance is under complex genetic control, with each contributing gene having a smaller individual effect than those conveying qualitative resistance. This characteristic decreases selection pressure, thereby slowing the counter-evolution of the pathogen, but also makes it difficult for breeders to select the required traits in practical breeding programs. Extensive genetic studies of quantitative resistance during the last two decades have identified several relevant genes and have contributed to our understanding of the genetic control of quantitative resistance. Such studies will accelerate marker-assisted gene pyramiding of resistance alleles to confer strong, non-race-specific, and environmentally stable resistance to blast disease; this achievement would contribute substantially to global food security.

Grain shape is an important trait for improving rice yield. A number of quantitative trait loci (QTLs) for this trait have been identified by using primary F2 mapping populations and recombinant inbred lines, in which QTLs with a small effect are harder to detect than they would be in advanced generations. In this study, we developed two advanced mapping populations (chromosome segment substitution lines [CSSLs] and BC4F2 lines consisting of more than 2000 individuals) in the genetic backgrounds of two improved cultivars: a japonica cultivar (Koshihikari) with short, round grains, and an indica cultivar (IR64) with long, slender grains. We compared the ability of these materials to reveal QTLs for grain shape with that of an F2 population. Only 8 QTLs for grain length or grain width were detected in the F2 population, versus 47 in the CSSL population and 65 in the BC4F2 population. These results strongly suggest that advanced mapping populations can reveal QTLs for agronomic traits under complicated genetic control, and that DNA markers linked with the QTLs are useful for choosing superior allelic combinations to enhance grain shape in the Koshihikari and IR64 genetic backgrounds.

The volume that the root system can occupy is associated with the efficiency of water and nutrient uptake from soil. Genetic improvement of root length, which is a limiting factor for root distribution, is necessary for increasing crop production. In this report, we describe identification of two quantitative trait loci (QTLs) for maximal root length, QUICK ROOTING 1 (QRO1) on chromosome 2 and QRO2 on chromosome 6, in cultivated rice (Oryza sativa L.). We measured the maximal root length in 26 lines carrying chromosome segments from the long-rooted upland rice cultivar Kinandang Patong in the genetic background of the short-rooted lowland cultivar IR64. Five lines had longer roots than IR64. By rough mapping of the target regions in BC4F2 populations, we detected putative QTLs for maximal root length on chromosomes 2, 6, and 8. To fine-map these QTLs, we used BC4F3 recombinant homozygous lines. QRO1 was mapped between markers RM5651 and RM6107, which delimit a 1.7-Mb interval on chromosome 2, and QRO2 was mapped between markers RM20495 and RM3430-1, which delimit an 884-kb interval on chromosome 6. Both QTLs may be promising gene resources for improving root system architecture in rice.

A quantitative trait locus (QTL) analysis was conducted on grain appearance in Emi-no-kizuna, a rice cultivar that has superior and stable appearance of the brown rice grain and high tolerance to high temperature stress, by using F3 lines derived from Emi-no-kizuna and Tomohonami. The investigation was performed 2013 and 2015. In summer 2013 the air temperature was higher and the larger differences in grain appearance were observed. In the QTL analysis, a highly contributing locus, qGA8, was detected at the end of the short arm of chromosome 8. Because trends of reduced the occurrence of white immature kernel and increased the percentage of perfect grain were observed in Emi-no-kizuna genotype in both years, qGA8 is likely to be an important QTL that is dominant in the superior grain appearance of Emi-no-kizuna. Also, qGA8 was linked to a QTL associated with days to heading. Another QTL, qGA7, associated with grain appearance was detected on chromosome 7 in 2013. Because no negative correlation was found between the genotype of qGA8 and thousand grain weight, it should be possible to breed cultivars that possess Emi-no-kizuna genotype qGA8 without a decrease in thousand grain weight.

