Confirmation of Novel Quantitative Trait Loci for Seed Dormancy at Different Ripening Stages in Rice

Seed dormancy contributes resistance to pre-harvest sprouting.Effects on respective quantitative trait loci (QTLs) for dormancy should be assessed by using fresh seeds before germinability altered through storage.We investigated QTLs related to seed dormancy using backcross inbred lines derived from a cross between Nipponbare and Kasalath.Four putative QTLs for seed dormancy were detected immediately after harvest using composite interval mapping.These putative QTLs were mapped near C1488 on chromosome 3 (qSD-3.1),R2171 on chromosome 6 (qSD-6.1),R1245 on chromosome 7 (qSD-7.1) and C488 on chromosome 10 (qSD-10.1).Kasalath alleles promoted dormancy for qSD-3.1,qSD-6.1 and qSD-7.1,and the respective proportions of phenotypic variation explained by each QTL were 12.9%,9.3% and 8.1%.We evaluated the seed dormancy harvested at different ripening stages during seed development using chromosome segment substitution lines (CSSLs) to confirm gene effects.The germination rates of CSSL27 and CSSL28 substituted with the region including qSD-6.1 were significantly lower than those of Nipponbare and other CSSLs at the late ripening stage.Therefore,qSD-6.1 is considered the most effective novel QTL for pre-harvest sprouting resistance among the QTLs detected in this study.     

基于单片段代换系玉米子粒性状的QTL定位

利用lx9801为遗传背景的昌7-2单片段代换系为基础材料,通过一年两点的田间试验,对粒重、粒长、粒宽和粒厚进行QTL分析。在两个试验点共检测到31个玉米子粒性状的QTL,包括12个粒重QTL、7个粒长QTL、8个粒宽QTL和4个粒厚QTL。其中,有2个粒重QTL、2个粒长QTL和1个粒厚QTL在两试验点同时检测到,可分别解释5.53%、9.09%、4.55%、4.18%和7.70%的表型变异。     

QTL Mapping for Rice RVA Properties Using High-Throughput Re-sequenced Chromosome Segment Substitution Lines

The rapid visco analyser （RVA） profile is an important factor for evaluation of the cooking and eating quality of rice. To improve rice quality, the identification of new quantitative trait loci （QTLs） for RVA profiling is of great significance. We used a japonica rice cultivar Nipponbare as the recipient and indica rice 9311 as the donor to develop a population containing 38 chromosome segment substitution lines （CSSLs） genotyped by a high-throughput re-sequencing strategy. In this study, the population and the parent lines, which contained similar apparent amylose contents, were used to map the QTLs of RVA properties including peak paste viscosity （PKV）, hot paste viscosity （HPV）, cool paste viscosity （CPV）, breakdown viscosity （BKV）, setback viscosity （SBV）, consistency viscosity （CSV）, peak time （PET） and pasting temperature （PAT）. QTL analysis was carried out using one-way analysis of variance and Dunnett＇s test, and stable QTLs were identified over two years and under two environments. We identified 10 stable QTLs： qPKV2-1, qSBV2-1; qPKV5-1, qHPV5-1, qCPV5-1; qPKV7-1, qHPV7-1, qCPV7-1, qSBV7-1; and qPKV8-1 on chromosomes 2, 5, 7 and 8, respectively, with contributions ranging from -95.6% to 47.1%. Besides, there was pleiotropy in the QTLs on chromosomes 2, 5 and 7.     

用培矮64S/日本晴F2群体对水稻6个农艺性状的QTL定位

 用水稻测序品种培矮64S和日本晴配组建立了由180个单株组成的F2群体，构建了含137个SSR标记的连锁遗传图谱，对水稻的分蘖数、有效分蘖数、分蘖率、株高、剑叶长和穗长等6个相关农艺性状进行了QTL定位分析。共检测到14个QTL，分布在第1、2、4、5、6、7染色体的11个区间。检测到1个控制株高的主效QTL(qPH1 2),位于第1染色体，其表型贡献率为24.0%；1个控制剑叶长的主效QTL(qFL4）,位于第4染色体，其表型贡献率为30.5%。对所定位QTL的价值、QTL在染色体上的区域分布等进行了探讨。     

