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.     

基于染色体片段置换系群体检测水稻株型性状QTL

株型是由多个形态和生理性状集成的复合性状,它与水稻产量密切相关。挖掘优异株型等位基因或QTL,对水稻超高产育种具有重要意义。本研究利用籼稻昌恢121和粳稻Koshihikari构建的208个染色体片段置换系(chromosome segment substitution lines, CSSLs),在3个环境下,对控制株高、剑叶形态和分蘖数的QTL进行检测,共鉴定到35个株型性状QTL,分布于11条染色体上(除9号染色体以外),解释表型变异2.00%～22.86%。值得关注的是qPH-1-1、qFLW-6和qFLA-3均能在3个环境下被检测到,其中qFLW-6为1个新鉴定到的剑叶宽QTL。对qPH-1-1和qFLA-3位点进行鉴定,验证了这2个位点等位基因的加性效应和环境稳定性。本研究为株型性状QTL的进一步精细定位、克隆及分子辅助聚合育种奠定了基础。     

A major gene for low temperature germinability in rice (Oryza sativa L.)

Low temperature-induced retardation of seedling growth is a common problem in temperate rice-growing areas, at high altitudes of tropical and sub-tropical areas, and in areas with a cold irrigation water supply. Studies on low temperature germinability have revealed complex inheritance of the trait. The purpose of this study was to identify the gene(s) for low temperature germinability using Italica Livorno as a donor parent. The frequency distributions for the germination rate at 15 °C in the F₂ and BC₁F₁ populations showed continuous segregation, suggesting that low temperature germinability was under polygenic control. Since some lines in the BC₁F₁ population showed vigorous low temperature germinability similar to that of Italica Livorno, backcrosses until the BC₅F₁ generation was carried out using Hayamasari as the recurrent parent. Clear segregations of low temperature germinability were observed in the BC₅F₁ and BC₅F₂ populations. The distribution of low temperature germinability fitted a single-gene segregation, indicating that a single dominant gene with a large effect was transferred to Hayamasari. This gene is tentatively symbolized as Ltg(t). Low temperature germinability of near isogenic lines for Ltg(t) was similar to that of Italica Livorno. Ltg(t) should greatly contribute to the improvement and manipulation of low temperature germinability in rice breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

增加穗粒数的水稻染色体代换系Z747鉴定及相关性状QTL定位

增加穗粒数对水稻高产品种培育至关重要。其遗传基础复杂,由多基因控制。水稻染色体片段代换系可以将多基因控制的复杂性状分解,因而是理想的遗传研究材料。本研究通过高代回交和自交结合分子标记辅助选择方法,鉴定了一个以日本晴为受体、西恢18为供体亲本的、含有15个代换片段的增加穗粒数的水稻染色体片段代换系Z747,平均代换长度为4.49 Mb。与受体日本晴相比, Z747的每穗总粒数、一次枝梗数、二次枝梗数、穗长和粒长显著增加,粒宽显著变窄、结实率显著降低,但结实率仍为81%。进一步以日本晴和Z747杂交构建的次级F2群体鉴定出46个相关性状的QTL,分布于水稻1号、2号、3号、5号、6号、9号、11号和12号染色体上。其中qGPP12、qPH-3-1、qPH-3-2等12个QTL可能与已克隆的基因等位, qSPP9等34个可能是新鉴定的QTL。Z747的每穗总粒数由2个具有增加粒数效应的QTL (qSPP3和qSPP5)和1个具有减少粒数效应的QTL (qSPP9)控制。研究结果对主效QTL的精细定位和克隆、以及有利基因的单片段代换系培育有重要意义。     

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.     

