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.     

Genetic relationship between grain chalkiness,protein content,and paste viscosity properties in a backcross inbred population of rice

A backcross inbred line population derived from a cross between Koshihikari and Kasalath was used to dissect the genetic relationship among chalkiness, protein content, and paste viscosity properties in rice in three environments. A total of 11 traits (or parameters) were analyzed, including percentage of grains with chalkiness (PGWC), protein content (PC) and protein index (PI), and eight parameters from the viscosity profile. PGWC, PC and PI were significantly correlated with the paste viscosity parameters. We identified 39 QTLs in three environments; ten QTL clusters emerged. Eight QTLs were consistently detected across the three environments and further confirmed using a set of chromosome segment substitution lines (CSSLs) where Kasalath was used as the donor parent and Koshihikari as the recurrent parent. One and two major clusters on chromosome 6 corresponded to the Wx and Alk loci, respectively. The former was responsible for PGWC and most of the viscosity parameters, and the latter for PI and some viscosity parameters. Particularly, QTL qPI-6.1 was linked with both the Wx and Alk loci. The co-locations of QTLs for PGWC and viscosity parameters and the linkage of qPI-6.1 and qBDV-6 at the Wx locus could be largely responsible for the phenotypic correlations between these traits.     

The QTL Analysis of RuBisCO in Flag Leaves and Non-Structural Carbohydrates in Leaf Sheaths of Rice Using Chromosome Segment Substitution Lines and Backcross Progeny F2 Populations

Abstract

In rice (Oryza sativa L.), the maintenance of high photosynthetic rate of flag leaves and the carbon remobilization from leaf sheaths after heading is a critical physiological component affecting the yield. To clarify the genetic basis of RuBisCO content of the flag leaf, a major determinant of photosynthetic rate, and non-structural carbohydrate (NSC) concentration in the third leaf sheath at heading, we carried out quantitative trait loci (QTL) analysis with 39 Koshihikari/Kasalath chromosome segment substitution lines (CSSLs) and backcross progeny F₂ population derived from target CSSL holding the QTL/Koshihikari in the field. QTLs for RuBisCO content and NSC concentration at heading were detected between R2447-C1286 and R2447-R716 on chromosome 10, respectively, by comparing Koshihikari with four CSSLs for chromosome 10 (SL-229, -230, -231 and -232). The progeny QTL for RuBisCO content and for NSC concentration at heading qRCH-10 and qNSCLSH-10-1, respectively, were detected at similar marker intervals between RM8201 and RM5708. In addition, QTLs for RuBisCO content at 14 d after heading, qRCAH-10-1 and qRCAH-10-2, were detected in regions different from that of qRCH-10. No QTL for NSC concentration at 14 d after heading was detected between RM8201 and R716, the region analyzed in this study. The QTLs qRCH-10 and qRCAH-10-1 for RuBisCO content would have additive effects. These QTLs for RuBisCO content and NSC concentration newly found using CSSLs and their backcross progeny F₂ population should be useful for better understanding the genetic basis of source and temporary-sink functions in rice and for genetic improvement of Koshihikari in terms of their functions.     

QTLs for malting flavour component associated with pre-harvest sprouting susceptibility in barley (Hordeum vulgare L.)

Lipoxygenase (LOX) is a key factor affecting quality of beer in terms of foam stability and flavour. Low LOX content is a desirable trait for malting quality. A doubled haploid (DH) population was made from a cross of Australian malting barley Stirling and Canadian malting barley Harrington and mapped with 513 molecular markers. The 120 DH lines with their parents were planted in field trials and the harvested grains were micro-malted for analysis of LOX content in two consecutive years. LOX content was controlled by both genetic effects and environment conditions. Three QTLs were consistently detected. One QTL flanked by the markers E6216 and SCssr03907 at the telomere region of chromosome 5HL contributed 39% of genetic variation in LOX content. The second QTL close to the centromere region of chromosome 5H accounted for 17% of genetic variation. A minor QTL on chromosome 2H explained 6% of genetic variation but was significant in both years. The Australian variety Stirling contributed to higher LOX content for the three QTLs. The two QTLs mapped at chromosome 5H for LOX content coincided with the QTLs for seed dormancy/pre-harvest sprouting from the same population. The pre-harvest sprouting susceptible alleles were associated with low LOX content, which indicated that the low LOX QTL from the Canadian malting barleys are only useful in the barley growing areas where the pre-harvest sprouting risk is low. New genetic sources for low LOX should be exploited in different germplasm with different mechanisms.     

