Fine mapping of qSS‐9, a major and stable quantitative trait locus,for seed storability in rice (Oryza sativa L.)

Seed storability in rice (Oryza sativa L.) is an important agronomic trait. We previously showed a quantitative trait locus of seed storability, qSS‐9, on chromosome 9 in a backcross population of ‘Koshihikari’ (japonica) / ‘Kasalath’ (indica) // ‘Koshihikari’. In this study, fine mapping of the chromosomal location of qSS‐9 was performed. Effect of ‘Kasalath’ allele of qSS‐9 was validated using a chromosome segment substitution line, SL36, which harboured the target quantitative trait loci (QTL) from ‘Kasalath’ in the genetic background of ‘Nipponbare’ under different ageing treatments in different environments. Subsequently, an F₂ population from a cross between ‘Nipponbare’ and SL36 was used for fine mapping of qSS‐9. Simultaneously, four subnear isogenic lines (sub‐NILs) that represented different recombination breakpoints across the qSS‐9 region were developed from F₃ progeny. Finally, the qSS‐9 locus was located between the Indel markers Y10 and Y13, which delimit a region of 147 kb in the ‘Nipponbare’ genome. These results provide a springboard for map‐based cloning of qSS‐9 and possibilities for breeding rice varieties with strong seed storability.     

利用回交重组自交系群体检测3个水稻光合功能相关性状QTL

利用98个家系组成的日本晴(粳稻)/Kasalath（籼稻）//日本晴回交重组自交系(backcross inbred lines, BILs) 群体(BC₁F₁₀),研究水稻光合功能相关的数量性状基因座（QTL）。基于水稻抽穗后7 d叶片全氮含量（TLN）、叶绿素a/b比值（Chl.a/b）和叶绿素含量（Chl）,共检测到8个QTL,其LOD值为2.61 ~ 6.42     

Detection and analysis of QTLs for ferrous iron toxicity tolerance in rice, Oryza sativa L.

A mapping population of 96 BC₁F₉lines (Backcross Inbred Lines: BILs),derived by a single-seed descent method rom a backcross of Nipponbare (japonica) / Kasalath (indica // Nippon are, was used to detect quantitative trait loci (QTLs) for leaf bronzing index (LBI), stem dry weight (SDW), tiller number (TN) and root dry weight (RDW) under Fe²⁺ stress condition in rice. Two parents and 96 BILs were phenotyped for the traits by growing them in Fe²⁺ toxicity nutrient solution. A total of four QTLs were detected on chromosome 1 and 3, respectively, with LOD of QTLs ranging from 3.17 to 7.03. One QTL controlling LBI, DW, N and RDW was located at the region of C955-C885 on chromosome 1, and their contributions to whole variation were 20.5%, 36.9%, 43.9% and 38.8%,respectively. The QTL located at the region of C955-C885 on chromosome 1 may be important to ferrous iron toxicity tolerance in rice. Another QTL for SDW and RDW was located at the region of C25-C515 on chromosome 3, with respective contributions of 47.9% and 35.0% to whole variation. Further, two QTLs on chromosome 1 were located for RDW at the region of R2329-R210 and for TN at the region of R1928-C178. Comparing with the other mapping results, the QTL located at the region of C955-C885 on chromosome 1 was identical with the results reported previously. There is a linkage between a TL detected under Fe²⁺ stress condition for stem and root dry weight and a QTL detected under phosphorus-deficiency condition for dry weight on chromosome 3. This revised version was published online in July 2006 with corrections to the Cover Date.     

QTL analysis for brown spot resistance in American rice cultivar ‘Dawn’

Rice brown spot (BS), caused by Bipolaris oryzae, causes yield loss and deterioration of grain quality. Using single-nucleotide polymorphism (SNP) markers, we conducted quantitative trait locus (QTL) analysis of BS resistance in backcross inbred lines (BILs) from a cross between an American rice cultivar, ‘Dawn’ (resistant), and ‘Koshihikari’ (susceptible). Four QTLs for BS resistance were detected in a three-year field evaluation, and ‘Dawn’ contributed the resistance alleles at all QTLs. The QTL with the greatest effect, qBSR6-kd, explained 15.1% to 20.3% of the total phenotypic variation. Although disease score and days to heading (DTH) were negatively correlated in all three years, qBSR6-kd was located near a QTL for DTH at which the ‘Dawn’ allele promoted heading. Another BS resistance QTL (qBSR3.1-kd) was unlinked to the QTLs for DTH. Therefore, these two QTLs are likely to be useful for breeding BS-resistant varieties without delaying heading. The other two BS resistance QTLs (qBSR3.2-kd and qBSR7-kd) were located near DTH QTLs at which the ‘Dawn’ alleles delayed heading. The QTLs reported here will be good candidates for developing BS-resistant cultivars.     

