Identification of RAPD and SCAR markers linked to northern leaf blight resistance in waxy corn (Zea mays var. ceratina)

Exserohilum turcicum causes northern corn leaf blight (NCLB), an important disease occurring in maize producing areas throughout the world. Currently, the development of cultivars resistant to E. turcicum seems to be the most efficient method to control NCLB damage. Marker-assisted selection (MAS) enables breeders to improve selection efficiency. The objective of this work was to identify random amplified polymorphic DNA (RAPD) and sequence characterized amplified region (SCAR) markers associated with NCLB resistance. Bulked segregant analysis (BSA) was used to search for RAPD markers linked to NCLB resistance genes, using F₂ segregating population obtained by crossing a susceptible inbred ‘209W’ line with a resistant inbred ‘241W’ line. Two hundred and twenty-two decamer primers were screened to identify four RAPD markers: OPA07₅₂₁, OPA16₄₅₇, OPB09₅₂₀, and OPE20₅₃₆ linked to NCLB resistance phenotype. These markers were converted into dominant SCAR markers: SCA07₄₉₆, SCA16₄₂₀, SCB09₄₆₄, and SCE20₄₂₉, respectively. The RAPD and SCAR markers were developed successfully to identify NCLB resistant genotypes in segregating progenies carrying NCLB resistant traits. Thus, the markers identified in this study should be applicable for MAS for the NCLB resistance in waxy corn breeding programs.     

Identification of QTL controlling adventitious root formation during flooding conditions in teosinte (Zea mays ssp. huehuetenangensis) seedlings

Adventitious root formation (ARF) at the soil surface is one of the most important adaptations to soil flooding or waterlogging. Quantitative trait loci (QTL) controlling ARF under flooding condition were identified in a 94 F₂ individual population by crossing maize (Zea mays L., B64) × teosinte (Z. mays ssp. huehuetenangensis). A base-map was constructed using 66 SSR and 42 AFLP markers, covering 1,378 cM throughout all ten maize chromosomes. The ARF capacity for seedlings was determined by evaluating the degree of root formation at the soil surface following flooding for 2 weeks. ARF showed continuous variation in the F₂ population. Interval mapping and composite interval mapping analyses revealed that the QTL for ARF was located on chromosome 8 (bin 8.05). Utilising a selective genotyping strategy with an additional 186 F₂ population derived from the same cross combination and 32 AFLP primer combinations, regions on chromosomes 4 (bin 4.07) and 8 (bin 8.03) were found to be associated with ARF. Z. mays ssp. huehuetenangensis contributed all of the QTL detected in this study. Results of the study suggest a potential for transferring waterlogging tolerance to maize from Z. mays ssp. huehuetenangensis.     

QTL mapping of a partial resistance to the corn delphacid‐transmitted viruses in Lepidopteran‐resistant maize line Mp705

A partial resistance to maize mosaic virus (MMV) and maize stripe virus (MStV) was mapped in a RILs population derived from a cross between lines MP705 (resistant) and B73 (susceptible). A genetic map constructed from 131 SSR markers spanned 1399 cM with an average distance of 9.6 cM. A total of 10 QTL were detected for resistance to MMV and MStV, using composite interval mapping. A major QTL explaining 34–41% of the phenotypic variance for early resistance to MMV was detected on chromosome 1. Another major QTL explaining up to 30% of the phenotypic variation for all traits of resistance to MStV was detected in the centromeric region of chromosome 3 (3.05 bin). After adding supplementary SSR markers, this region was found to correspond well to the one where a QTL of resistance to MStV already was located in a previous mapping study using an F₂ population derived from a cross between Rev81 and B73. These results suggested that these QTL of resistance to MStV detected on chromosome 3 could be allelic in maize genome.     

