A large-effect QTL for rice grain yield under upland drought stress on chromosome 1

Drought is a major abiotic stress factor limiting rice production in rainfed areas. In this study we identified a large-effect quantitative trait locus (QTL) associated with grain yield under stress in five different populations on chromosome 1. The effect of this QTL was further confirmed and characterized in five backcross populations in a total of sixteen stress and non-stress trials during 2006 and 2008. In all the stress trials (eight in total) qDTY1.1 showed strong association with grain yield explaining on average 58% of the genetic variation in the trait. Homozygotes for the tolerant parent allele (Apo) yielded on average 27% more than the susceptible parent allele (IR64) homozygotes. Using an Apo/^3*IR64 population, the peak of this QTL (qDTY1.1) was mapped to an interval between RM486 and RM472 at 162.8?cM at a LOD score of 9.26. qDTY1.1 was strongly associated with plant height in all the environments; this was probably due to the presence of the sd1 locus in this genomic region. In a Vandana/^3*IR64 population segregating for sd1, a strong relation between plant height and yield under stress was observed. The observed relation between increased height and drought tolerance is likely due to tight linkage between qDTY1.1 and sd1 and not due to pleiotrophy of sd1. Thus there is a possibility of combining reduced plant height and drought tolerance in rice. The large and consistent effect of qDTY1.1 across several genetic backgrounds and environments makes it a potential strong candidate for use in molecular breeding of rice for drought tolerance.     

Quantitative trait locus analysis for rice panicle and grain characteristics

 The development of molecular genetic maps has accelerated the identification and mapping of genomic regions controlling quantitative characters, referred to as quantitative trait loci or QTLs. A molecular map derived from an F₂ population of a tropical japonica×indica cross (Labelle/Black Gora) consisted of 116 restriction fragment length polymorphism (RFLP) markers. Composite interval mapping was used to identify the QTLs controlling six panicle and grain characteristics. Two QTLs were identified for panicle size at LOD>3.0, with one on chromosome 3 accounting for 16% of the phenotypic variation. Four loci controlling spikelet fertility accounted for 23% of the phenotypic variation. Seven, four, three and two QTLs were detected for grain length, breadth, shape and weight, respectively, with the most prominent QTLs being on chromosomes 3, 4, and 7. Grain shape, measured as the ratio of length to breadth, was mostly controlled by loci on chromosomes 3 and 7 that coincided with the most important QTLs identified for length and breadth, respectively. A model including three loci accounted for 45% of the phenotypic variation for this trait. The identification of economically important QTLs will be useful in breeding for improved grain characteristics. Received: 18 July 1997 / Accepted: 9 December 1997     

Identification and characterization of large-effect quantitative trait loci for grain yield under lowland drought stress in rice using bulk-segregant analysis

An F_4:5 population of 490 recombinant inbred lines (RILs) from the cross Apo/^2*Swarna was used to detect quantitative trait loci (QTL) with large effects on grain yield under drought stress using bulk-segregant analysis (BSA). Swarna is an important rainfed lowland rice variety grown on millions of hectares in Asia, but is highly susceptible to drought and aerobic soil conditions. Apo is an aerobic-adapted variety with moderate tolerance to drought. Two rice microsatellite (RM) markers, RM324, and RM416, located on chromosomes 2 and 3, respectively, were shown via BSA to be strongly associated with yield under lowland drought stress. The effects of these QTL were tested in a total of eight hydrological environments over a period of 3 years. The QTL linked to RM416 (DTY _3.1 ) had a large effect on grain yield under severe lowland drought stress, explaining about 31% of genetic variance for the trait (P < 0.0001). It also explained considerable variance for yield under mild stress in lowland conditions and aerobic environments. To our knowledge this is the first reported QTL that has a large effect on yield in both lowland drought and aerobic environments. The QTL linked to RM324 (DTY _2.1 ) had a highly significant effect on grain yield in lowland drought stress (R ² = 13–16%) and in two aerobic trials. The effect of these QTL on grain yield was verified to be not mainly due to phenology differences. Effects of DTY _3.1 on yield under stress have been observed in several other rice mapping populations studied at IRRI. Results of this study indicate that BSA is an effective method of identifying QTL alleles with large effects on rice yield under severe drought stress. The Apo alleles for these large-effect QTL for grain yield under drought and aerobic conditions may be immediately exploited in marker-assisted-breeding to improve the drought tolerance of Swarna.     