Identification of Quantitative Trait Loci Associated with Aluminum Tolerance in Rice (Oryza Sativa L.)

Summary Quantitative trait loci (QTL) analysis for Al tolerance was performed in rice using a mapping population of 98 BC₁F₁₀ lines (backcross inbred lines: BILs), derived from a cross of Al-tolerant cultivar of rice (Oryza sativa L. cv. Nipponbare) and Al-sensitive cultivar (cv. Kasalath). Three characters related to Al tolerance, including root elongation under non-stress conditions (CRE), root elongation under Al stress (SRE) and the relative root elongation (RRE) under Al stress versus non-stress conditions, were evaluated for the BILs and the parents at seedling stage. A total of seven QTLs for the three traits were identified. Among them, three putative QTLs for CRE (qCRE-6, qCRE-8 and qCRE-9) were mapped on chromosomes 6, 8 and 9, respectively. One QTL for SRE (qSRE-4) was identified on chromosome 4. Three QTLs (qRRE-5, qRRE-9 and qRRE-10) for RRE were detected on chromosomes 5, 9, 10 and accounted for 9.7–11.8% of total phenotypic variation. Interestingly, the QTL qRRE-5 appears to be syntenic with the genomic region carrying a major Al tolerance gene on chromosome 6 of maize. Another QTL, qRRE-9, appears to be similar among different rice populations, while qRRE-10 is unique in the BIL population. The common QTLs for CRE and RRE indicate that candidate genes conferring Al tolerance in the rice chromosome 9 may be associated with root growth rates. The existence of QTLs for Al tolerance was confirmed in substitution lines for corresponding chromosomal segments. These results also provide the possibilities of enhancing Al tolerance in rice through using marker-assisted selection (MAS) and pyramiding QTLs.     

水稻品种倒伏指数QTL分析

利用由98个株系组成的Nipponbare（粳）/ Kasalath（籼）∥Nipponbare 回交重组自交系群体,对水稻倒伏指数与株高、茎粗及单株生物量等性状进行了相关分析,并对水稻倒伏相关性状QTL进行了分子标记定位和遗传效应分析。结果表明,株高、茎粗、单株生物量3性状与倒伏指数的相关系数均达1%显著水平。利用基于性状-标记多元线性     

Mapping of quantitative trait loci controlling seedling vigor in rice (Oryza sativa L.) under submergence

Submergence-induced suppression of seedling vigor is a serious constraint particularly in the direct seeding rice cultivation system. To identify quantitative trait loci (QTLs) associated with seedling vigor in rice under submergence, a mapping population of 98 Backcross Inbred Lines derived from a cross of Nipponbare/Kasalath//Nipponbare was used. Phenotypic evaluation of seedling vigor under submergence was based on shoot length (SHL), root length (RTL) and shoot fresh weight (SFW) using a test tube bioassay method. Thirty-two putative QTLs were detected among which 7 were for SHL, 11 for RTL and 14 for SFW. Phenotypic evaluation was also made of the parental lines and a set of 54 chromosome segment substitution lines in which Nipponbare segments were substituted for by their homologous Kasalath segments covering the entire rice genome. Two QTLs with more than 10 % contribution to the total phenotypic variance were verified for SHL, and at least one for RTL and six for SFW on chromosomes 1, 3, 4, 6 and 7 at the 1 % significance level. Among these, all but two showed reductions in one, two or all three traits. Our present and previous results suggest that the Nipponbare genome has a potential to improve seedling vigor under submergence and that japonica germplasms can be used to breed for this important trait in indica rice.     

