Detection and verification of QTLs associated with heat-induced quality decline of rice (Oryza sativa L.) using recombinant inbred lines and near-isogenic lines

Decline in the apparent quality of rice (Oryza sativa L.) grain due to high temperatures during ripening recently became a major concern in many areas in Japan. The occurrence of white-back kernels (WBK) is one of the main problems of heat-induced quality decline. We identified QTLs associated with the occurrence of WBK using recombinant inbred lines (RILs) and verified their effects using near-isogenic lines (NILs). The QTL analysis used F₇ and F₈ RILs derived from ‘Hana-echizen’ (HE), which is tolerant to high temperature, × ‘Niigata-wase’ (NW), which is sensitive to high temperature. Four QTLs were identified on chromosomes 3, 4, 6, and 9 (qWB3, qWB4, qWB6 and qWB9). To verify the effects of qWB6 and qWB9, we developed two NILs in which qWB6 or both were introduced from HE into the NW background. The HE allele at qWB6 significantly decreased WBK under multiple environments. The combination of qWB6 and qWB9 in an F₂ population derived from a cross between a NIL and NW showed that the NW allele at qWB9 significantly decreased WBK if the qWB6 allele was HE. These results will be of value in marker-assisted selection for the breeding of rice with tolerance to heat-induced quality decline.     

水稻耐冷性鉴定及定位研究概况

为了更好研究水稻耐冷机理,对水稻耐冷基因进行定位克隆,通过分子育种来选育水稻耐冷品种,由此对水稻耐冷研究现状进行分析。对水稻发芽期、芽期、苗期、孕穗期的耐冷鉴定方法,室内与大田鉴定方法进行比较分析,对各时期耐冷指标的应用及相关定位研究进行总结。从生理生化方面阐述了水稻耐冷机理的研究,分析水稻在受到冷害后水稻膜相的改变,酶活的改变情况。并概括了近年来水稻在冷害敏感时期的耐冷性数量性状QTL定位研究方面取得的进展,研究的群体有用近等基因系,重组自交系,DH系,其中多数利用近等基因系来进行研究。其中研究集中在芽期、发芽期的耐冷QTLs定位,苗期、孕穗期的QTLs研究较少。介绍了第一个水稻芽期耐冷基因克隆情况。并展望水稻耐冷研究方面今后的方向,很长一段时期内,研究依然集中在QTLs的定位上,水稻耐冷基因的克隆正在逐步开展,耐冷机理的研究应关注蛋白间的互作。     

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.     

Detection of QTLs to reduce cadmium content in rice grains using LAC23/Koshihikari chromosome segment substitution lines

To advance the identification of quantitative trait loci (QTLs) to reduce Cd content in rice (Oryza sativa L.) grains and breed low-Cd cultivars, we developed a novel population consisting of 46 chromosome segment substitution lines (CSSLs) in which donor segments of LAC23, a cultivar reported to have a low grain Cd content, were substituted into the Koshihikari genetic background. The parental cultivars and 32 CSSLs (the minimum set required for whole-genome coverage) were grown in two fields with different natural levels of soil Cd. QTL mapping by single-marker analysis using ANOVA indicated that eight chromosomal regions were associated with grain Cd content and detected a major QTL (qlGCd3) with a high F-test value in both fields (F = 9.19 and 5.60) on the long arm of chromosome 3. The LAC23 allele at qlGCd3 was associated with reduced grain Cd levels and appeared to reduce Cd transport from the shoots to the grains. Fine substitution mapping delimited qlGCd3 to a 3.5-Mbp region. Our results suggest that the low-Cd trait of LAC23 is controlled by multiple QTLs, and qlGCd3 is a promising candidate QTL to reduce the Cd level of rice grain.     

Identification of QTLs for rice grain size and shape of Iranian cultivars using SSR markers

Grain size is a main component of rice appearance quality. In this study, we performed the SSR mapping of quantitative trait loci (QTLs) controlling grain size (grain length and breadth) and shape (length/breadth ratio) using an F₂ population of a cross between two Iranian cultivars, Domsephid and Gerdeh, comprising of 192 individuals. A linkage map with 88 markers was constructed, which covered 1367.9 cM of the rice genome with an average distance of 18 cM between markers. Interval mapping procedure was used to identify the QTLs controlling three grain traits, and QTLs detected were further confirmed using composite interval mapping. A total of 11 intervals carrying 18 QTLs for three traits were identifed, that included five QTLs for grain length, seven QTLs for grain breadth, and six QTLs for grain shape. A major QTL for grain length was detected on chromosome 3, that explained 19.3% of the phenotypic variation. Two major QTLs for grain breadth were mapped on chromosomes 3 and 8, which explained 34.1% and 20% of the phenotypic variation, respectively. Another two major QTLs were identified for grain shape on chromosomes 3 and 8, which accounted for 27.1% and 20.5% of the phenotypic variance, respectively. The two QTLs that were mapped for grain shape coincided with the major QTLs detected for grain length and grain breadth. Intrestingly, gs2 QTL specific to grain shape was detected on chromosome 2 that explained 15% of the phenotypic variation.     

