Quantitative trait loci analysis in Theobroma cacao using molecular markers. Inheritance of polygenic resistance to Phytophthora palmivora in two related cacao populations

Two related segregating populations of Theobroma cacao L. were analysed for their resistance to Phytophthora palmivora. The first F₁ population was obtained by crossing two susceptible cacao clones of Catongo (a highly homozygous genotype) and Pound 12(a highly heterozygous genotype) and the second population was obtained by backcrossing a single F₁ tree with Catongo. The genetic maps obtained for each population were compared. The F₁ map includes 162 loci and the backcross has 140 loci. The two maps, F₁ and BC₁, exhibit high co-linear loci organization covering respectively, 772 and 944 cM.Phytophthora resistance was assessed by measuring the size increase of a lesion at five (DL5)and ten days (DL10) after pod inoculation. Six different QTL were detected in the F₁ and BC₁ populations. One QTL was found in both populations, and appeared to be a major component of disease resistance, and explaining nearly 48% of the phenotypic variance in the F₁ population. The absence of some yield QTL detection in the BC₁ in comparison with the F₁ population is due to the lack of transmission of the favouring alleles for these QTL from the single F₁ tree used for the backcross. The phenotypic variance explained by the action of the quantitative trait alleles indicated that genetic factors of both major and minor effects were involved in the control of the character studied. QTL conferring increased resistance to Phytophthorawere identified in both susceptible parents, suggesting the presence of transgressive traits and the possibility of selection in cacao. Pleiotropic and epistatic effects for the QTL were also detected. Finally, the use of marker assisted selection (MAS) in cacao breeding programs is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Experimental evaluation of several cycles of marker-assisted selection in maize

A program was initiated in 1994 to compare the efficiency of marker-assisted selection (MAS) and conventional phenotypic selection. A population of 300 F_3:4 families was generated from the cross between two maize inbred lines F2 and F252 and selected on an index combining grain yield and grain moisture at harvest. This population was characterised for 93 RFLP markers and evaluated as testcrosses in a large range of environments. Three methods of selection were applied (i) two cycles of conventional phenotypic selection; (ii) two cycles of MAS based on an index combining phenotypic values and QTL genetic values and (iii) one cycle of combined MAS followed by two cycles of selection based only on the QTL effects estimated in the first generation. The different populations were characterised for RFLP markers. The evolution of allele frequencies showed that selection on only-markers was very efficient for fixing QTL alleles found favourable in the initial population. This evolution was quite different from that observed for phenotypic selection or combined MAS. Genetic gain was evaluated and found significant for each method of selection. Nevertheless, the difference between phenotypic selection and combined MAS was not significant. The two additional cycles of MAS on only-markers did not improve significantly the genetic value of the population. Moreover, the genetic variance of this population remained high, despite most of the QTL initially detected were almost fixed. The results suggest that the QTL effects estimated in the initial population were not stable due to epistasis and/or QTL by environment interactions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Increased grain protein content and its association with agronomic and end-use quality in two hard red spring wheat populations derived from Triticum turgidum L. var. dicoccoides

Suitability of wheat (Triticum aestivum L.) for many food products depends on its unique protein. Elevated grain protein content (GPC) and its quality influences the bread making properties of wheat. The objective of this study was to examine the association of elevated GPC with agronomic and end-use quality in two hard red spring wheat recombinant inbred (RI) populations derived from wild emmer (Triticum turgidum L. var. dicoccoides). The two hard red spring populations (ND683/Bergen and Glupro/Bergen) were developed using a single-seed-descent method. ND683 and ‘Glupro’ are high in GPC (180 g kg^-1), presumably due to the introgression of gene(s) from Triticum turgidum L. var. dicoccoides and ‘Bergen’ is low in GPC (145 g kg^-1). From each of the two populations 12 high- and 12 low-GPC RI lines (F_5:7) were selected for replicated testing at two North Dakota (ND) locations in 1995. In both populations, the high-GPC lines had significantly (p < 0.05) higher values compared to the low-GPC lines for mean GPC and water absorption. Mean grain yield of the high-GPC lines was not significantly different from the low-GPC lines in both populations. In the ND683/Bergen population, the high-GPC lines had significantly (p < 0.05) higher values than the low-GPC lines for mean plant height, days to heading, and flour extraction. GPC was significantly (p < 0.05)and negatively associated with test weight and also significantly (p < 0.01) and positively associated with water absorption in the Glupro/Bergen population. In these populations, results suggested that it may be possible to select lines that combine higher GPC and acceptable yield level, but later in maturity and taller in plant height. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of high yielding IR64 × <Emphasis Type="Italic">Oryza rufipogon</Emphasis> (Griff.) introgression lines and identification of introgressed alien chromosome segments using SSR markers

