利用RIL和CSSL群体检测水稻种子休眠性QTL

利用由梗稻品种Asominori与籼稻品种IR24的杂交组合衍生的重组自交F10。家系(Recombinant Inbred Lines,RIL)群体及其衍生的染色体片段置换系(Chromosome Segment Substitution Lines,CSSL)群体,进行了种子休眠性QTL的检测和遗传效应分析。其中CSSL群体有2个,即CSSLl(以Asominori为背景,置换片段来自IR24)和CSSL2(以IR24为背景,置换片段来自Asominori)。在RIL群体上共检测到3个种子休眠性QTL,分别位于第3、6和9染色体上;在CSSL1群体中检测到分布在第1、3和7染色体上的3个休眠性QTL;而在CSSl2群体上检测到的3个QTL则分别位于第1、2和7染色体上。同时在两套CSSL群体上,分别检测到位于第1、7染色体上位置相近且效应一致的休眠性QTL,分析表明其所在的Asominori片段含对种子休眠性的增效基因,相应的IB24段含有减效基因。     

Identification of Indica rice chromosome segments for the improvement of Japonica inbreds and hybrids

Exploitation of heterosis has brought significant advance in plant breeding and agricultural production, although its genetic basis is still poorly understood. In this study, a total of 66 chromosome segment substitution (CSS) lines, derived from a cross between japonica rice inbred line Asominori (as the recurrent parent) and indica rice inbred line IR24 (as the donor parent), were used to investigate the genetic basis of heterosis in indica × japonica inter-subspecific rice hybrids. Each CSS line was crossed with the background parent Asominori, and the heterosis of F(1) hybrids was estimated by comparing the F(1) performance with its two parental lines. Field experiments were carried out across six different environments to evaluate yield and yield-related traits in the 66 CSS lines and their 66 corresponding F(1) hybrids. Quantitative trait loci (QTL) analyses were conducted using a likelihood ratio test based on the stepwise regression. Thirty-six QTL were identified with significant effects in CSSL, 21 with significant effects in hybrids and 13 with significant effects in both. On the basis of average dominance degree, of all the 70 QTL affecting yield-related agronomic traits, 28.6% (20) showed an overdominance, 35.7% (25) a partial dominance and 30% (21) an additive effect, indicating that all effects contribute to trait variation in japonica-indica rice hybrids. Effects of these QTL were examined to identify Indica rice chromosome segments of interest for the improvement of japonica inbred lines and hybrids.     

利用粳稻染色体片段置换系群体检测水稻抗亚铁毒胁迫有关性状QTL

潜育性水稻田广泛分布于中国、斯里兰卡、印度、印度尼西亚、塞拉里昂、利比亚、尼日利亚、哥伦比亚和菲律宾等国,其中我国南方稻区就有近700万公顷低产潜育性水稻田。该类水稻田还原性强,矿质营养失调,尤以Fe^2 过量积累,对水稻生长发育产生不良的逆境胁迫作用。培育抗亚铁毒的水稻品种是简便、经济有效地提高稻谷产量的重要途径之一。该文利用由粳稻品种Asominori与籼稻品种IR24杂交衍生的Asominori染色体片段置换系(Chromosome Segment Substitution Lines,CSSLs)群体为材料,检测与抗亚铁毒胁迫有关性状QTL。共检测到与抗亚铁毒胁迫有关性状QTL14个,各QTL的LOD值为2．72～6．63。其中检测到与抗亚铁毒胁迫直接有关的性状叶片棕色斑点指数QTL3个,分别位于第3、9、11染色体C515～XNpb279、R2638～C1263和G1465～C950之间,对应的贡献率分别为16．45％、11．16％和28．02％;与其他已发表的定位结果比较发现,位于第三染色体C515～XNpb279间控制叶片棕色斑点指数的QTL与水稻功能图谱上控制叶绿素含量的QTL的位置一致;表明在亚铁毒胁迫条件下,水稻在其叶片表面出现棕色斑点,叶片衰老,产生一些叶绿素降解物或衍生物,以提高叶片细胞对亚铁等重金属毒害的耐受力。另外,在第11染色体G1465～C950之间检测到了控制叶片棕色斑点指数、茎干重和根干重QTL1个,为主效QTL。在第6染色体XNpb386～XNpb342之间检测到控制茎干重、株高、根长和根干重QTL1个,是否与水稻抗亚铁毒有关需要进一步研究。本研究旨在通过定位与抗亚铁毒有关的QTL,借助与之紧密连锁的分子标记有效地聚合这些QTL,培育出抗亚铁毒性强的水稻新种质材料。     

