Mapping quantitative trait loci influencing panicle‐related traits from Chinese common wild rice (Oryza rufipogon) using introgression lines

Panicle‐related traits are important agronomic traits which directly associated with grain yield. In this study, we investigated quantitative trait loci (QTLs) associated with panicle‐related traits using a set of 265 introgression lines (ILs) of common wild rice (Oryza rufipogon Griff.) in the background of Indica high‐yielding cultivar Guichao 2 (O. sativa L.). A total of 39 QTLs associated with panicle‐related traits including panicle length (PL), primary branch number (PBN), secondary branch number (SBN), spikelet number per panicle (SPP) and spikelet density (SD), were detected in the ILs with single‐point analysis. The alleles of 20 QTLs derived from wild rice showed positive effects, and some QTLs, such as, QPl1b for PL, QPbn8 for PBN, QSd4 and QSd11b for SD and QSpp4 for SPP showed larger positive effects, providing good candidates and useful information for marker‐aided improvement of yield potential of rice. Most of the QTLs controlling SPP, SBN and SD were located in cluster or closely linked on chromosomes, and the directions of their additive effects were consistent, which explained the genetic basis of significant correlations between their phenotypic characters.     

Fine mapping of a QTL for the number of spikelets per panicle by using near‐isogenic lines derived from an interspecific cross between Oryza sativa and Oryza minuta

We constructed a high‐resolution physical map for the qSPP7 QTL for spikelets per panicle (SPP) on rice chromosome 7 across a 28.6‐kb region containing four predicted genes. Using a series of BC₇F₄ near‐isogenic lines (NILs) derived from a cross between the Korean japonica cultivar ‘Hwaseongbyeo’ and Oryza minuta (IRGC Acc. No. 101144), three QTLs for the number of SPP, grains per panicle and primary branches were identified in the cluster (P ≤ 0.01). All three QTLs were additive, and alleles from the O. minuta parent were beneficial in the ‘Hwaseongbyeo’ background. qSPP7 was mapped to a 28.6‐kb region between the two simple sequence repeat (SSR) markers RM4952 and RM21605. The additive effect of the O. minuta allele at qSPP7 was 23 SPP, and 43.6% of the phenotypic variance was explained by the segregation of the SSR marker RM4952. Colocalization of the three QTLs suggested that this locus was associated with panicle structure and had pleiotropic effects. The NIL populations and molecular markers are useful for cloning qspp7.     

Identification and application of major quantitative trait loci for panicle length in rice (Oryza sativa) through single‐segment substitution lines

Panicle length (PL), an important yield‐related trait, strongly affects yield components, such as grain number, grain density and rice quality. More than 200 panicle length quantitative trait loci (PL QTLs) are identified, but only a small number are applied in rice breeding. In this study, we performed QTL analysis for PL using 42 single‐segment substitution lines (SSSLs) derived from nine donors in the genetic background of HJX74. Fourteen QTLs and five heterosis QTLs (HQTLs) for PL were recognised. Three QTLs and four HQTLs acted positively, and the other eleven QTLs and one HQTL acted negatively. By scanning the single heterozygous background region of the F₂ population with large‐genetic‐effect SSSLs, we mapped PL loci qPL6‐2 and qPL7‐1 to different locations on chromosomes 6 and 7, respectively, in three consecutive years of independent trials. The genetic effects of these QTLs were further assessed. qPL6‐2 demonstrated the most positive additive effect (QTL), whereas qPL7‐1 achieved the most positive dominant effect (HQTL) for PL. These results indicated that the pyramiding of PL QTLs might increase grain yield and facilitate the application of the beneficial allele in hybrid rice breeding.     

Identification and validation of a yield-enhancing QTL cluster in rice (Oryza sativa L.)

