Natural Variation in the Flag Leaf Morphology of Rice Due to a Mutation of the NARROW LEAF 1 Gene in Oryza sativa L.

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 H(233)-type products function differently from and compete with R(233)-type products

Detection of quantitative trait loci controlling pre-harvest sprouting resistance by using backcrossed populations of japonica rice cultivars

Backcrossed inbred lines (BILs) and a set of reciprocal chromosome segment substitution lines (CSSLs) derived from crosses between japonica rice cultivars Nipponbare and Koshihikari were used to detect quantitative trait loci (QTLs) for pre-harvest sprouting resistance. In the BILs, we detected one QTL on chromosome 3 and one QTL on chromosome 12. The QTL on the short arm of chromosome 3 accounted for 45.0% of the phenotypic variance and the Nipponbare allele of the QTL increased germination percentage by 21.3%. In the CSSLs, we detected seven QTLs, which were located on chromosomes 2, 3 (two), 5, 8 and 11 (two). All Nipponbare alleles of the QTLs were associated with an increased rate of germination. The major QTL for pre-harvest sprouting resistance on the short arm of chromosome 3 was localized to a 474-kbp region in the Nipponbare genome by the SSR markers RM14240 and RM14275 by using 11 substitution lines to replace the different short chromosome segments on chromosome 3. This QTL co-localized with the low-temperature germinability gene qLTG3-1. The level of germinability under low temperature strongly correlated with the level of pre-harvest sprouting resistance in the substitution lines. Sequence analyses revealed a novel functional allele of qLTG3-1 in Nipponbare and a loss-of-function allele in Koshihikari. The allelic difference in qLTG3-1 between Nipponbare and Koshihikari is likely to be associated with differences in both pre-harvest sprouting resistance and low-temperature germinability

野生ダイズ染色体断片置換系統群を用いたダイズの重要農業形質の遺伝解析

この博士論文は内容の要約のみの公開（または一部非公開）になっています筑波大学 (University of Tsukuba)201

Discerning combining ability loci for divergent environments using chromosome segment substitution lines (CSSLs) in pearl millet

Pearl millet is an important crop for arid and semi-arid regions of the world. Genomic regions associated with combining ability for yield-related traits under irrigated and drought conditions are useful in heterosis breeding programs. Chromosome segment substitution lines (CSSLs) are excellent genetic resources for precise QTL mapping and identifying naturally occurring favorable alleles. In the present study, testcross hybrid populations of 85 CSSLs were evaluated for 15 grain and stover yield-related traits for summer and wet seasons under irrigated control (CN) and moisture stress (MS) conditions. General combining ability (GCA) and specific combining ability (SCA) effects of all these traits were estimated and significant marker loci linked to GCA and SCA of the traits were identified. Heritability of the traits ranged from 53–94% in CN and 63–94% in MS. A total of 40 significant GCA loci and 36 significant SCA loci were identified for 14 different traits. Five QTLs (flowering time, panicle number and panicle yield linked to Xpsmp716 on LG4, flowering time and grain number per panicle with Xpsmp2076 on LG4) simultaneously controlled both GCA and SCA, demonstrating their unique genetic basis and usefulness for hybrid breeding programs. This study for the first time demonstrated the potential of a set of CSSLs for trait mapping in pearl millet. The novel combining ability loci linked with GCA and SCA values of the traits identified in this study may be useful in pearl millet hybrid and population improvement programs using marker-assisted selection (MAS)

Identification of rice chromosome segment substitution line Z322-1-10 and mapping QTLs for agronomic traits from the F₃ population

Chromosome segment substitution lines (CSSLs) are powerful tools to combine naturally occurring genetic variants with favorable alleles in the same genetic backgrounds of elite cultivars. An elite CSSL Z322-1-10 was identified from advanced backcrosses between a japonica cultivar Nipponbare and an elite indica restorer Xihui 18 by SSR marker-assisted selection (MAS). The Z322-1-10 line carries five substitution segments distributed on chromosomes 1, 2, 5, 6 and 10 with an average length of 4.80 Mb. Spikilets per panicle, 1000-grain weight, grain length in the Z322-1-10 line are significantly higher than those in Nipponbare. Quantitative trait loci (QTLs) were identified and mapped for nine agronomic traits in an F3 population derived from the cross between Nipponbare and Z322-1-10 using the restricted maximum likelihood (REML) method in the HPMIXED procedure of SAS. We detected 13 QTLs whose effect ranging from 2.45% to 44.17% in terms of phenotypic variance explained. Of the 13 loci detected, three are major QTL (qGL1, qGW5-1 and qRLW5-1) and they explain 34.68%, 44.17% and 33.05% of the phenotypic variance. The qGL1 locus controls grain length with a typical Mendelian dominance inheritance of 3:1 ratio for long grain to short grain. The already cloned QTL qGW5-1 is linked with a minor QTL for grain width qGW5-2 (13.01%) in the same substitution segment. Similarly, the previously reported qRLW5-1 is also linked with a minor QTL qRLW5-2. Not only the study is important for fine mapping and cloning of the gene qGL1, but also has a great potential for molecular breeding

Identification and Validation of QTLs for Yield and Yield Components under Long-Term Salt Stress Using IR64 CSSLs in the Genetic Background of Koshihikari and Their Backcross Progenies

Unraveling the complex genetic bases and mechanisms underlying salt tolerance is of great importance for developing salt-tolerant varieties. In this study, we evaluated 42 chromosome segment substitution lines (CSSLs) carrying chromosome segments from IR64 on the genetic background of Koshihikari under salt stress. Two CSSLs, SL2007 and SL2038, produced higher plant dry weight and grain yield than did Koshihikari under the stress condition. These CSSLs also showed lower Na+ and Cl- accumulation in the leaf and whole plant at the full heading stage, which might be related to the higher grain yield and yield components. To understand the genetic control of its grain yield and yield components, a SL2007/Koshihikari F-2 population was generated for quantitative trait locus (QTL) analysis. Six QTLs for grain yield and yield-related traits were detected on chromosome 2. Using near-isogenic lines (NILs) from a SL2007/Koshihikari F-5 population, qSTGY2.2 was delimited to a 2.5 Mb region and novel qSTPN2 was delimited to a 0.6 Mb region. We also detected a novel QTL, qSTGF2, for grain filling, which was considered an important contributor to grain yield under salt stress in this CSSL. Our results provide insights into mechanisms conferring grain yield under salinity stress and new genetic resources for cloning and breeding