Genetic analysis and validation of quantitative trait loci associated with reproductive-growth traits and grain yield under drought stress in a doubled haploid line population of rice (Oryza sativa L.)

Drought is a major constraint for rice production and yield stability in rainfed ecosystems, especially when it occurs during the reproductive stage. Combined genetic and physiological analysis of reproductive-growth traits and their effects on yield and yield components under drought stress is important for dissecting the biological bases of drought resistance and for rice yield improvement in water-limited environments. A subset of a doubled haploid (DH) line population of CT9993-5-10-1-M/IR62266-42-6-2 was evaluated for variation in plant water status, phenology, reproductive-growth traits, yield and yield components under reproductive-stage drought stress and irrigated (non-stress) conditions in the field. Since this DH line population was previously used in extensive quantitative trait loci (QTLs) mapping of various drought resistance component traits, we aimed at identifying QTLs for specific reproductive-growth and yield traits and also to validate the consensus QTLs identified earlier in these DH lines using meta-analysis. DH lines showed significant variation for plant water status, reproductive-growth traits, yield and yield components under drought stress. Total dry matter, number of panicles per plant, harvest index, panicle harvest index, panicle fertility, pollen fertility, spikelet fertility and hundred grain weight had significant positive correlations with grain yield under drought stress. A total of 46 QTLs were identified for the various traits under stress and non-stress conditions with phenotypic effect ranging from 9.5 to 35.6% in this study. QTLs for panicle exsertion, peduncle length and pollen fertility, identified for the first time in this study, could be useful in marker-assisted breeding (MAB) for drought resistance in rice. A total of 97 QTLs linked to plant growth, phenology, reproductive-growth traits, yield and its components under non-stress and drought stress, identified in this study as well as from earlier published information, were subjected to meta-analysis. Meta-analysis identified 23 MQTLs linked to plant phenology and production traits under stress conditions. Among them, four MQTLs viz., 1.3 for plant height, 3.1 for days to flowering, 8.1 for days to flowering or delay in flowering and 9.1 for days to flowering are true QTLs. Consensus QTLs for reproductive-growth traits and grain yield under drought stress have been identified on chromosomes 1 and 9 using meta-QTL analysis in these DH lines. These MQTLs associated with reproductive-growth, grain yield and its component traits under drought stress could be useful targets for drought resistance improvement in rice through MAB and/or map-based positional analysis of candidate genes.     

Dissecting Genetic Basis of Deep Rooting in Dongxiang Wild Rice

Deep rooting is an important trait in rice drought resistance. Genetic resources of deep-rooting varieties are valuable in breeding of water-saving and drought-resistant rice. In the present study, 234 BC₂F₇ backcross introgression lines were derived from a cross of Dongye 80 (an accession of Dongxiang wild rice as the donor parent) and R974 (an indica restorer line as the recurrent parent). A genetic linkage map containing 1 977 bin markers was constructed by ddRADSeq for QTL analysis. Thirty-one QTLs for four root traits (the number of deep roots, the number of shallow roots, the total number of deep roots and the ratio of deep roots) were assessed on six rice chromosomes in two environments (2020 Shanghai and 2021 Hainan). Two of the QTLs, qDR5.1 and qTR5.2, were located on chromosome 5 in a 70-kb interval. They were detected in both environments. qDR5.1 explained 13.35% of the phenotypic variance in 2020 Shanghai and 12.01% of the phenotypic variance in 2021 Hainan. qTR5.2 accounted for 10.88% and 10.93% of the phenotypic variance, respectively. One QTL (qRDR2.2) for the ratio of deep roots was detected on chromosome 2 in a 210-kb interval and accounted for 6.72% of the phenotypic variance in 2020. The positive effects of these three QTLs were all from Dongxiang wild rice. Furthermore, nine and four putative candidate genes were identified in qRDR2.2 and qDR5.1/qTR5.2, respectively. These findings added to our knowledge of the genetic control of root traits in rice. In addition, this study will facilitate the future isolation of candidate genes of the deep-rooting trait and the utilization of Dongxiang wild rice in the improvement of rice drought resistance.     

