QTL analysis and mapping of pi21, a recessive gene for field resistance to rice blast in Japanese upland rice

Field resistance is defined as the resistance that allows effective control of a parasite under natural field conditions and is durable when exposed to new races of that parasite. To identify the genes for field resistance to rice blast, quantitative trait loci (QTLs) conferring field resistance to rice blast in Japanese upland rice were detected and mapped using RFLP and SSR markers. QTL analysis was carried out in F₄ progeny lines from the cross between Nipponbare (moderately susceptible, lowland) and Owarihatamochi (resistant, upland). Two QTLs were detected on chromosome 4 and one QTL was detected on each of chromosomes 9 and 12. The phenotypic variation explained by each QTL ranged from 7.9 to 45.7% and the four QTLs explained 66.3% of the total phenotypic variation. Backcrossed progeny lines were developed to transfer the QTL with largest effect using the susceptible cultivar Aichiasahi as a recurrent parent. Among 82 F₃ lines derived from the backcross, resistance segregated in the expected ratio of resistant 1 : heterozygous 2 : susceptible 1. The average score for blast resistance measured in the field was 4.2 ± 0.67, 7.5 ± 0.51and 8.2 ± 0.66, for resistant, heterozygous and susceptible groups, respectively. The resistance gene, designated pi21, was mapped on chromosome 4 as a single recessive gene between RFLP marker loci G271 and G317 at a distance of 5.0 cM and 8.5 cM, respectively. The relationship to previously reported major genes and QTLs conferring resistance to blasts, and the significance of marker-assisted selection to improve field resistance, are discussed. Received: 8 June 2000 / Accepted: 24 November 2000     

QTL mapping in rice

In the past 10 years, interest in applying the tools of molecular genetics to the problem of increasing world rice production has resulted in the generation of two highly saturated, molecular linkage maps of rice, and the localization of numerous genes and quantitative trait loci (QTLs). Primary studies have identified QTLs associated with disease resistance, abiotic stress tolerance and yield potential of rice in a range of ecosystems. The ability to identify, manipulate and potentially clone individual genes involved in quantitatively inherited characters, combined with the demonstrated conservation of numerous linkage blocks among members of the grass family, emphasizes the contribution of map-based genetic analyses both to applied and to basic crop research.     

Phenotypic screening and molecular analysis of blast resistance in fragrant rice for marker assisted selection

Experiments were conducted to identify blast-resistant fragrant genotypes for the development of a durable blast-resistant rice variety during years 2012–2013. The results indicate that out of 140 test materials including 114 fragrant germplasms, 25 differential varieties (DVs) harbouring 23 blast-resistant genes, only 16 fragrant rice germplasms showed comparatively better performance against a virulent isolate of blast disease. The reaction pattern of single-spore isolate of Magnaporthe oryzae to differential varieties showed that Pish, Pi9, Pita-2 and Pita are the effective blast-resistant genes against the tested blast isolates in Bangladesh. The DNA markers profiles of selected 16 rice germplasms indicated that genotype Chinigura contained Pish, Pi9 and Pita genes; on the other hand, both BRRI dhan50 and Bawaibhog contained Pish and Pita genes in their genetic background. Genotypes Jirakatari, BR5, and Gopalbhog possessed Pish gene, while Uknimodhu, Deshikatari, Radhunipagol, Kalijira (3), Chinikanai each contained the Pita gene only. There are some materials that did not contain any target gene(s) in their genetic background, but proved resistant in pathogenicity tests. This information provided valuable genetic information for breeders to develop durable blast-resistant fragrant or aromatic rice varieties in Bangladesh.     

Targeted molecular mapping of a major wheat QTL for Fusarium head blight resistance using wheat ESTs and synteny with rice.

A major QTL for resistance to Fusarium head blight (FHB) in wheat, Qfhs.ndsu-3BS, has been identified and verified by several research groups. The objective of this study was to increase the marker density in this QTL region using STS (sequence-tagged site) markers developed from wheat expressed sequence tags (ESTs) near Qfhs.ndsu-3BS. Because wheat chromosome 3BS and rice chromosome 1S are syntenous, the sequences of P1-derived artificial chromosome (PAC) and (or) bacterial artificial chromosome (BAC) clones covering the sub-distal portion of rice chromosome 1S were used as queries for a BLASTn search to identify wheat ESTs most likely near Qfhs.ndsu-3BS. Sixty-eight out of 79 STS primer pairs designed from wheat ESTs amplified PCR products from the genomic DNA of Triticum aestivum 'Chinese Spring'. Twenty-eight STS markers were localized on chromosome 3BS by aneuploid analysis. Six out of the nine STS markers that could be mapped in the T. aestivum 'Sumai 3'/T. aestivum 'Stoa' population had higher R2 and LOD values for this QTL than the most significant marker reported previously. Therefore, leveraging genome sequence information available in rice for wheat genetics is an effective strategy to develop DNA markers for Qfhs.ndsu-3BS, and this strategy may have broad applications for targeted mapping of other traits in cereal crops.     