利用单片段代换系鉴定水稻株高及其构成因素的QTL

 通过t测验比较了单片段代换系与受体亲本华粳籼74之间的表型差异，对以6个水稻品种为供体的52个单片段代换系代换片段上株高及其构成因素的QTL进行了鉴定。以P≤0.001为阈值，在14个代换片段上共鉴定出24个QTL，包括10个株高QTL、2个穗长QTL、4个倒1节间长QTL、5个倒2节间长QTL、3个倒3节间长QTL，这些QTL分布于水稻的9条染色体上。QTL加性效应值为-4.08～3.98 cm，加性效应百分率为-19.35%～10.43%。     

基于玉米87-1综3单片段代换系的穗长QTL分析

以优良玉米自交系综3为供体、87-1为受体,利用回交和SSR标记辅助选择相结合的方法,获得了51份纯合的染色体单片段代换系(Single Segment Substitution Line,SSSL),每个SSSL内只含有一个来源于供体的染色体片段。51个代换片段不均匀分布在玉米10条染色体上,代换片段长度在1.75～172.45 cM之间,平均长度为24.44cM。覆盖玉米基因组的总长度为933.90 cM,覆盖率为40.20%。2008年在郑州利用17份SSSL材料及在三亚利用39份SSSL材料对玉米穗长进行了1年2点的表型鉴定,共检测到20个穗长QTL,加性效率百分率在-12.10%～19.18%之间。结果表明,这些单片段代换系存在较大的遗传变异,是用于玉米复杂性状的QTL鉴定、基因克隆及功能分析的理想材料。     

Genetic and physiological analysis of tolerance to acute iron toxicity in rice

Background

Fe toxicity occurs in lowland rice production due to excess ferrous iron (Fe²⁺) formation in reduced soils. To contribute to the breeding for tolerance to Fe toxicity in rice, we determined quantitative trait loci (QTL) by screening two different bi-parental mapping populations under iron pulse stresses (1,000 mg L⁻¹ = 17.9 mM Fe²⁺ for 5 days) in hydroponic solution, followed by experiments with selected lines to determine whether QTLs were associated with iron exclusion (i.e. root based mechanisms), or iron inclusion (i.e. shoot-based mechanisms).

Results

In an IR29/Pokkali F₈ recombinant inbred population, 7 QTLs were detected for leaf bronzing score on chromosome 1, 2, 4, 7 and 12, respectively, individually explaining 9.2-18.7% of the phenotypic variation. Two tolerant recombinant inbred lines carrying putative QTLs were selected for further experiments. Based on Fe uptake into the shoot, the dominant tolerance mechanism of the tolerant line FL510 was determined to be exclusion with its root architecture being conducive to air transport and thus the ability to oxidize Fe²⁺ in rhizosphere. In line FL483, the iron tolerance was related mainly to shoot-based mechanisms (tolerant inclusion mechanism). In a Nipponbare/Kasalath/Nipponbare backcross inbred population, 3 QTLs were mapped on chromosomes 1, 3 and 8, respectively. These QTLs explained 11.6-18.6% of the total phenotypic variation. The effect of QTLs on chromosome 1 and 3 were confirmed by using chromosome segment substitution lines (SL), carrying Kasalath introgressions in the genetic background on Nipponbare. The Fe uptake in shoots of substitution lines suggests that the effect of the QTL on chromosome 1 was associated with shoot tolerance while the QTL on chromosome 3 was associated with iron exclusion.

Conclusion

Tolerance of certain genotypes were classified into shoot- and root- based mechanisms. Comparing our findings with previously reported QTLs for iron toxicity tolerance, we identified co-localization for some QTLs in both pluse and chronic stresses, especially on chromosome 1.     

Fine Mapping of QTLs for Stigma Exsertion Rate from Oryza glaberrima by Chromosome Segment Substitution

Stigma exsertion is an important trait for outcrossing ability in rice. Stigma exsertion rate (SER) of male sterile lines (MSLs) is a key factor affecting F₁-seed production in hybrid rice. In this study, seven QTLs for SER were detected on five chromosomes using a set of single-segment substitution lines (SSSLs) derived from O. glaberrima. Three of the QTLs were mapped in the estimated intervals of 92.5–333.0 kb. qSER-5 was located in a substitution segment of 92.5 kb. qSER-1b and qSER-8b were respectively limited to 333.0 kb and 107.5 kb by secondary substitution mapping. qSER-1b and qSER-3 had bigger additive effects of 11.5% and 11.9%, respectively, while the other five QTLs had smaller additive effects from 5.7% to 8.6%. Open reading frames were identified in the regions of qSER-5 and qSER-8b in O. sativa and O. glaberrima genomes. Fine mapping of the QTLs laid a foundation for the cloning of genes, and QTLs for SER will be used to develop MSLs with strong ability of outcrossing.     