基于高密度SNP遗传图谱的粳稻芽期耐低温QTL鉴定

水稻直播由于省时、省工和节约成本而备受农户关注。然而,芽期耐冷性不强致使现行推广的许多优良水稻品种不适于直播生产。因此,挖掘鉴定芽期耐冷位点,为后续的辅助育种提供基因资源就日益受到重视。本研究利用丽江新团黑谷和沈农265构建的重组自交系群体及其重测序构建的包含2818个bin标记的遗传图谱对水稻芽期的耐冷性进行QTL定位分析。共检测到5个芽期耐冷QTL,分布在水稻的1号、3号、9号和11号染色体上,增效等位基因均来自耐冷亲本丽江新团黑谷。这些QTL的LOD值的范围从3.05到24.01,表型贡献率为8.0%～53.5%。其中,表型贡献率最大的主效QTL是qCTB11b,位于11号染色体长臂端的21.24 Mb～22.03 Mb之间,物理图谱区间为790 kb。随后利用"选择作图"的策略进行了QTL验证和累加效应分析,明确了可以通过QTL的累加聚合实现芽期耐冷能力的遗传改良,聚合的增效QTL越多,耐冷能力提升越明显。上述研究结果不仅可以增强人们对芽期水稻耐冷能力遗传基础的认识和理解,也可以为后续直播品种的遗传改良提供理论依据和技术指导。     

Two novel QTLs regulate internode elongation in deepwater rice during the early vegetative stage

Deepwater rice possesses internode elongation ability to avoid drowning under deepwater conditions. Previous studies identified three QTLs regulating internode elongation ability on chromosomes 1, 3 and 12 using different populations. However, these QTLs only induce internode elongation in response to deepwater conditions from the 7-leaf stage and not during the early leaf stage. In this study, we detected two novel QTLs, qTIL2 and qTIL4 regulating deepwater response at the early leaf stage using an F₂ population derived from the cross between NIL1-3-12 carrying the three QTLs regulating deepwater response in T65 (O. sativa ssp. japonica) genetic background and C9285 (O. sativa ssp. indica, deepwater rice). Plants of the BC₂F₂ population derived from NIL1-3-12/C9285 and the RILs of T65/Bhadua (O. sativa ssp. indica, deepwater rice) possessing these QTLs as well as the three QTLs previously identified also showed internode elongation during the early leaf stage. These results indicate that qTIL2 and qTIL4 regulate early internode elongation and function in coordination with the three major QTLs under deepwater conditions. The results presented here would not only help define the mechanism of deepwater response in rice but also contribute in the breeding of deepwater tolerant rice that is adapted to various water depths.     

QTL mapping and effect confirmation for anaerobic germination tolerance derived from the japonica weedy rice landrace PBR

As one of the main rice establishment methods, direct seeding has the advantages of saving labour and reducing costs for farmers. However, the expansion of direct seeding has been constrained by poorly levelled fields, heavy rainfall and poor drainage after sowing. The development of cultivars with anaerobic germination tolerance could improve or overcome these shortcomings of most rice under flooding conditions. In this study, a japonica-type rice with anaerobic germination tolerance was suggested as a new genetic source and QTL analysis was carried out to improve adaptation to direct seeding. Three QTLs, qAG1, qAG3 and qAG11, were identified, explaining 13.44%, 6.71% and 14.52% of the phenotypic variance, respectively. In an evaluation of the most effective QTL combination, two QTL combinations (qAG1^NP + qAG3^PBR + qAG11^PBR and qAG1^NP + qAG11^PBR) showed stable survival rates under submergence of filed condition. Based on the results, we will characterize the candidate genes within the detected target regions in further research, and the lines with advantageous alleles will be used to develop reliable cultivars to support the wide adoption of direct-seeding practices in japonica rice.     