Modern rice varieties that ushered in the green revolution brought about dramatic increase in rice production worldwide but at the cost of genetic diversity at the farmers’ fields. The wild species germplasm can be used for broadening the genetic base and improving productivity. Mining of alleles at productivity QTL from related wild species under simultaneous backcrossing and evaluation, accompanied by molecular marker analysis has emerged as an effective plant breeding strategy for utilization of wild species germplasm. In the present study, a limited backcross strategy was used to introgress QTL associated with yield and yield components from Oryza rufipogon (acc. IRGC 105491) to cultivated rice, O. sativa cv IR64. A set of 12 BC₂F₆ progenies, selected from among more than 100 BC₂F₅ progenies were evaluated for yield and yield components. For plant height, days to 50% flowering and tillers/plant, the introgression lines did not show any significant change compared to the recurrent parent IR64. For yield, 9 of the 12 introgression lines showed significantly higher yield (19–38%) than the recurrent parent IR64. Four of these lines originating from a common lineage showed higher yield due to increase in grain weight and another three also from a common lineage showed yield increase due to increase in grain number per panicle. For analyzing the introgression at molecular level all the 12 lines were analyzed for 259 polymorphic SSR markers. Of the total 259 SSR markers analyzed, only 18 (7.0%) showed introgression from O. rufipogon for chromosomes 1, 2, 3, 5, 6 and 11. Graphical genotypes have been prepared for each line and association between the introgression regions and the traits that increased yield is reported. Based on marker trait association it appears that some of the QTL are stable across the environments and genetic backgrounds and can be exploited universally.     

Molecular genetic analysis of flour color using a doubled haploid population in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Flour color is an important trait in the assessment of flour quality for the production of many end products. In this study, quantitative trait loci (QTLs) with additive effects, epistatic effects, and QTL × environment (QE) interactions for flour color in bread wheat (Triticum aestivum L.) were studied, using a set of 168 doubled haploid (DH) lines derived from a Huapei 3 × Yumai 57 cross. A genetic map was constructed using 283 simple sequence repeats (SSR) and 22 expressed sequence tags (EST)-SSR markers. The DH and parents were evaluated for flour color in three environments. QTL analyses were performed using QTLNetwork 2.0 software based on a mixed linear model approach. A total of 18 additive QTLs and 24 pairs of epistatic QTLs were detected for flour color, which were distributed on 19 of the 21 chromosomes. One major QTL, qa1B, closely linked to barc372 0.1 cM, could account for 25.64% of the phenotypic variation of a* without any influence from the environments. So qa1B could be used in the molecular marker-assisted selection (MAS) in wheat breeding programs. The results showed that both additive and epistatic effects were important genetic basis for flour color, and were also sometimes subject to environmental modifications. The information obtained in this study should be useful for manipulating the QTLs for flour color by MAS in wheat breeding programs. Kun-Pu Zhang and Guang-Feng Chen contributed equally to this study.     

Identification of RAPD markers linked to the rust (<Emphasis Type="Italic">Uromyces fabae</Emphasis>) resistance gene in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>)

Summary Two RAPD markers linked to gene for resistance (assayed as pustule number cm⁻² leaf area) to rust [Uromyces fabae (Pers.) de Bary] in pea (Pisum sativum L.) were identified using a mapping population of 31 BC₁F₁ [HUVP 1 (HUVP 1 × FC 1] plants, FC 1 being the resistant parent. The analysis of genetics of rust resistance was based on the parents, F₁, F₂, BC₁F₁ and BC₁F₂ generations. Rust resistance in pea is of non-hypersensitive type; it appeared to be governed by a single partially dominant gene for which symbol Ruf is proposed. Further, this trait seems to be affected by some polygenes in addition to the proposed oligogene Ruf. A total of 614 decamer primers were used to survey the parental polymorphism with regard to DNA amplification by polymerase chain reaction. The primers that amplified polymorphic bands present in the resistant parent (FC 1) were used for bulked segregant analysis. Those markers that amplified consistently and differentially in the resistant and susceptible bulks were separately tested with the 31 BC₁F₁ individuals. Two RAPD makers, viz., SC10-82₃₆₀ (primer, GCCGTGAAGT), and SCRI-71₁₀₀₀ (primer, GTGGCGTAGT), flanking the rust resistance gene (Ruf) with a distance of 10.8 cM (0.097 rF and LOD of 5.05) and 24.5 cM (0.194 rF and a LOD of 2.72), respectively, were identified. These RAPD markers were not close enough to Ruf to allow a dependable maker-assisted selection for rust resistance. However, if the two makers flanking Ruf were used together, the effectiveness of MAS would be improved considerably.     

