水稻抗穗瘟基因的分子定位

 本试验以中156和谷梅2号为亲本建立F₈重组自交系群体,应用稻瘟病菌株92-183对群体的叶瘟和穗瘟抗性表现进行了分析。结果表明,叶瘟和穗瘟抗性遗传控制机制存在明显差异。应用DNA标记将一个兼抗叶瘟和穗瘟的基因定位于水稻第6染色体,其抗性等位基因来源于谷梅2号。而且,该基因在病区表现出较强的效应。     

Molecular mapping of the blast resistance genes Pi2-1 and Pi51(t) in the durably resistant rice 'Tianjingyeshengdao'

Tianjingyeshengdao' (TY) is a rice cultivar with durable resistance to populations of Magnaporthe oryzae (the causal agent of blast) in China. To understand the genetic basis of its resistance to blast, we developed a population of recombinant inbred lines from a cross between TY and the highly susceptible 'CO39' for gene mapping analysis. In total, 22 quantitative trait loci (QTLs) controlling rice blast resistance were identified on chromosomes 1, 3, 4, 5, 6, 9, 11, and 12 from the evaluation of four disease parameters in both greenhouse and blast nursery conditions. Among these QTLs, 19 were contributed by TY and three by CO39. Two QTL clusters on chromosome 6 and 12 were named Pi2-1 and Pi51(t), respectively. Pi2-1 was detected under both growth chamber and natural blast nursery conditions, and explained 31.24 to 59.73% of the phenotypic variation. Pi51(t) was only detected in the natural blast nursery and explained 3.67 to 10.37% of the phenotypic variation. Our results demonstrate that the durable resistance in TY is controlled by two major and seven minor genes. Identification of the markers linked to both Pi2-1 and Pi51(t) in this study should be useful for marker-aided selection in rice breeding programs as well as for molecular cloning of the identified resistance genes.     

小麦山农4143抗黑胚病遗传分析及抗性遗传位点检测

黑胚病是小麦生产的重要籽粒病害,麦根腐平脐蠕孢(Bipolaris sorokiniana)是黑胚病的主要致病菌.为分析小麦抗黑胚病遗传规律并检测抗性位点,本研究以抗黑胚病小麦品系山农4143与感病品系宛原白1号的F7代重组自交系(RIL)群体为材料,于2018～2019年在3个试验点种植,采用“孢子液喷洒、套袋保湿”...     

Molecular mapping of the new blast resistance genes Pi47 and Pi48 in the durably resistant local rice cultivar Xiangzi 3150

The indica rice cultivar Xiangzi 3150 (XZ3150) confers a high level of resistance to 95% of the isolates of Magnaporthe oryzae (the agent of rice blast disease) collected in Hunan Province, China. To identify the resistance (R) gene(s) controlling the high level of resistance in this cultivar, we developed 286 F(9) recombinant inbred lines (RILs) from a cross between XZ3150 and the highly susceptible cultivar CO39. Inoculation of the RILs and an F(2) population from a cross between the two cultivars with the avirulent isolate 193-1-1 in the growth chamber indicated the presence of two dominant R genes in XZ3150. A linkage map with 134 polymorphic simple sequence repeat and single feature polymorphism markers was constructed with the genotype data of the 286 RILs. Composite interval mapping (CIM) using the results of 193-1-1 inoculation showed that two major R genes, designated Pi47 and Pi48, were located between RM206 and RM224 on chromosome 11, and between RM5364 and RM7102 on chromosome 12, respectively. Interestingly, the CIM analysis of the four resistant components of the RILs to the field blast population revealed that Pi47 and Pi48 were also the major genetic factors responsible for the field resistance in XZ3150. The DNA markers linked to the new R genes identified in this study should be useful for further fine mapping, gene cloning, and marker-aided breeding of blast-resistant rice cultivars.     