Improving rice yield and quality by QTL pyramiding

To facilitate marker-assisted transfer of desirable genes for improvement of yield traits, we used a set of backcross recombinant inbred lines (BRIL) derived from two elite parental lines, ‘Zhenshan97’ and ‘93-11’, to resolve a quantitative trait loci (QTL) cluster for heading date and yield-related traits in rice. Four main-effect QTL (qHD6.1, qHD6.2, qHD7, and qHD8) and four epistatic QTL affecting heading date in the BRIL were detected in two experimental trials. The major QTL (qHD8) was confirmed in three heterogeneous inbred families (HIF) that segregated for this target region, and narrowed down to a 20-kb segment in a large HIF-derived population. qHD8 was found to interact with qHD7 and had a pleiotropic effect responsible for heading date and yield components. To test usability of the identified QTL in rice improvement, we further developed near-isogenic lines (NIL) containing one or more target genes by marker-assisted transfer of ‘93-11’ alleles at qHD8, qHD7, and qHD6.1, and the GS3 gene for grain size into ‘Zhenshan97’. The pyramid line NIL(qHD8 + GS3) had higher yield potential, longer grains, and a more suitable heading date than ‘Zhenshan97’. Comparison of the NIL showed existence of epistasis between alleles at different loci and background effect on qHD8, which are very important for pyramiding of desirable alleles at the target QTL. These results will be particularly useful not only to understand the genetic basis of yield-related traits but also to improve the efficiency of marker-assisted selection for favorable loci in rice breeding programs.     

Differential responses of sorghum genotypes to drought stress revealed by physio-chemical and transcriptional analysis

Molecular Biology Reports - Sorghum is an essential food crop for millions of people in the semi-arid regions of the world, where its production is severely limited by drought stress. Drought in...     

Fine mapping and candidate gene analysis of a major QTL for panicle structure in rice

Key message

A gene not only control tiller and plant height, but also regulate panicle structure by QTL dissection in rice.

Abstract

An ideal panicle structure is important for improvement of plant architecture and rice yield. In this study, using recombinant inbred lines (RILs) of PA64s and 93-11, we identified a quantitative trait locus (QTL), designated qPPB3 for primary panicle branch number. With a BC₃F₂ population derived from a backcross between a resequenced RIL carrying PA64s allele and 93-11, qPPB3 was fine mapped to a 34.6-kb genomic region. Gene prediction analysis identified four putative genes, among which Os03g0203200, a previously reported gene for plant height and tiller number, Dwarf 88 (D88)/Dwarf 14 (D14), had three nucleotide substitutions in 93-11 compared with PA64s. The T to G substitution resulted in one amino acid change from valine in 93-11 to glycine in PA64s. Real-time PCR analysis showed expression level of D88 was higher in 93-11 than PA64s. The expression of APO1 and IPA1 increased, while GN1a and DST decreased in 93-11 compared with PA64s. Therefore, D88/D14 is not only a key regulator for branching, but also affects panicle structure.     

Fine mapping of QTLs for rice grain yield under drought reveals sub-QTLs conferring a response to variable drought severities

Fine-mapping studies on four QTLs, qDTY(2.1), qDTY(2.2), qDTY(9.1) and qDTY(12.1), for grain yield (GY) under drought were conducted using four different backcross-derived populations screened in 16 experiments from 2006 to 2010. Composite and bayesian interval mapping analyses resolved the originally identified qDTY(2.1) region of 42.3 cM into a segment of 1.6 cM, the qDTY(2.2) region of 31.0 cM into a segment of 6.7 cM, the qDTY(9.1) region of 32.1 cM into two segments of 9.4 and 2.4 cM and the qDTY(12.1) region of 10.6 cM into two segments of 3.1 and 0.4 cM. Two of the four QTLs (qDTY(9.1) and qDTY(12.1)) having effects under varying degrees of stress severity showed the presence of more than one region within the original QTL. The study found the presence of a donor allele at RM262 within qDTY(2.1) and RM24334 within qDTY(9.1) showing a negative effect on GY under drought, indicating the necessity of precise fine mapping of QTL regions before using them in marker-assisted selection (MAS). However, the presence of sub-QTLs together in close vicinity to each other provides a unique opportunity to breeders to introgress such regions together as a unit into high-yielding drought-susceptible varieties through MAS.     