A single segment substitution line population for identifying traits relevant to drought tolerance and avoidance

A population of chromosome segment substitution lines was developed using KDML105 as the recurrent parent and one of DH212 (IR68586-F2-CA-143) or DH103 (IR68586-F2-CA-31) as the donor parent. The donor parents are part of a doubled haploid population from a cross between CT9993, an upland japonica accession, and IR62266, a lowland indica accession. Multiple QTL that are relevant to drought avoidance, drought tolerance and yield traits under drought stress were mapped in this doubled haploid population and the segments selected for the chromosome segment substitution lines were chosen to capture these QTL. The chromosome segment substitution line population was phenotyped under irrigated and mild drought stress conditions, which identified that many yield traits under drought stress had been introduced into the chromosome segment substitution lines.     

Marker-assisted selection and evaluation of the QTL for stigma exsertion under japonica rice genetic background

Stigma exsertion is one of the important traits which contribute to the efficient improvement of commercial seed production in hybrid rice. In order to understand the genetic factors involved in the stigma exsertion of an indica variety—IR24—a QTL analysis was conducted using the F₂ population between a japonica variety—Koshihikari—and a breeding line showing exserted stigma selected from the backcross population between IR24 as a donor and japonica varieties. As a result, a highly significant QTL (qES3), which had been predicted in the recombinant inbred population of IR24, was confirmed at the centromeric region on chromosome 3. qES3 increases about 20% of the frequency of the exserted stigmas at the IR24 allele and explains about 32% of the total phenotypic variance. A QTL near-isogenic line for qES3 increased the frequency of the exserted stigma by 36% compared to that of Koshihikari in a field evaluation, which suggests that qES3 is a promising QTL for the development of a maternal line for hybrid rice. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users. Maiko Miyata and Toshio Yamamoto contributed equally to this study.     

水稻耐亚铁毒QTLs的定位

亚铁毒是潜育性水稻土中限制水稻产量的主要因子。利用龙杂8503／IR64的F2和等价的F3群体,在营养液中培养来定位耐亚铁毒的QTLs。通过构建101SSR标记的遗传连锁图谱来确定耐亚铁毒QTLs的位置和特性。借助叶片棕色斑点指数、株高和最大根长3个性状,利用营养液在水稻苗期来评价F2单株、F3群体和亲本龙杂8503、IR64,共检测到叶片棕色斑点指数、株高和最大根长的QTLs20个,分布在水稻的10条染色体上,表明这些性状受多基因控制。控制叶片棕色斑点指数的QTLs分别定位在第1染色体的RM315-RM212、第2染色体的RM6-RM240和第4染色体的RM252-RM451之间。与前人的研究结果比较发现：1）位于第4染色体RM252-RM451之间的控制叶片棕色斑点指数的QTL与水稻功能图谱上控制叶绿素含量减少的QTL的位置一致。另一个位于第1染色体的RM315-RM212之间的控制叶片棕色斑点指数的QTL与水稻功能图谱上位于C178-R2635之间控制叶绿素含量的QTL连锁。2）位于第2染色体RM6-RM240之间的第3个控制叶片棕色斑点指数的QTL与位于RZ58-CD0686的控制钾吸收的QTL连锁。     

Quantitative trait loci (QTL) analysis for rice grain width and fine mapping of an identified QTL allele gw-5 in a recombination hotspot region on chromosome 5