小麦SSR和SNP标记在Taichung29×白芒麦重组自交系群体中的偏分离分析

为明确小麦SSR和SNP标记在Taichung29×白芒麦重组自交系群体中的偏分离现象及产生原因,提高遗传图谱构建的质量,以我国小麦农家品种白芒麦与感病品种Taichung29杂交构建的包含181个F8代重组自交系(recombinant inbred line,RIL)为试验材料,利用筛选得到的110个SSR标记和6 859个SNPs多态性标记进行偏分离分析。结果显示,共有1 069个标记位点表现出偏分离,占标记总数的15.3%。其中,683个标记偏向于白芒麦,占偏分离标记数的63.9%;386个标记偏向于Taichung29,占偏分离标记数的36.1%。偏分离位点在染色体上分布大多数是成簇出现,有76.3%的偏分离标记形成偏分离热点区域(segregation distortion region,SDR),共检测到了74个SDR,分布在15条染色体上,其中B染色体组包含有最多的SDR,偏分离方向为白芒麦。抗病亲本白芒麦和感病亲本Taichung29的基因型在RIL群体中分布比例为1.03∶1.00,接近1∶1的理论比例,标记在RIL群体整体分离比基本平衡,显示出偏分离主要是由遗传因素引起。     

Molecular characterization of a powdery mildew resistance gene in wheat cultivar suwon 92

ABSTRACT Powdery mildew, caused by Blumeria graminis f. sp tritici, is an important foliar disease of wheat worldwide. Pyramiding race-specific genes into a single cultivar and combining race-specific resistance genes with durable resistance genes are the preferred strategies to improve the durability of powdery mildew resistance. The objectives of this study were to characterize a powdery mildew resistance gene in Suwon 92 and identify gene-specific or tightly linked molecular markers for marker-assisted selection (MAS). A population of recombinant inbred lines (RILs) was derived by single seed descent from a cross between Suwon 92 and a susceptible cultivar, CI 13227. The RILs were screened for adult-plant infection type of powdery mildew and characterized with amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. The linked markers explained 41.3 to 69.2% of the phenotypic variances measured in 2 years. A morphological marker, hairy glume, was also associated with powdery mildew resistance in Suwon 92, and explained 43 to 51% of the phenotypic variance. The powdery mildew resistance gene in Suwon 92 was located on the short arm of chromosome 1A where Pm3 was located. Two gene-specific markers were developed based on the sequence of the cloned Pm3b gene. These two markers, which were mapped at the same locus in the peak region of the LOD score for the RIL population, explained most of the phenotypic variance for powdery mildew resistance in the RIL population. The powdery mildew resistance in Suwon 92 is most likely conditioned by the Pm3 locus. The gene markers developed herein can be directly used for MAS of some of the Pm3 alleles in breeding programs.     

Genetic analysis and molecular mapping of quantitative trait loci in common bean against Pythium ultimum

Pythium ultimum is a soil pathogen that can cause seed decay and damage to roots in common bean. In this study, the response of a set of 40 common bean genotypes to P. ultimum and inheritance of the resistance in the 92 F? recombinant inbred lines (RIL) developed from a cross between Xana and Cornell 49242 was investigated by using emergence rate and seedling vigor. Emergence of the 40 genotypes showed a significant association between white seed coat and response to this pathogen. Among these, 11 common bean genotypes, all with colored seeds, exhibited a high percentage of emergence and seedling vigor not significantly different (P > 0.05) to noninoculated plants. Response of the RIL population revealed both qualitative and quantitative modes of inheritance. A major gene (Py-1) controlling the emergence rate was mapped in the region of the gene P, a basic color gene involved in control of seed coat color, located on LG 7. Using the RIL subpopulation with colored seeds, a significant quantitative trait loci (QTL) associated with the emergence rate (ER3(XC)) and another with seedling vigor (SV6(XC)) were identified on the LG 3 and 6, respectively. QTL SV6(XC) was mapped in the region of the gene V, another gene involved the genetic control of color. QTLs associated with seed traits were mapped in the same relative position as regions involved in responses to P. ultimum suggesting the possible implication of avoidance mechanisms in the response to this pathogen.     

Analysis of quantitative trait Loci for resistance to southern leaf blight in juvenile maize

ABSTRACT A set of 192 maize recombinant inbred lines (RILs), derived from a cross between the inbred lines Mo17 and B73, were evaluated as 3-week-old seedlings in the greenhouse for resistance to southern leaf blight, caused by Cochliobolus heterostrophus race O. Six significant (LOD >3.1) quantitative trait loci (QTL) were identified for disease resistance, located on chromosomes 1, 2, 3, 6, 7, and 8. Results were compared with a previous study that had used the same RIL population and pathogen isolate, but had examined resistance in mature rather than juvenile plants. There was a very weak but significant correlation between the overall resistance phenotypes of the RILs scored as mature and juvenile plants. Two QTL were found in similar positions on chromosomes 1 and 3 at both growth stages. Other QTL were specific to one growth stage or the other. Twenty-three of these RILs, together with the parental lines, were inoculated in the greenhouse with four C. heterostrophus isolates. Results indicated that the quantitative resistance observed was largely isolate non-specific.     