Construction of chromosomal segment substitution lines and genetic dissection of introgressed segments associated with yield determination in the parents of a super‐hybrid rice

Heterosis is a phenomenon whereby hybrids of inbred lines produce favourable phenotypes that exceed those of their parents. Traits of interest are higher yield and stronger stress tolerance. The two‐line super‐hybrid rice ‘Liangyoupei9’ (LYP9) shows superiority to both its elite inbred line ‘93‐11’ and ‘Pei'ai64s’ (‘PA64s’) parents and conventional hybrids. However, the genetic basis of its hybrid vigour, especially yield determination, remains elusive. In the present study, a set of 156 chromosome segment substitution lines (CSSLs) carrying overlapping segments from ‘PA64s’ in a genetic background of ‘93‐11’ were constructed and planted in six environments. Three major agronomic traits, viz. panicle length (PL), heading date (HD) and plant height (PH), and five yield‐related traits, viz. grain weight per panicle (GWP), number of grains per panicle (GPP), 1000‐grain weight (TGW), seed set (SS) and number of panicles of per plant (PPP), were evaluated over 3 years. Quantitative trait loci (QTL) analysis was conducted using a likelihood ratio test based on stepwise regression. Forty‐six putative QTL distributed on 11 chromosomes were detected in more than one year. Remarkably, GWP of four CSSLs carrying positive yield QTL outperformed the recurrent parent ‘93‐11’ by more than 15%, in at least two environments. These results indicate that CSSLs are effective in identifying yield‐associated traits, and lines harbouring such QTL will be rich in resources for future molecular breeding programmes.     

Quantitative trait loci mapping for flag leaf traits in rice using a chromosome segment substitution line population

To study the genetic basis of rice flag leaf morphology, quantitative genetic analysis was conducted in a population of 37 chromosome segment substitution lines (CSSLs) of indica elite variety ‘Habataki’ in the background of japonica cultivar ‘Sasanishiki’ across three different environments. The CSSLs showed normal distribution with transgressive segregation, indicating that these four traits are controlled by polygenes. Moreover, analyses of variance showed that these traits were highly influenced by the growing environment, which are typical for polygenic quantitative traits. Seven quantitative trait loci (QTLs) on four chromosomes were detected in total: four for flag leaf width, one for flag leaf area and two for flag leaf angle. Two key QTLs, qFLW4 and qFLAG5 controlling flag leaf width and angle, respectively, were identified in all three environments. These QTLs could provide useful information for marker‐assisted selection in improving the performance of plant architecture with regard to leaf angle and area. Moreover, developed CSSLs with these QTLs information are also useful research materials to reveal the importance of leaf morphology in relation to grain yield.     

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.     

Induction of photoperiod sensitive genetic male steriles for use in hybrid rice seed production

The most notable photoperiod sensitive genetic male sterile in rice is the Nongken 58S line in China. Two sources of environmentally sensitive genetic male sterile mutants were found in California. The photoperiod-sensitive genetic male steriles (PGMS) have the potential for reducing hybrid rice breeding from the present 3-line system to a 2-line system. The long-range objectives of the present project were to induce an array of genetic male steriles in rice, and then to evaluate these at different day lengths in order to find steriles that become fertile in selected environments. Three genetic male steriles were selected from gamma irradiation of southern US cultivars. Inheritance of these mutants was studied through progeny tests in M₅ to M₇ generations and through fertility characterization of pollen and spikelets. Seed set on steriles under open pollination ranged from a low of 8.1% for Cypress 1819 to a high of 32.9% for Kaybonnet 1789. This revised version was published online in August 2006 with corrections to the Cover Date.     

优良水稻染色体片段代换系Z746的鉴定及重要农艺性状QTL定位及其验证

粒型、株高及穗部组成性状与产量形成密切相关,是水稻重要农艺性状,但遗传基础复杂。染色体片段代换系是用于复杂性状遗传研究的良好材料。本研究鉴定了一个以日本晴为受体、西恢18为供体亲本的水稻优良染色体片段代换系Z746。Z746携带来自西恢18的7个代换片段,平均代换长度为3.99 Mb,其株高、粒长和穗部性状均与受体存在显著差异。进一步通过日本晴与Z746杂交构建的次级F2群体共检测到36个相关QTL,分布于2号、3号、4号、6号和11号染色体。其中5个可能与已克隆基因等位,如qPH3-1等, 8个可被多次检出,表明这些是遗传稳定的主效QTL。Z746的粒长主要由4个QTL(qGL3、qGL4、qGL2、qGL6)控制,其中qGL3和qGL4对粒长变异的贡献率分别为60.28%和27.47%。株高由5个QTL控制,穗长由4个QTL控制,每穗粒数由2个QTL控制,千粒重由2个QTL控制。然后以MAS在F3共选出8个单片段代换系,并以此在F4进行了相关QTL验证,共有24个QTL可被8个单片段代换系(SSSL)检出,重复检出率为66.7%,表明这些QTL遗传稳定。本研究为目的 QTL的进一步...     