Rice grain width and shape play a crucial role in determining grain quality and yield. The genetic basis of rice grain width was dissected into six additive quantitative trait loci (QTL) and 11 pairs of epistatic QTL using an F(7) recombinant inbred line (RIL) population derived from a single cross between Asominori (japonica) and IR24 (indica). QTL by environment interactions were evaluated in four environments. Chromosome segment substitution lines (CSSLs) harboring the six additive effect QTL were used to evaluate gene action across eight environments. A major, stable QTL, qGW-5, consistently decreased rice grain width in both the Asominori/IR24 RIL and CSSL populations with the genetic background Asominori. By investigating the distorted segregation of phenotypic values of rice grain width and genotypes of molecular markers in BC(4)F(2) and BC(4)F(3) populations, qGW-5 was dissected into a single recessive gene, gw-5, which controlled both grain width and length-width ratio. gw-5 was narrowed down to a 49.7-kb genomic region with high recombination frequencies on chromosome 5 using 6781 BC(4)F(2) individuals and 10 newly developed simple sequence repeat markers. Our results provide a basis for map-based cloning of the gw-5 gene and for marker-aided gene/QTL pyramiding in rice quality breeding.     

不同遗传背景及环境中水稻（Oryza sativa L.）穗长的QTLs和上位性分析

以粳稻Ａｚｕｃｅｎａ为父本与灿稻ＩＲ６４杂交发展的一双单倍体（ＤＨ） 本,与灿稻ＩＲ１５５２杂交发展的一重组自交系（ＲＩ）群体为材料,应用分子标记图说对２个群体在大田答舅栽２个环境下的穗长进行ＱＴＬｓ及上位性效应分析,ＤＨ群体中共检测６个穗长ＱＴＬｓ,位于第１、４长染色体上的３个ＱＴＬｓ,,在２个环境中稳定表达,未检测一闰性效应,加性效应为穗长遗传主效应,ＲＩ群体中,共检测到３个穗长ＱＴＬｓ及６对     

Mapping of QTLs associated with cytosolic glutamine synthetase and NADH-glutamate synthase in rice (Oryza sativa L.).

Ninety-eight backcross inbred lines (BC1F6) developed between Nipponbare, a japonica rice, and Kasalath, an indica rice were employed to detect putative quantitative trait loci (QTLs) associated with the contents of cytosolic glutamine synthetase (GS1; EC 6.3.1.2) and NADH-glutamate synthase (NADH-GOGAT; EC 1.4.1.14) in leaves. Immunoblotting analyses showed transgressive segregations toward lower or greater contents of these enzyme proteins in these backcross inbred lines. Seven chromosomal QTL regions for GS1 protein content and six for NADH-GOGAT protein content were detected. Some of these QTLs were located in QTL regions for various biochemical and physiological traits affected by nitrogen recycling. These findings suggested that the variation in GS1 and NADH-GOGAT protein contents in this population is related to the changes in the rate of nitrogen recycling from senescing organs to developing organs, leading to changes in these physiological traits. Furthermore, a structural gene for GS1 was mapped between two RFLP markers, C560 and C1408, on chromosome 2 and co-located in the QTL region for one-spikelet weight. A QTL region for NADH-GOGAT protein content was detected at the position mapped for the NADH-GOGAT structural gene on chromosome 1. A QTL region for soluble protein content in developing leaves was also detected in this region. Although fine mapping is required to identify individual genes in the future, QTL analysis could be a useful post-genomic tool to study the gene functions for regulation of nitrogen recycling in rice.     

Differentiation in wild-type allele of the sd1 locus concerning culm length between indica and japonica subspecies of Oryza sativa (rice)

A dwarfing gene (allele) sd1-d has been intensively utilized to develop short-culm indica varieties in southeast Asia up to now. Before the first sd1-d-carrying variety IR8 was released, rice researchers had recognized the general tendency that culm length is higher in indica varieties than in temperate-japonica ones. Inter-subspecific difference of the tall (wild-type) allele SD1 at the sd1 locus was examined on the common genetic background, using five isogenic lines developed by substituting sd1-d of the recurrent parent IR36 by SD1s of two indica varieties, two temperate-japonica varieties and one tropical-japonica variety. The two indica -donor isogenic lines had longer culms than the three japonica-donor isogenic lines consistently in two different environmental conditions. Moreover, nonsynonymous single-nucleotide polymorphism between the two subspecies was detected at two sites in Exon 1 and Exon 3 of the sd1 locus. It is demonstrated that the inter-subspecific differentiation of SD1 contributes height difference between indica and japonica. The indica-originating and japonica-originating alleles at the sd1 locus were designated as SD1-in(t) and SD1-ja(t), respectively.     