Pikahei-1(t) is the strongest quantitative trait locus (QTL) for blast resistance in upland rice cv. Kahei, which has strong field resistance to the rice blast disease. A high-quality bacterial artificial chromosome library was used to fine-map Pikahei-1(t) within ~300 kb on the 31-Mb region on rice chromosome 4. Of the 42 predicted open reading frames, seven resistance gene analogs (RGAs) with the nucleotide-binding site and leucine-rich repeat (NBS-LRR) domain were identified. Among these, RGA1, 2, 3, 5, and 7, but not RGA4 and 6, were found to be expressed in Kahei and monogenic lines containing Pikahei-1(t). Blast inoculation of transgenic rice lines carrying the genomic fragment of each RGA revealed that only RGA3 was associated with blast resistance. On the basis of these results, we concluded that RGA3 is the Pikahei-1(t) and named it Pi63. Pi63 encoded a typical coiled-coil-NBS-LRR protein and showed isolate-specificity. These results suggest that Pi63 behaves like a typical Resistance (R) gene, and the strong and broad-spectrum resistance of Kahei is dependent on natural pyramiding of multiple QTLs. The blast resistance levels of Pi63 were closely correlated with its gene expression levels, indicating a dose-dependent response of Pi63 function in rice resistance. Pi63 is the first cloned R gene in the R gene cluster on rice chromosome 4, and its cloning might facilitate genomic dissection of this cluster region.

Rice blast, caused by Pyricularia oryzae Cavara (Magnaporthe grisea (Hebert) Barr), is one of the most severe global rice diseases. The Philippine rice landrace ‘Ingngoppor-tinawon’ (IT) displays not only high field resistance to rice blast but also high tolerance to lodging, despite its very long culms. These characteristics make IT suitable for use in breeding animal feed varieties as whole-crop silage (WCS). To characterize the blast resistance of IT, quantitative trait locus (QTL) analysis of the resistance using simple sequence repeat (SSR) markers was conducted in F3 progenies derived from crossing IT with a susceptible lowland variety, ‘Koshihikari’. A QTL for blast field resistance (qBFR4) was identified on chromosome 4, and the resistance allele was derived from IT. The QTL explained 73.5% of the phenotypic variation for blast resistance. qBFR4 mapped to almost the same position as that of the blast field-resistance gene Pi39. Inoculation assays revealed that IT (containing qBFR4) showed resistance characteristics that differed from those of Mineharuka (containing Pi39), making it very likely that resistance derived from IT is distinct from Pi39. IT showed broad resistance to Japanese isolates and race-specific resistance to some Philippine isolates.

In this study, we investigated the genetic basis of seed shattering and dormancy in Hokuriku 193 and bred an NIL improved these traits. Analysis of an F3 population from Hokuriku 193 × Koshihikari revealed a general correspondence between seed shattering and genotypes at the qSH1 locus, suggesting a strong influence of this locus on the seed shattering in Hokuriku 193. An F2 population from [ms-bo] Nekken 2 × Hokuriku 193 was also analyzed to identify quantitative trait loci (QTLs) for seed dormancy as measured by germination rate in the first December and March after seed harvest. The results revealed a concurrence QTLs of on chromosomes 1, 3, and 6 (qSDo1, qSDo3, qSDo6). In particular, qSDo1 and qSDo6 were considered regions worthy of active modification because they were QTL regions that promoted seed dormancy when carrying Hokuriku 193 genome regions around. SSDo_NIL, a near isogenic line (NIL) derived from Hokuriku 193 by introgressing Nekken 2 alleles only at the qSH1 locus and qSDo1, did not shatter, and its germination rate was significantly higher than that of Hokuriku 193. Yield performance was similar between SSDo_NIL and Hokuriku 193, suggesting that improvement of seed shattering and dormancy does not affect yield.