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.     

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。     

水稻灌浆期耐热害的数量性状基因位点分析

 利用由98个家系组成的Nipponbare / Kasalath // Nipponbare回交重组自交系群体及其分子连锁图谱，以粒重感热指数\[（适温粒重-高温粒重）/适温粒重×100\]为评价指标，采用混合线性模型的QTL定位方法，对水稻灌浆期耐热性的主效、上位性数量性状基因位点及其与环境的互作进行分析。共检测到3个灌浆期耐热性主效QTL，分别位于第1、4和7染色体上，LOD值为8.16、11.08和12.86，贡献率8.94%、17.25%和13.50%。其中位于第4染色体标记C1100-R1783之间的QTL，没有显著的上位性和环境互作效应，表明在不同环境和遗传背景中的表达较为稳定，在水稻耐热性育种中可能具有较大的利用价值，其耐热性等位基因来自亲本Kasalath，高温热害时可减少粒重损失3.31%。位于第1染色体标记R1613-C970之间的QTL和第7染色体标记C1226-R1440之间的QTL，耐热性等位基因来自亲本Nipponbare，分别可减少粒重损失2.38%和2.92%。这两个QTL均具有与环境的互作效应，其中第7染色体上的QTL还和其他基因位点有互作。检测到8对加性×加性上位性互作QTL，分布于第1、2、3、5、7、8、10和12染色体上。没有检测到上位性QTL与环境的互作效应。     

Identification of Rice QTLs for Important Agronomic Traits with Long-Kernel CSSL-Z741 and Three SSSLs

Rice kernel shape affects kernel quality (appearance) and yield (1000-kernel weight) and therefore is an important agronomic trait, but its inheritance is complicated. We identified a long-kernel rice chromosome segment substitution line (CSSL), Z741, derived from Nipponbare as a recipient and Xihui 18 as a donor parent. Z741 has six substitution segments distributed on rice chromosomes 3, 6, 7, 8 and 12 with an average replacement length of 5.82 Mb. Analysis of a secondary F₂ population from a cross between Nipponbare and Z741 identified 20 QTLs for important agronomic traits. The kernel length of Z741 is controlled by a major QTL (qKL3) and a minor QTL (qKL7). Candidate gene prediction and sequencing indicated that qKL3 may be an allele of OsPPKL1, which encodes a protein phosphatase implicated in brassinosteroid signaling, and qKL7 is an unreported QTL. Finally, we validated eight QTLs (qKL3, qKL7, qRLW3-1, qRLW7, qPH3-1, qKWT3, qKWT7 and qNPB6) using three selected single- segment substitution lines (SSSLs), S1, S2 and S3. Also, we detected five QTLs (qKL6, qKW3, qKW7, qKW6 and qRLW6) in S1, S2 and S3, which were not found in the Nipponbare/Z741 F₂ population. However, qNPB3, qNPB7 and qPL3 QTLs were not validated by the three SSSLs in 2019, suggesting that minor QTLs are susceptible to environmental factors. These results lay the foundation for studying the biodiversity of kernal length and molecular breeding of different kernel types.     