QTL Detection for Rice Grain Shape Using Chromosome Single Segment Substitution Lines

Rice grain shape is one of the important factors affecting grain quality and yield,but it is liable to be influenced by genetic backgrounds and environments.The chromosome single segment substitution lines(SSSLs) in rice have been considered as ideal populations to identify the quantitative trait loci(QTLs).In this study,22 QTLs affecting rice grain shape were detected to be distributed on eight chromosomes except chromosomes 6,9,11 and 12 by using SSSLs.Among them,seven QTLs conditioned grain length,six co...     

利用重测序的水稻染色体片段代换系定位控制稻米淀粉黏滞性谱QTL

 稻米RVA谱是评价稻米蒸煮与食用品质的重要指标之一,发掘新的控制稻米RVA谱QTL对稻米品质改良具有重要意义。利用以粳稻品种日本晴为受体、籼稻品种9311为供体并经高通量重测序的染色体片段代换系群体为材料,在两年两点的环境下对该群体中控制RVA谱特征值的QTL进行了定位分析。通过单因素方差分析和Dunnett多重比较, 分析染色体片段代换系与受体亲本之间相关RVA谱特征值的差异,以两年两点都能检测到的显著差异位点作为稳定表达的QTL。共检测到10个稳定表达的QTL,包括控制峰值黏度的4个QTL qPKV2.1、qPKV5.1、qPKV7.1和qPKV8.1,控制热浆黏度的2个QTL qHPV5.1和qHPV7.1,控制冷胶黏度的2个QTL qCPV5.1和qCPV7.1及控制消减值的2个QTL qSBV2.1和qSBV7.1。在两年两点的检测中10个稳定表达QTL的贡献率介于-31.8%～53.2%,这10个QTL分布在第2、5、7和8染色体上,其中位于第2、5和7染色体上的位点存在一因多效性。     

基于水稻单片段代换系的粒形QTL定位

 粒形是重要的农艺性状,既影响水稻产量也影响稻米品质,是典型的数量性状,易受遗传背景和环境因素的影响。染色体单片段代换系降低了遗传背景的干扰,是鉴定QTL的新型遗传材料。利用单片段代换系为材料,共鉴定出了22个与粒形相关的QTL,包括7个粒长QTL,6个粒宽QTL,5个谷粒长宽比QTL和4个粒厚QTL,分布于除第6、9、11和12染色体外的8条染色体上。这些结果为粒形QTL的克隆和分子育种奠定了基础。     

利用单片段代换系群体定位玉米株型性状QTL

以优良玉米自交系许178背景的综3单片段代换系群体为基础材料,通过一年两点的田间试验,对株高、穗位高、总叶片数、穗上叶片数、叶面积等植株性状进行QTL分析。结果表明,利用显著性检验方法在两点共检测到5个株型相关性状的72个QTL,包括23个株高QTL、12个穗位高QTL、6个总叶片数QTL、15个穗上叶片数QTL、16个叶面积QTL,其中,有10个株高QTL、3个穗位高QTL、3个总叶片数QTL、2个穗上叶片数QTL和1个叶面积QTL在2个试验环境中同时检测到。     

基于重测序的染色体片段代换系定位水稻抽穗期QTL

【目的】水稻的抽穗期是决定水稻产量及其适用性的重要农艺性状之一,是由多基因控制的数量性状。染色体片段代换系减少了个体间遗传背景的干扰,已经成为定位和克隆复杂性状QTL的重要材料。【方法】本研究以9311为受体,日本晴为供体构建的128个重测序的染色体片段代换系群体为试验材料,利用多元回归,结合Bin-map图谱,【结果】鉴定到6个在南京、扬州不同年份间稳定表达的抽穗期QTL,其中,qHD2.1被定位在第2染色体上的759 848 bp区间内;qHD2.2被定位在第2染色体上的45 286 bp区间内;qHD 3.1被定位在第3染色体上的147 931 bp区间内;qHD5.1被定位在第5染色体上的213 351 bp区间内;qHD5.2被定位在第5染色体上的442 305 bp区间内;qHD8.1被定位在第8染色体上的538 176 bp区间内。【结论】本研究为精细定位并克隆相应QTL,进而探明抽穗期QTL的分子调控机制奠定了基础。     