Mechanism and molecular markers associated with rust resistance in a chickpea interspecific cross (Cicer arietinum × Cicer reticulatum)

A gene that controls resistance to chickpea rust (Uromyces ciceris-arietini) has been identified in a recombinant inbred line (RIL) population derived from an interspecific cross between Cicer arietinum (ILC72) × Cicer reticulatum (Cr5-10), susceptible and resistant to rust, respectively. Both parental lines and all RILs displayed a compatible interaction but differed in the level of infection measured as Disease Severity (DS) and Area Under the Disease Progress Curve (AUDPC). Histological studies of the seedlings of resistant parental Cr5-10 line revealed a reduction in spore germination, appressorium formation, number of haustoria per colony and colony size, with little host cell necrosis, fitting the definition of partial resistance. A Quantitative Trait Locus (QTL) explaining 31% of the total phenotypic variation for DS in seedlings and 81% of the AUDPC in adult plants in the field was located on linkage group 7 of the chickpea genetic map. The AUDPC displayed a bimodal distribution with high frequency of susceptible lines and both the AUDPC and markers showed the same distorted segregation. Consequently, it was hypothesised that a single dominant gene (proposed as Uca1/uca1) controlled resistance to rust in adult plants. This allowed us to locate the gene on the genetic linkage map. Two Sequence Tagged Microsatellite Sites (STMS) markers, TA18 and TA180 (3.9 cM apart) were identified that flank the resistance gene. These findings could be the starting point for a Marker-Assisted Selection (MAS) programme for rust resistance in chickpea.     

Identification and Molecular Mapping of a Gene Conferring Resistance to Pyrenophora tritici-repentis Race 3 in Tetraploid Wheat

ABSTRACT Race 3 of the fungus Pyrenophora tritici-repentis, causal agent of tan spot, induces differential symptoms in tetraploid and hexaploid wheat, causing necrosis and chlorosis, respectively. This study was conducted to examine the genetic control of resistance to necrosis induced by P. tritici-repentis race 3 and to map resistance genes identified in tetraploid wheat (Triticum turgidum). A mapping population of recombinant inbred lines (RILs) was developed from a cross between the resistant genotype T. tur-gidum no. 283 (PI 352519) and the susceptible durum cv. Coulter. Based on the reactions of the Langdon-T. dicoccoides (LDN[DIC]) disomic substitution lines, chromosomal location of the resistance genes was determined and further molecular mapping of the resistance genes for race 3 was conducted in 80 RILs of the cross T. turgidum no. 283/Coulter. Plants were inoculated at the two-leaf stage and disease reaction was assessed 8 days after inoculation based on lesion type. Disease reaction of the LDN(DIC) lines and molecular mapping on the T. turgidum no. 283/Coulter population indicated that the gene, designated tsn2, conditioning resistance to race 3 is located on the long arm of chromosome 3B. Genetic analysis of the F(2) generation and of the F(4:5) and F(6:7) families indicated that a single recessive gene controlled resistance to necrosis induced by race 3 in the cross studied.     

Identification and characterization of a new blast resistance gene located on rice chromosome 1 through linkage and differential analyses

ABSTRACT The Chinese native cv. Q14 expresses a high level of resistance to many isolates of Pyricularia grisea collected from Japan, Thailand, and China. Q14 was crossed to an indica-susceptible cultivar, Q61. To rapidly determine the chromosomal location of the major resistance gene present in the cultivar, a linkage analysis using microsatellite markers was performed in the F(2) population segregating 3R:1S (resistant/susceptible) through bulked-segregant analysis (BSA) in combination with recessiveclass analysis (RCA). A total of 189 microsatellite markers selected from each chromosome equally (with approximately 10 centimorgans) were tested with the BSA approach. Only two markers, RM151 and RM259, located on chromosome 1 showed positive and negative polymorphisms, respectively, for a resistance gene segregating in the population. To confirm the polymorphic markers, a total of 155 viable susceptible individuals were tested with the RCA approach. The markers RM151 and RM259 were found to link to the resistance gene with recombination frequencies of 11.9 +/- 2.8% and 9.7 +/- 8.0%, respectively. For further characterization of the resistance gene, 3 resistance genes mapped on chromosome 1, as well as 15 major resistance genes that might be employed in the breeding program, were selected for differential tests with 85 Chinese isolates. The resistance gene identified in this research conveys reactions distinct from those conditioned by the 18 resistance genes. This new resistance gene tentatively was designated Pi27(t).     