Marker-assisted introgression of quantitative trait loci associated with plant regeneration ability in anther culture of Rice (Oryza sativa L.)

A marker-assisted selection (MAS) breeding program was used to improve the plant regenerability of indica rice. A significant quantitative trait loci (QTL) that is associated with the capacity for green plant regeneration in the anther culture of rice was mapped on chromosome 10 using recombinant inbred (RI) population from Milyang 23/Gihobyeo. The marker that was chosen to follow the QTL region was used in MAS. This marker co-segregated with the regeneration ability in F2 individuals that were derived from MGRI 079/IR 36. In order to clarify the relationship between this marker and plant regenerability, the backcross population was screened with a RFLP marker. The capacity of plant regeneration of the backcross population was clearly distinguished by the marker genotype. The development of near isogenic line (NILs) with high regenerability through MAS will save time, labor, and cost in indica rice breeding.     

水稻条纹叶枯病抗性位点的检测和效应分析

利用111个家系组成的热研2号(Oryza sativa subsp. japonica ‘Reyan2’)/ Mi lyang23(Oryza sativa subsp. indica ‘Mi lyang23’)重组自交系(recombinant inbred l ines, RIL)群体(F7), 采用重病区田间自然接种方法, 以病情指数作为条纹叶枯病的表型值, 鉴定了2个亲本及111个RIL家系对条纹叶枯病的抗性。使用QTL Cartographer 软件复合区间作图法, 对水稻(Oryza a sativa)条纹叶枯病抗性基因进行了QTL分析。结果检测到2个抗水稻条纹叶枯病的QTL, 分别位于第2和第11染色体上, 其中第11染色体上的QTL贡献率为19.58%, 表明这是一个主效的QTL, 该QTL及其附近的分子标记, 可以用于水稻条纹叶枯病抗性分子标记辅助育种。     

水稻萌发耐淹性的遗传分析

水稻(Oryza sativa)萌发耐淹性受到复杂的网络调控, 其分子机制不同于苗期耐淹性的相关机制, 萌发耐淹性的强弱影响着直播稻的成苗率。通过对256份水稻核心种质的萌发耐淹性评估, 发现粳稻和籼稻之间的萌发耐淹性差异并不显著, 都存在广泛的遗传变异。利用以粳稻R0380为供体亲本, 籼稻RP2334为轮回亲本的170个高代回交自交系构建含146个分子标记的连锁图谱, 以低氧胚芽鞘长度为性状指标, 通过复合区间作图法检测到影响萌发耐淹性的4个QTLs(quantitative trait loci), 分别定位于第2(2个)、3(1个)和8号(1个)染色体。贡献率最大的QTL为qGS2.2, 其值为17.34%, 增效等位基因来自轮回亲本籼稻RP2334; 其余3个QTLs的增效等位基因均来自供体亲本粳稻R0380, 贡献率分别为12.86%、9.37%和14.60%。     

Response of rice to Al stress and identification of quantitative trait Loci for Al tolerance

Rice (Oryza sativa L.) shows the highest tolerance to Al toxicity among small-grain cereal crops, however, the mechanisms and genetics responsible for its high Al tolerance are not yet well understood. We investigated the response of rice to Al stress using the japonica variety Koshihikari in comparison to the indica variety Kasalath. Koshihikari showed higher tolerance at various Al concentrations than Kasalath. The Al content in root apexes was less in Koshihikari than in Kasalath, suggesting that exclusion mechanisms rather than internal detoxification are acting in Koshihikari. Al-induced secretion of citrate was observed in both Koshihikari and Kasalath, however, it is unlikely to be the mechanism for Al tolerance because there was no significant difference in the amount of citrate secreted between Koshihikari and Kasalath. Quantitative trait loci (QTLs) for Al tolerance were mapped in a population of 183 backcross inbred lines (BILs) derived from a cross of Koshihikari and Kasalath. Three putative QTLs controlling Al tolerance were detected on chromosomes 1, 2 and 6. Kasalath QTL alleles on chromosome 1 and 2 reduced Al tolerance but increased tolerance on chromosome 6. The three QTLs explained about 27% of the phenotypic variation in Al tolerance. The existence of QTLs for Al tolerance was confirmed in substitution lines for corresponding chromosomal segments.     