玉米株高主效QTL qPH2.4的定位分析

课题组前期以玉米自交系郑58为轮回亲本,以昌7-2为供体亲本,通过分子标记辅助选择获得染色体单片段代换系Z12和W16。Z12和W16在bin2.07区域均含有1个来源于昌7-2的染色体片段,株高均显著高于轮回亲本郑58。利用郑58和Z12为材料构建F2分离群体,基于重测序和混合分离分析(BSA)策略,将株高主效QTL qPH2.4定位于第2染色体13.95 Mb(201 457 953~215 022 157 bp)的区域内。利用在目标区间内筛选出的20对多态性分子标记对包含743个单株的郑58×Z12 F2分离群体和包含1 720个单株的郑58×W16 F2分离群体进行基因型分析,结合田间株高数据进行QTL定位,将株高主效QTL qPH2.4定位在InDel分子标记ph-18和ph-19之间,区间的遗传距离为0.57 cM,物理距离为626 kb。参考B73基因组(RefGen_v4)注释信息,该区间内存在17个注释基因,其中,包含可以调控油菜素内酯信号的基因Zm00001d006677。     

利用染色体片段置换系群体定位和分析水稻粒重和粒型QTL

[目的]挖掘水稻粒重和粒型相关性状QTL,对于解析水稻籽粒遗传机理具有重要作用.[方法]本研究以籼稻9311为受体、粳稻日本晴为供体构建的染色体片段置换系(Chromosome Segment Substitution Lines,CSSLs)群体为材料,在4个环境下对控制稻谷与糙米的粒重和粒型QTL进行了定位分析.[...     

Identification of QTLs for Plant Height and Its Components by Using Single Segment Substitution Lines in Rice （Oryza sativa）

QTLs for plant height and its components on the substituted segments of fifty-two single segment substitution lines （SSSLs） in rice were identified through t-test （P〈0.001） for comparison between each SSSL and recipient parent Huajingxian 74. On the 14 substituted segments, 24 QTLs were detected, 10 for plant height, 2 for panicle length, 4 for length of the first internode from the top, 5 for length of the second internode from the top and 3 for length of the third internode from the top, respectively. All these QTLs were distributed on nine rice chromosomes except chromosomes 5, 9 and 11. The additive effect ranged from -4.08 to 3.98 cm, and the additive effect percentages varied from -19.35% to 10.43%.     

Identification of QTLs for Improvement of Plant Type in Rice (Oryza sativa L.) Using Koshihikari / Kasalath Chromosome Segment Substitution Lines and Backcross Progeny F2 Population

Abstract

Thirty-nine chromosome segment substitution lines (CSSLs) population derived from a Koshihikari / Kasalath cross was used for quantitative trait locus (QTL) analysis of plant type in rice (Oryza sativa L.). Putative rough QTLs (26.2～60.3cM of Kasalath chromosomal segments) for culm length, plant height, panicle number, chlorophyll content of flag leaf blade at heading and specific leaf weight, were mapped on the several chromosomal segments based on the comparison of CSSLs with Koshihikari in the field experiment for 3 years. In order to verify and narrow QTLs detected in CSSLs, we conducted QTL analyses using F₂ populations derived from a cross between Koshihikari and target CSSL holding a putative rough QTL. The qPN-2, QTL for panicle number was mapped on chromosome 2. In traits of flag leaf, the qCHL-4-1 and qCHL-4-2 for chlorophyll content was mapped on chromosome 4, and the qSLW-7 for specific leaf weight on chromosome 7. All QTLs were detected in narrow marker intervals, compared with rough QTLs in CSSLs. The qPN-2, qCHL-4-1 and qCHL-4-2 had only additive effect. On the other hand, the qSLW-7 showed over-dominance. It could be emphasized that QTL analysis in the present study with the combination of CSSLs and backcross progeny F₂ population can not only verify the rough QTLs detected in CSSLs but also estimate allelic effects on the QTL.     

