Transgressive variants for red pericarp grain with high yield potential derived from Oryza rufipogon × Oryza sativa: Field evaluation, screening for blast disease, QTL validation and background marker analysis for agronomic traits

Five transgressive variants (advanced breeding lines from BC₂F₅ and BC₂F₆ generation) were derived from a cross between the wild relative, O. rufipogon Griff. and O. sativa L. subsp. indica cv. MR219, a popular high yielding Malaysian rice cultivar. The aim of the study was to evaluate the pericarp colour of the grains along with yield potential and to validate quantitative trait loci (QTLs) for agronomic traits. The variants were screened against blast disease. Background marker analysis was also done for the promising variants. The field trials were carried out at a single location (due to containment purposes) over two seasons using randomized complete block design (RCBD) with three replications. A trait-based marker analysis was used to identify QTLs for validation in BC₂F₅ generation. Analysis of variance (ANOVA) showed that the seasonal factors influenced different agronomic traits. Variant G33 produced significantly (p < 0.05) higher yield (5.20 t/ha) than the control, MR219 (4.53 t/ha). Eighteen QTLs for different agronomic traits were identified in BC₂F₂ population in a previous study. Among them 14 QTLs were found in BC₂F₅ population of the present study. The yield of variant G33 was influenced by several QTLs viz. qGPL-1, qSPL-1-2, qSPL-8 and qYLD-4, which were introgressed from the donor parent revealed by background marker analysis using BC₂F₇ generation. Percentage (99%) of red pericarp grain of G33 and G34 in BC₂F₅ and BC₂F₆ generations indicated the stability of pericarp colour which was transferred from the wild relative. Variant G33 showed resistance against two pathotype of blast disease (Magnaporthe oryzae). Among the evaluated variants, G33 could be considered for inclusion in the cultivar development program for red rice with high yield potential and resistance to blast disease. This study demonstrated that the alleles from wild relative could improve the yield and yield related traits through allelic interaction, even though the phenotypic traits were inferior to the recurrent parent.     

Characterizing the Saltol Quantitative Trait Locus for Salinity Tolerance in Rice

This study characterized Pokkali-derived quantitative trait loci (QTLs) for seedling stage salinity tolerance in preparation for use in marker-assisted breeding. An analysis of 100 SSR markers on 140 IR29/Pokkali recombinant inbred lines (RILs) confirmed the location of the Saltol QTL on chromosome 1 and identified additional QTLs associated with tolerance. Analysis of a series of backcross lines and near-isogenic lines (NILs) developed to better characterize the effect of the Saltol locus revealed that Saltol mainly acted to control shoot Na⁺/K⁺ homeostasis. Multiple QTLs were required to acquire a high level of tolerance. Unexpectedly, multiple Pokkali alleles at Saltol were detected within the RIL population and between backcross lines, and representative lines were compared with seven Pokkali accessions to better characterize this allelic variation. Thus, while the Saltol locus presents a complex scenario, it provides an opportunity for marker-assisted backcrossing to improve salt tolerance of popular varieties followed by targeting multiple loci through QTL pyramiding for areas with higher salt stress.     

Introgression of Two Drought QTLs into FUNAABOR-2 Early Generation Backcross Progenies Under Drought Stress at Reproductive Stage

FUNAABOR-2 is a popular Ofada rice variety grown in a large area under rainfed upland condition across western states of Nigeria. We used the combination of phenotypic and marker-assisted selection(MAS) to improve grain yield of FUNAABOR-2 under drought stress(DS) at the reproductive stage via introgression of two drought quantitative trait loci(QTLs), qDTY12.1 and qDTY2.3. Foreground selection was carried out using peak markers RM511 and RM250, associated with qDTY12.1 and qDTY2.3, respectively, followed by recombinant selection with RM28099 and RM1261 distally flanking qDTY12.1. Furthermore, BC1 F2-derived introgressed lines and their parents were evaluated under DS and non-stress(NS) conditions during the 2015–2016 dry season. Overall reduction of grain yield under DS compared to NS was recorded. Introgressed lines with qDTY12.1 and qDTY2.3 combinations showed higher yield potential compared to lines with single or no QTL under DS, indicating significant positive interactions between the two QTLs under the FUNAABOR-2 genetic background. Pyramiding of qDTY12.1 and qDTY2.3 in the FUNAABOR-2 genetic background led to higher grain yield production under DS and NS.     