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.     

QTL mapping for resistance of maize to grey leaf spot

Grey leaf spot (GLS) is a global maize leaf disease that seriously endangers maize production. Discovering and utilizing genetic loci for GLS resistance would be useful for breeding new varieties with improved resistance. In this study, 233 F_2:3 families (produced from the susceptible inbred line 08‐641 × the resistant inbred line 446) were used for quantitative trait locus (QTL) mapping of resistance to GLS. Five GLS resistance QTLs were detected on chromosomes 1, 2, 3, 4, and 6, which explained 6.7%‐21.3% of the phenotypic variation. The QTLs, qRgls.CH‐4, qRgls.CH‐1, qRgls.CH‐2, and qRgls.CH‐6, were stably expressed in the four environments, and all loci for GLS resistance were derived from the resistant parent, 446. The additive effects of qRgls.CH‐4, qRgls.CH‐1, and qRgls.CH‐6 were significantly greater than their single dominant effects, which may be beneficial for GLS resistance breeding. The QTL qRgls.CH‐6, located in bins 6.02–6.05, did not overlap with any previously reported resistance QTL and thus was identified here for the first time. QTL analysis of PI (leaf performance index) detected three leaf function QTLs on chromosomes 4, 8, and 9 were related to GLS resistance and explained 4.8%‐6.2% of the phenotypic variation. Among them, qPI.CH‐4 was significantly stronger expressed in several environments; this allele associated with increased leaf function came from the resistant parent, 446, and its interval overlapped with that of qRgls.CH‐4. Furthermore, both qRgls.CH‐4 and qPI.CH‐4 were located in a hotspot area for GLS resistance in bins 4.05‐4.06, indicating that GLS resistance was significantly related to leaf performance and that GLS significantly reduced leaf photosynthetic performance.     

Significance testing for QTL mapping by marker difference regression

Marker difference regression, also known as joint mapping or simple marker regression, regresses differences of means for marker classes (’marker differences’) on their expected value, which is a function of the QTL effect and position. The genome is scanned by a likelihood ratio statistic, which conditionally on the QTL position follows a chi-squared distribution. The unconditional distribution is a chi-squared process. This paper proposes a quick method for controlling the genome-wise Type-I error rate. The method is shown by simulation to work satisfactorily for a number of settings.     

Fine mapping of qSB-11 LE , the QTL that confers partial resistance to rice sheath blight

Sheath blight (SB), caused by Rhizoctonia solani kühn, is one of the most serious global rice diseases. No major resistance genes to SB have been identified so far. All discovered loci are quantitative resistance to rice SB. The qSB-11^LE resistance quantitative trait locus (QTL) has been previously reported on chromosome 11 of Lemont (LE). In this study, we report the precise location of qSB-11 ^LE . We developed a near isogenic line, NIL-qSB11^TQ, by marker-assisted selection that contains susceptible allele(s) from Teqing (TQ) at the qSB-11 locus in the LE genetic background. NIL-qSB11^TQ shows higher susceptibility to SB than LE in both field and greenhouse tests, suggesting that this region of LE contains a QTL contributing to SB resistance. In order to eliminate the genetic background effects and increase the accuracy of phenotypic evaluation, a total of 112 chromosome segment substitution lines (CSSLs) with the substituted segment specific to the qSB-11 ^LE region were produced as the fine mapping population. The genetic backgrounds and morphological characteristics of these CSSLs are similar to those of the recurrent parent LE. The donor TQ chromosomal segments in these CSSL lines contiguously overlap to bridge the qSB-11 ^LE region. Through artificial inoculation, all CSSLs were evaluated for resistance to SB in the field in 2005. For the recombinant lines, their phenotypes were evaluated in the field for another 3 years and during the final year were also evaluated in a controlled greenhouse environment, showing a consistent phenotype in SB resistance across years and conditions. After comparing the genotypic profile of each CSSL with its phenotype, we are able to localize qSB-11 ^LE to the region defined by two cleaved-amplified polymorphic sequence markers, Z22-27C and Z23-33C covering 78.871 kb, based on the rice reference genome. Eleven putative genes were annotated within this region and three of them were considered the most likely candidates. The results of this study will greatly facilitate the cloning of the genes responsible for qSB-11 ^LE and marker-assisted breeding to incorporate qSB-11 ^LE into other rice cultivars.     