A gene in European wheat cultivars for resistance to an African isolate of Mycosphaerella graminicola

Genes for specific resistance to European and American isolates of Mycosphaerella graminicola , the causal agent of septoria tritici blotch (STB) of wheat, have been identified and mapped in various cultivars and breeding lines and are distributed throughout the genome. The location of a gene for resistance to an Ethiopian isolate, IPO88004, which is currently the most widespread resistance present in European wheat cultivars, is reported. The resistance was mapped in the Swiss cultivar Arina which, besides high partial resistance to STB, also has specific resistance to IPO323, controlled by Stb6 and to IPO88004. An F5 recombinant inbred population from a cross between Arina and the susceptible cultivar Forno was tested in whole seedling trials. Using multiple QTL mapping (MQM), a gene for resistance to M. graminicola isolate IPO88004 in cv. Arina was located to chromosome 6AS. The gene is named Stb15 . Seedling tests on a double haploid population of cvs Arina × Riband indicated that the UK wheat cv. Riband also has Stb15 or another gene for specific resistance to IPO88004 allelic or closely linked to Stb15 .     

Molecular marker mapping of leaf rust resistance gene lr46 and its association with stripe rust resistance gene yr29 in wheat

ABSTRACT Leaf and stripe rusts, caused by Puccinia triticina and P. striiformis, respectively, are globally important fungal diseases of wheat that cause significant annual yield losses. A gene that confers slow rusting resistance to leaf rust, designated as Lr46, has recently been located on wheat chromosome 1B. The objectives of our study were to establish the precise genomic location of gene Lr46 using molecular approaches and to determine if there was an association of this locus with adult plant resistance to stripe rust. A population of 146 F(5) and F(6) lines produced from the cross of susceptible 'Avocet S' with resistant 'Pavon 76' was developed and classified for leaf rust and stripe rust severity for three seasons. Using patterns of segregation for the two diseases, we estimated that at least two genes with additive effects conferred resistance to leaf rust and three to four genes conferred resistance to stripe rust. Bulked segregant analysis and linkage mapping using amplified fragment length polymorphisms with the 'Avocet' x 'Pavon 76' population, F(3) progeny lines of a single chromosome recombinant line population from the cross 'Lalbahadur' x 'Lalbahadur (Pavon 1B)', and the International Triticeae Mapping Initiative population established the genomic location of Lr46 at the distal end of the long arm of wheat chromosome 1B. A gene that is closely linked to Lr46 and confers moderate levels of adult plant resistance to stripe rust is identified and designated as Yr29.     

Identification of a Novel Fusarium Head Blight Resistance Quantitative Trait Locus on Chromosome 7A in Tetraploid Wheat

ABSTRACT Fusarium head blight (FHB) caused by Fusarium graminearum is one of the most destructive diseases of durum (Triticum turgidum sp. durum) and common wheat (T. aestivum). Promising sources of FHB resistance have been identified among common (hexaploid) wheats, but the same is not true for durum (tetraploid) wheats. A previous study indicated that chromosome 7A from T. turgidum sp. dicoccoides accession PI478742 contributed significant levels of resistance to FHB. The objectives of this research were to develop a genetic linkage map of chromosome 7A in a population of 118 recombinant inbred lines derived from a cross between the durum cv. Langdon (LDN) and a disomic LDN-T. turgidum sp. dicoccoides PI478742 chromosome 7A substitution line [LDN-DIC 7A(742)], and identify a putative FHB resistance quantitative trait locus (QTL) on chromosome 7A derived from LDN-DIC 7A(742). The population was evaluated for type II FHB resistance in three greenhouse environments. Interval regression analysis indicated that a single QTL designated Qfhs.fcu-7AL explained 19% of the phenotypic variation and spanned an interval of 39.6 cM. Comparisons between the genetic map and a previously constructed physical map of chromosome 7A indicated that Qfhs.fcu-7AL is located in the proximal region of the long arm. This is only the second FHB QTL to be identified in a tetraploid source, and it may be useful to combine it with the QTL Qfhs.ndsu-3AS in order to develop durum wheat germ plasm and cultivars with higher levels of FHB resistance.     