Identification and validation of QTLs for cold tolerance at the booting stage and other agronomic traits in a rice cross of a Japanese tolerant variety,Hananomai, and a NERICA parent,WAB56-104

In Africa, cold temperatures occur in the highlands of East and Southern Africa and in some areas of the Sahel region of West Africa leading to substantial rice yield losses. Cold tolerance (CT) at booting stage on basis of spikelet fertility after cold water irrigation was evaluated using F₂ population derived from a cross between temperate japonica, Hananomai, and tropical japonica, WAB56-104. Two Quantitative trait loci (QTLs) for CT were detected on chromosome 8 and 10 with enhanced effects on the trait coming from Hananomai and WAB56-104 allele, respectively. The QTLs explained 30% and 33% of phenotypic variation in spikelet fertility, respectively. CT was negatively correlated with panicle number (r = ?0.35, p < 0.01) and positively correlated with panicle weight (r = 0.61, p < 0.001). Selected BC₁F₄ and BC₁F₅ genotypes having homozygous alleles for both CT QTLs exhibited higher spikelet fertility under cold stress. The identified QTLs will be useful in the development of cold-tolerant varieties for production in high altitude areas through marker-assisted selection.     

Quantitative trait loci mapping and meta-analysis across three generations for popping characteristics in popcorn

Popping characteristics play a determinant role in the utilization of popcorn (Zea mays L.). In this study, the RIL population with 258 recombinant inbred lines was evaluated to detect quantitative trait loci (QTLs) for three popping characteristics (PF, popping fold; PV, popping volume; PR, popping rate) under four environments. Meta-analysis was used to integrate detected QTLs across three generations (RIL, F_2:3 and BC₂F₂) derived from the same cross. All eleven QTLs were detected for three traits, on chromosomes 1, 2, 4, 6 and 10 for PF, on chromosomes 1, 4, 6, 7 and 10 for PV, and on chromosomes 1, 4, 6 and 10 for PR. Three, 1, 3, 6 and 6 QTL were detected in the same marker intervals in 4, 3, 2, 1 cases, respectively. Four QTLs at bins 1.05–1.06, 1.08–1.09 and 7.03–7.04 were commonly detected in the same or near bins in all three generations. Six and 2 QTLs showed consistency across RIL/F_2:3 or RIL/BC₂F₂ generations respectively. Nine meta-QTLs (mQTL) were detected on chromosomes 1, 4, 6, 7, 8 and 10. Except mQTL7-1, only related with PV, other mQTLs included two or three traits, reflecting pleiotropic or tightly linkaged QTLs for popping characteristics. The QTL influencing all the three popping traits at bins 1.05–1.06 were also detected in other previous researches using different populations, which could be put into use in marker assisted breeding for popping characteristics in popcorn.     

Quantitative trait loci mapping and meta-analysis across three generations for popping characteristics in popcorn

Popping characteristics play a determinant role in the utilization of popcorn (Zea mays L.). In this study, the RIL population with 258 recombinant inbred lines was evaluated to detect quantitative trait loci (QTLs) for three popping characteristics (PF, popping fold; PV, popping volume; PR, popping rate) under four environments. Meta-analysis was used to integrate detected QTLs across three generations (RIL, F_2:3 and BC₂F₂) derived from the same cross. All eleven QTLs were detected for three traits, on chromosomes 1, 2, 4, 6 and 10 for PF, on chromosomes 1, 4, 6, 7 and 10 for PV, and on chromosomes 1, 4, 6 and 10 for PR. Three, 1, 3, 6 and 6 QTL were detected in the same marker intervals in 4, 3, 2, 1 cases, respectively. Four QTLs at bins 1.05–1.06, 1.08–1.09 and 7.03–7.04 were commonly detected in the same or near bins in all three generations. Six and 2 QTLs showed consistency across RIL/F_2:3 or RIL/BC₂F₂ generations respectively. Nine meta-QTLs (mQTL) were detected on chromosomes 1, 4, 6, 7, 8 and 10. Except mQTL7-1, only related with PV, other mQTLs included two or three traits, reflecting pleiotropic or tightly linkaged QTLs for popping characteristics. The QTL influencing all the three popping traits at bins 1.05–1.06 were also detected in other previous researches using different populations, which could be put into use in marker assisted breeding for popping characteristics in popcorn.     