两种供氮水平下玉米穗部性状的QTL定位

以优良杂交种豫玉22两亲本Z3和87-1为基础构建一套F8家系的重组自交系群体为研究材料,在正常供氮和低氮两种氮水平下进行田间试验,利用复合区间作图法对玉米穗长、穗行数、行粒数、百粒重和单穗粒数进行QTL定位分析。两种氮水平下共定位到24个玉米穗部性状的QTL位点,其中正常供氮条件下定位到13个QTL,低氮水平下定位到11个QTL,集中分布在第1(8个QTL)、第5(6个QTL)和第8(5个QTL)染色体上。两种氮水平下共位或紧密连锁的QTL位点较少,表明玉米穗部性状在低氮水平下的遗传机制发生很大改变。研究发现,第1染色体umc1122/bnlg1556位点是一个控制低氮水平下玉米单穗粒数的主效QTL,单个QTL可解释19.7%的表型变异,该位点还同时影响低氮水平下玉米穗长、穗行数和百粒重的表型。与前人定位结果比较发现,该位点所在的染色体区域是一个产量及氮效率相关性状的QTL富集区,对此位点附近进行相关分子标记辅助选择,可能会在玉米氮高效分子育种上有所突破。     

Analysis of QTLs for Flag Leaf Shape and Its Response to Elevated CO2 in Rice （Oryza sativa）

To understand the responses of flag leaf shape in rice to elevated CO2 environment and their genetic characteristics,quantitative trait loci(QTLs)for flag leaf shape in rice were mapped onto the molecular marker linkage map of chromosome segment substitution lines(CSSLs)derived from a cross between a japonica variety Asominori and an indica variety IR24 under free air carbon dioxide enrichment(FACE,200μmol/mol above current levels)and current CO2 concentration(Ambient,about 370μmol/mol).Three flag-leaf traits,flag-leaf length(LL),width(LW)and the ratio of LL to LW (RLW),were estimated for each CSSL and their parental varieties.The differences in LL,LW and RLW between parents and in LL and LW within IR24 between FACE and Ambient were significant at 1%level.The continuous distributions and transgressive segregations of LL,LW and RLW were also observed in CSSL population,showing that the three traits were quantitatively inherited under both FACE and Ambient.A total of 16 QTLs for the three traits were detected on chromosomes 1,2,3,4,6,8 and 11 with LOD(Log10-likelihood ratio)scores ranging from 3.0 to 6.7.Among them,four QTLs (qLL-6*,qLL-8*,qLW-4*,and qRLW-6*)were commonly detected under both FACE and Ambient.Therefore,based on the different responses to elevated CO2 in comparison with current CO2 level,it can be suggested that the expressions of several QTLs associated with flag-leaf shape in rice could be induced by the high CO2 level.     

Genetic analysis of grain protein content,grain hardness and dough rheology in a hard×hard bread wheat progeny

Kernel texture (hard vs soft grain) and more subtle within-class variations are known to have a large influence on end-use properties, mostly through the proportion of damaged starch and subsequent water requirement. In this study, quantitative trait loci (QTL) affecting kernel texture and dough rheology in a progeny from a cross between two ‘medium-hard’ wheat cultivars were identified and compared to the QTL locations for both traits. One hundred and sixty-five F₇ recombinant inbred lines were studied in three environments in 1999. Kernel texture was estimated by both near infrared reflectance (NIR: Hard_NIR) and the single-kernel characterisation system (SKCS: Hard_SKCS). Dough rheology, taken as a predictor of end-use quality, was estimated by the empirical parameters measured by Chopin alveograph. The genetic map for this population consisted of 254 loci quite evenly distributed over the wheat genome. Considering only the QTL which were stable (detected in the three locations) and robust (through bootstrap resampling), five genomic regions were found to influence Hard_NIR, but only two of them are significant for Hard_SKCS, which was probably due to a less representative measure of the phenomenon. Eight QTLs were found for rheological traits. Some QTLs for dough rheology co-located with those for hardness or grain protein content, particularly on chromosomes 3A and 5B and close to the unlinked marker Xgwm130. According to trait, individual QTLs explained from 5.4 to 26.6% of the phenotypic variation and when taken together up to 46.0% of the variation.