Severe lodging has occurred in many improved rice varieties after the recent strong typhoons in East and Southeast Asian countries. The indica variety Takanari possesses strong culm characteristics due to its large section modulus, which indicates culm thickness, whereas the japonica variety Koshihikari is subject to substantial bending stress due to its thick cortical fibre tissue. To detect quantitative trait loci (QTLs) for lodging resistance and to eliminate the effects of genetic background, we used reciprocal chromosome segment substitution lines (CSSLs) derived from a cross between Koshihikari and Takanari. The oppositional effects of QTLs for section modulus were confirmed in both genetic backgrounds on chromosomes 1, 5 and 6, suggesting that these QTLs are not affected by the genetic background and are controlled independently by a single factor. The candidate region of a QTL for section modulus included SD1. The section modulus of NIL-sd1 was lower than that of Koshihikari, whereas the section modulus of NIL-SD1 was higher than that of Takanari. This result indicated that those regions regulate the culm thickness. The reciprocal effects of the QTLs for cortical fibre tissue thickness were confirmed in both genetic backgrounds on chromosome 9 using CSSLs.

To investigate the genetic background responsible for blast resistance in Oryza sativa L. ‘Hokuriku 193’, QTL analysis was conducted using the F3 lines from the cross [ms-bo] Nekken 2 × Hokuriku 193 that were artificially infected with rice blast fungus (Magnaporthe grisea). QTLs were detected on chromosomes 1, 4, 6 and 12 that correlated with greater blast resistance in the Hokuriku 193-type lines. Notably, the QTL on chromosome 12 had a major effect and localized to the same region where Pi20(t), a broad-spectrum blast resistance gene, is positioned, suggesting strongly that the blast resistance of Hokuriku 193 was controlled by Pi20(t). Also, QTL analysis of the lines found to have no Pi20(t) detected two QTLs on chromosome 4 (qBR4-1 and qBR4-2) and one QTL on chromosome 6 (qBR6), of which qBR4-2 and qBR6 correlated with higher percentages of resistant plants in the Hokuriku 193-type lines. The blast susceptibility of BR_NIL (a NIL of Hokuriku 193 from which Pi20(t) was eliminated) was greater than that of Hokuriku 193, suggesting that elimination of Pi20(t) may markedly increase blast susceptibility. The disease severity of BR_NIL was mild, which might be the effect of qBR4-2 and/or qBR6.

In rice (Oryza sativa L.), damage from diseases such as brown spot, caused by Bipolaris oryzae, and bacterial seedling rot and bacterial grain rot, caused by Burkholderia glumae, has increased under global warming because the optimal temperature ranges for growth of these pathogens are relatively high (around 30 °C). Therefore, the need for cultivars carrying genes for resistance to these diseases is increasing to ensure sustainable rice production. In contrast to the situation for other important rice diseases such as blast and bacterial blight, no genes for complete resistance to brown spot, bacterial seedling rot or bacterial grain rot have yet been discovered. Thus, rice breeders have to use partial resistance, which is largely influenced by environmental conditions. Recent progress in molecular genetics and improvement of evaluation methods for disease resistance have facilitated detection of quantitative trait loci (QTLs) associated with resistance. In this review, we summarize the results of worldwide screening for cultivars with resistance to brown spot, bacterial seedling rot and bacterial grain rot and we discuss the identification of QTLs conferring resistance to these diseases in order to provide useful information for rice breeding programs., PDF 12 page