QTL analysis of floral traits of rice (Oryza Sativa L.) under well-watered and drought stress conditions

Three floral traits, spikelet number per panicle (SNP), percentage of single exserted stigma (PSES) and dual exserted stigma (PDES) of a RI population with 185 lines under water stress and non-stress conditions for two years were investigated in a drought tolerance screening facility. ANOVA results showed high significance between years, lines, and water stress treatments, together with interactions among them in pairs. High phenotypic correlation was found between PSES and PDES (r=0.5424***). Based on a linkage map of 203 SSR markers, when under well-watered condition, six QTLs (qSNP-3b, qSNP-4, qSNP-11 qSNP-2, qSNP-5 andqSNP-9) were detected for SNP. Half of them had significant Q × E interactions. Three QTLs (qPSES-1, qPSES-2, qPSES-5) were found to influence PSES, including one locus (qPSES-2) having Q × E interaction. And three QTLs (qPDES-2, qPDES-5 andqPDES-8) were also detected to influence PDES.qPDES-5 was found to have Q × E interaction. The contribution rate of a single QTL varied from 0.80% to 8.83% for additive effect, and 1.86% to 15.25% for Q × E interactions. Under drought stress, six QTLs (qSNP-3a, qSNP-4, qSNP-7a, qSNP-7b, qSNP-8 andqSNP-9) were associated with SNP, includingqSNP-3a andqSNP-4 with Q × E interaction. Three QTLs (qPSES-1, qPSES-10 andqPSES-12) were located on rice chromosome 1, 10 and 12 for PSES. Four QTLs (qPDES-1a, qPDES-1b, qPDES-4 andqPDES-9) were detected for PDES, includingqPDES-9 with Q × E interaction. The additive effect of single QTL can only explain 1.16% to 5.84% of total variance while Q × E interaction of four loci can explain 4.25% to 11.54% of total variance for each locus. There were one to nine pairs of epistatic QTLs influencing SNP and stigma exsertion. The contribution rates of additive and epistatic effects seemed to be in a low magnitude for most cases (0.76%≈9.92%) while a few QTLs or QTL pairs explained more than 10% of total variance. Some main effect QTL and epistasis were commonly detected among PSES and PDES, explaining the high positive correlation between them. Few QTLs were detected under both water stress and non-stress conditions, indicating that drought had severe impact on the genetic behaviors of both spikelet number and stigma exsertion.     

Meta-analysis of quantitative trait loci for grain yield and component traits under reproductive-stage drought stress in an upland rice population

A recombinant inbred population developed from a cross between high-yielding lowland rice (Oryza sativa L.) subspecies indica cv. IR64 and upland tropical rice subspecies japonica cv. Cabacu was used to identify quantitative trait loci (QTLs) for grain yield (GY) and component traits under reproductive-stage drought stress. One hundred fifty-four lines were grown in field trials in Indonesia under aerobic conditions by giving surface irrigation to field capacity every 4 days. Water stress was imposed for a period of 15 days during pre-flowering by withholding irrigation at 65 days after seeding. Leaf rolling was scored at the end of the stress period and eight agronomic traits were evaluated after recovery. The population was also evaluated for root pulling force, and a total of 201 single nucleotide polymorphism markers were used to construct the molecular genetic linkage map and QTL mapping. A QTL for GY under drought stress was identified in a region close to the sd1 locus on chromosome 1. QTL meta-analysis across diverse populations showed that this QTL was conserved across genetic backgrounds and co-localized with QTLs for leaf rolling and osmotic adjustment (OA). A QTL for percent seed set and grains per panicle under drought stress was identified on chromosome 8 in the same region as a QTL for OA previously identified in three different populations.     

Multivariate statistical analysis of grain yield and agronomic characters in Durum wheat

Summary Correlation, stepwise multiple regression and factor analyses were conducted on grain yield and a number of agronomic characters in the parental, F ₁ and F ₂ families originating from a 10 X 10 diallel cross in durum wheat. For the F ₁ diallel, the correlation analysis indicated that the number of spikes and kernels per plant and 1,000 kernel weight had the highest correlations with grain yield; for the F ₂ diallel, the number of spikes and kernels per meter, 1000 kernel weight and plant height showed most striking correlations with same.Stepwise multiple regression analysis indicated that, for the F ₁ diallel, number of kernels per plant, 1000 kernel weight and days to maturity were the most potent predictor variables for grain yield, accounting for 78% of its variability. For the F ₂ diallel, the number of kernels and number of spikes per meter, 1000 kernel weight and number of kernels per spike were the most potent predictors for grain yield, accounting for 91 % of its variability. Five common factors were extracted which explained 98.8% and 98.1% of the total variance in the F ₁ and F ₂ diallel, respectively. However, the importance of each of the five factors and the characters which loaded highly on each of them differed from generation to generation.Contribution No. 323 from Plant Science Department, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada.     