QTL mapping rolling,stomatal conductance and dimension traits of excised leaves in the Bala × Azucena recombinant inbred population of rice

The identification of markers linked to genes contributing to drought resistance promises opportunities to breed high yielding rice varieties for drought prone areas. Several studies using different mapping populations have previously identified quantitative trait loci (QTLs) for traits theoretically related to drought resistance. A mapping population of 176 F₆ recombinant inbred lines (RILs) derived from two upland rice varieties with contrasting aboveground drought avoidance traits (Bala and Azucena) with a linkage map of 157 markers was used to map QTLs for aboveground leaf morphological and physiological traits related to drought avoidance. Plants were grown for 6 weeks under controlled environmental conditions with three replications. Leaves were excised and placed on a balance. The rate of leaf rolling and water loss was recorded, after which leaf area, dry weight and specific leaf area were characterized. A simple method of estimating time to stomatal closure was employed. A total of 13 QTLs were detected for leaf morphological traits, three for initial transpiration and four for the proportion of water loss required to reach a specific advanced state of leaf rolling. No QTLs were detected for time of stomatal closure or speed of leaf rolling, nor for either water loss or transpiration at stomatal closure despite clear parental differences and moderate heritabilities in most of these traits. The co-location of QTLs for traits measured here and for drought avoidance previously reported from field experiments on chromosome 1, 3 and 5 link the genetics of drought resistance to leaf dimensions and physiology. However, a physiological explanation for a QTL for drought avoidance on chromosome 7 remains elusive.     

水稻生理特性与抗旱性的相关分析及QTL定位

利用籼稻品种IR64和粳稻品种Azucena杂交产生的包含110个加倍单倍体株系的群体,在干旱胁迫和正常水分条件下,连续在2004年和2005年于抽穗期分别测定了叶片水势、相对含水量、叶绿素含量(SPAD值)、游离脯氨酸含量、气孔导度和蒸腾速率,并于成熟期取样,计算抗旱系数。与正常水分状况下相比,干旱胁迫条件下叶片的游离脯氨酸含量的增加达极显著水平,干旱胁迫条件下叶片的相对含水量、水势、叶绿素含量和气孔导度的降低均达显著或极显著水平。相关分析表明,在干旱胁迫条件下,叶片相对含水量、叶片水势与抗旱系数呈显著或极显著正相关。 利用175个RFLP标记构建的遗传连锁图谱分析了与抗旱性相关的叶片生理指标,共检测到与抗旱性相关的6个生理指标的7个加性QTL,31对上位性QTL,其中有2个主效QTL、9对上位性QTL存在环境互作效应。在两种水分条件下检测到的QTL结果有较大差异,说明干旱胁迫对控制与抗旱性相关的叶片生理性状基因的表达有显著的影响。在6个抗旱相关生理指标中,检测到的控制叶片气孔导度和水势的QTL较多,有3个加性QTL和8对上位性QTL控制气孔导度,有8对上位性QTL控制水势。     

Q-TARO: QTL Annotation Rice Online Database

Over the past two decades, genetic dissection of complex phenotypes of economic and biological interest has revealed the chromosomal locations of many quantitative trait loci (QTLs) in rice and their contributions to phenotypic variation. Mapping resolution has varied considerably among QTL studies owing to differences in population size and number of DNA markers used. Additionally, the same QTLs have often been reported with different locus designations. This situation has made it difficult to determine allelic relationships among QTLs and to compare their positions. To facilitate reliable comparisons of rice QTLs, we extracted QTL information from published research papers and constructed a database of 1,051 representative QTLs, which we classified into 21 trait categories. This database (QTL Annotation Rice Online database; Q-TARO, http://qtaro.abr.affrc.go.jp/) consists of two web interfaces. One interface is a table containing information on the mapping of each QTL and its genetic parameters. The other interface is a genome viewer for viewing genomic locations of the QTLs. Q-TARO clearly displays the co-localization of QTLs and distribution of QTL clusters on the rice genome.     