QTL epistatic analysis for yield components with single‐segment substitution lines in rice

Epistasis is an important genetic component in determining the phenotype of complex quantitative trait. In this article, 12 single‐locus heterozygotes and 66 double‐locus heterozygotes were developed and then were applied to assay QTL epistasis for four yield‐associated traits under two planting densities. Of 264 (66 × 4) tested interactions, 130 (49.2%) were significant at the p < .05 level. QTL with the same effect directions had higher probabilities of interactions. The negative epistasis at least included one positive effect QTL but the positive epistasis one negative QTL. The detected epistasis was sensitive to planting density. Epistasis also exhibited pleiotropic effects.     

通过分子标记辅助选择将耐储藏主效QTL qSS-9~(Kas)转入宁粳4号提高其种子贮藏能力

利用置换系SL36作为qSS-9位点上Kasalath等位基因的供体亲本,各方面农艺性状较好的宁粳4号作为轮回亲本,通过回交和自交,并使用4个与qSS-9紧密连锁的分子标记Y-10、Y-11、Y-14、Y-13进行基因型的检测筛选,对宁粳4号进行耐贮性的遗传改良,获得了既耐贮藏、大多农艺性状又接近于宁粳4号的较为稳定的优良新品系,克服了宁粳4号耐贮藏性差的弱点。获得的新品系种子在人工老化和自然老化条件下均表现出发芽率明显提高、丙二醛含量显著降低、TTC染色效果更明显,表明转入耐储藏主效QTL Qss-9~(Kas)的宁粳4号新品系耐贮性显著提高。     

Quantitative trait loci mapping and stability for low temperature germination ability of rice

A high rate of germination at low temperatures is necessary for economic yields to be maintained. In this paper, the genetic control of low temperature germination ability (LTG) was assessed by the measurement of germination rate (GR), germination rate index (GI) and mean germination time (MGT), and genetically mapped using a set of recombinant inbred lines, derived from a cross between the japonica cultivar 'Asominori' and the indica cultivar 'IR24'. Putative quantitative trait loci (QTL) were validated by testing in two related sets of chromosome segment substitution lines (CSSL). In this genetic background, LTG is under the control of a number of QTL, each of relatively small effect, and is spread over six chromosomes. The most stable of these QTL was for GR, mapping to a segment of chromosome 11 which also carries a QTL for GI. On chromosome 2, qGR-2 not only controlled GR, but also was associated with GI and MGT. Significant differences in LTG were detected between 'Asominori' and some CSSL harbouring qGR-2 or qGR-11 .     

两系杂交稻干物质积累与运转特性研究

根据湘西山区生态环境特点,分析比较了4个两系杂交中稻与2个三系杂交中稻的干物质积累与运转情况,结果表明：两系杂交组合前中期叶片、茎鞘养分积累较多,后期向穗部转移亦比较充分,供试组合剑叶展开期平均干重为39.7g/蔸,齐穗期茎鞘叶平均干重为41.0g/蔸,分别比三系杂交组合高14.41%和23.88%;茎鞘叶干物质向穗部的平均运转量为16.4 g/蔸,比三系杂交组合高18.84%;平均运转率为40.00%,与三系杂交组合平均值40.18%相近;对产量的贡献率为43.27%,比三系杂交组合平均值高近5个百分点。     