Identification of quantitative trait loci for flowering-related traits in the D genome of synthetic hexaploid wheat lines

The gene pool of Aegilops tauschii, the D-genome donor of common wheat (Triticum aestivum L.), can be easily accessed in wheat breeding, but remains largely unexplored. In our previous studies, many synthetic hexaploid wheat lines were produced through interspecific crosses between the tetraploid wheat cultivar Langdon and various A. tauschii accessions. The synthetic hexaploid wheat lines showed wide variation in many characteristics. To elucidate the genetic basis of variation in flowering-related traits, we analyzed quantitative trait loci (QTL) affecting time to heading, flowering and maturity, and the grain-filling period using four different F₂ populations of synthetic hexaploid wheat lines. In total, 10 QTLs located on six D-genome chromosomes (all except 4D) were detected for the analyzed traits. The QTL on 1DL controlling heading time appeared to correspond to a flowering time QTL, previously considered to be an ortholog of Eps-A ^m 1 which is related to the narrow-sense earliness in einkorn wheat. The 5D QTL for heading time might be a novel locus associated with wheat flowering, while the 2DS QTL appears to be an allelic variant of the photoperiod response locus Ppd-D1. Some of the identified QTLs seemed to be novel loci regulating wheat flowering and maturation, including a QTL controlling the grain filling period on chromosome 3D. The exercise demonstrates that synthetic wheat lines can be useful for the identification of new, agriculturally important loci that can be transferred to, and used for the modification of flowering and grain maturation in hexaploid wheat.     

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。     

控制水稻品种Koshihikari抽穗期的数量性状位点

利用Koshihikari × Kasalath BILs群体及相应的Kasalath/Koshihikari CSSLs群体,在江苏南京和海南陵水两个环境下对抽穗期的QTL定位分析。结果表明,在两地重复检测到3个QTL,分别位于第3、6和8染色体上;在qHd-3和qHd-8位点,来自Koshihikari等位基因能够提早抽穗,在qHd-6位点,来自Koshihikari等位基因能够延迟抽穗;第3染色体qHd-3和第7染色体非加性QTL之间存在上位性互作,通过重组自交系和置换系相互验证发现,qHd-3 所在的标记区间与W008的Kasalath插入片段位置大体一致,qHd-8所在的标记区间与W023、W024的Kasalath插入片段相吻合,qHd-7-1所在的标记区间位于W20的Kasalath片段之内,表明确实存在这3个位点。同时还对Koshihikari × 桂朝2号RILs抽穗期的QTL定位分析,在南京和海南分别检测到3个加性抽穗期QTL,1对非加性抽穗期QTL存在互作。     

Identification of quantitative trait loci for resistance to bending-type lodging in rice (Oryza sativa L.)

Bending-type lodging is one of the most important factors affecting the yield and grain quality of rice. This study identified quantitative trait loci (QTLs) for physical strength of the upper culms, and evaluated QTL effects on lodging resistance. In 2010 and 2011, QTLs for breaking strength, length, and diameter of the top three internodes were identified by analyzing chromosomal segment substitution lines (CSSLs) developed from ‘Koshihikari’ and ‘Kasalath’. The QTL analysis indicated that ‘Kasalath’ had two types of QTLs: one to strengthen specific internodes and one to simultaneously improve the physical strengths of plural internodes or the top three internodes. A QTL for breaking strengths of the top three internodes (bsuc11) was detected on chromosome 11 in both years. This QTL did not overlap with that for internode length. To evaluate the effects of bsuc11 on lodging resistance, this study selected three CSSLs with bsuc11 and analyzed the breaking strengths of the top three internodes after heading and the pushing resistance of the lower part. Internodes of ‘Koshihikari’ showed decreased breaking strengths after grain filling, while those of CSSLs with bsuc11 did not show this decrease in breaking strength. The pushing resistance of the lower part at the fully ripe stage was the same in ‘Koshihikari’ and CSSLs with bsuc11. These results suggested that bsuc11 could be a target to improve the physical strength of the upper culms to resist bending-type lodging, and that the physical strengths of upper and lower parts are controlled by different genetic factors in rice.     

Identification of qRL7, a major quantitative trait locus associated with rice root length in hydroponic conditions

Root system development is an important target for improving yield in rice. Active roots that can take up nutrients more efficiently are essential for improving grain yield. In this study, we performed quantitative trait locus (QTL) analyses using 215 recombinant inbred lines derived from a cross between Xieqingzao B (XB), a maintainer line with short roots and R9308, a restorer line with long roots. Only a QTLs associated with root length were mapped on chromosomes 7. The QTL, named qRL7, was located between markers RM3859 and RM214 on chromosome 7 and explained 18.14–18.36% of the total phenotypic variance evaluated across two years. Fine mapping of qRL7 using eight BC₃F₃ recombinant lines mapped the QTL to between markers InDel11 and InDel17, which delimit a 657.35 kb interval in the reference cultivar Nipponbare. To determine the genotype classes for the target QTL in these BC₃F₃ recombinants, the root lengths of their BC₃F₄ progeny were investigated, and the result showed that qRL7 plays a crucial role in root length. The results of this study will increase our understanding of the genetic factors controlling root architecture, which will help rice breeders to breed varieties with deep, strong and vigorous root systems.     