Improvement of rice grain yield (YD) is an important goal in rice breeding. YD is determined by its related traits such as spikelet fertility (SF), 1,000-grain weight (TGW), and the number of spikelets per panicle (SPP). We previously mapped quantitative trait loci (QTLs) for SPP and TGW using the recombinant inbred lines (RILs) derived from the crosses between Minghui 63 and Teqing. In this study, four QTLs for SF and four QTLs for YD were detected in the RILs. Comparison of the locations of QTLs for these three yield-related traits identified one QTL cluster in the interval between RM3400 and RM3646 on chromosome 3. The QTL cluster contained three QTLs, SPP3a, SF3 and TGW3a, but no YD QTL was located there. To validate the QTL cluster, a BC₄F₂ population was obtained, in which SPP3a, SF3 and TGW3a were simultaneously mapped to the same region. SPP3a, SF3 and TGW3a explained 36.3, 29.5 and 59.0 % of phenotype variance with additive effect of 16.4 spikelets, 6 % SF and 1.8 g grain weight, respectively. In the BC₄F₂ population, though the region has opposite effects on TGW and SPP/SF, a YD QTL YD3 identified in this cluster region can increase 4.6 g grains per plant, which suggests this QTL cluster is a yield-enhancing QTL cluster and can be targeted to improve rice yield by marker aided selection.     

Comparison of quantitative trait loci for 1,000-grain weight and spikelets per panicle across three connected rice populations

The ability to detect quantitative trait loci (QTLs) in a bi-allelic population is often limited. The power of QTL detection and identification of the most beneficial allele at each QTL could be greatly improved by comparing QTLs among different populations derived from connecting multi-parents. In this study, three sets of connected recombinant inbred lines (RILs) derived from the crosses between Zhenshan 97 and Minghui 63 (PZM), Zhenshan 97 and Teqing (PZT), and Minghui 63 and Teqing (PMT), respectively, were used. QTL analyses for the number of spikelets per panicle (SPP) and 1,000-grain weight (TGW) were performed in PZT, and five SPP QTLs on chromosomes 1, 6, and 7 and two TGW QTLs on chromosome 1 were detected. QTL for SPP was also identified in PMT, and six QTLs were detected on chromosomes 1, 2, 3, 6, and 7 in this population. In an earlier study, we identified five SPP QTLs and four TGW QTLs in PMT and nine TGW QTLs in PZM. Comparison of the QTL mapping results of these two studies showed that one QTL was common to the three populations, 11 QTLs were detected in two populations, and six QTLs were found in only one population. Comparison of genetic effect and the action direction of the QTLs detected in the three populations showed that additive effects of QTLs estimated in different populations were also expressed additively among three parental alleles. Additive effects of SPP7a estimated in three near-isogenic line F₂ populations supported this finding. Based on these results, we suggest that pyramiding the most beneficial alleles among the three parents could efficiently improve rice yield.     

Molecular dissection of the primary sink size and its related traits in rice

The genetic mechanism underlying the relationship between three traits of the primary sink size ‐ spikelet number per panicle (SNP), panicle number per plant (PN), and 1000‐grain weight (GWT), and their 10 component traits in rice was dissected in 292 F₁₃ recombinant inbred lines using a complete linkage map. A total of 43 genomic regions on 12 rice chromosomes were found to contain quantitative trait loci (QTLs) affecting the sink size traits, which revealed several important aspects of the genetic basis of sink capacity in rice. First, QTLs for SNP, PN and GWT were largely independent. Secondly, most QTLs affecting SNP and GWT showed close characteristics in both genomic locations and directions of effects to QTLs for their components, suggesting that pleiotropy, rather than linkage, was the primary genetic basis of the correlated panicle and grain traits. Thirdly, some QTLs affecting component traits did not contribute to SNP or GWT. In these cases, two or more QTLs with opposite effects on their component traits were detected, which could be due to either linkage or pleiotropy. Fourthly, some QTLs had large effects on panicle number (QPn4), panicle branching and length (QPbn3a, QPbn3b and QPb14), grain length and volume (QG13, QG15 and QGv2), and grain shape (QGs1 and QGs7), which were consistently detected in the related rice mapping populations and in different environments, providing good candidates and useful information for marker‐aided improvement of sink size and yield potential of rice.     