A proposed mechanism for loss of heterozygosity in rice hybrids

A phenomenon, loss of heterozygosity (LOH), was discovered in hybrid plants involving a selected plant (named AMR) of the Chinese rice cultivar ‘ZhongxinNo. 1’ as one parent. In these hybrids and some of their progenies, somatic variations were manifested by molecular genotypes and/or morphological phenotypes in vegetative parts of the same plant. Random amplified polymorphic DNA (RAPD) markers for the parents have been followed through the F₃ generation. RAPD markers were uniformly present or absent in all plants within some or all F₂ panicle rows derived from F₁ hybrids involving AMR. In contrast, RAPD markers segregated in the Mendelian manner for dominant markers in panicle rows derived from control hybrids. Certain F₂ panicle rows from F₁hybrids involving the special rice became fixed for all assayed RAPD markers. Genotype fixation was confirmed by molecular assays and field observations of the F₃ progenies. We propose a new biological mechanism, called ‘assortment mitosis,’ as being responsible for the observed phenomenon. The use of this mechanism in plant hybrids allows the development of uniform progenies as early as the F₂ generation. Therefore, the time required to obtain fixed non-parental type progenies for subsequent performance trials can be drastically shortened. Utilizing this mechanism in plant breeding represents a new approach and requires the modification of conventional plant breeding procedures. This revised version was published online in July 2006 with corrections to the Cover Date.     

Fine mapping of the genetic locus L1 conferring black pods using a chromosome segment substitution line population of soybean

The colour of plant organs is a useful trait in crop breeding. The pod colours of soybeans primarily include black, brown and tan types, which are controlled by two classical genetic loci, L1 and L2. Most wild soybeans have black pods, which reflect a possible role in adaptation to the natural environment. Here, an improved chromosome segment substitution line (CSSL) population SojaCSSLP3 was established to identify the L1 gene. The segment on the 19th chromosome represented by the SSR marker Satt313 was found to link with locus L1. The region was further delimited three times with increased SSR and InDel markers using a population derived from a heterozygous plant of CSSL124 from SojaCSSLP3. The L1 gene was finally located in a 184.43‐kb region between SSR_19p09 and Indel_19P7. Thirteen putative genes in this region were analysed with qRT‐PCR. The expression level of Glyma19 g27460, which is a member of the SANT superfamily with a MYB DNA‐binding domain, was significantly upregulated in black pods and was recognized to be the most likely candidate for the L1 gene.     

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.     

基于染色体片段置换系对水稻粒形及千粒重QTL检测与稳定性分析

粒形及千粒重是水稻产量的重要影响因素,通过挖掘这些性状的优异基因,对水稻超高产育种具有重要意义。本研究利用1套以籼稻恢复系昌恢121为背景亲本,粳稻越光为供体亲本构建的染色体片段代换系为材料,在3个环境下对水稻粒形及千粒重进行QTL检测及稳定性分析,共检测到59个QTL,分布于1号、2号、3号、4号、5号、6号、7号、10号、11号和12号染色体上,贡献率为0.77%～36.26%,其中发现10个QTL多效位点。值得关注的是qGW2-1、qGW2-2、qGW3-1、qGW3-2、qGL3和qGL12这6个QTL能在3个环境中重复检测到,其中qGW3-1为新鉴定的QTL位点。这些结果为进一步开展水稻粒形基因的精细定位、克隆和分子辅助育种奠定了一定的理论基础。     

利用东乡普通野生稻染色体片段置换系定位产量相关性状QTL

以东乡普通野生稻和日本晴为亲本构建的染色体片段置换系为研究材料, 2019年分别在北京、山东临沂和江西南昌对分蘖数、穗粒数和粒形等11个产量相关性状进行多环境鉴定,结合染色体片段置换系基因型数据定位水稻产量相关性状QTL。3个环境共检测到68个QTL,包括株高4个、穗长5个、分蘖数2个、一次枝梗数7个、一次枝梗粒数8个、二次枝梗数8个、二次枝梗粒数10个、每穗粒数6个、千粒重7个、粒长8个和粒宽3个; LOD值介于2.50～12.66之间,贡献率变幅为4.67%～27.79%,15个QTL的贡献率大于15%;24个QTL与已报道位点/基因位置重叠,44个QTL为新发现位点; 6个QTL在2个环境能被检测到, 1个QTL qTGW2能在3个环境检测到,且是还未报道的新位点。最后,利用BSA法验证了qPH7、qPBPP8-2和qGW10三个QTL的可靠性。本研究将为后续产量相关性状基因克隆以及进一步解析其遗传基础和分子调控机制奠定基础。