Detection of QTLs for traits associated with pre-harvest sprouting resistance in bread wheat (Triticum aestivum L.)

Pre-harvest sprouting (PHS) is one of the serious problems for wheat production, especially in rainy regions. Although seed dormancy is the most critical trait for PHS resistance, the control of heading time should also be considered to prevent seed maturation during unfavorable conditions. In addition, awning is known to enhance water absorption by the spike, causing PHS. In this study, we conducted QTL analysis for three PHS resistant related traits, seed dormancy, heading time and awn length, by using recombinant inbred lines from ‘Zenkouji-komugi’ (high PHS resistance) × ‘Chinese Spring’ (weak PHS resistance). QTLs for seed dormancy were detected on chromosomes 1B (QDor-1B) and 4A (QDor-4A), in addition to a QTL on chromosome 3A, which was recently cloned as TaMFT-3A. In addition, the accumulation of the QTLs and their epistatic interactions contributed significantly to a higher level of dormancy. QDor-4A is co-located with the Hooded locus for awn development. Furthermore, an effective QTL, which confers early heading by the Zenkouji-komugi allele, was detected on the short arm of chromosome 7B, where the Vrn-B3 locus is located. Understanding the genetic architecture of traits associated with PHS resistance will facilitate the marker assisted selection to breed new varieties with higher PHS resistance.     

Identification of coincident QTL for days to heading,spike length and spikelets per spike in <Emphasis Type="Italic">Lolium perenne</Emphasis> L.

Flowering time is a trait which has a major influence on the quality of forage. In addition, flowering and subsequent seed yields are important traits for seed production by grass breeders. In this study, we have identified quantitative trait loci (QTL) for flowering time and morphological traits of the flowering head in an F₁ mapping population in Lolium perenne L (perennial ryegrass), a number of which have not previously been identified in L. perenne mapping studies. QTL for days to heading (DTH) were mapped in both outdoor and glasshouse experiments, revealing three and five QTL for DTH which explained 53% and 42% of the total phenotypic variation observed, respectively. Two QTL for DTH were detected in both environments, although they had contrasting relative magnitudes in each environment. One QTL for spike length and three QTL for spikelets per spike were also identified explaining, a total of 32 and 33% of the phenotypic variance, respectively. Furthermore, the QTL for spike length and spikelets per spike generally coincided with QTL for days to heading, implying co-ordinate regulation by underlying genes. Of particular interest was a region harbouring overlapping QTL for days to heading, spike length and spikelets per spike on the top of linkage group 4, containing the major QTL for spike length identified in this population.     

Development and evaluation of pyramiding lines carrying early or late heading QTLs in the indica rice cultivar ‘IR64’

The heading date is an important trait for determining regional and climatic adaptability in rice. To expand the adaptability of the indica rice cultivar ‘IR64’, we pyramided multiple early or late heading quantitative trait locus (QTLs) in the ‘IR64’ genetic background by crossing previously developed near-isogenic lines (NILs) with a single QTL for early or late heading. The effects of pyramiding QTLs were observed in three different climatic zones of the Philippines, Madagascar, and Japan. The early heading pyramiding lines (PYLs) headed 6.2 to 12.8 days earlier than ‘IR64’ while the late heading PYLs headed 18.8 to 27.1 days later than ‘IR64’. The PYLs tended to produce low grain yield compared to ‘IR64’. The low yield was not improved by combining SPIKE, which is a QTL that increases the number of spikelets per panicle. Conversely, ‘IR64-PYL(7+10)’ carrying Hd5 and Hd1 headed earlier, produced more tillers, and more panicles per m² than ‘IR64’, and mitigated the yield decrease in early heading. These results suggest that the effects of pyramided QTLs on heading date were consistent across various environments and PYLs could be used to enhance the adaptation of ‘IR64’ in other rice growing environments.     