Molecular mapping of a major QTL conferring resistance to SCMV based on immortal RIL population in maize

Sugarcane mosaic virus (SCMV) is one of devastating pathogens in maize (Zea mays L.), and causes serious yield loss in susceptible cultivars. An effective solution to control the virus is utilizing resistant genes to improve the resistance of susceptible materials, whereas the basic work is to analyze the genetic basis of resistance. In this study, maize inbred lines Huangzao4 (resistant) and Mo17 (susceptible) were used to establish an F₉ immortal recombinant inbred line (RIL) population containing 239 RILs. Based on this segregation population, a genetic map was constructed with 100 simple sequence repeat (SSR) markers selected from 370 markers, and it covers 1421.5 cM of genetic distance on ten chromosomes, with an average interval length of 14.2 cM. Analysis of the genetic map and resistance by mapping software indicated that a major quantitative trait locus (QTL) was between bin6.00 and bin6.01 on chromosome 6, linked with marker Bnlg1600 (0.1 cM of interval). This QTL could account for 50.0% of phenotypic variation, and could decrease 27.9% of disease index.     

甜玉米果皮厚度QTL的定位及上位性互作

果皮厚度是影响甜玉米口感的一个重要因素。发掘果皮厚度的基因资源、了解玉米果皮厚度的遗传机制,是指导其育种的基础。本研究以日超-1(薄果皮,56.57μm)×1021(厚果皮,100.23μm)的190个BC1F2家系为作图群体,分别采用2种遗传模型检测QTL。基于复合区间作图(CIM)共检测到3个影响果皮厚度的QTL,位于3.01、6.01、8.05区段,分别解释8.6%、16.0%和7.2%的表型变异,其中3.01和8.05处QTL以加性效应为主;基于混合线性CIM模型(MCIM)共检测到5个影响果皮厚度的QTL,其中除8.05处QTL为加性QTL外,另有2对加×加上位性互作QTL,1对是2.01和6.05处QTL之间的互作,另1对则是5.06和6.01处QTL间的互作。这2对互作QTL分别解释了6.63%和12.48%的表型变异率。本结果表明,加性效应和上位性互作效应等都在果皮厚度的形成和遗传中起重要作用。能够检测QTL上位互作的MCIM模型更适用于果皮厚度QTL定位。本研究还在其中4个QTL的区域内分别检索到胚乳中色素合成以及细胞转变的相关候选基因,这些基因的表达是否与果皮厚度的变异有关值得进一步研究。     

QTL analysis of resistance to Fusarium head blight in wheat using a 'Frontana'-derived population

Fusarium head blight (FHB or head scab) has become a major limiting factor for sustainable wheat (Triticum aestivum L.) production around the world. For quantitative trait loci (QTL) analysis of resistance to FHB, F₃ plants and F_{3 : 5} lines, derived from a ‘Frontana’ (moderately resistant)/‘Seri82’ (susceptible) cross, were spray‐inoculated in 2001 and 2002, respectively. Artificial inoculations were carried out under field conditions. Of 273 SSR and AFLP markers, 250 could be mapped and they yielded 42 linkage groups, covering a genetic distance of 1931 cM. QTL analysis was based on the constructed linkage map and area under the disease progress curve (AUDPC). The analyses revealed three consistent QTLs associated with FHB resistance on chromosomes 1BL, 3AL and 7AS explaining 7.9%, 7.7% and 7.6% of the phenotypic variation, respectively, above 2 years. The results confirmed the previously described resistance QTL of ‘Frontana’ on chromosome 3AL. A combination of ‘Frontana’ resistance with ‘Sumai‐3’ resistance may lead to lines with augmented resistance expression.     

Molecular mapping of QTL for southwestern corn borer resistance, plant height and flowering in tropical maize

Development of multiple insect resistance in tropical maize represents a major effort of the maize breeding programme at CIMMYT. Resistance to the southwestern corn borer (SWCB) is polygenically controlled with primarily additive gene action. Our main objective was to identify quantitative trait loci (QTL) involved in resistance to SWCB. Other objectives were to detect QTL in the same population for plant height, female flowering, and the anthesis-silking interval (ASI). A population of 472 F₂ individuals derived from a cross between the susceptible line Ki3 and the resistant inbred CML139, was restriction fragment length polymorphism (RFLP) genotyped using 110 maize probes. F₃ families were rated for leaf-feeding damage after artificial infestation at one location in three consecutive years. Height and flowering were measured in protected trials in two locations. QTL analyses were conducted using joint composite interval mapping. Seven QTL on chromosomes 3, 5, 6, 8, and 9 explained 30% of the phenotypic variance (σ²_p) for SWCB damage. Most QTL alleles conferring resistance were contributed from CML139. QTL showed dominance, partial dominance and additive gene action. Eleven QTL dispersed across the genome were determined to affect plant height and explained 43% of σ²_p. Four of these were in close proximity to loci with qualitative effects on plant height. Thirteen QTL (50% of sigma;²_p) were identified for days to female flowering and nine (30% of σ²_p) for ASI. Our results, along with those from other mapping studies at CIMMYT, are allowing us to formulate marker-assisted selection schemes to complement the breeding efforts for such complex traits as borer resistance.     