几个水稻株系的亲和性测定及微卫星标记分析

利用典型的籼稻南京11号、典型的粳稻秋光对16个水稻株系的亲和性进行了测定,测交结果表明,NLl73、NL15、NL61、NL67、NL82和NL279表现亲籼性,NL208表现亲粳性,NL78表现一定广亲和性,其余株系与籼、粳测验种杂交育性都不高。根据测交结果选择不同育性的株系用分布于12条染色体上的前人证明籼粳稻间有多态性的56个微卫星标记进行分析,在微卫星标记聚类分析基础上计算梗型性判别值Dj,并据此把NL173、NLl5、NL78、NL58判别为偏籼型,而NL208为偏粳型。综合亲和性测交结果和分子标记分析结果发现,在亲和性测交中表现亲籼的2个株系NL173、NL15分子标记判别为偏籼类型,杂交亲和性表现亲粳的株系NL208分子标记判断为偏粳类型,表现广亲和特性的株系NL78被分子标记判别为偏籼类型,与测验种杂交育性均不高的NL58分子标记判别为偏籼型。     

Genetic analysis of rice CMS-WA fertility restoration based on QTL mapping

 The inheritance of fertility restoration of rice cytoplasmic male sterility of the wild abortive type was studied by means of QTL mapping. The two segregating populations examined showed high frequencies of highly sterile and highly fertile progenies, but a low frequency of partially sterile and partially fertile progenies. The distributions suggested that fertility restoration was mainly controlled by major genes. Based on a linkage map constructed with 57 RFLP and 61 AFLP markers on a B₁F₁ population, composite interval mapping (CIM) revealed that the fertility was restored by the additive effects of two restorer loci located on chromosome 10. One QTL, tightly linked to RFLP marker C1361 in the middle of the long arm of chromosome 10, explained 71.5% of the phenotypic variance. The second QTL was located between RFLP markers R2309 and RG257 on the short arm and explained 27.3% of the phenotypic variance. Similar results were obtained using the simple interval mapping (SIM) methods. Recived: 8 January 1998/Accepted: 22 April 1998     

Inheritance of faba bean resistance to Broomrape,genetic diversity and QTL mapping analysis

Molecular Biology Reports - Six faba bean parents and their F1 and F2 generations were used in this investigation to study the genetic system controlling resistance of faba bean (Vicia faba L.) to...     

QTL mapping of resistance to Sporisorium reiliana in maize

We mapped and characterized quantitative trait loci (QTL) for resistance to Sporisorium reiliana. A population of 220 F₃ families produced from the cross of two European elite inbreds (D32, D145) was evaluated with two replications at a French location with high natural incidence of S. reiliana and at a Chinese location employing artificial inoculation. The 220 F₃ families were genotyped with 87 RFLP and seven SSR markers. Using composite interval mapping, we identified two different sets of 3 and 8 QTL for the French and the Chinese locations explaining 13% and 44% of respectively. Individual QTL explained up to 14% of σ^² _p. The 11 QTL mapped to eight maize chromosomes and displayed mostly additive or partial dominant gene action. Significant digenic epistatic interactions were detected for one pair of these QTL. Only a few QTL for S. reiliana were in common with QTL for resistance to Ustilago maydis and Puccinia sorghi, identified at a German location for the same population. Consequently, in our materials resistance to these three fungal pathogens of maize seems to be inherited independently. Received: 14. December 1998 / Accepted: 30 January 1999     

QTL mapping of resistance to leaf rust in Salix

Genetic mapping of quantitative trait loci (QTL) for resistance to Melampsora leaf rust was performed in two willow families: the progeny from a backcross between Salix viminalis and a hybrid S. viminalis × Salix schwerinii (population S₁), and the F₁ progeny of a cross between two S. viminalis (population S₃). Disease levels were scored in the field for three consecutive years. In the laboratory, five different rust strains were sprayed onto leaf disks and the following resistance components were scored: latent period, diameter and number of uredinia, and flecking. One major QTL and 14 smaller were identified in the S₁ host population. One rust strain, that represents a Melampsora form with limited incidence on S. viminalis, showed lower aggressiveness to the S₁ host population together with a different pattern in QTLs. In the S₃ host population, we detected 13 QTLs for rust resistance, of which two were located to the same genomic regions as those found for the S₁ population. We showed that the strongest QTL co-segregated with a gene homologous to a known Toll/interleukin receptor-nucleotide binding site-leucine-rich repeat resistance gene in poplar. The importance of the identified QTLs is discussed in relation to breeding for durable resistance.     

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.     