Fine mapping the QTL qFS‐chr.23, using a CSIL

In an earlier advanced‐backcross quantitative trait locus (QTL) analysis of an interspecific cross of Gossypium hirsutum cv. ‘Xinluzhong 36’(‘XLZH36’) and G. barbadense cv. ‘Xinhai 21’(‘XH21’), a QTL for fibre strength in the chromosome segment introgression line IL23‐09 was analysed. Single marker analysis revealed that the markers on chro.23 were associated with fibre strength. Using composite interval mapping with the F₂ population (1296 plants), a QTL for fibre strength was detected on chro. 23. The QTL explained 8.9% and 15.9% of phenotypic variances in the F₂ and F_2 : 3 generations, respectively. Substitution mapping suggested that the QTL was located at a physical distance of 23.4 kb between the markers BNL1414 and the single nucleotide polymorphism (SNP) locus D09_43776813 C‐G. We designated this QTL as qFS‐chr.23 (quantitative trait locus for fibre strength on chro.23). This work provides a valuable genetic resource for the breeding of high fibre quality in cotton and will facilitate future efforts for map‐based cloning.     

水稻芽性状耐冷性的QTL分析

低温发芽和芽期耐冷性是影响水稻芽生长发育的两个重要因素.本试验利用182个越光(粳型)/Kasalath(籼型)//越光回交重组自交株系(backcross recombinant inbred lines,BILs),对2个芽性状耐冷性(低温发芽和芽期耐冷性)进行QTL(quantitative trait loci)定位和相关性分析.结果表明,BIL群体中这2个芽性状耐冷性均呈连续分布,属于数量性状遗传,两性状间的相关性不显著;控制低温发芽的4个QTL分布于第1、7、9、11染色体上,其贡献率为6.72～12.78%;芽期耐冷性相关的4个QTL分布在第4、6(2个QTL)和11染色体上,贡献率为6.61%～14.93%;其中第11染色体上的2个QTL位于相同区域内,并且其增强耐冷性等位基因均来自耐冷性较差亲本Kasalath.本研究结果及其中检测到的QTLs两侧的连锁分子标记可为水稻芽性状耐冷性分子育种提供理论依据.     

Variation in heading date conceals quantitative trait loci for other traits of importance in breeding selection of rice

To identify quantitative trait loci (QTLs) associated with the primary target traits for selection in practical rice breeding programs, backcross inbred lines (BILs) derived from crosses between temperate japonica rice cultivars Nipponbare and Koshihikari were evaluated for 50 agronomic traits at six experimental fields located throughout Japan. Thirty-three of the 50 traits were significantly correlated with heading date. Using a linkage map including 647 single-nucleotide polymorphisms (SNPs), a total of 122 QTLs for 38 traits were mapped on all rice chromosomes except chromosomes 5 and 9. Fifty-eight of the 122 QTLs were detected near the heading date QTLs Hd16 and Hd17 and the remaining 64 QTLs were found in other chromosome regions. QTL analysis of 51 BILs having homozygous for the Koshihikari chromosome segments around Hd16 and Hd17 allowed us to detect 40 QTLs associated with 27 traits; 23 of these QTLs had not been detected in the original analysis. Among the 97 QTLs for the 30 traits measured in multiple environments, the genotype-by-environment interaction was significant for 44 QTLs and not significant for 53 QTLs. These results led us to propose a new selection strategy to improve agronomic performance in temperate japonica rice cultivars.     

Advanced-backcross QTL analysis in spring barley: IV. Localization of QTL × nitrogen interaction effects for yield-related traits

The advanced backcross quantitative trait locus (AB-QTL) analysis has proven its usefulness to identify and localize favourable alleles from exotic germplasm and to transfer those alleles into elite varieties. In a balanced design with up to six environments and two nitrogen fertilization (N treatment) levels, a 4-factorial mixed model analysis of variance (ANOVA) was used to identify QTL main effects, QTL × environment interaction effects and QTL × N treatment interaction effects in the spring barley BC₂DH population S42. The yield-related traits studied were number of ears per m², days until heading, plant height, thousand grain weight (TGW) and grain yield. In total, 82 QTLs were detected for all traits. This finding was compared to a previous QTL study of the same population S42, where the current field data was reduced to one half through restriction of the analysis to the standard N treatment level (von Korff et al., Theor Appl Genet 112: 1221–1231, 2006). These authors located 54 QTLs for the same traits by applying a 3-factorial mixed model similar to the current model but excluding the factor N treatment. We found that QTL × environment interaction, alone or in combination, accounted for 24 of the newly uncovered QTLs, whereas QTL × N treatment interaction was of lesser importance with six new cases in total. A valuable QTL interacting with N treatment has been identified on chromosome 7H where lines carrying the wild barley allele were superior in number of ears per m² in either N treatment. We conclude that in population S42 the extension of the phenotype data set and the inclusion of N treatment into the mixed model increased the power of QTL detection by providing an additional replication rather than by revealing specific N treatment QTLs.