水稻外观品质的数量性状基因位点分析

利用由98个家系组成的Nipponbare(粳)／Kasalath(秒)∥Nipponbare回交重组自交系(backcross inbred lines,BILs)群体(BC1F9)及其分子连锁图谱,采用复合区间作图的方法,在2个不同年份对粒长、粒宽、粒形、垩白率、垩白大小、垩白度和透明度等7个稻米外观品质性状的数量性状基因位点(Quantiative trait loci,QTL)进行了定位分析。共定位到33个四QTLs,单个性状QTL数目在4-7个之间,以垩白率最多,为7个;粒长和垩白大小次之,为5个;其他性状均为4个,表明该组合外观品质是由多基因控制的数量性状。单个QTL对性状变异解释率粒长为6．2％-15．2％,粒宽为8．3％-32．5％,长宽比为6．8％-19．8％,垩白率为6．4％-28．5％,垩白大小为6．1％-16．9％,垩白度为9．3％-17．2％,透明度为5．6％-25．2％．QTL在染色体上成集中分布的特点,第3染色体C1488-C563、第5染色体R830-R3166和R1436-R2289、第6染色体R2147-R2171均有3个以上的QTLs分布。比较2年的检测结果表明,外观品质性状的QTL定位都受环境影响,但不同性状受影响的程度差异很大。粒长和粒形的QTL定位受环境影响很小,垩白率、垩白大小和垩白度的QTL定位受环境影响很大。     

QTL-based analysis of leaf senescence in an indica/japonica hybrid in rice (Oryza sativa L.)

In order to identify quantitative trait loci (QTLs) for leaf senescence and related traits in rice (Oryza sativa L.), we developed two backcross populations, indica/japonica// japonica and indica/japonica//indica, using IR36 as the indica parent and Nekken-2 as the japonica parent. The QTLs were mapped using a set of simple sequence-repeat markers (SSRs) in the BC₁F₁ population. Senescence was characterized in these plants by measuring the leaf chlorophyll content 25 days after flowering (DAF), the reduction in chlorophyll content (the difference between the chlorophyll content at flowering and at 25 DAF), and the number of late-discoloring leaves per panicle at 25 DAF in five plants from each BC₁F₂ line. These plants were moved into a temperature-controlled growth cabinet at the time of flowering and allowed to mature under identical conditions. Eleven QTLs were detected in the two populations. The major of QTLs for senescence were found on the short arm of chromosome 6 and on the long arm of chromosome 9. Of these, one QTL on chromosome 6 and two on chromosome 9 were verified by confirming the effects of the genotypes on the phenotypes of the BC₁F₃ lines. The japonica parent was found to contribute to late senescence at all but one QTL. Based on a comparison of the effects of heterozygotes and homozygotes on the phenotypic values of each QTL genotype, we concluded that the differential senescence observed in the indica-japonica hybrid was not due to over-dominance; rather, it was the result of partial-dominance genes that were donated from either of the parents.     