Genetic Diversity Analysis of Rice Germplasm in Tripura State of Northeast India Using Drought and Blast Linked Markers

We genotyped 74 rice germplasms including Tripura's local landraces, improved varieties, cultivars and breeding lines and other rice varieties using molecular markers for genetic diversity, drought QTLs, and blast resistance genes. The number of alleles per locus ranged from 2 to 5 with an average of 2.9. The polymorphic information content value per locus ranged from 0.059 (RM537) to 0.755 (RM252) with an average of 0.475. Cluster analysis based on 30 simple sequence repeat markers revealed 5 clusters and also indicated the presence of variability within the rice accessions. The drought QTL qDTY2.1 was found in 56.0% of germplasms and qDTY1.1 was detected in only 6.8% of the germplasms. Out of seven rice blast resistance genes screened, only two rice varieties, RCPL-1-82 and Buh Vubuk (Lubuk), were positive for four blast resistance genes while only Releng possessed two blast resistance genes. Among 74 rice germplasms, only three accessions, Releng, RCPL1-82 and Buh Vubuk (Lubuk), possessed both drought-related QTLs and blast resistance genes. Overall, the 74 indigenous rice genotypes showed low level of genetic diversity, which is in contrast to high level of genetic diversity among rice varieties in northeast India, where highlights the good farming practice, conservation of germplasms and the limitation of molecular markers employed in this study. The presence of both drought related QTLs and blast resistance genes in some of the germplasms can be useful in future breeding programmes.     

水稻胚芽鞘长度与抗旱性的关系及QTL定位

对由水稻品种珍汕97B和旱稻品种IRAT109构建的重组自交系195个株系的胚芽鞘长度及抗旱系数的研究表明,水分胁迫下水稻重组自交系群体的胚芽鞘长度与抗旱系数的相关系数为0.2206**。应用由213个SSR标记构建的遗传连锁图对控制胚芽鞘长度和抗旱系数的QTL进行了定位。检测到胚芽鞘长度和抗旱系数的主效QTL各为13个和5个,单个QTL对表型的贡献率为2.28%～22.65%;在第9染色体上两者的QTL出现在相同的分子标记区间（RM160-RM215）。检测到胚芽鞘长度和抗旱系数的互作位点分别为17对和3对,影响胚芽鞘长度的互作位点联合贡献率为5835%;影响抗旱系数的互作位点联合贡献率为11.93%。控制胚芽鞘长度和抗旱系数的QTL分别与其他研究中控制根系性状（深根干质量、根深、根长、根数等）的QTL位于相同的标记区间。     

Upland rice grown in soil-filled chambers and exposed to contrasting water-deficit regimes: II. Mapping quantitative trait loci for root morphology and distribution

Root morphological characteristics are known to be important in the drought resistance of some rice (Oryza sativa L.) varieties. The identification of quantitative trait loci (QTLs) associated with root morphology and other drought resistance-related traits should help breeders produce more drought resistant varieties. Stability in the expression of root growth QTL across rooting environments is critical for their use in breeding programs. A greenhouse experiment in which a mapping population of 140 recombinant inbred lines and the parental varieties Bala and Azucena were grown in glass-sided soil chambers and evaluated for root growth and water uptake was conducted. In each of 2 years, two treatments were used; an early water-deficit (WD0) in which seeds were sown into wet soil but received no more water, and a late water-deficit (WD49) in which the plants were watered for 49 days and then received no water for a week. The major differences between treatments and years in dry matter partitioning and root growth traits are reported elsewhere. Here, the identification of QTLs for root growth traits by composite interval mapping is described. At LOD>3.2, there were six QTLs for the weight of roots below 90 cm and maximum root length, 11 for root to shoot ratio, 12 for the number of roots past 100 cm, and 14 for root thickness. A total of 24 regions were identified as containing QTLs (these regions often contained several QTLs identified for different root traits). Some were revealed only in individual experiments and/or for individual traits, while others were common to different traits or experiments. Seven QTLs, on chromosomes 1, 2, 4, 7, 9 (two QTLs) and 11, where considered particularly noteworthy. The complex results are discussed in the context of previously reported QTLs for root growth in other populations, the interaction between QTL with the environment and the value of QTLs for breeding.     