Mutations in the F-box gene LARGER PANICLE improve the panicle architecture and enhance the grain yield in rice

Panicle architecture is one of the most important agronomical traits that directly contribute to grain yield in rice (Oryza sativa L.). We report herein an in-depth characterization of two allelic larger panicle (lp) mutants that show significantly increased panicle size as well as improved plant architecture. Morphological analyses reveal that panicles of two mutants produced more inflorescence branches, especially the primary branches, and contained more grains. Moreover, mutant plants also display more lodging resistance than the wild type. The grain yield per plant in mutants is also increased, suggesting that mutant plants have useful potential for high grain yield in rice breeding. Map-based cloning reveals that LARGER PANICLE (LP) encodes a Kelch repeat-containing F-box protein. RNA in situ hybridization studies display that LP expression was enriched in the branch primordial region. Subcellular localization analyses demonstrate that LP is an endoplasmic reticulum (ER) localized protein, suggesting that LP might be involved in ER-associated protein degradation (ERAD). Using yeast two-hybrid assay and bimolecular fluorescence complementation analysis, we confirm that LP is an F-box protein and could interact with rice SKP1-like protein in an F-box domain-dependent manner. Quantitative real-time PCR results show that OsCKX2, which encodes cytokinin oxidase/dehydrogenase, is down-regulated evidently in mutants, implying that LP might be involved in modulating cytokinin level in plant tissues. These results suggest that LP plays an important role in regulating plant architecture, particularly in regulating panicle architecture, thereby representing promising targets for genetic improvement of grain production plants.     

Variation in monoculture and in mixture for grain yield and other characters in spring barley

Fourteen north-west European spring barley cultivars were grown alone or in binary mixtures sampled according to a partial diallel scheme. On the basis of the association between cultivars in mixture and monoculture, three groups of characters were distinguished. Group A characters, plant height, ear weight/tiller, grain yield/tiller, number of grains/tiller and 1000-grain weight, showed strong positive associations between performance in monoculture and mixture. Group B characters, number of tillers/plant and harvest index showed incomplete positive associations, while for group C characters, dry matter/plant, ear weight/plant, grain yield/plant and number of grains/plant, associations were weak or non-existent. Compound characters in group C showed less genetic variation in monoculture and lower general competitive effects in mixture than component characters in groups A or B. These results clearly indicate that while selection for grain yield and other characters on a per plant basis (group C characters) is confounded by intergenotypic competition, characters such as the yield components number of grains/tiller and 1000-grain weight (group A characters) are hardly affected in this range of cultivars. Selection for opposing group A characters may start in the F₂ generation, while any selection for group B and C characters should be delayed until later generations. The merits of indirect selection for yield using visual assessment of yield components are discussed. Separate analyses obtained by the inclusion of spring wheat cv. Timmo in monoculture and in the set of mixtures indicated that the use of spring wheat plants to minimise intergenotypic competition ranges from superfluous (group A characters) to useless (group C characters). A large degree of mixture advantage and the lack of complementary dominance and suppression between competitor and associate was attributed to the relatively low density of plants in the experiment which, though suitable for single plant selection, is not typical of normal seed rates for cropping.     

QTL analysis for rice grain length and fine mapping of an identified QTL with stable and major effects

Grain length in rice plays an important role in determining rice appearance, milling, cooking and eating quality. In this study, the genetic basis of grain length was dissected into six main-effect quantitative trait loci (QTLs) and twelve pairs of epistatic QTLs. The stability of these QTLs was evaluated in four environments using an F₇ recombinant inbred line (RIL) population derived from the cross between a Japonica variety, Asominori, and an Indica variety, IR24. Moreover, chromosome segment substitution lines (CSSLs) harboring each of the six main-effect QTLs were used to evaluate gene action of QTLs across eight environments. A major QTL denoted as qGL-3a, was found to express stably not only in the isogenic background of Asominori but also in the recombinant background of Asominori and IR24 under multiple environments. The IR24 allele at qGL-3a has a positive effect on grain length. Based on the test of advanced backcross progenies, qGL-3a was dissected as a single Mendelian factor, i.e., long rice grain was controlled by a recessive gene gl-3. High-resolution genetic and physical maps were further constructed for fine mapping gl-3 by using 11 simple sequence repeat (SSR) markers designed using sequence information from seven BAC/PAC clones and a BC₄F₂ population consisting of 2,068 individuals. Consequently, the gl-3 gene was narrowed down to a candidate genomic region of 87.5 kb long defined by SSR markers RMw357 and RMw353 on chromosome 3, which provides a basis for map-based cloning of this gene and for marker-aided QTL pyramiding in rice quality breeding.     