稻米直链淀粉含量和胶稠度对高温耐性的QTL分析

利用由98个家系组成的Nipponbare/Kasalath//Nipponbare回交重组自交系群体,以直链淀粉含量耐热指数（高温下直链淀粉含量/适温下直链淀粉含量×100）和胶稠度耐热指数（高温下胶稠度/适温下胶稠度×100）为评价指标,采用混合线性模型的QTL定位方法,在南昌和南京两个试验地点对水稻蒸煮食用品质性状的高温耐性QTL进行了检测。两个性状在两个试验地点共检测到9个QTLs,其中直链淀粉含量高温耐性QTL 3个,胶稠度高温耐性QTL 6个。两个性状中共有3个QTLs在两个地点同时被检测到。其中位于第6染色体上与Wx基因相同的染色体区域和第8染色体G1073-R727区域分别是控制直链淀粉含量和胶稠度高温耐性的重要区域。     

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 Cooking and Eating Quality of Rice Grain

Rice is one of the major crops served as the staple food of more than 50% of the world’s population. Recently, more attention has been paid to rice quality, especially the cooking and eating quality than achieving the higher yield. The amylose content (AC), gelatinization temperature (GT) and gel consistency (GC) are the three major rice characteristics directly related to cooking and eating quality, responsible for the physical and chemical characteristics of the starch in the endosperm [1…     

水稻抽穗期基因的精细定位、克隆和生物学功能分析

介绍了水稻抽穗期QTL研究的进展,在相同亲本日本晴/Kasalath衍生的不同类型的多个群体中,共检测到15个QTL;应用高世代回交后代,精细定位了其中8个QTL;将在初步定位时同一区间检测到的1个控制种子休眠期QTL(Sdr1)和1个抽穗期QTL (Hd8),分解为两个紧密连锁的基因;将经过精细定位表明可能具有双重功能的单个孟德尔因子Hd3,分解为两个功能不同的紧密连锁的基因Hd3a和Hd3b;根据QTL近等基因系的光周期反应以及这些座位间上位性互作的研究,明确了其中6个QTL的生物学功能;应用图位法克隆了其中3个QTL,研究了它们的表达和调控,并与拟南芥的同源基因进行比较。为水稻其他数量性状以及其他作物数量性状的遗传学研究,提供了一个范例。     

Mapping Quantitative Trait Loci for Palatability of Milled Rice

Quantitative trait loci (QTLs) controlling palatability in rice were identified using a set of 98 backcross inbred lines (BILs) population derived from a cross between a japonica variety Nipponbare and an indica variety Kasalath. The palatability scores of the population measured by RQ1/Plus Rice Analyzer, showed a continuous and transgressive segregative distribution with a range from 66 to 92. Four putative QTLs for palatability, qPAL-5, qPAL-7, qPAL-8a and qPAL-8b, were detected on chromosome 5, 7 and 8, and they accounted 7.83, 7.03, 11.58 and 7.19% of the total phenotypic variation, respectively. Three alleles qPAL-5, qPAL-7 and qPAL-8b from Kasalath increased the palatability score, whereas only one Nipponbare allele qPAL-8a increased the score. Eight transgressive lines in palatability were selected to make a comparison between phenotypic and genotypic classes. The result explained the possibility of positive QTLs pyramiding through marker-assisted selection of highly palatable rice.     

Chromosome Segment Substitution Lines: A Powerful Tool for the Introgression of Valuable Genes from Oryza Wild Species into Cultivated Rice (O. sativa)

Wild species of rice (genus Oryza) contain many useful genes but a vast majority of these genes remain untapped to date because it is often difficult to transfer these genes into cultivated rice (Oryza sativa L.). Chromosome segment substitution lines (CSSLs) and backcross inbred lines (BILs) are powerful tools for identifying these naturally occurring, favorable alleles in unadapted germplasm. In this paper, we present an overview of the research involving CSSLs and BILs in the introgression of quantitative trait loci (QTLs) associated with the improved performance of rice including resistance to various biotic and abiotic stresses, and even high yield from wild relatives of rice and other unadapted germplasm into the genetic background of adapted rice cultivars. The CSSLs can be used to dissect quantitative traits into the component genetic factors and evaluate gene action as single factors (monogenic loci). CSSLs have the potential to uncover new alleles from the unadapted, non-productive wild rice accessions, develop genome-wide genetic stocks, and clone genes identified in QTL studies for functional genomics research. Recent development of high-density single-nucleotide polymorphism (SNP) arrays in rice and availability of custom-designed medium- and low-density SNP arrays will enhance the CSSL development process with smaller marker-defined segment introgressions from unadapted germplasm.     