Quantitative trait loci (QTLs) for resistance to rice blast offer a potential source of durable disease resistance in rice. However, few QTLs have been validated in progeny testing, on account of their small phenotypic effects. To understand the genetic basis for QTL-mediated resistance to blast, we dissected a resistance QTL, qBR4-2, using advanced backcross progeny derived from a chromosome segment substitution line in which a 30- to 34-Mb region of chromosome 4 from the resistant cultivar Owarihatamochi was substituted into the genetic background of the highly susceptible Aichiasahi. The analysis resolved qBR4-2 into three loci, designated qBR4-2a, qBR4-2b, and qBR4-2c. The sequences of qBR4-2a and qBR4-2b, which lie 181 kb apart from each other and measure, 113 and 32 kb, respectively, appear to encode proteins with a putative nucleotide-binding site (NBS) and leucine-rich repeats (LRRs). Sequence analysis of the donor allele of qBR4-2a, the region with the largest effect among the three, revealed sequence variations in the NBS-LRR region. The effect of qBR4-2c was smallest among the three, but its combination with the donor alleles of qBR4-2a and qBR4-2b significantly enhanced blast resistance. qBR4-2 comprises three tightly linked QTLs that control blast resistance in a complex manner, and thus gene pyramiding or haplotype selection is the recommended strategy for improving QTL-mediated resistance to blast disease through the use of this chromosomal region. Electronic supplementary material The online version of this article (doi:10.1007/s00122-012-1852-4) contains supplementary material, which is available to authorized users.

One of the major challenges in genetics has been to identify the nucleotide polymorphisms responsible for phenotypic variation. Through intensive analysis, several major quantitative trait loci (QTLs) for agronomic traits in rice have been identified and the underlying candidate genes have been delimited. Advanced mapping populations, including chromosome segment substitution lines, have enhanced the power of genetic analysis to detect QTL alleles, even those with minor effects. Recent examples of marker-assisted selection have proven the potential of this strategy for crop improvement. The genome-wide discovery of single nucleotide polymorphisms (SNPs), even among closely related cultivars, has enhanced the power of allele mining in a wide range of rice breeding materials. An array-based SNP genotyping system can be used to visualize pedigree haplotypes in breeding materials, including landraces and modern cultivars. All of these technologies are accelerating the genetic dissection of complex agronomic traits and further improvement of rice.

Over the past two decades, genetic dissection of complex phenotypes of economic and biological interest has revealed the chromosomal locations of many quantitative trait loci (QTLs) in rice and their contributions to phenotypic variation. Mapping resolution has varied considerably among QTL studies owing to differences in population size and number of DNA markers used. Additionally, the same QTLs have often been reported with different locus designations. This situation has made it difficult to determine allelic relationships among QTLs and to compare their positions. To facilitate reliable comparisons of rice QTLs, we extracted QTL information from published research papers and constructed a database of 1,051 representative QTLs, which we classified into 21 trait categories. This database (QTL Annotation Rice Online database; Q-TARO, http://qtaro.abr.affrc.go.jp/) consists of two web interfaces. One interface is a table containing information on the mapping of each QTL and its genetic parameters. The other interface is a genome viewer for viewing genomic locations of the QTLs. Q-TARO clearly displays the co-localization of QTLs and distribution of QTL clusters on the rice genome.

The elucidation of Sorghum bicolor intraspecific genetic diversity is of interest in studies on evo- lutionary forces under domestication and geographical distribution, and has application in the de- sign of programs for conservation, management and uses of genetic resources in breeding pro- grams. Since molecular markers enable assessment of genetic relationship, various types of mark- ers have been developed in many investigations of crops. This work aimed to assess the level of genetic diversity in grain sorghum collected from different regions in Asia including Central Asia and Africa. For the purpose, 14 rice cDNA-STS and 10 insertion/deletion (Indel) markers have been used. The majority of genes tagged by these markers were genes of complex crucial en- zymes of cellular metabolism and stress-inducible proteins. Based on the markers, 48 sorghum ac- cessions selected from the collection in NIAS Genebank (Tsukuba, Japan), and those from the All-Russian Research Institute of Plant Industry (St. Petersburg, Russia) were analyzed using clus- ter and principal coordinate analyses. The results demonstrated the existence of three major geo- graphical centers for sorghum evolution and differentiation. Two of the centers are found in the African continent where sorghum is suggested to have been domesticated. Asian sorghums are grouped in a geographical cluster distinctive from the African groups. More precisely, there are separate sub-groups of accessions originating from East Asia and Central Asia, respectively. This is the first report of the Central Asian sorghum cultivars forming a unique genetic group. The va- riety groups demonstrated here may provide important information on phylogenic differentiation of this crop in Asia and Africa. This work demonstrated that rice cDNA-STS and Indel markers are useful instruments for evaluation of sorghum genetic diversity.