灌浆温度和氮肥及其互作效应对稻米贮藏蛋白组分的影响

灌浆结实期温度与氮肥施用量是影响稻米品质的两个重要生态因子,尤其是与稻米蛋白含量及米饭食味关系密切。本文以多个水稻主栽品种为材料,通迆灌浆结实期的人工控温试验、大田长期定位点的施氮处理试验和盆栽条件下的温氮两因素复合处理试验,探讨了水稻灌浆结实期温度对稻米贮藏蛋白含量与组分影响及其有别于氮肥处理效应的差异觃律,幵分析了温度与氮肥两个因素对稻米贮藏蛋白及其组分影响的交互作用特点。结果表明,高温胁迫和增施氮肥均引起水稻籽粒总蛋白及其谷蛋白组分含量(%)的显著增加,但两者对稻米醇溶蛋白影响却存在明显差别。其中,高温处理引起醇溶蛋白含量显著下降,提高稻米谷蛋白/醇溶蛋白比值,而增施氮肥引起稻米谷蛋白和醇溶蛋白含量明显增加,但对谷蛋白/醇溶蛋白比值与贮藏蛋白各亚基的组成比例影响相对较小。在高温处理下,谷蛋白的57kD前体亚基组分含量有所提高,而37kD酸性亚基和22kD碱性亚基随温度处理的差异变化却因品种而异,且高温处理对水稻籽粒蛋白绝对含量(mg grain^–1)的影响程度也进没有其对蛋白相对含量(%)的影响明显。高氮×高温处理组合对稻米总蛋白与谷蛋白含量的影响程度显著大...     

水稻耐冷性遗传及基因定位研究进展

综述了水稻芽期、苗期、孕穗期和开花灌浆期耐冷性的遗传研究概况和耐冷性基因定位的研究进展,并对今后水稻耐冷性研究提出了建议和展望。     

Development and characterization of chromosome segment substitution lines derived from backcross between japonica donor rice cultivar Yukihikari and japonica recipient cultivar Kirara397

Grain yield-related traits and grain quality-related traits are important for rice cultivars. The quantitative trait loci (QTLs) involved in controlling the natural variation in these traits among closely related cultivars are still unclear. The present study describes the development of a novel chromosome segment substitution line (CSSL) population derived from a cross between the temperate japonica cultivars Yukihikari and Kirara397, which are grown in Hokkaido, the northernmost limit for rice cultivation. Days to heading, culm length, panicle length, panicle number, brown grain weight per plant, thousand brown grain weight, brown grain length, brown grain width, brown grain thickness, apparent amylose content, and protein content were evaluated. Panicle length, brown grain length and amylose content differed significantly in the parental cultivars. Thirty-five significant changes in the evaluated traits were identified in the CSSLs. A total of 28 QTLs were located on chromosomes 1, 2, 3, 4, 5, 6, 8, 9, 10, 11 and 12. These findings could be useful for breeding rice cultivars in the northernmost limit for rice cultivation.     

Detection and characterization of epistasis between QTLs on plant height in rice using single segment substitution lines

Hua-jing-xian 74 and its 12 single segment substitution lines (SSSLs) in rice were used as crossing parents to construct a half diallel crossing population. A total number of 91 materials were grown under three planting densities. By analysis of average plant height (PH) over all environments 10 SSSLs were detected with significant additives and 6 SSSLs with significant dominances. These SSSLs were further tested under different densities respectively, indicating that some of single locus effects were sensitive to densities and the conditions under the density of 16.7 cm × 16.7 cm maybe inhibited the expressing of these PH QTLs. Qualitative and quantitative analyses of each four participating genotypes indicated that digenic interactions among these QTLs were prevalent. Of 66 tested interactions, about 42.4% were epistatic (P < 5%). Although some QTLs hadn’t single locus effects, they were possible to form digenic interactions. A significant finding was that the detected epistases were mostly negative. Additionally, these epistases were also found being sensitive to planting densities, the conditions under the density of 10 cm × 16.7 cm perhaps promoted the expressing of epistatic interactions among PH QTLs.     