QTL analysis for panicle characteristics in temperate japonica rice

To understand the genetic background of panicle characteristics in temperate japonica rice (Oryza sativa L.), we genetically analyzed DH lines derived from a cross between two temperate japonica rice cultivars, ‘Akihikari’ and ‘Koshihikari’,in 1996 and 1997. Four traits of panicle characteristics, number of primary branches per panicle (NPB), number of secondary branches per panicle (NSB), average number of spikelets on one primary branch (NSP)and average number of spikelets on one secondary branch (NSS), in 212 DH lines were measured, and the interval mapping of QTLs for these traits was carried out using169 DNA markers with an LOD threshold of2.5. Five, three and one putative QTLs for NPB, NSB and NSS were identified,respectively, and no QTLs relating to NSP appeared. The percentages in total phenotypic variation explained by all putative QTLs for NPB were 35.5%: and43.8% in 1996 and 1997, respectively. All putative QTLs for NSB accounted for 35.5%and 27.5% of total phenotypic variation in1996 and 1997, respectively. The QTLs identified in this study will be useful intemperate japonica rice breeding for improved spikelet yield. This revised version was published online in August 2006 with corrections to the Cover Date.     

利用已测序水稻品种分析其农艺性状基因座

水稻重要农艺性状的基因定位研究在育种上具有重要意义。2004年在海南陵水县种植两个完全测序水稻品种日本晴与9311的F₂群体及双亲，分别考察了其单株分蘖数、穗数、有效分蘖数、穗长、主穗长、抽穗期、株高和剑叶8个农艺性状3次重复的平均值。用已构建的连锁遗传图谱（Nipponbare/9311-F₂遗传图谱）及Excel 2000和Mapmaker/QTL 1.1b软件对这8个性状间的相互关系和基因位点进行了分析。结果在LOD>2.0和P＜0.005的条件下共检测到41个QTLs，它们分布在水稻所有染色体上，单个QTL对性状表型贡献率11.0%～46.4%，其中大于20%的有22个。对选用已测序材料为亲本构建图谱来探讨水稻农艺性状的分子基础及其育种意义进行了讨论。     

Novel quantitative trait loci for yield and yield related traits identified in Basmati rice (Oryza sativa)

Increasing crop productivity is one of the prime goals of crop breeding research. Rice grain yield is a complex quantitative trait governed by polygenes. Although several QTLs governing grain yield traits have been reported and limited attempts have been made to map QTLs for grain yield parameters in Basmati rice. A population from the cross Sonasal and Pusa Basmati 1121 comprising 352 RILs was generated through the single seed descent method. A total of 12 QTLs governing yield and yield-related traits were mapped on six chromosomes, namely, 1, 2, 3, 7, 8 and 9, of which five QTLs were novel. We identified a novel and robust epistatic QTL (qPH1.1 and qPL1.1) governing plant height and panicle length, flanked by the markers RM5336-RM1 on chromosome 1. The gene encoding brassinosteroid insensitive 1-associated receptor kinase 1 precursor is the putative candidate gene underlying this epistatic QTL. Another novel QTL, qNT3.1, governing tiller number was bracketed to a region of .77 Mb between the markers RM15247 and RM15281 on chromosome 3. Of the 57 annotated gene models, Os03g0437600 encoding alpha/beta-fold hydrolase, a homologous to AtKai2 is a putative candidate gene underlying the novel QTL qNT3.1. The other QTLs such as qDFF1.1 governing days to 50% flowering co-localizes with the gene Ghd7, QTL for plant height qPH1.2 co-localizes with the gene sd1, the QTLs for panicle length co-localizes with FUWA and DEP2, the QTL for tiller number co-localizes with OsRLCK57 and QTLs for thousand-grain weight co-localize with the major gene GS3. The QTLs identified in the current study can be effectively used in marker-assisted selection for developing Basmati rice varieties with a higher yield.     

Identification of a novel QTL for the number of spikelets per panicle using a cross between indica‐ and japonica‐type high‐yielding rice cultivars in Japan

Yield is a complex trait. To improve it, the accumulation of the favourable alleles of valuable genes is required for each yield‐related trait. In this study, we used two high‐yielding rice cultivars developed in Japan, indica‐type ‘Takanari’ and japonica‐type ‘Momiroman’, for a genetic analysis of the sink capacity‐related traits. An F₂ population showed transgressive segregation for the number of spikelets per panicle. Quantitative trait locus (QTL) analysis detected four QTLs for the trait. Two of the QTLs were most likely identical to previously cloned GN1a and APO1, and their Takanari alleles had positive effects. The Momiroman alleles of the other two QTLs had positive effects, and one of these QTLs was most likely identical to SPIKE/GPS. The QTL on the long arm of chromosome 3 appeared to be novel; it clustered with QTLs for grain length and days‐to‐heading. Substitution mapping revealed that the close linkage of QTLs caused the clustering. These results suggest that the combination of the favourable alleles of detected QTLs could lead to greater sink capacity than that of the parental cultivars.     