Identification of a major QTL affecting resistance to brown spot in tobacco (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L.) via linkage and association mapping methods

Brown spot (BS) is a destructive foliar disease in tobacco (Nicotiana tabacum L.) and is caused by Alternaria alternata. BS poses a serious threat to tobacco production worldwide. To develop molecular markers that are tightly linked to BS resistance for marker-assisted selection (MAS), F₂, F_2:3 and BC₃F_2:3 populations were developed from a cross between a source of BS resistance Jingyehuang (JYH) and a BS susceptible flue-cured variety NC82. One hundred eighty-one F₂ individuals, 180 F_2:3 lines and 256 BC₃F_2:3 lines were evaluated for field resistance under different environments and quantitative trait loci (QTL) were identified by linkage mapping. A major QTL was mapped on chromosome15; this QTL explained 8.6–18.0% of the phenotypic variation under different conditions. Furthermore, 219 accessions were evaluated for their responses to BS at two sites, and association mapping (AM) was used to verify the chromosomal region harboring the major QTL. The AM results showed that six significant marker-trait associations were detected at two sites. Among these markers, the marker Indel53 within the specific chromosomal region exhibited the most significant association with resistance to BS and explained 20.0 and 21.5% of the phenotypic variation at the two sites, respectively. An approximately 2-Mb physical interval at the locus of marker Indel53 contained 31 predicted genes; quantitative real-time PCR results suggested that two of these genes (Nitab 4.5_0000264g0050.1 and Nitab 4.5_0000264g0130.1) were probable candidate genes for resistance to BS. In summary, our results suggested that the novel major QTL from tobacco variety JYH for resistance to BS provided partial effective resistance against A. alternata and was useful for MAS of resistance to BS in tobacco breeding.     

Localization of QTL for basal root thickness in <Emphasis Type="Italic">japonica</Emphasis> rice and effect of marker-assisted selection for a major QTL

Root traits are key components of plant adaptation to drought environment. By using a 120 recombined inbred lines (RILs) rice population derived from a cross between IRAT109, a japonica upland rice cultivar and Yuefu, a japonica lowland rice cultivar, a complete genetic linkage map with 201 molecular markers covering 1,833.8 cM was constructed and quantitative trait loci (QTLs) associated with basal root thickness (BRT) were identified. A major QTL, conferring thicker BRT, located on chromosome 4, designated brt4, explained phenotypic variance of 20.6%, was selected as target QTL to study the effects of marker-assisted selection (MAS) using two early segregating populations derived from crosses between IRAT109 and two lowland rice cultivars. The results showed that the flanking markers of brt4 were genetically stable in populations with different genetic backgrounds. In the two populations under upland conditions, the difference between the means of BRT of plants carrying positive and negative favorable alleles at brt4 flanking markers loci was significant. Phenotypic effects of BRT QTL brt4 were 5.05–8.16%. When selected plants for two generations were planted at Beijing and Hainan locations under upland conditions, MAS effects for BRT QTL brt4 were 4.56–18.56% and 15.46–26.52% respectively. The means of BRT for the homozygous plants were greater than that of heterozygous plants. This major QTL might be useful for rice drought tolerance breeding. L. Liu and P. Mu are contributed equally to this work.     

Microsatellite Marker Identification of a Triticum Aestivum -Aegilops Umbellulata Substitution Line with Powdery Mildew Resistance