Difference of optimum germination temperature of seeds of intact and dehusked japonica rice during seed development

The effects of incubation temperature on germination of intact and dehusked japonica rice that had been harvested every 5 days from 10 days after anthesis (DAA) to 70 DAA were surveyed. At 30 ^°C incubation, the germination percentage of intact seeds at 25 DAA exceeded that of dehusked seeds. This tendency persisted until 65 DAA, when around 95% of both intact and dehusked seeds germinated. Germination percentages of intact seeds incubated at 20 ^°C were lower than seeds incubated at 30 ^°C for all DAA. However, germination percentages of dehusked seeds incubated at 20 ^°C were consistently higher than at 30 ^°C. Germination percentages of intact and dehusked seeds were similar at 20 ^°C incubation. Germination of intact seeds under 34/11 ^°C (16/8 h) was higher than at 30 ^°C. In contrast, germination of dehusked seeds under 34/11 ^°C (16/8 h) was lower than at 30°C. Inhibitory effects of removing husk on germination were observed from 15 to 70 DAA. Almost all of the intact seeds at 45 DAA germinated when imbibed 35, 30 and 25 ^°C, and germination percentages decreased with decreasing incubation temperatures from 20 to 15 ^°C. By contrast, germination percentage of dehusked seeds was highest in 20 ^°C incubation temperature; further increase or decrease of incubation temperature reduced germination percentage. The optimum germination temperature of intact and dehusked seeds of japonica rice differs during seed formation, such that the extent of germination inhibition by husk removal depends on the incubation temperature.     

水稻黑条矮缩病抗性QTL定位

发掘水稻黑条矮缩病的抗性基因有助于抗病品种的选育,减少黑条矮缩病对水稻生产的危害。本研究构建了包含222个家系的L5494/IR36重组自交系群体。对该群体进行黑条矮缩病的田间诱发鉴定,抗性亲本IR36发病率为28.70%,感病亲本L5494发病率为84.26%,群体发病率范围为11.21%～89.81%。利用134对分子标记构建覆盖12条染色体的遗传连锁图谱,总遗传距离为1475.97 cM,平均标记间距为11.1 cM。利用QTL IciMapping 4.0对抗黑条矮缩病QTL进行分析,共检测到4个QTL,其中第1、第2、第9染色体上QTL的表型贡献率分别为12.64%、16.00%和8.43%,抗病等位基因来自抗病亲本IR36;第6染色体上QTL的表型贡献率为10.82%,抗病等位基因来自感病亲本L5494。在此基础上,利用93-11为供体、日本晴为背景的近等基因系材料,在qRBSDV-1定位区间内检测到来自93-11的抗性QTL。本研究结果为水稻黑条矮缩病抗性基因定位及分子标记辅助选择育种提供借鉴。     

Effects of allelic variations in starch synthesis-related genes on grain quality traits of Korean nonglutinous rice varieties under different temperature conditions

Influences of allelic variations in starch synthesis-related genes (SSRGs) on rice grain quality were examined. A total of 187 nonglutinous Korean rice varieties, consisting of 170 Japonica and 17 Tongil-type varieties, were grown in the field and in two greenhouse conditions. The percentages of head rice and chalky grains, amylose content, alkali digestion value, and rapid visco-analysis characteristics were evaluated in the three different environments. Among the 10 previously reported SSRG markers used in this study, seven were polymorphic, and four of those showed subspecies-specific allele distributions. Six out of the seven polymorphic SSRG markers were significantly associated with at least one grain quality trait (R² > 0.1) across the three different environments. However, the association level and significance were markedly lower when the analysis was repeated using only the 170 Japonica varieties. Similarly, the significant associations between SSRG allelic variations and changes in grain quality traits under increased temperature were largely attributable to the biased allele frequency between the two subpopulations. Our results suggest that within Korean Japonica varieties, these 10 major SSRG loci have been highly fixed during breeding history and variations in grain quality traits might be influenced by other genetic factors.     

Identification of QTLs controlling low‐temperature tolerance during the germination stage in cucumber (Cucumis sativus L.)

Poor seed germination of cucumber at suboptimal temperatures is a great concern for growers wishing to take advantage of the early market. The development of low‐temperature‐tolerant varieties would be aided by understanding the inheritance of the trait and mapping its locus. In this study, a set of 140 recombinant inbred lines (RILs) derived from a cross between 65G (low‐temperature‐tolerant) and 02245 (low‐temperature‐sensitive) was used to identify the QTLs linked with low‐temperature tolerance. A linkage map was developed using 135 simple sequence repeat (SSR) markers and five insertion–deletion (Indel) markers, and three QTLs were identified, qLTG1.1, qLTG2.1 and qLTG4.1. qLTG1.1, the major one for germination rate, germination energy and germination index, explained more than 50% of the observed phenotypic variability. The major QTL for radicle length, qLTG4.1, explained 13.8% of the phenotypic variability. The results showed that qLTG1.1 and qLTG4.1 play an important role in low‐temperature tolerance during seed germination of cucumber and provide a basis for further fine mapping to determine the molecular mechanism for this trait.     