Identification of QTLs controlling quantitative characters in rice using RFLP markers

Summary Segregation of plant height (PH), tiller number (TN), panicle number (PN), average panicle length per plant (PL), average primary branch number per panicle per plant (PBN) and 1000 grain weight (1000G) were specific in an F₂ population derived from a cross of Palawan, a tall Javanica variety, and IR42, an Indica semidwarf variety. One hundred and four informative RFLP markers covering all 12 chromosomes were used for detecting putative QTLs controlling the traits. Orthogonal contrasts and interval mapping analysis were used for the analysis. QTL detected for PH on the region of chromosome 1, where semidwarfing gene sd-1 locus is located, seems to be a multiple allelic locus. An additional QTL for PH was identified on chromosome 2. Two QTLs for TN were detected on chromosomes 4 and 12. The QTL on chromosome 4 seemed also to govern the variation in PN. Four QTLs were found for the other traits, two of them for PL were located on chromosomes 6 and 2, one for PBN on chromosome 6 and the other for 1000G on chromosome 1. Additive gene actions were found to be predominant, except one QTL for PH and one QTL for PL, but partial or incomplete dominance also existed for the QTLs detected.     

Fine mapping of a major quantitative trait locus, <Emphasis Type="Italic">qFLL6.2</Emphasis>, controlling flag leaf length and yield traits in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Fine mapping of a quantitative trait locus, qFLL6.2, controlling flag leaf length (FLL) and yield traits in rice was conducted using four sets of near isogenic lines (NILs) that were developed from a common residual heterozygote at F₇ generation of the indica rice cross Zhenshan 97/Milyang 46. Each of the NIL sets consisted of 40 lines that are S₁ progenies of ten maternal homozygotes, ten paternal homozygotes, and 20 heterozygotes differing in a portion of the 1.19-Mb interval RM3414–RM6917 on the short arm of rice chromosome 6. Analysis of phenotypic differences among the three genotypic groups in each NIL set delimited qFLL6.2 to a 62.1-kb region flanked by simple sequence repeat marker RM3414 and sequence-tagged site marker Si2944. This QTL explained 52.73% of the phenotypic variance, and the Zhenshan 97 allele increased FLL by 2.40 cm. Based on data collected from homozygous lines of three of the NIL sets, qFLL6.2 was shown to have major effects on all the three yield traits analyzed, including the number of spikelets per panicle, the number of filled grains per panicle, and grain weight per panicle. A comparison of the different groups revealed that the effect of qFLL6.2 was highly consistent across different genetic backgrounds and environments, providing a good candidate for map-based cloning and investigating the source–sink relationship in rice.     

Trait identification and QTL validation for reproductive stage drought resistance in rice using selective genotyping of near flowering RILs

Drought resistance is becoming an indispensable character for rice improvement due to the dwindling global water resources. Genetic improvement for drought resistance is achieved through physiological dissection and genetic analysis of independent component traits associated with crop productivity under stress. A subset mapping population of 93 near flowering recombinant inbred lines with uniform phenology was constituted for genetic analysis of reproductive stage drought resistance. The population was phenotyped for 22 physio-morphological traits under two contrasting water regimes imposed at reproductive stage. Broad sense heritabilities of morphological traits were lower under stress than irrigated. Predominant association of plant height, panicle exsertion and harvest index with grain yield were observed under stress. The sustenance of panicle exsertion through maintaining growth during moisture stress was found as a significant trait associated with the grain yield through minimizing spikelet sterility. Selective genotyping was carried out with 23 polymorphic microsatellite markers of the established target genomic regions for drought resistance. The study validated the association of a QTL region on the long arm of chromosome 1 with plant height, panicle length, panicle exsertion, biological yield and stomatal conductance under stress. This region, flanked by markers RM246 and RM315, was known to possess the semi-dwarf gene, sd-1. Role of another major interval lying between RM256 and RM149 on chromosome 8 in defining the drought resistance could be established through identification of QTLs associated with leaf rolling, panicle exsertion, plant height, panicle length, senescence and biological yield under moisture stress condition. Few other QTLs were also identified.     