Mapping quantitative trait loci associated with root growth in upland rice (Oryza sativa L.) exposed to soil water-deficit in fields with contrasting soil properties

A mapping population of 114 lines from Bala × Azucena was grown under drought stress at two field sites with contrasting soil physical properties. Drought was imposed between 35 and 65 days after sowing (DAS) and root density at 35 cm depth was measured 70 DAS. Leaf rolling, leaf drying and relative water content were recorded as indicators of drought avoidance. Root density correlated with indicators of drought avoidance. Two significant and two putative quantitative trait loci (QTLs) for root density and 28 QTLs for drought avoidance were identified. Most QTLs did not agree between sites. There was also reasonable agreement between leaf-drying QTLs and previously reported root-growth QTLs detected under controlled conditions (in contrast to a previous screen on soil with a higher penetration resistance). These data also reveal QTL × environment interaction, which will need to be understood more clearly if progress towards breeding for drought resistance via alterations of root morphology is to be achieved.     

利用重组自交系分析水稻稻曲病抗性位点及效应

利用157个家系组成的大关稻/IR28重组自交系群体,采用高效引发稻曲病人工接种方法,以病情指数作为稻曲病的表型值,鉴定了亲本及157个重组自交系群体对水稻稻曲病的抗性。利用QTL Cartographer 软件,对水稻稻曲病抗性基因进行检测分析。检测到qFsr1、qFsr4、qFsr10、qFsr11和qFsr12共 5个QTL位点,分别位于第1、4、10、11和12染色体上,贡献率为9.8%~22.5%。根据抗性位点加性效应方向,在qFsr1、qFsr10、qFsr11和qFsr12位点上,亲本IR28存在抗稻曲病的增效等位基因,大关稻具有减效等位基因;而qFsr4位点抗性效应来源于大关稻。     

Root plasticity under fluctuating soil moisture stress exhibited by backcross inbred line of a rice variety,Nipponbare carrying introgressed segments from KDML105 and detection of the associated QTLs

In rainfed lowland rice ecosystem, rice plants are often exposed to alternating recurrences of waterlogging and drought due to erratic rainfall. Such soil moisture fluctuation (SMF) which is completely different from simple or progressive drought could be stressful for plant growth, thereby causing reduction in yield. Root plasticity is one of the key traits that play important roles for plant adaptation under such conditions. This study aimed to evaluate root plasticity expression and its functional roles in dry matter production and yield under SMF using Nipponbare, KDML 105 and three backcross inbred lines (BILs) and to identify QTL(s) associated with root traits in response to SMF at two growth stages using Nipponbare/KDML105 F₂ plants. A BIL, G3-3 showed higher shoot dry matter production and yield than Nipponbare due to its greater ability to maintain stomatal conductance concomitant with greater root system development caused by promoted production of nodal and lateral roots under SMF. QTLs were identified for total nodal root length, total lateral root length, total root length, number of nodal roots, and branching index under SMF at vegetative and reproductive stages. The QTLs detected at vegetative and reproductive stages were different. We discuss here that relationship between root system of G3-3 and the detected QTLs. Therefore, G3-3 and the identified QTLs could be useful genetic materials in breeding program for improving the adaptation of rice plants in target rainfed lowland areas.     

Quantitative Trait Loci for Heading Date and Their Relationship with Genetic Control of Yield Traits in Rice (Oryza sativa)

Grain yield and heading date are key factors determining the commercial potential of a rice variety. Mapping of quantitative trait loci (QTLs) in rice has been advanced from primary mapping to gene cloning, and heading date and yield traits have always attracted the greatest attention. In this review, genomic distribution of QTLs for heading date detected in populations derived from intra-specific crosses of Asian cultivated rice (Oryza sativa) was summarized, and their relationship with the genetic control of yield traits was analyzed. The information could be useful in the identification of QTLs for heading date and yield traits that are promising for the improvement of rice varieties.     