Translocation efficiencies and allocation of nitrogen,phosphorous and potassium in rice as affected by silicon fertilizer under high daytime temperature

High temperature has become a bottleneck limiting rice production in many rice‐growing districts. Silicon is considered as a beneficial element for rice development, being involved in mitigating adversity stress. In order to ascertain how high temperature and silicon affect nitrogen (N), phosphorous (P) and potassium (K) translocation efficiencies and allocation in rice plants, a field experiment with split plot design was conducted in two consecutive years. Silicon fertilizer treatments, including applying silicon fertilizer and without applying silicon fertilizer, were regarded as main plots. Temperature treatments, including high daytime temperature (HDT) and normal temperature (NT), were assigned as subplots. The results indicated that, as compared to NT, HDT reduced the translocation efficiencies of N, P and K in leaves and stems plus sheaths except for the K translocation efficiency in stems plus sheaths. Moreover, HDT decreased grain yield and the allocation rates of N, P and K in panicles at maturity. Under HDT, the application of silicon fertilizer obviously enhanced the N translocation efficiency of leaves and stems plus sheaths, and the K translocation efficiency of leaves. The application of silicon fertilizer increased grain yield and the allocation rates of N and K in panicles at maturity under HDT. Correlation analysis showed that rice grain yield was positively significantly correlated with N, P and K translocation efficiencies of leaves and their allocation rates in panicles at maturity. Conversely, grain yield was negatively related to the N and P allocation rates in leaves and stems plus sheaths at maturity. These results imply that HDT generated adverse effect on the translocation efficiency of nutrition in rice plants, which might be another damage induced by high temperature to the formation of rice grain yield. Additionally, silicon fertilizer could play a key role in positively regulating the N and K translocation efficiencies and allocation rates in rice under HDT.     

江淮优良食味高产中熟常规粳稻品种的特征

从103个中熟常规粳稻品种(品系)中,根据不同食味值和产量水平筛选出具有代表性的3种类型(味优高产、味优中产、味中高产),系统比较各类型品质和产量,以探究江淮优质食味与高产协同的品种特征。结果表明,味优高产类型品种加工品质与味中高产类型品种无显著差异。与味中高产类型相比,味优高产类型品种垩白粒率、垩白面积比、垩白度分别高82.06%、56.34%和93.28%,蛋白质含量、直链淀粉含量分别低14.21%、39.78%,胶稠度高8.73%,消减值和回复值分别低282.11%、37.88%。在产量方面,味优高产类型比味优中产类型高26.73%,其高产原因主要是具有较高的每穗粒数和结实率。与味优中产类型相比,味优高产类型单位面积穗数低22.26%,每穗粒数与结实率分别高42.12%、6.18%,成穗率高4.2%,抽穗期与成熟期叶面积指数分别高5.47%、16.94%,叶面积衰减率低7.25%,抽穗至成熟期干物质积累量和积累比例分别高24.07%和15.50%,着粒密度高40.33%。综上所述,味优高产类中熟常规粳稻的特征是,出糙率和整精米率都达国标1级;透明度由3级至5级不等;蛋白质含量在8%左右;直链淀粉含量在10%左右;胶稠度在75 mm以上; RVA谱消减值在-300 cP以下,回复值在600 cP以下。在产量方面其单位面积穗数310万穗hm–2左右,每穗粒数140个左右,抽穗后叶面积指数、干物质积累量与积累比例都能维持较高水平。     

Transgressive segregation due to linked QTLs for grain characteristics of rice

Summary It has been theoretically proposed that multiple linked quantitative trait loci (QTLs) play a role in the accumulation of hidden variation within and between populations. In this study, the genetic bases for grain characteristics were examined by comparing two accessions representing the two rice subspecies by QTL analysis. Grain dimensions are known to be quantitative traits and to be diagnostic between these two subspecies. To enhance the power to detect QTL with small effects, after transferring a segment of chromosome 6 from an Indica type into a Japonica type of rice by repeated backcrosses, the introgressed segment was dissected by making recombinant inbred lines (RILs) which were expected to have different sizes of the introgressed segment in the same genetic background. The resulting RILs showed distinct transgression of the grain characteristics examined. Multiple QTLs controlling each of the length and breadth of seeds were detected on the introgressed segment, and showed positive and negative additive effects as well as epistatic interactions. The present study confirmed that transgressive segregation resulted from a breakdown of linkage and that the detection of QTLs was highly dependent upon the genetic effects of the neighboring QTLs, indicating the need for caution in interpreting QTL effects.