Mapping of rhizoctonia root rot resistance genes in sugar beet using pathogen response‐related sequences as molecular markers

Rhizoctonia root and crown rot caused by the fungus Rhizoctonia solani is a serious disease of sugar beet. An F_2:3 population from a cross between a resistant and a susceptible parent has been tested for R. solani resistance and a genetic map has been constructed from the corresponding F₂ parents. The map encompasses 38 expressed sequence tags (ESTs) with high similarity to genes which are involved in resistance reactions of plants (R‐ESTs) and 25 bacterial artificial chromosomes (BACs) containing nucleotide binding site (NBS)‐motifs typical for disease resistance genes. Three quantitative trait loci (QTL) for R. solani resistance were found on chromosomes 4, 5 and 7 collectively explaining 71% of the total phenotypic variation. A number of R‐ESTs were mapped in close distance to the R. solani resistance QTL. In contrast, the NBS‐BACs mapped to chromosomes 1, 3, 7 and 9 with two major clusters of NBS‐BACs on chromosome 3. No linkage between NBS‐BACs and R. solani resistance QTL was found. The data are discussed with regard to using R‐ESTs and NBS markers for mapping quantitative disease resistances.     

Inheritance of resistance to Fusarium head blight in the wheat lines 'CJ 9306' and 'CJ 9403'

Fusarium head blight (FHB or scab) caused by Fusarium graminearum is a worldwide serious disease in wheat. Exploitation and genetic studies of elite resistance sources can speed up the development of resistant cultivars. To characterize the inheritance of host plant resistance in two new lines, ‘CJ 9306’ and ‘CJ 9403’, developed from a recurrent selection programme in China, six generations P₁, P₂, F₁, F₂, B₁ and B₂ of four crosses and 137 F_{6 : 7} recombinant inbred lines (RILs) from one cross were evaluated in the greenhouse for scab resistance using single‐floret inoculation. The data of area under disease progress curve (AUDPC) in F₂, backcross (BC) and RIL populations exhibited mono‐modal distributions without clear‐cut demarcations and skewing towards resistance. An additive–dominance model was well‐fitted, additive effects played a predominating role, and dominance effects were also significant. Continuous distributions with two major peaks and one minor peak for the number or percentage of scabby spikelets (NSS or PSS) in segregating populations implied the existence of major genes or quantitative trait loci (QTL) for resistance. The estimates of broad‐sense and narrow‐sense heritabilities based on the six‐generation experiment were 56–76% and 26–67% respectively. The estimates of broad‐sense heritabilities based on anova with RILs were 89–90%. These two improved lines with excellent scab resistance and good agronomic traits are of interest for wheat breeding and production.     

Dent corn genetic background influences QTL detection for grain yield and yield components in high-oil maize

Despite the well-recognized importance of grain yield in high-oil maize (Zea mays L.) breeding and production, few studies have reported the application of QTL mapping of such traits. An inbred line of high-oil maize designated ‘GY220’ was crossed with two dent maize inbred lines to generate two connected F_2:3 populations with 284 and 265 F_2:3 families. Our main objective was to evaluate the influence of genetic background on QTL detection of grain yield traits through comparisons between the F_2:3 populations. The field experiments were conducted during the spring in Luoyang and summer in Xuchang, Henan, China. Two genetic linkage maps were constructed with a genetic distance of 2111.7 and 2298.5 cM using 185 and 173 polymorphic SSR markers, respectively. In total, 18 and 15 QTL were detected for six grain yield traits in the two populations. Only one common QTL marker was shared between the two populations. A QTL cluster associated with five traits was identified at bin 1.05–1.06, including the shared QTL for 100GW, which demonstrated the largest effect (16.7%). Among the detected QTL, 12 digenic interactions were identified. Our results reflect the substantial influence of dent maize genetic background on QTL detection of grain yield traits.     