Proteomic analysis of rice leaves during drought stress and recovery

Three-week old plants of rice (Oryza sativa L. cv CT9993 and cv IR62266) developed gradual water stress over 23 days of transpiration without watering, during which period the mid-day leaf water potential declined to approximately -2.4 MPa, compared with approximately -1.0 MPa in well-watered controls. More than 1000 protein spots that were detected in leaf extracts by proteomic analysis showed reproducible abundance within replications. Of these proteins, 42 spots showed a significant change in abundance under stress, with 27 of them exhibiting a different response pattern in the two cultivars. However, only one protein (chloroplast Cu-Zn superoxide dismutase) changed significantly in opposite directions in the two cultivars in response to drought. The most common difference was for proteins to be up-regulated by drought in CT9993 and unaffected in IR62266; or down-regulated by drought in IR62266 and unaffected in CT9993. By 10 days after rewatering, all proteins had returned completely or largely to the abundance of the well-watered control. Mass spectrometry helped to identify 16 of the drought-responsive proteins, including an actin depolymerizing factor, which was one of three proteins detectable under stress in both cultivars but undetectable in well-watered plants or in plants 10 days after rewatering. The most abundant protein up-regulated by drought in CT9993 and IR62266 was identified only after cloning of the corresponding cDNA. It was found to be an S-like RNase homologue but it lacked the two active site histidines required for RNase activity. Four novel drought-responsive mechanisms were revealed by this work: up-regulation of S-like RNase homologue, actin depolymerizing factor and rubisco activase, and down-regulation of isoflavone reductase-like protein.     

Proteomic analysis of rice leaf sheath during drought stress

Drought is one of the most severe limitations on the productivity of rainfed lowland and upland rice. To investigate the initial response of rice to drought stress, changes in protein expression were analyzed using a proteomic approach. Two-week-old rice seedlings were exposed to drought conditions from 2 to 6 days, and proteins were extracted from leaf sheaths, separated by two-dimensional polyacrylamide gel electrophoresis and stained with Coomassie brilliant blue. After drought stress for 2 to 6 days, 10 proteins increased in abundance and the level of 2 proteins decreased. The functional categories of these proteins were identified as defense, energy, metabolism, cell structure, and signal transduction. In addition to drought stress, accumulations of protein were analyzed under several different stress conditions. The levels of an actin depolymerizing factor, a light harvesting complex chain II, a superoxidase dismutase and a salt-induced protein were changed by drought and osmotic stresses, but not cold or salt stresses, or abscisic acid treatment. The effect of drought stress on protein in the leaf sheaths of drought-tolerant rice cultivar was also analyzed. The light harvesting complex chain II and the actin depolymerizing factor were present at high levels in a drought-tolerant rice cultivar before stress application. With drought stress, actin depolymerizing factor was expressed in leaf blades, leaf sheaths, and roots. These results suggest that actin depolymerizing factor is one of the target proteins induced by drought stress.     

The use of phenotypic correlations and factor analysis in determining characters for grain yield selection in chickpea (Cicer arietinum L.)

To our knowledge, this is the first report on the use of factor analysis in determining characters for yield selection in chickpea (Cicer arietinum L.). The present investigation was undertaken to evaluate yield criteria in chickpea using phenotypic correlations and factor analysis. Factor 1 composed of biological yield, reaction to ascochyta blight (Ascochyta rabiei (Pass.) Labr.), plant height, grain yield and harvest index. Factor 2 consisted of branches and pods per plant. Factor 3 encompassed of only the grain weight. The total factors explained 92.9% of the total variance caused in the characters. The grain yield was positively and statistically significant correlated with biological yield, harvest index, plant height, branches and pods per plant, while it was negatively and statistically significant related with reaction to ascochyta blight and grain weight. Biological yield, harvest index, plant height and reaction to ascochyta blight instead of many selection criteria should previously be evaluated in selection to increase the grain yield in chickpea breeding programs. Pods per plant should be handed together with and branches per plant. Apart from the other selection criteria, the grain weight should solely be evaluated to select large grained genotypes.     

Comparative analysis of panicle proteomes of two upland rice varieties upon hyper-osmotic stress

Drought is a major environmental factor that limits the yield of rice dramatically. Upland rice is now regarded as a promising rice cultivar in water saving agriculture. Two varieties of upland rice Zhonghan 3 and IR29 were used to compare the physiological and proteomic responses to hyper-osmotic stress induced by 15% polyethyleneglycol (PEG) at the reproductive stage. Osmotic stress affected the growth development and caused the loss of production especially the grain yield. IR29 was more tolerant to PEG than Zhonghan 3 as shown by less yield loss under osmotic stress conditions. Comparative proteomic analysis of the panicle suggested that the up-regulation of glycolysis related proteins and defense proteins may contribute to the better osmotic tolerance in IR29.