Marker-Assisted Selection of Xa21 Conferring Resistance to Bacterial Leaf Blight in indica Rice Cultivar LT2

Bacterial leaf blight of rice (BLB), caused by Xanthomonas oryzae pv. oryzae, is one of the most destructive diseases in Asian rice fields. A high-quality rice variety, LT2, was used as the recipient parent. IRBB21, which carries the Xa21 gene, was used as the donor parent. The resistance gene Xa21 was introduced into LT2 by marker-assisted backcrossing. Three Xoo races were used to inoculate the improved lines following the clipping method. Eleven BC₃F₃ lines carrying Xa21 were obtained based on molecular markers and agronomic performance. The 11 lines were then inoculated with the three Xoo races. All the 11 improved lines showed better resistance to BLB than the recipient parent LT2. Based on the level of resistance to BLB and their agronomic performance, five lines (BC₃F₃ 5.1.5.1, BC₃F₃ 5.1.5.12, BC₃F₃ 8.5.6.44, BC₃F₃ 9.5.4.1 and BC₃F₃ 9.5.4.23) were selected as the most promising for commercial release. These improved lines could contribute to rice production in terms of food security.     

Seed Length Controlled by Same Locus in Four Different AA GenomeSpecies of Genus Oryza简

To broaden the genetic basis and overcome the yield plateau in Asian cultivated rice, the exploitation and utilization of favorable alleles from rice species with the AA genome has become important and urgent in modern breeding programs. Four different interspecific populations were used to detect quantitative trait locus (QTL) for seed length, including a BC₄F₂ population derived from Oryza glumaepatula crossed with Dianjingyou 1 (a japonica cultivar), a BC₄F₂ population derived from O. nivara crossed with Dianjingyou 1, a BC₇F₁ population derived from a cross between O. longistaminata and RD23 (an indica cultivar), and a BC₈F₁ population derived from a cross between O. glaberrima and Dianjingyou 1. The QTLs for seed length in four different populations were termed as SL-3a, SL-3b, SL-3c and SL-3d, respectively. They had good collinearity and accounted for 49% to 60% of the phenotypic variations. Sequencing data indicated that four QTLs were different alleles of GS3 which were responsible for the seed length variation between O. sativa and its four AA genome relatives. These results will be valuable for confirming the evolution of GS3 and also be helpful for rice breeding.     

Improvement of Upland Rice Variety by Pyramiding Drought Tolerance QTL with Two Major Blast Resistance Genes for Sustainable Rice Production

Varalu is an early maturing rice variety widely grown in the rainfed ecosystem preferred for its grain type and cooking quality. However, the yield of Varalu is substantially low since it is being affected by reproductive drought stress along with the blast disease. The genetic improvement of Varalu was done by introgressing a major yield QTL, qDTY_12.1, along with two major blast resistance genes i.e. Pi54 and Pi1 through marker-assisted backcross breeding. Both traits were transferred till BC₂ generation and intercrossing was followed to pyramid the two traits. Stringent foreground selection was carried out using linked markers as well as peak markers (RM28099, RM28130, RM511 and RM28163) for the targeted QTL (qDTY_12.1), RM206 for Pi54 and RM224 for Pi1. Extensive background selection was done using genome-wide SSR markers. Six best lines (MSM-36, MSM-49, MSM-53, MSM-57, MSM-60 and MSM-63) having qDTY_12.1 and two blast resistance genes in homozygous condition with recurrent parent genome of 95.0%-96.5% having minimal linkage drag of about 0.1 to 0.7 Mb were identified. These lines showed yield advantage under drought stress as well as irrigated conditions. MSM-36 showed better performance in the national coordinated trials conducted across India, which indicated that improved lines of Varalu expected to replace Varalu and may have an important role in sustaining rice production. The present study demonstrated the successful marker-assisted pyramiding strategy for introgression of genes/QTLs conferring biotic stress resistance and yield under abiotic stress in rice.     