Validation of a major QTL for scab resistance with SSR markers and use of marker-assisted selection in wheat

The objectives of this study were to validate the major quantitative trait locus (QTL) for scab resistance on the short arm of chromosome 3B in bread wheat and to isolate near‐isogenic lines for this QTL using marker‐assisted selection (MAS). Two resistant by susceptible populations, both using ‘Ning7840’ as the source of resistance, were developed to examine the effect of the 3BS QTL in different genetic backgrounds. Data for scab resistance and simple sequence repeat (SSR) markers linked to the resistance QTL were analyzed in the F_2:3 lines of one population and in the F_3:4 lines of the other. Markers linked to the major QTL on chromosome 3BS in the original mapping population (‘Ning7840’/‘Clark’) were closely associated with scab resistance in both validation populations. Marker‐assisted selection for the QTL with the SSR markers combined with phenotypic selection was more effective than selection based solely on phenotypic evaluation in early generations. Marker‐assisted selection of the major QTL during the seedling stage plus phenotypic selection after flowering effectively identified scab resistant lines in this experiment. Near‐isogenic lines for this 3BS QTL were isolated from the F₆ generation of the cross ‘Ning7840’/‘IL89‐7978’ based on two flanking SSR markers, Xgwm389 and Xbarc147. Based on these results, MAS for the major scab resistance QTL can improve selection efficiency and may facilitate stacking of scab resistance genes from different sources.     

QTL detection for grain oil and starch content and their associations in two connected F<Subscript>2:3</Subscript> populations in high-oil maize

High-oil maize (Zea mays L.) has special value in animal feed and human food. Two hundred and eight-four and 265 F_2:3 families developed from two crosses between one high-oil maize inbred and two normal dent maize inbreds were evaluated for grain oil and starch contents under two environments. Using composite interval mapping, 1–6 QTL for each trait were detected under each environment and in combined analysis in both populations. Only one common QTL across two environments in each population and across two populations were found for starch content. Among the detected QTL, nine digenic interactions with small effects were identified. Comparison of single-trait QTL and the results of multiple-trait QTL mapping showed that oil content might be complicatedly correlated with starch content. Although single-trait QTL with the same parental effects for both traits and oil-starch QTL were all detected at the same genetic bin 6.04 as the cloned high-oil QTL (qHO6) with no unfavorable effects on grain weight, our results did reflect the difficulty to realize simultaneous improvement on grain oil and starch contents. Of course, these results should be validated in further experiments under more environments using RILs, NILs and other permanent populations.     

Identification of QTLs for niacin concentration in maize kernels

A genetically induced increase in the niacin concentration of maize kernels is important for the improvement of kernels nutrition. In this paper, the correlation, heredity and variation of free niacin concentration and total niacin concentration in maize kernels were analysed. The results showed that a low or moderate correlation existed between free niacin concentration and total niacin concentration, and that the heritabilities of the two traits were approximately 70%. Quantitative trait loci (QTL) analysis showed that qFNA4a‐HC and qTNA4a‐HC were detected to have a common marker, umc1294 (Bin4.02), in different populations. In population A, qFNA8b and qTNA8a were detected in two generations or two sites, qTNA8c‐F₂ and qFNA8b‐HC have a common interval. These QTLs explained 9.7%–17.3% of phenotypic variation. In population B, qFNA4a, explaining 9.3%–11.2% of phenotypic variation, was detected in two environments. Two marker intervals, umc1294‐bnlg490 (Bin4.02‐4.03) and umc1959‐umc1562 (Bin8.05), harbored the QTL for free niacin concentration and total niacin concentration. Furthermore, among a total of 78 QTLs obtained from all datasets, 19, 9 and 9 QTLs were located in Bin8.05‐8.06, Bin4.02‐4.04 and Bin10.01‐10.03, respectively.     