Identification of a quantitative trait locus for high-temperature adult-plant resistance against Puccinia striiformis f. sp. hordei in 'Bancroft' barley

Sustainable control of plant diseases can be achieved by developing cultivars with durable resistance. 'Bancroft' barley has durable high-temperature, adult-plant (HTAP) resistance to stripe rust caused by Puccinia striiformis f. sp. hordei. The objectives of this study were to determine the inheritance of the HTAP resistance in Bancroft, develop molecular markers for the HTAP resistance using the resistance gene analog polymorphism (RGAP) technique, map the HTAP resistance quantitative trait locus or loci (QTL) on barley chromosomes, and determine the usefulness of the RGAP markers in other barley cultivars for marker-assisted selection. The parents and F(4) recombinant inbred lines (RIL) and the parents and F(5) RIL were evaluated in 2004 and 2005 in one and three field sites, respectively, in Washington State. Infection type (IT) and disease severity (DS) were recorded three times at each location during each growing season. Area under the disease progress curve (AUDPC) was calculated for each parent and RIL based on the DS data. Genetic analyses of IT data of the parents, F(1), and F(2) tested in the adult-plant stage under controlled high-temperature cycle in the greenhouse and the parents, F(4), and F(5) RIL in the field indicated that one dominant gene controlled the HTAP resistance in Bancroft. Using 119 F(5:6) RIL and IT data, a linkage map on chromosome arm 3HL was constructed with eight RGAP markers and three simple sequence repeat (SSR) markers. Using the QTL analysis, a QTL for HTAP resistance was mapped with the DS and AUDPC data on the same chromosome location as with the IT data. The QTL explained >70% of the total phenotypic variation for the DS and AUDPC. The heritability of the HTAP resistance based on the AUDPC data was 76%. The two markers most close to the QTL peak detected polymorphisms in 84 and 88% of 25 barley genotypes that do not have the Bancroft HTAP resistance when used individually, and detected polymorphism in 100% of the genotypes when used in combination, indicating that the markers could be used in incorporating the HTAP resistance into these barley genotypes to improve the level and durability of resistance to stripe rust.     

福建稻瘟菌毒性类型组成及其对水稻几个Pi基因的毒性频率

 稻瘟病是福建省水稻生产中的重要病害之一,系统掌握稻瘟菌毒性类型组成和变化动态及其与主要抗病基因的互作特点,是制定抗病品种选育与合理利用的依据。本研究根据稻瘟病菌与6个CO39近等基因系品种互作亲和性的结果,将1995~2001年从福建采集分离的398个有效单孢菌株区分为26个毒性类型,其中毒性类型I34.1出现频率最高,为优势毒性类型,出现频率较高的还有I20.1、I04.1、I24.1、I0.1、I30.1等;结果还发现福建稻瘟菌群体对Pi1和Pi2毒性频率较低,分别为7.53%和11.31%,特别是对Pi1和Pi2基因联合毒性频率仅2.76%,说明在水稻抗瘟育种中可以考虑将Pi1和Pi2基因累加利用。     

籼稻恢复系桂R106对稻瘟病的抗性评价及遗传分析

通过与‘丽江新团黑谷’及13个已知抗性基因品种杂交和接种广西稻瘟病菌系鉴定,研究了籼稻恢复系‘桂R106’对稻瘟病的抗性及遗传。结果表明,‘桂R106’对广西565个稻瘟病菌系的抗谱达98.11%,对广西6个主要优势菌系表现全抗,对菌系03E-11和03E-23的抗性均受一对显性基因控制。等位性分析确认,‘桂R106’中抗菌系03E-11的抗病基因与Pi-ta、Pi-12(t)、Pi-3、Pi-5、Pi-7、Pi-a、Pi-at、Pi-sh、Pi-i、Pi-km、Pi-19(t)、Pi-ks、Pi-b这13个已知基因不等位,认为它可能是一个未知的新基因。     