QTL Analysis for Grain Iron and Zinc Concentrations in Two O. nivara Derived Backcross Populations

Identification of quantitative trait loci (QTLs) for grain mineral elements can assist in faster and more precise development of micronutrient dense rice varieties through marker-assisted breeding. In the present study, QTLs were mapped for Fe and Zn concentrations in two BC₂F₃ mapping populations derived from the crosses of O. sativa cv Swarna with two different accessions of O. nivara. In all, 10 and 8 QTLs were identified for grain Fe and Zn concentrations in population 1, and 7 and 5 QTLs were identified in population 2, respectively. Eighty percent of the QTLs detected in both populations were derived from O. nivara. Five QTLs for Fe and three QTLs for Zn explained more than 15% phenotypic variance either in interval or composite interval mapping. The locations of O. nivara derived QTLs such as qFe2.1, qFe3.1, qFe8.2 and qZn12.1 were consistently identified in both the populations. Epistatic interaction was observed only between RM106 and RM6 on chromosome 2 and between RM22 and RM7 on chromosome 3 for Fe concentration in population 1. Sixteen candidate genes for metal homeostasis were found to co-locate with 10 QTLs for Fe and Zn concentrations in both the populations. Most of the Fe and Zn QTLs were found to co-locate with QTLs for grain yield and grain quality traits. Some of the major effect QTLs identified can be used to improve rice grain Fe and Zn concentrations.     

Molecular Mapping of Sterility QTLs qSS-3, qSS-6a and qSS-7 as Single Mendelian Factors via NIL strategy

Hybrid sterility between Oryza glaberrima and O. sativa seriously hampers the introgression of favorable genes from each other. In order to further understand this issue, identification and isolation of hybrid sterility QTLs as single Mendelian factors are an effective strategy. A genetic map was constructed using a BC₁F₁ population derived from a cross between an O. sativa japonica cultivar and an O. glaberrima accession. Four main-effect QTLs for pollen sterility were detected in the BC₁F₁. Five BC₈F₁ advanced backcross populations were developed via successive backcrosses based on phenotype and molecular selections. The BC₈F₁ populations showed bimodal distribution for pollen fertility and could be classified into semi-sterile and fertile types, fitting single Mendilian factor inheritance ratios. Three QTLs detected in the BC₁F₁ corresponding to qSS-3, qSS-6a and qSS-7 were mapped on chromosomes 6, 3 and 7, respectively, as single Mendilian factors.     

Mapping of QTLs for Germination Characteristics under Non-stress and Drought Stress in Rice

Identification of genetic factors controlling traits associated with seed germination under drought stress conditions, leads to identification and development of drought tolerant varieties. Present study by using a population of F2：, derived from a cross between a drought tolerant variety, Gharib （indica） and a drought sensitive variety, Sepidroud （indica）, is to identify and compare QTLs associated with germination traits under drought stress and non-stress conditions. Through QTL analysis, using composite interval mapping, regarding traits such as germination rate （GR）, germination percentage （GP）, radicle length （RL）, plumule length （PL）, coleorhiza length （COL） and coleoptile length （CL）, totally 13 QTLs were detected under pole drought stress （-8 MPa poly ethylene glycol 6000） and 9 QTLs under non-stress conditions. Of the QTLs identified under non-stress conditions, QTLs associated with COL （qCOL-5） and GR （qGR-1） explained 21.28% and 19.73% of the total phenotypic variations, respectively Under drought stress conditions, QTLs associated with COL （qCOL-3） and PL （qPL-5） explained 18.34% and 18.22% of the total phenotypic variations, respectively. A few drought-tolerance-related QTLs identified in previous studies are near the QTLs detected in this study, and several QTLs in this study are novel alleles. The major QTLs like qGR-1, qGP-4, qRL-12 and qCL-4 identified in both conditions for traits GR, GP, RL and CL, respectively, should be considered as the important and stable trait-controlling QTLs in rice seed germination. Those major or minor QTLs could be used to significantly improve drought tolerance by marker-assisted selection in rice.     