水稻抽穗期QTL及其与产量性状遗传控制的关系

 产量和抽穗期是评价水稻品种应用价值的基本性状。水稻QTL分析已经历了从初定位到基因克隆的发展过程,并以抽穗期和产量性状最受重视。总结了亚洲栽培稻抽穗期QTL的基因组分布,分析了这些QTL与产量性状遗传控制的关系,为进一步筛选和鉴定具有较高育种应用潜力的水稻抽穗期QTL和产量性状QTL提供参考。     

小麦穗粒数QTL整合与元分析

小麦穗粒数是由多基因控制的复杂数量性状。为发掘控制小麦穗粒数(KNS)的真实主效数量性状位点(quantitative trait loci, QTL)，本研究利用生物信息学手段，借助小麦高密度分子标记遗传图谱，对来自不同遗传作图群体的控制小麦穗粒数的163个QTL位点进行图谱整合、映射和元分析。结果表明，目标性状QTL在小麦21条染色体上不均匀分布，在2B染色体上最多，在7D染色体上最少；建立控制小麦KNS的QTL一致性图谱，最终获得35个一致性QTL(meta quantitative trait loci, MQTL)位点及其紧密连锁的候选分子标记，置信区间最小可达到0.55 cM。     

水稻抗条纹叶枯病数量性状座位分析

为探明水稻品种窄叶青8号抗条纹叶枯病的数量性状座位,构建了窄叶青8号/武育粳3号F2群体的分子图谱,采用人工接种和田间自然接种两种鉴定方法,以病情指数比率为表型值,对每个F2单株衍生的F2∶3家系进行了抗条纹叶枯病鉴定。整个群体的病情指数比率均呈偏向于抗性亲本的连续性分布,表明条纹叶枯病抗性受数量性状基因的控制。进一步的QTL分析发现,两种鉴定方法所检测到的QTL完全不同,人工接种（强迫饲毒）方法仅检测到1个抗性基因位点qSTV7,其增强抗性的等位基因来源于窄叶青8号,而田间自然接种方法检测到2个抗性基因位点qSTV5和qSTV1,其增强抗性的等位基因分别来源于窄叶青8号和武育粳3号,暗示抗性亲本窄叶青8号可能携带耐病毒基因和抗灰飞虱基因,而感病亲本武育粳3号经遗传重组后,其抗性基因也得以表现。比较前人研究结果,发现检测到的QTL为新的抗条纹叶枯病基因位点,这些基因不同于抗条纹叶枯病主基因Stvb i,可为防止单一基因广泛使用造成的遗传脆弱性,提供新的抗性基因资源。     

Quantitative Trait Loci for Resistance to Stripe Disease in Rice （Oryza sativa）

In order to map the quantitative trait loci for rice stripe resistance, a molecular linkage map was constructed based on the F2 population derived from a cross between Zhaiyeqing 8 and Wuyujing 3. Reactions of the two parents, F1 individual and 129 F2:3 lines to rice stripe were investigated by both artificial inoculation at laboratory and natural infection in the field, and the ratios of disease rating index were scored. The distribution of the ratios of disease rating index in Zhaiyeqing 8/Wuyujing 3 F2:3 population ranged from 0 to 134.08 and from 6.25 to 133.6 under artificial inoculation at laboratory and natural infection in the field, respectively, and showed a marked bias towards resistant parent (Zhaiyeqing 8), indicating that the resistance to rice stripe was controlled by quantitative trait loci (QTL). QTL analysis showed that the QTLs detected by the two inoculation methods were completely different. Only one QTL, qSTV7, was detected under artificial inoculation, at which the Zhaiyeqing 8 allele increased the resistance to rice stripe, while two QTLs, qSTV5 and qSTV1, were detected under natural infection, in which resistant alleles came from Zhaiyeqing 8 and Wuyujing 3, respectively. These results showed that resistant parent Zhaiyeqing 8 carried the alleles associated with the resistance to rice stripe virus and the small brown planthopper, and susceptible parent Wuyujing 3 also carried the resistant allele to rice stripe virus. In comparison with the results previously reported, QTLs detected in the study were new resistant genes to rice stripe disease. This will provide a new resistant resource for avoiding genetic vulnerability for single utilization of the resistant gene Stvb-i.     