Mapping a new source of resistance to powdery mildew in mungbean

Both restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP) analyses were employed to map a new source of resistance to powdery mildew in mungbean. Disease scores of an F₂ population derived from the cross between a moderately resistant breeding line VC1210A and a susceptible wild relative (Vigna radiata var. sublobata, accession TC1966) showed a continuous distribution and was treated as a quantitative trait. Although no significant quantitative trait loci (QTL) that can explain the variation was detected by QTL analysis based on the reconstructed RFLP linkage map, new marker loci associated with resistance were discovered by AFLP analysis. The RFLP loci detected by two of the cloned AFLP bands are associated with resistance and constitute a new linkage group. A major resistance quantitative trait locus was found on this linkage group that accounted for 64.9% of the variation in resistance to powdery mildew. One of the probes developed in this study has the potential to assist in breeding for powdery mildew resistance in mungbean.     

Dissection of the genetic architecture for grain quality‐related traits in three RIL populations of maize (Zea mays L.)

To manipulate the composition of the maize kernel to meet future needs, an understanding of the molecular regulation of kernel quality‐related traits is required. In this study, the quantitative trait loci (QTL) for the concentrations of grain protein, starch and oil were identified using three sets of RIL populations in three environments. The genetic maps and the initial QTL were integrated using meta‐analyses. A total of 38 QTL were identified, including 15 in population 1, 12 in population 2 and 11 in population 3. The individual effects ranged from 2.87% to 13.11% of the phenotypic variation, with seven QTL each contributing over 10%. One common QTL was found for the concentrations of grain protein and starch in bin 3.09 in the three environments and the three RIL populations. Of the 38 initial QTL, 22 were integrated into eight mQTL by meta‐analysis. mQTL3 and mQTL8 of the key mQTL in which the initial QTL displayed R² > 10% included six and three initial QTL for grain protein and starch concentrations from two or three populations, respectively. These results will provide useful information for marker‐assisted selection to improve the quality of the maize kernel.     

玉米大斑病广谱抗性外引自交系的发掘与抗病基因初步鉴定

从2014—2016年连续3年对43份来自美国、法国、俄罗斯和德国的玉米资源进行了抗大斑病人工接种鉴定, 筛选到高抗玉米大斑病材料7份, 抗病材料1份, 中抗材料6份, 抗性材料占鉴定总材料的比例为32.6%。利用F₂群体, 对7份高抗材料进行了大斑病抗性遗传分析, 抗感植株分离比例和适合性测验证明, 自交系A04、F02、F05和R01对大斑病的抗性可能是由一对显性基因控制的。抗谱分析表明, 自交系A04、F02、F05和R01携带的抗大斑病基因不同于Ht1、Ht2、Ht3和HtN, 可能是新的抗病基因。该研究结果可为今后我国玉米大斑病抗性种质的引进及改良提供重要参考。     

Comparison of QTLs mapped in RILs and their test-cross progenies of tropical maize for insect resistance and agronomic traits

Quantitative trait loci (QTL) affecting resistance to south-western corn borer Diatraea grandiosella (SWCB) and sugarcane borer Diatraea saccharalis (SCB) have been identified previously in F_2:3 lines and recombinant inbred lines (RILs) of tropical maize using restriction fragment length polymorphism (RFLP) analyses. Our objective was to determine whether QTLs identified in these generations are also expressed in test crosses (TC) of RILs. A population of 166 TC progenies was developed by crossing RILs from the cross CML131 (susceptible) × CML67 (resistant) with the unrelated, susceptible tester line CML216. Resistance to first-generation SWCB, measured as leaf-feeding damage (LFD) under artificial infestation, and other agronomic traits were evaluated in two environments for the TC progenies and three environments for 183 RILs. The correlation between line per se and TC performance was low for LFD and intermediate for most agronomic traits. Estimates of the genotypic variance and heritabilities were smaller in the TC progenies than in the RILs for all traits. Quantitative trait loci were identified using an RFLP linkage map with 136 loci. For LFD, four QTLs were detected in the TC progenies, of which two were in common with nine QTLs previously mapped in the RILs. Few QTLs for agronomic traits were common to the two types of progeny, because of the low consistency of QTL positions for all traits in RIL and TC progenies, the use of TC progenies should be considered in QTL mapping studies as the first step for marker-assisted selection in hybrid breeding.     