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.     

Development of elite breeding lines conferring Bph18 gene-derived resistance to brown planthopper (BPH) by marker-assisted selection and genome-wide background analysis in japonica rice (Oryza sativa L.)

Brown planthopper (BPH) is a serious threat to rice production. In this study, we have used the novel resistance gene Bph18 derived from Oryza australiensis and incorporated it into an elite japonica cultivar, Junambyeo, which is highly susceptible to BPH. The Bph18 gene was introduced by marker-assisted backcross (MAB) breeding into Junambyeo. The backcrossed progenies were evaluated for desirable agronomic and grain quality traits and the selection of improved breeding lines while simultaneously evaluating BPH resistance by bioassays in the greenhouse and foreground selection. Of the 26 advanced backcross breeding lines (ABL), four lines showed agronomic traits similar to those of the recurrent parent, with strong resistance to BPH. Molecular genotyping of the four ABL revealed the conversion of genotypes closely resembling the genotype of Junambyeo. The percentage of donor chromosome segments in ABL decreased from 12.3% in the BC₂ to 9.4%, 8.4% and 5.3% in BC₃, BC₄ and BC₅ generations, respectively. ABL retained small sizes of the donor chromosome segments on chromosomes 1, 2, 10, 11 and 12 but the genomes of ABL2, ABL3 and ABL4 were homosequential to the recurrent parent on chromosomes 3, 4, 5, 6, 7 and 9 without donor chromosome segment introgression. The ABL1 and ABL2 retained only some small segments of the donor genome on chromosomes 9 and 8, respectively. Fine structure analysis of the Bph18 flanking region between RM511 and RM1584 markers on chromosome 12 showed a progressive elimination of donor-derived chromosome segments from BC₂ to BC₅ generations. The percentage of O. australiensis derived chromosome segment substitution in the recurrent parent background decreased from 28% of the donor parent to 6.7%, 3.9%, 3.4% and 3.4% in BC₂, BC₃, BC₄ and BC₅ generations, respectively. However, it was revealed that the O. australiensis-derived chromosome segment (1320 kb) in ABL containing the Bph18 gene was consistently maintained irrespective of advances in backcross generations. BPH resistant elite breeding lines with agronomic and grain quality traits similar to those of the recurrent parent were successfully developed by foreground and background analysis in japonica background without linkage drag.     

Genetic Control of Germination Ability under Cold Stress in Rice

An F₉ recombinant inbred lines (RIL) population, derived from a cross between IR28 (Oryza sativa L. spp. indica) and Daguandao (O. sativa L. spp. japonica), was used to construct a molecular linkage map and to identify germination ability including the traits of imbibition rate, germination rate, germination index, root length, shoot length and seed vigor at 14°C for 23 d. A composite interval mapping approach was applied to conduct genetic analysis for germination ability. The frequency distributions of the germination ability traits under the cold stress in the RIL population showed continuous segregation, suggesting they were quantitative traits controlled by several genes. A total of seven QTLs were identified on chromosomes 4, 6 and 9, including two for imbibition rate (qIR-6, qIR-9), one for germination rate (qGR-4), two for germination index (qGI-4-1, qGI-4-2) and two for root length (qRL-4-1, qRL-4-2). There were no detected QTLs controlling shoot length and seed vigor. The phenotypic variance explained by a single QTL ranged from 9.1% to 37.0%, and two major QTLs, qIR-6 and qGI-4-2, accounted for over 30% of the phenotypic variance. The expressions of QTLs were developmentally regulated and growth stage-specific. Most of the QTLs observed here were located in the regions similar to the QTLs for rice cold tolerance reported previously, indicating that these QTLs were reliable. However, qRL-4-2 is not reported before.     