Root biology and genetic improvement for drought avoidance in rice

Rice root growth encompasses a remarkable genetic diversity in terms of growth patterns, architecture, and environmental adaptations. In order to harness this valuable diversity for improving rice response to drought, an understanding of key root traits and effective drought response mechanisms is necessary. A trait-based approach with precise understanding of the target environment, including temporal and spatial heterogeneity, is a possible path toward the use of roots and dehydration avoidance traits for improved drought resistance in rice. The ability to grow deep roots is currently the most accepted target trait for improving drought resistance, but genetic variation has been reported for a number of traits that may affect drought response. Here, we review variation in rice root response to drought from a physiological perspective in terms of morphology and function with respect to the different growth environments (upland and lowland) commonly used by farmers. Recent advances in linking physiology and breeding are also presented.     

Genetic Relationship and Structure Analysis of Root Growth Angle for Improvement of Drought Avoidance in Early and Mid-Early Maturing Rice Genotypes

Deeper rooting 1(Dro1) and Deeper rooting 2(Dro2) are the QTLs that contribute considerably to root growth angle assisting in deeper rooting of rice plant. In the present study, a set of 348 genotypes were shortlisted from rice germplasm based on root angle study. Screening results of the germplasm lines under drought stress identified 25 drought tolerant donor lines based on leaf rolling, leaf drying, spikelet fertility and single plant yield. A panel containing 101 genotypes was constituted based on screening results and genotyped using Dro1 and Dro2 markers. Structure software categorized the genotypes into four sub-populations with different fixation index values for root growth angle. The clustering analysis and principal coordinate analysis could differentiate the genotypes with or without deeper rooting trait. The dendrogram constructed based on the molecular screening for deep rooting QTLs showed clear distinction between the rainfed upland cultivars and irrigated genotypes. Eleven genotypes, namely Dular, Tepiboro, Surjamukhi, Bamawpyan, N22, Dinorado, Karni, Kusuma, Bowdel, Lalsankari and Laxmikajal, possessed both the QTLs, whereas 67 genotypes possessed only Dro1. The average angle of Dro positive genotypes ranged from 82.7° to 89.7°. These genotypes possessing the deeper rooting QTLs can be taken as donor lines to be used in marker-assisted breeding programs.     

栽培稻抗旱性研究的现状与策略

 水资源短缺正成为制约我国农业发展的重要因素。培育抗旱的栽培稻品种并实现水稻旱作，不但可在很大程度上节约水资源，而且有利于增产稳产，节约能源和减少环境污染。抗旱性包括逃旱性、避旱性、耐旱性和复原抗旱性。形态生理学的研究揭示出大量的与栽培稻抗旱性有关的形态特征和生理特性，如根系和叶片性状、生育期、渗透调节、脱落酸含量与栽培稻抗旱性密切相关，且已利用分子标记对上述性状进行了基因定位（QTL）研究。旱稻品种改良也已取得重大进展。在进行抗旱品种改良的基础上，通过引进相应的栽培技术，节水种植，实现水稻旱作，并达到稳产与增产的目的，是抗旱性研究的战略目标。在增产、稳产和优质的前提下，以培育耐旱性极强的水稻（或旱稻）为中心，建立有代表性的抗旱性研究基地, 进一步加强稻属抗旱基因资源的发掘和创新、抗旱生理学和遗传学的研究、利用现代生物技术实现不同物种间抗旱基因的转移、建立节水种植栽培技术新体系是目前抗旱性研究的主要内容。