Consistent Quantitative Trait Loci in Pea for Partial Resistance to Aphanomyces euteiches Isolates from the United States and France

ABSTRACT Development of pea cultivars resistant to Aphanomyces root rot, the most destructive root disease of pea worldwide, is a major disease management objective. In a previous study of a mapping population of 127 recombinant inbred lines (RILs) derived from the cross 'Puget' (susceptible) x '90-2079' (partially resistant), we identified seven genomic regions, including a major quantitative trait locus (QTL), Aph1, associated with partial resistance to Aphanomyces root rot in U.S. fields (21). The objective of the present study was to evaluate, in the same mapping population, the specificity versus consistency of Aphanomyces resistance QTL under two screening conditions (greenhouse and field, by comparison with the previous study) and with two isolates of Aphanomyces euteiches originating from the United States and France. The 127 RILs were evaluated in the greenhouse for resistance to pure culture isolates SP7 (United States) and Ae106 (France). Using the genetic map previously described, a total of 10 QTL were identified for resistance in greenhouse conditions to the two isolates. Among these were Aph1, Aph2, and Aph3, previously detected for partial field resistance in the United States. Aph1 and Aph3 were detected with both isolates and Aph2 with only the French isolate. Seven additional QTL were specifically detected with one of the two isolates and were not identified for partial field resistance in the United States. The consistency of the detected resistance QTL over two screening environments and isolates is discussed with regard to pathogen variability, and disease assessment and QTL detection methods. This study suggests the usefulness of three consistent QTL, Aph1, Aph2, and Aph3, for marker-assisted selection.     

Identification of a new resistance gene to a chinese blast fungus isolate in the Japanese rice cultivar aichi asahi

ABSTRACT Genetic analysis of the rice cultivar Aichi Asahi and some other Japanese cultivars for the high resistance to the blast fungus isolate CHNOS58-3-1 from China was performed. All the Japanese differential cultivars were resistant to the isolate except for 'Pi No. 4', which showed moderate resistance. Analysis of the F(2) population of a cross of the susceptible cultivar Reiho and the resistant cultivar Aichi Asahi indicated that the resistance of 'Aichi Asahi' to the isolate was conferred by one dominant gene. To identify the gene in other Japanese differential cultivars, AK lines, which were derived from a cross of 'Aichi Asahi' x 'K59' and assumed to harbor no known genes except for the new one, were used for the allelism tests. The new, completely dominant resistance gene was detected in 14 differential cultivars, but not in 'Pi No. 4', 'Yashiro-mochi', and 'K1', and was designated as Pi19(t). Pi19(t) was allelic or closely linked to Pita(2) on chromosome 12. Pi19(t) was extensively distributed among Japanese traditional local cultivars.     

In the eye of the beholder: the effect of rater variability and different rating scales on QTL mapping

The agronomic importance of developing durably resistant cultivars has led to substantial research in the field of quantitative disease resistance (QDR) and, in particular, mapping quantitative trait loci (QTL) for disease resistance. The assessment of QDR is typically conducted by visual estimation of disease severity, which raises concern over the accuracy and precision of visual estimates. Although previous studies have examined the factors affecting the accuracy and precision of visual disease assessment in relation to the true value of disease severity, the impact of this variability on the identification of disease resistance QTL has not been assessed. In this study, the effects of rater variability and rating scales on mapping QTL for northern leaf blight resistance in maize were evaluated in a recombinant inbred line population grown under field conditions. The population of 191 lines was evaluated by 22 different raters using a direct percentage estimate, a 0-to-9 ordinal rating scale, or both. It was found that more experienced raters had higher precision and that using a direct percentage estimation of diseased leaf area produced higher precision than using an ordinal scale. QTL mapping was then conducted using the disease estimates from each rater using stepwise general linear model selection (GLM) and inclusive composite interval mapping (ICIM). For GLM, the same QTL were largely found across raters, though some QTL were only identified by a subset of raters. The magnitudes of estimated allele effects at identified QTL varied drastically, sometimes by as much as threefold. ICIM produced highly consistent results across raters and for the different rating scales in identifying the location of QTL. We conclude that, despite variability between raters, the identification of QTL was largely consistent among raters, particularly when using ICIM. However, care should be taken in estimating QTL allele effects, because this was highly variable and rater dependent.