Mapping QTLs for root morphology of a rice population adapted to rainfed lowland conditions

In the rainfed lowlands, rice ( Oryza sativa L.) develops roots under anaerobic soil conditions with ponded water, prior to exposure to water stress and aerobic soil conditions that arise later in the season. Constitutive root system development in anaerobic soil conditions has been reported to have a positive effect on subsequent expression of adaptive root traits and water extraction during progressive water stress in aerobic soil conditions. We examined quantitative trait loci (QTLs) for constitutive root morphology traits using a mapping population derived from a cross between two rice lines which were well-adapted to rainfed lowland conditions. The effects of phenotyping environment and genetic background on QTLs identification were examined by comparing the experimental data with published results from four other populations. One hundred and eighty-four recombinant inbred lines (RILs) from a lowland indica cross (IR58821/IR52561) were grown under anaerobic conditions in two experiments. Seven traits, categorized into three groups (shoot biomass, deep root morphology, root thickness) were measured during the tillering stage. Though parental lines showed consistent differences in shoot biomass and root morphology traits across the two seasons, genotype-by-environment interaction (GxE) and QTL-by-environment interaction were significant among the progeny. Two, twelve, and eight QTLs for shoot biomass, deep root morphology, and root thickness, respectively, were identified, with LOD scores ranging from 2.0 to 12.8. Phenotypic variation explained by a single QTL ranged from 6% to 30%. Only two QTLs for deep root morphology, in RG256-RG151 in chromosome 2 and in PC75M3-PC11M4 in chromosome 4, were identified in both experiments. Comparison of positions of QTLs across five mapping populations (the current population plus populations from four other studies) revealed that these two QTLs for deep root morphology were only identified in populations that were phenotyped under anaerobic conditions. Fourteen and nine chromosome regions overlapped across different populations as putative QTLs for deep root morphology and root thickness, respectively. PC41M2-PC173M5 in chromosome 2 was identified as an interval that had QTLs for deep root morphology in four mapping populations. The PC75M3-PC11M4 interval in chromosome 4 was identified as a QTL for root thickness in three mapping populations with phenotypic variation explained by a single QTL consistently as large as 20-30%. Three QTLs for deep root morphology were found only in japonica/indica populations but not in IR58821/IR52561. The results identifying chromosome regions that had putative QTLs for deep root morphology and root thickness over different mapping populations indicate potential for marker-assisted selection for these traits.     

与水稻籼粳分化有关的穗颈维管束性状基因的分子标记定位

水稻穗颈维管束数及其与穗一次枝梗数之比（V／R）是与籼粳分化有关的重要性状,采用籼粳交（圭630／02428）杂种F1花药培养获得的DH群体,对水稻穗颈维管束数,穗一次枝梗数及V／R比进行了QTL分析,检测到3个控制穗颈维管束数的QTL;其中,效应最大的qVB-8的贡献率为31．1％,加性效应值为1．96％,增效等位基因来自灿稻亲本圭630,2个控制一次枝梗数的QTL效应较小,但分别与控制穗颈维管束数的2个QTL同位,检测到影响V／R比的3个QTL,其中,效应最大的qV/R-1的贡献率为25．3％,被定位于第1染色体上,与落粒性基因sh-2紧密连锁（亦或为一因多效）。此外,还检测到4对和2对分别控制穗颈维管束数和V／R比的互作QTL。结果分析表明,水稻穗颈维管束数与穗一次枝梗数受不同的多基因系统控制,但这2个多基因系统的某些位点在基因组中具有同位性;在第1染色体上,控制V／R比,且效应最大的qV/R-1所在的染色体区段在水稻籼粳分化过程中可能具有重要作用。     

Mapping of QTLs associated with cold tolerance during the vegetative stage in rice

Low-temperature stress is an important factor affecting the growth and development of rice (Oryza sativa L.) in temperate and high-elevation areas. Cold stress may cause various seedling injuries, delayed heading and yield reduction due to spikelet sterility. In this study, 181 microsatellite marker loci were used to identify quantitative trait loci (QTLs) associated with cold tolerance at the vegetative stage in 191 recombinant inbred lines (RILs) derived from a cross of a cold-tolerant temperate japonica cultivar (M-202) with a cold-sensitive indica cultivar (IR50). Different temperature regimes were applied in growth chambers on 191 RILs. The temperature regimes imposed in the growth chamber simulated cold-stress injuries at the seedling and late vegetative stages. In this study a major QTL was identified on chromosome 12, designated as qCTS12a, that was closely associated with cold-induced necrosis and wilting tolerance, and accounted for 41% of the phenotypic variation. A number of QTLs with smaller effects were also detected on eight rice chromosomes.