A novel Rice QTL <Emphasis Type="Italic">qOPW11</Emphasis> Associated with Panicle Weight Affects Panicle and Plant Architecture

Background

The improvement of rice yield is a crucial global issue, but evaluating yield requires substantial efforts. Rice yield comprises the following indices: panicle number (PN), grain number per panicle (GN), 1000-grain weight, and percentage of ripened grain. To simplify measurements, we analyzed one panicle weight (OPW) as a simplified yield index that integrates GN, grain weight, and percentage of ripened grain, and verified its suitability as a proxy for GN and grain weight in particular.

Results

Quantitative trait locus (QTL) analysis using 190 recombinant inbred lines derived from Koshihikari (large panicle and small grain) and Yamadanishiki (small panicle and large grain), japonica cultivars detected three QTLs on chromosomes 5 (qOPW5), 7 (qOPW7) and 11 (qOPW11). Of these, qOPW5 and qOPW11 were detected over two years. qOPW5 and qOPW7 increased OPW, and qOPW11 decreased it at Yamadanishiki alleles. A chromosome segment substitution line (CSSL) with a genomic segment from Yamadanishiki substituted in the Koshihikari genetic background harboring qOPW5 increased grain weight. qOPW11 had the largest genetic effect of QTLs, which was validated using a CSSL. Substitution mapping using four CSSLs revealed that qOPW11 was located in the range of 1.46 Mb on chromosome 11. The CSSL harboring qOPW11 decreased primary and secondary branch numbers, culm length, and panicle length, and increased PN.

Conclusions

In this study, three QTLs associated with OPW were detected. The CSSL with the novel and largest QTL, qOPW11, differed in some traits associated with both panicle and plant architecture, indicating different functions for the meristem in the vegetative versus the reproductive stages. qOPW5 coincided with an identified QTL for grain width and grain weight, suggesting that qOPW5 was affected by rice grain size. OPW can be considered a useful trait for efficient detection of QTLs associated with rice yield.

     

利用回交重组自交系定位稻米赖氨酸含量的基因座位

利用日本晴（粳稻）/Kasalath（籼稻）//日本晴组合衍生的98个回交重组自交系（BILs）株系和具有245个RFLP标记的遗传图谱,在浙江和海南2个环境条件下,开展了精米赖氨酸含量的QTL定位。精米赖氨酸含量在浙江和海南均表现连续变异和超亲分离。在第6染色体发现2个具有显著遗传主效应的QTL(qLYS6 1和qLYS6 2）,表型贡献率分别为2708%和4756%。qLYS6 1 还具有显著的环境互作效应。qLYS6 1的增效基因来自Kasalath,而qLYS6 2的增效基因来自日本晴。未检测到显著的上位性效应。     