Identification and verification of quantitative trait loci for eating and cooking quality of rice (Oryza sativa)

This study was conducted to identify new quantitative trait loci (QTLs) that have stable effects for eating and cooking quality (ECQ) of rice. Three recombinant inbred line populations of indica rice were each planted in two years. Three traits for ECQ, amylose content (AC), gel consistency (GC) and alkali spreading value (ASV), were measured for QTL analysis. A total of 13 QTLs were detected, including four for AC, six for ASV and three for GC. Two QTLs, qGC4 in the interval RM16252–RM335 on the short arm of chromosome 4 and qGC6.2 in the Alk region, were validated in a population derived from a residual heterozygote that was homozygous at the major locus Wx. In the absence of segregation at the Wx locus, qGC4 and qGC6.2 had additive effects of 2.46 and 8.18 mm, respectively, offering a potential for improving GC property of rice varieties. Comparison between qGC4 and previous results suggests that qGC4 is likely a new QTL for GC, providing a candidate for gene cloning and functional characterization.     

Introgression of a quantitative trait locus for spikelets per panicle from Oryza minuta to the O. sativa cultivar Hwaseongbyeo

A new QTL for spikelets per panicle (SPP) was detected on the long arm of chromosome 7 in an F₂ population derived from a cross between the japonica cultivar Hwaseongbyeo and WH29001. WH29001, an advanced backcross line was developed by introgressing chromosomal segments from an accession of O. minuta (2n = 48, BBCC, Acc. no. 101141 ) into the O. sativa subsp. japonica cv. Hwaseongbyeo. The O. minuta allele increased SPP in the Hwaseongbyeo background despite the fact that O. minuta was the small panicle parent. Using F₃ and F₄ progenies, spp7 was validated and mapped to a 2.3 Mb region in the interval between the SSR markers RM445 and RM21615 based on the japonica genome sequence. A yield trial using F₄ lines indicated that the lines carrying an O. minuta chromosome segment across the entire spp7 target region out‐yielded its sister lines containing Hwaseongbyeo chromosome in the target region and Hwaseongbyeo by 14.3% and 15.9%, respectively. Increase in SPP in WH29001 was mainly because of the increase in primary branches per panicle. The locus, spp7 is of particular interest because of its independence from undesirable height and flowering time. SSR markers tightly linked to the spp7 will facilitate cloning of the gene underlying this QTL as well as marker‐assisted selection for variation in SPP in an applied breeding program.     

水稻芽期与幼苗前期耐碱性状QTL定位

利用包含120个株系的籼粳交来源(春江06/TN1)的加倍单倍体群体, 在Na₂CO₃胁迫下, 以发芽期和幼苗前期的相对发芽势等10个性状作为耐碱性评价指标, 进行水稻耐碱性的QTL定位。相关性分析表明, 相对发芽势和相对发芽率显著正相关, 相对苗高、相对根数和相对根长之间显著正相关。采用QTLNetwork统计软件共定位到14个加性QTL和13个上位性QTL。在第3染色体RM251~RM3280间有2个QTL, 在第7染色体RM3286~RM1279区域有3个QTL; 在第1、2和7染色体同一位置同时检测到2个上位性QTL, 在第12染色体RM1246~RM5199之间集中了4个上位性QTL, 耐碱数量基因表现出一因多效或紧密连锁现象。耐碱性盐QTL可能包括两类, 一类与K+、Na+等离子胁迫有关, 另一类与高pH胁迫有关。不同类型的水稻品种都具有一些耐碱基因, 可以通过有性杂交和分子标记辅助选择的方法选育优良的耐碱品种。     