玉米不育系回交转育群体背景选择效果的研究

以玉米细胞质不育系cms-合344为供体亲本、辐63018为轮回亲本,构建BC_1和BC_2群体,研究利用SSR标记进行遗传背景选择加速回交转育不育系的效果。选用两亲本间多态性好的并覆盖玉米10条染色体的60个SSR标记进行轮回亲本的遗传背景分析。结果表明,对两群体遗传背景回复率分析,86株BC_1群体背景回复率平均值为74.41%,63株BC_2群体背景回复率平均值为87.36%,BC_1群体表型与分子标记辅助选择相似度均较高的株系有9株,BC_2有8株。对两群体进行不同标记数目分析,两群体中使用40个标记与总数60个标记所计算的背景回复率基本相同,相关系数分别达到0.96和0.95以上。     

利用玉米选择导入系进行营养生长期抗旱性基因组区段定位

采用我国优异骨干自交系黄早四与掖478的回交群体(BC₂S₁和BC₃S₁)的抗旱性选择导入系群体,通过SSR标记检测群体中等位基因的导入频率,利用卡方检验对等位基因导入频率进行检测,同时结合表型-基因型方差分析(ANOVA)的方法,对抗旱群体进行抗旱性相关位点的分析。结果表明：利用卡方测验检测到31个供体等位基因导入,利用方差分析检测到98个位点导入,其中有17个位点是重合的,这些导入位点可能是抗旱相关的重要的候选基因组区段。     

Quantitative Trait Loci for Mercury Tolerance in Rice Seedlings

Mercury (Hg) is one of the most toxic heavy metals to living organisms and its conspicuous effect is the inhibition of root growth.However,little is known about the molecular genetic basis for root growth under excess Hg2+ stress.To map quantitative trait loci (QTLs) in rice for Hg2+ tolerance,a population of 120 recombinant inbred lines derived from a cross between two japonica cultivars Yuefu and IRAT109 was grown in 0.5 mmol/L CaCl2 solution.Relative root length (RRL),percentage of the seminal root length in +HgCl2 to-HgCl2,was used for assessing Hg2+ tolerance.In a dose-response experiment,Yuefu had a higher RRL than IRAT109 and showed the most significant difference at the Hg2+ concentration of 1.5 μmol/L.Three putative QTLs for RRL were detected on chromosomes 1,2 and 5,and totally explained about 35.7% of the phenotypic variance in Hg2+ tolerance.The identified QTLs for RRL might be useful for improving Hg2+ tolerance of rice by molecular marker-assisted selection.     

Identification of QTLs for Cadmium Tolerance During Seedling Stage and Validation of qCDSL1 in Rice

Cadmium(Cd)is a non-essential toxic metal that is harmful to plants.To investigate the genetic mechanism of Cd tolerance in rice,quantitative trait loci(QTLs)associated with Cd tolerance at the seedling stage were analyzed using a recombinant inbred line(RIL)population derived from a cross between PA64s and 93-11.A total of 36 QTLs associated with shoot length,root length,shoot dry weight,root dry weight and total dry weight were detected in Hangzhou and Lingshui of China.Among them,15 QTLs were identified under the control condition and 15 QTLs were identified under the Cd stress condition,and 6 QTLs for Cd tolerant coefficient were detected on chromosomes 1,3,7 and 9.The qCDSL1.1 and qCDSL1.2 were identified in Hangzhou and Lingshui,respectively,and had overlapping intervals on chromosome 1.To further confirm the effects of qCDSL1.1 and qCDSL1.2,we developed a chromosome segment substitution line(CSSL),CSSLqCDSL1,in 93-11 background harboring qCDSL1.1/qCDSL1.2 from PA64s.Compared to 93-11,CSSLqCDSL1 had increased shoot length under the Cd stress condition.These results pave the way for further isolation of those genes controlling Cd tolerance in rice and marker-assistant selection of rice elite varieties with Cd tolerance.