Mapping QTL for Heat-Tolerance at Grain Filling Stage in Rice

A mapping population of 98 lines (backcross inbred lines, BILs) derived from a backcross of Nipponbare/Kasalath// Nipponbare was planted at two experimental sites, Nanjing and Nanchang, and treated with high and optimal temperature during grain filling, respectively. The grain weight heat susceptibility index [GWHSI= (grain weight at optimum temperature-grain weight at high temperature) / grain weight at optimum temperature ×100] was employed to evaluate the tolerance of rice to heat stress. A genetic linkage map with 245 RFLP markers and a mixed linear-model approach was used to detect quantitative trait loci (QTLs) and their main effects, epistatic interactions and QTL×environment interactions (Q×E). The threshold of LOD score=2.0 was used to detect the significance of association between marker and trait. A total of 3 QTLs controlling heat tolerance during grain filling were detected, on chromosomes 1, 4 and 7, with LOD scores of 8.16, 11.08 and 12.86, respectively, and they explained the phenotypic variance of 8.94, 17.25 and 13.50 %, correspondingly. The QTL located in the C1100-R1783 region of chromosome 4 showed no QTL×environment interaction and epistatic effect, suggesting that it could be stably expressed in different environments and genetic backgrounds, and thus it would be valuable in rice breeding for heat tolerance improvement. This QTL allele, derived from Kasalath reduced 3.31% of the grain weight loss under heat stress. One located between R1613-C970 on chromosome 1 and the other between C1226-R1440 on chromosome 7, with additive effect 2.38 and 2.92%, respectively. The tolerance alleles of both these QTLs were derived from Nipponbare. Both of these QTLs had significant QTL×environment interactions, and the latter was involved in epistatic interaction also. Eight pairs of epistatic effect QTLs were detected, one pair each on chromosomes 1,2,3, 5, 7, 8, 10 and 12. The results could be useful for elucidating the genetic mechanism of heat-tolerance and the development of new rice varieties with heat tolerance during grain filling phase.     

QTL Mapping for Grain Size Traits Based on Extra-Large Grain Rice Line TD70

Grain size traits, including grain length, grain width and grain thickness, are controlled by quantitative trait loci （QTLs）. Many QTLs relating to rice grain size traits had been reported, but their control mechanisms have not yet been elucidated. A recombinant inbred line （RIL） population of 240 lines, deriving from a cross between TD70, an extra-large grain size japonica line with 80 g of 1000-grain weight, and Kasalath, a small grain size indica variety, were constructed and used to map grain size QTLs to a linkage map by using 141 SSR markers in 2010 and 2011. Five QTLs for grain length, six for grain width and seven for grain thickness were detected distributing over chromosomes 2, 3, 5, 7, 9 and 12. Seven QTLs, namely qGL3.1, qGW2, qGW2.2, qGW5.1, qGW5.2, qGT2.3 and qGT3.1, were detected in either of the two years and explained for 56.19%, 4.42%, 29.41%, 10.37%, 7.61%, 21.19% and 17.06% of the observed phenotypic variances on average, respectively. The marker interval RM1347-RM5699 on chromosome 2 was found common for grain length, grain width and grain thickness; qGL3.1 and qGT3.1 were mapped to the same interval RM6080-RM6832 on chromosome 3. All 18 QTL alleles were derived from the large grain parent TD70. Most of the QTLs mapped in the present study were found the same as the genes previously cloned （GW2, GS3 or qGL3, GW5 and GS5）, and several were the same as the QTLs （GS7 and qGL-7） previously mapped. Three QTLs, qGL2.2 on chromosome 2, qGW9 and qGT9 on chromosome 9, were first detected. These results laid a foundation for further fine mapping or cloning of these QTLs.     

利用染色体片段置换系群体检测水稻叶片形态QTL

 水稻叶片的形态改良是水稻株型育种和产量育种的重要目标之一。以9311/日本晴染色体片段置换系（CSSLs）群体为材料,定位了上3叶叶片长、宽、叶面积共9个性状QTL,分析了叶片性状与产量性状之间的相关性,同时定位了主穗重及产量构成因素（颖花数、千粒重、结实率）相关QTL。结果表明,CSSLs群体的叶片性状之间存在显著或极显著相关性;叶片性状与主穗重呈显著或极显著正相关,与主穗颖花数呈极显著正相关;叶片形态多数性状与结实率、千粒重没有显著相关性。两年共定位到20个叶片性状QTL,分布于第1、3、4、5、6、9、11共7条染色体的10个区间,贡献率为3.82%~14.61%,其中贡献率大于10%有6个,多个QTL成簇分布在相同区间,3个QTL在两年间重复检测到,8个QTL为前人未报道的新位点。两年共检测到16个与控制主穗产量相关的QTL,分布于第1、2、3、5、7、8、10共7条染色体13个区间,其中有7个主穗产量相关QTL所在5个区间与叶片形态14个QTL所在区间一致。     

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.