Genetic dissection of a wide range of naturally occurring variations in rice has significantly progressed by means of quantitative trait locus analysis. This genetic dissection has resulted in molecular cloning of genes and loci with biological and agronomic interest. The success of these analyses depends strongly on the plant materials used. In the last decade, many kinds of plant materials, and particularly advanced backcross populations, have been developed for the genetic analysis of traits of interest. Some of those materials have been deposited in the public domain in order to facilitate further analyses of rice genetics and molecular biology. In this review, we describe how such plant materials, including chromosome segment substitution lines (CSSLs) and introgression lines, could be used in genetic analysis, as well as the kinds of plant materials that could be developed and that are now available to the rice research community. Furthermore, we introduce our current activities related to large-scale development of CSSLs using diverse Asian rice accessions as donors.

To reveal the sequence diversity and population structure of Asian rice (Oryza sativa L.) cultivars, we surveyed genome-wide single-nucleotide polymorphisms (SNPs) in 140 diverse accessions. We identified 4357 SNPs distributed on the 12 chromosomes by sequencing PCR amplicons from the exons and introns of anonymous rice genes. We detected 4.87 SNPs per 1 kb genome-wide. By classifying the 140 accessions on the basis of these SNPs, we identified seven cultivar groups that reflected the geographical distribution of the accessions. Three cultivar groups were defined from tropical japonica that corresponded to previous categories, and three indica cultivar groups were also defined within indica. The linkage disequilibrium (LD) distance between SNPs was approximately 250 kb, except for a longer LD detected in the Indica I cultivar group (corresponding to the previously identified aus group). The allele frequency of the SNPs varied among cultivar groups, reflecting the level of genetic diversity in each group. These SNPs for the diverse accessions enhance our understanding of natural variation in rice.

Blast-Resistant Rice The durability of disease resistance for an agricultural crop reflects the extent to which the defense stands up to evolutionary dodges on the part of the pathogen. Pi21 , which is a quantitative trait locus (QTL) of rice, contributes to a particularly durable resistance to a fungal rice blast disease: Rice plants carrying the resistant allele have been in cultivation for more than a century, and yet the pathogen has been unable to find a way through the defense. Fukuoka et al. (p. 998 ; see the news story by Normile ) have now cloned the responsible Pi21 QTL allele and were able to separate Pi21 resistance from tightly linked reductions in grain quality, paving the way for more widespread use of this allele in rice breeding.

Association mapping can be a powerful tool for detecting quantitative trait loci (QTLs) without requiring line-crossing experiments. We previously proposed a Bayesian approach for simultaneously mapping multiple QTLs by a regression method that directly incorporates estimates of the population structure. In the present study, we extended our method to analyze ordinal and censored traits, since both types of traits are common in the evaluation of germplasm collections. Ordinal-probit and tobit models were employed to analyze ordinal and censored traits, respectively. In both models, we postulated the existence of a latent continuous variable associated with the observable data, and we used a Markov-chain Monte Carlo algorithm to sample the latent variable and determine the model parameters. We evaluated the efficiency of our approach by using simulated- and real-trait analyses of a rice germplasm collection. Simulation analyses based on real marker data showed that our models could reduce both false-positive and false-negative rates in detecting QTLs to reasonable levels. Simulation analyses based on highly polymorphic marker data, which were generated by coalescent simulations, showed that our models could be applied to genotype data based on highly polymorphic marker systems, like simple sequence repeats. For the real traits, we analyzed heading date as a censored trait and amylose content and the shape of milled rice grains as ordinal traits. We found significant markers that may be linked to previously reported QTLs. Our approach will be useful for whole-genome association mapping of ordinal and censored traits in rice germplasm collections.