Intersection tests for single marker QTL analysis can be more powerful than two marker QTL analysis

Background

It has been reported in the quantitative trait locus (QTL) literature that when testing for QTL location and effect, the statistical power supporting methodologies based on two markers and their estimated genetic map is higher than for the genetic map independent methodologies known as single marker analyses. Close examination of these reports reveals that the two marker approaches are more powerful than single marker analyses only in certain cases.Simulation studies are a commonly used tool to determine the behavior of test statistics under known conditions. We conducted a simulation study to assess the general behavior of an intersection test and a two marker test under a variety of conditions. The study was designed to reveal whether two marker tests are always more powerful than intersection tests, or whether there are cases when an intersection test may outperform the two marker approach.We present a reanalysis of a data set from a QTL study of ovariole number in Drosophila melanogaster.

Results

Our simulation study results show that there are situations where the single marker intersection test equals or outperforms the two marker test. The intersection test and the two marker test identify overlapping regions in the reanalysis of the Drosophila melanogaster data. The region identified is consistent with a regression based interval mapping analysis.

Conclusion

We find that the intersection test is appropriate for analysis of QTL data. This approach has the advantage of simplicity and for certain situations supplies equivalent or more powerful results than a comparable two marker test.

     

Identification and fine mapping of qRBSDV-6 MH , a major QTL for resistance to rice black-streaked dwarf virus disease

Rice black streaked-dwarf virus (RBSDV) disease is recently expanding in southern China and poses a serious threat to rice crops. Few studies related to the genetics and breeding of RBSDV resistance have been reported. We have previously mapped a number of quantitative trait loci (QTLs) for RBSDV resistance by using a recombinant inbred line population of ‘Zhenshan 97’ (ZS97, susceptible)/‘Minghui 63’ (MH63, resistant) with natural infection data in two locations. In the present study, we confirmed the presence of a number of resistant QTLs on chromosomes 6, 7, and 9 from MH63 by using the same population in four different locations. We then focused on a major QTL, qRBSDV-6 ^MH , on chromosome 6 and introduced it into a highly susceptible japonica rice variety, ‘Huaidao 5’, using MH63 as the donor via marker-assisted selection, to generate seven backcross inbred lines (BILs). Natural infection and artificial inoculation-based tests revealed that all of the BILs had a significantly higher resistance to RBSDV than the recurrent parent. These results demonstrate that qRBSDV-6 ^MH is a stable major resistance QTL of high breeding value. We also constructed a set of chromosome segment substitution lines (CSSLs) specific to the qRBSDV-6 ^MH region and these used as fine mapping population. Combining the genotypes of CSSLs with the phenotypes from natural infection data in a highly RBSDV epidemic area during two different sowing seasons, we were able to precisely map qRBSDV-6 ^MH to the markers S18 and S23 at a physical distance of 627.6 kb on the Nipponbare reference genome.     

水稻抗稻瘟病基因的标记辅助选择及定位克隆

水稻抗稻瘟病基因-稻瘟病菌无毒基因相互作用体系是当今植物分子病理学和抗病育种学研究领域的模式体系之一,其中抗病基因的分子定位与克隆及其标记辅助选择已成为该体系的重要内容。本文就这方面的研究进展作一简要综述,以期为水稻抗病育种提供有益的信息。     

QTL analysis of resistance to the rice blast pathogen in barley (Hordeum vulgare)

Barley is compatible with the rice blast pathogen (Pyricularia oryzae Cav.). Fiftyfour barley cultivars of diverse geographic origin and pedigree were inoculated with three isolates of the rice blast pathogen. All barley genotypes showed blast disease symptoms when inoculated at the seedling stage with each of the three isolates. However, one genotype showed quantitative resistance to all three isolates and three genotypes showed quantitative resistance to one or two of the isolates. By inoculating a set of doubled-haploid lines derived from the cross ’Harrington’ (susceptible) and ’TR306’ (resistant) with isolate Ken 54–20, we mapped quantitative trait loci (QTLs) determining seedling stage blast resistance. At all QTLs, TR306 contributed the resistance alleles. The four QTLs, when considered jointly, explained 43.6% of the phenotypic variation in blast symptom expression. A comparison of the blast resistance QTLs with other disease resistance QTLs reported in this population revealed a region on chromosome 4 (4H) with multiple disease resistance loci. It will be useful to capitalize on the syntenic relationship of rice and barley and to integrate information on species-specific resistance genes with information on the reaction of the two species to the same pathogen. Received: 7 January 2000 / Accepted: 22 September 2000     

QTL mapping for nitrogen-use efficiency and nitrogen-deficiency tolerance traits in rice

Plant and Soil - The improvement of nitrogen-deficiency tolerance (NDT) and nitrogen-use efficiency (NUE) traits is an important objective of many rice breeding programs. A better understanding of...