Evaluation of F2 and F3 plants introgressed with QTLs for clubroot resistance in cabbage developed by using SCAR markers

Clubroot disease caused by Plasmodiophora brassicae is one of the major diseases of Brassica crops, often devastating to the cultivation of cruciferous crops in temperate regions. In a previous study (Moriguchi et al. 1999) identified three major quantitative trait loci (QTLs) for clubroot resistance, each in a separate linkage group, in a population derived from a cross between a clubroot‐susceptible inbred cabbage line, Y2A and a resistant inbred kale line, K269. In this study, the original random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers were converted into sequence‐characterized amplified region (SCAR) markers to facilitate large‐scale marker‐assisted screening of clubroot resistance in cabbage breeding. Of 15 RAPD markers closely linked to the three QTLs, nine SCARs were developed as dominant markers after cloning and sequencing. In addition, two RAPD markers were converted into co‐dominant cleaved amplified polymorphic sequence (CAPS) markers, and one RFLP marker out of three tested was converted to a dominant SCAR marker. The effect of selection for resistance by the improved markers was evaluated in progeny plants in the F2 and F3. A total of 138 F2 plants were genotyped with nine SCARs and 121 well‐distributed makers consisting of 98 RAPD, 19 RFLP, two isozymes, and two morphological markers in order to estimate the level of resistance and the proportion of undesirable alleles from the kale in non‐target areas in each of the F2 populations. An F2 plant, YK118, had kale alleles at QTL1, QTL3 and QTL9. Three F2 plants, namely, YK107, YK25 and YK51 had kale alleles at only QTL1, QTL3 and QTL9, respectively. These F2 plants were selected for their low proportion of alleles derived from kale in non‐target regions. YK118, like the resistant kale parent, expressed very high resistance to three field isolates of Plasmodiophora brassicae, whereas the mean disease index in the F2 and F3 plants carrying only single QTLs was intermediate. The QTLs showed no differential response to the isolates. These plants with improved resistance will be useful as parental inbred lines for F1 hybrids.     

Quantitative trait loci mapping of forage stover quality traits in six mapping populations derived from European elite maize germplasm

Four forage maize stover quality traits were analysed including in vitro digestibility of organic matter (IVDOM), neutral detergent fibre (NDF), water‐soluble carbohydrates (WSC) and digestibility of NDF (DNDF). We mapped quantitative trait loci (QTL) in three DH (doubled haploid) populations (totally 250–720 DH lines): one RIL population (358 lines) and two testcross (TC) populations, based on field phenotyping at multiple locations and years for each. High phenotypic and genotypic correlations were found for all traits and significant (p < .01) at two locations, and NDF was negatively correlated with the other traits. QTL analyses were conducted by composite interval mapping. A total of 33, 23, 32 and 25 QTL were identified for IVDOM, NDF, WSC and DNDF, respectively, with three, four, five and two major QTL for each. Few consistent QTL for IVDOM, WSC and DNDF were detected in more than two populations. This study contributed to the identification of key QTL associated with forage maize digestibility traits and is beneficial for marker‐assisted breeding and fine mapping of candidate genes associated with forage maize quality.     

Application of Triticum monococcum for the improvement of triticale resistance to leaf rust (Puccinia triticina)

The aim of this work was to evaluate the leaf rust resistance introduced into introgressive triticale lines with Triticum monococcum genes, and to study the expression of these genes at the hexaploid level. The introgressive lines were developed by incorporating diploid wheat (T. monococcum s.s.) genes into hexaploid triticale LT 522/6 using the synthetic allotetraploid T. monococcum/Secale cereale (A^mA^mRR) as a bridging form. A group of 44 those lines, parental stocks and check cultivars were inoculated at the seedling stage (in a greenhouse) and at the adult‐plant stage (in the field) with four pathotypes of Puccinia triticina. At the seedling stage the assessment of infection type showed that four lines had resistance to all pathotypes as high as in the T. monococcum donor. Adult plant examinations showed some introgressive lines with complete resistance and also lines with partial resistance, expressed in area under the disease progress curve (AUDPC) calculations as slow rusting. Some lines comprise low AUDPC with complete resistance at seedling stage.