Summary Aegilops umbellulata acc. Y39 and Triticum carthlicum acc. PS5, immune to many powdery mildew isolates, were crossed to make an amphidiploid line Am9. The powdery mildew resistance of Am9 was transferred to common wheat cultivar Laizhou953 by crossing and backcrossing. In this study, the origin of powdery mildew resistance in a BC₃F_4:5 population derived from a cross of Am9 and Laizhou953 was identified. Microsatellite markers analysis showed that markers Xgwm257, Xgwm296, and Xgwm319, co-segregated with the powdery mildew resistance, whereas markers Xgwm210, Xgwm388/140, Xgwm388/170 and Xgwm526 were related to susceptibility and linked to resistance in repulsion. Of three markers related to resistance, Xgwm257 and Xgwm319 were codominant, whereas Xgwm296 was dominant. All three markers were Ae. umbellulata-specific indicating that resistance in the test population originated from Ae. umbellulata acc. Y39. The chromosome location and mapping of these linked microsatellite markers, the chromosome numbers of derived BC₃F_4:6 families, and chromosome pairing in F₁ plants from a cross of a homozygous resistant BC₃F_4:5 plant and Laizhou953, showed that wheat chromosome 2B was substituted by Ae. umbellulata chromosome 2U. This is the first gene conferring powdery mildew resistance transferred to wheat from Ae. umbellulata, and it should be a novel resistance gene to powdery mildew. It was temporarily designated PmY39.The first two authors made equal contributions     

Identification of AFLP and SCAR markers linked to the male fertility restorer gene of <Emphasis Type="Italic">pol</Emphasis> CMS (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

The pol cytoplasmic male-sterility system has been widely used as a component for utilization of heterosis in Brassica napus and offers an attractive system for study on nuclear–mitochondrial interactions in plants. Genetic analyses have indicated that one dominant gene, Rfp, was required to achieve complete fertility restoration. As a first step toward cloning of this restorer gene, we attempted molecular mapping of the Rfp locus using the amplified fragment length polymorphism (AFLP) technique combined with bulked segregant analysis (BSA) method. A BC₁ population segregating for Rfp gene was used for tagging. From the survey of 1,024 AFLP primer combinations, 13 linked AFLP markers were obtained and five of them were successfully converted into sequence characterized amplified region (SCAR) markers. A population of 193 plants was screened using these markers and the closest AFLP markers flanking Rfp were at the distances of 2.0 and 5.3 cM away, respectively. Further the AFLP or SCAR markers linked to the Rfp gene were integrated to one doubled-haploid (DH) population derived from the cross Quantum × No.2127-17 available in our laboratory, and Rfp gene was mapped on N18, which was the same as the previous report. These molecular markers will facilitate the marker-assisted selection (MAS) of pol CMS restorer lines.     

利用染色体片段代换系定位陆地棉株高QTL

以陆地棉中棉所36为轮回亲本和海岛棉海1为供体亲本, 构建染色体片段代换系。为了能检测到稳定的株高QTL,将三个代换系群体(BC₅F₃, BC₅F_3:4和BC₅F_3:5)在5个环境中种植,2009年和2010年分别在河南安阳种植BC₅F₃单株、BC₅F_3:4株行, 2011年分别在河南安阳、辽宁辽阳和新疆石河子种植BC₅F_3:4株系。结果表明,在不同群体环境中株高的超亲比例为53.43%~88.97%。从早期构建的总图距为5088.28 cM, 含有2280个SSR标记位点,覆盖26条染色体的遗传连锁图谱中筛选标记,对408个单株进行的SSR鉴定,结果检测到16个株高QTL,分布在10条染色体上。单个QTL解释的表型变异为7.35%~13.17%。有7个QTL在2个以上环境被检测到。与标记MUSS563紧密连锁的qPH-15-19在一个环境中被检测到,在前人的研究中也有报道。这些结果为进一步精细定位QTL、基因克隆、分子辅助选择等研究奠定基础。     

Identification of four genes for stable hybrid sterility and an epistatic QTL from a cross between Oryza sativa and Oryza glaberrima

To further understand the nature of hybrid sterility between Oryza sativa and Oryza glaberrima, quantitative trait loci (QTL) controlling hybrid sterility between the two cultivated rice species were detected in BC₁F₁ and advanced backcross populations. A genetic map was constructed using the BC₁F₁ population derived from a cross between WAB450-16, an O. sativa cultivar, and CG14, an O. glaberrima cultivar. Seven main-effect QTLs for pollen and spikelet sterility were detected in the BC₁F₁. Forty-four sterility NILs (BC₆F₁) were developed via successive backcrosses using pollen sterility plants as female and WAB450-16 as the recurrent parent. Seven NILs, in which the target QTL regions were heterozygous while the other QTL regions as well as most of the reminder of the genome were homozygous for the WAB450-16 allele, were selected as the QTL identification materials. BC₇F₁ for the seven NILs showed a continuous variation in pollen and spikelet fertility. The four identified pollen sterility QTLs were located one each on chromosomes 1, 3, 7 and 7. Pollen sterility loci qSS-3 and qSS-7a were on chromosomes 3 and 7, respectively, which coincides with the previously identified S19, and S20, while loci qSS-1 and qSS-7b on chromosomes 1 and 7L appear distinct from all previously reported loci. An epistatic interaction controlling the hybrid sterility was detected between qSS-1 and qSS-7a.     