QTL mapping of sheath blight resistance in a deep-water rice cultivar

Sheath blight, caused by the pathogen Rhizoctonia solani Kühn, is one of the most serious diseases of rice and leads to severe yield loss worldwide. A recombinant inbred line (RIL) population consisting of 121 lines was constructed from a cross between HH1B and RSB03, the latter of which is a deep-water rice variety. Five traits were used to evaluate sheath blight resistance, namely disease rating (DR), lesion length (LL), lesion height (LH), relative lesion length [RLL, the ratio of LL to plant height (PH)], and relative LH (RLH, the ratio of LH to PH). Using the RIL population and 123 molecular markers, we identified 28 quantitative trait loci (QTLs) for the five traits in two environments. These QTLs are located on nine chromosomes and most of them are environment specific. A major QTL for DR (qSBR1) on chromosome 1 was identified with contributions of 12.7% at Shanghai and 42.6% at Hainan, and it collocated with a QTL for PH. The allele at this locus from RSB03 enhances sheath blight resistance and increases PH. Another QTL for DR on chromosome 7 was adjacent to QTLs for heading date (HD) and four other disease traits. RSB03 also carries the resistant allele at this locus and shortens HD. The susceptible parent, HH1B, provides the resistance allele at the locus qSBR8, where QTLs for four other disease traits were identified. QTL mapping results showed that most QTLs for LL, LH, RLL, and RLH are collocated with QTLs for DR. Three QTLs for DR are independent from HD, PH, and four other disease traits, while four QTLs are closely related to HD and PH. Four QTLs for LL, LH, RLL, and RLH are independent from DR, HD, and PH, while there is only one region harboring QTLs for these four traits and HD. Correlation analysis and QTL mapping results indicated that LL, LH, RLL, and RLH might be important indices, like DR, for evaluating the level of resistance to rice sheath blight.     

Comparison and analysis of main effects,epistatic effects,and QTL × environment interactions of QTLs for agronomic traits using DH and RILs populations in rice

Two genetic linkage maps based on doubled haploid (DH) and recombinant inbred lines (RILs) populations, derived from the same indica-japonica cross ‘Samgang × Nagdong’, were constructed to analyze the quantitative trait loci (QTLs) affecting agronomic traits in rice. The segregations of agronomic traits in RILs population showed larger variations than those in DH population. A total of 10 and 12 QTLs were identified on six chromosomes using DH population and seven chromosomes using RILs population, respectively. Three stable QTLs including pl9.1, ph1.1, and gwp11.1 were detected through different years. The percentages of phenotypic variation explained by individual QTLs ranged from 8 to 18% in the DH population and 9 to 33% in the RILs population. Twenty-three epistatic QTLs were identified in the DH population, while 21 epistatic QTLs were detected in the RILs population. Epistatic interactions played an important role in controlling the agronomic traits genetically. Four significant main-effect QTLs were involved in the digenic interactions. Significant interactions between QTLs and environments (QE) were identified in two populations. The QTLs affecting grain weight per panicle (GWP) were more sensitive to the environmental changes. The comparison and QTLs analysis between two populations across different years should help rice breeders to comprehend the genetic mechanisms of quantitative traits and improve breeding programs in marker-assisted selection (MAS).     

一个具有主效晚抽穗基因的水稻染色体片段代换系的鉴定、形态分析及Ehd4-2定位

抽穗期是决定水稻品种种植地区和季节适应性的重要农艺性状,鉴定抽穗期基因对水稻生产具有重要意义。本研究采用高代回交和SSR标记辅助选择相结合的方法获得了1个以日本晴为受体亲本、西恢18为供体亲本的含有1个控制晚抽穗表型的主效单基因的水稻染色体片段代换系Z315。Z315携带来自西恢18的5个代换片段,分布于第1、第3、第6和第7染色体上,平均代换片段长度为7.39 Mb。Z315的叶绿素含量、株高、穗长、倒一节间长、倒二叶长、倒三叶长、有效穗数、每穗实粒数和总粒数均显著高于受体日本晴,暗示其代换片段可能携带这些性状的QTL。进一步利用日本晴与Z315杂交产生的F1和F2群体对晚抽穗基因进行遗传分析和分子定位。该晚抽穗表型受1对隐性核基因控制,最终将该基因定位于第3染色体RM14283和RM6349之间,物理距离为233 kb。对该区间进行候选基因预测和测序,发现1个与抽穗相关的编码锌指蛋白的基因LOC_Os03g02160在日本晴和Z315间存在差异,该基因可能与Ehd4等位,称作Ehd4-2。由于染色体片段代换系除代换片段外与受体亲本一致,因此本研究无论对进一步分离其他QTL还是进行基因聚合育种均具有较大利用价值。