Mapping genes for grain protein concentration and grain yield on chromosome 5B of Triticum turgidum (L.) var. dicoccoides

Grain protein concentration (GPC) is an important quality factor in durum wheat [Triticum turgidum (L.) var. durum]. Due to the strong environmental influence on GPC, molecular markers linked to quantitative trait loci (QTL) affecting GPC have the potential to be valuable in wheat breeding programs. Various quantitative traits in a population of 133 recombinant inbred chromosome lines were studied in replicated trials at three locations in North Dakota. Segregation for GPC, 1000-kernel weight, gluten strength, heading date, and plant height was observed. By relating phenotypic data to a linkage map obtained from the same population, three QTL affecting GPC, and one affecting yield were identified. The genotypic coefficients of determination for both traits were high.     

QTL mapping of fiber quality in an elite hybrid derived-RIL population of upland cotton

Xiangzamian 2 (XZM2) was the most widely cultivated cotton hybrid planted as F₁ hybrids and as selfed F₂ seeds in China before the release of transgenic Bt hybrids. By crossing two parents of XZM2, Gossypium hirsutum cv. Zhongmiansuo12 (ZMS12) and G. hirsutum acc. 8891, and through subsequent selfings, we obtained F₈ and F₉ populations of 180 recombinant inbred lines (RILs). A RIL population was cultivated in two cotton-growing regions in China for 2 years. The purpose of the present research was to detect quantitative trait loci (QTL) for fiber quality and provide information applicable to cotton breeding. A genetic map was constructed mainly using SSR markers. QTL controlling fiber quality traits were determined at the single-locus and two-locus levels, and genotype-by-environment interactions were analyzed. Among the main-effect QTL, a fiber length QTL qFL-D2-1 and a reflectance QTL qFR-D2-1 were simultaneously detected at two growing regions in 2 years, which suggested a high degree of stability in different environments, and might be of particular value for a marker-assisted selection (MAS) program. The results suggested that epistatic effects, as well as additive effects, of QTL play important roles in fiber quality in these RILs. In our research, the phenomenon of QTL clusters was detected in the cotton genome.     

Identification and mapping of a quantitative trait locus controlling extreme late bolting in Chinese cabbage (Brassica rapa L. ssp. pekinensis syn. campestris L.) using bulked segregant analysis

DNA markers linked to a locus controlling an extreme late bolting trait, which was originally found in a local cultivar of a non-heading leafy vegetable,‘Osaka Shirona Bansei’ (Brassica rapa L. ssp. pekinensis syn. campestris L.) were identified using bulked segregant analysis. A doubled haploid (DH) line, DH27, which is a progeny of ‘Osaka Shirona Bansei’, shows extreme late bolting, and bolts without vernalization. DH27 was crossed with a normal bolting DH line, G309. The plantlets of the parents, F₁ and F₂, were vernalized and then grown in a greenhouse. The bolting time of F₂ plants showed a continuous distribution from 19 to 231 days after vernalization (DAV), suggesting the effects of a few major genes and polygenes. Possible linkage markers for this trait were screened by modified bulked segregant analysis (BSA). The BSA using four bulks suggested that a 530-bp RAPD band RA1255C was linked to a locus controlling the bolting trait. The RAPD band was cloned and used as a probe to detect RFLP. The fragment detected a single locus, BN007-1,the segregation of which in the F₂ population matched that of RA1255C. Three other RAPDs were found to be linked to BN007-1. A quantitative trait locus(QTL) affecting the bolting time was detected around BN007-1 using MAPMAKER/QTL. Since the difference between bolting times of both the parental genotypes in the F₂ was 138 days, these markers may be useful for a marker-assisted selection (MAS) in the breeding program for late bolting or bolting-resistant cultivars in B. rapa crops. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Inheritance of root traits and phosphorus uptake in common bean (Phaseolus vulgaris L.) under limited soil phosphorus supply

Heterosis is an important way to improve yield and quality for many crops. Hybrid rice and hybrid maize contributed to enhanced productivity which is essential to supply enough food for the increasing world population. The success of hybrid rice in China has led to a continuous interest in hybrid wheat, even when most research on hybrid wheat has been discontinued in other countries for various reasons including low heterosis and high seed production costs. The Timopheevii cytoplasmic male sterile system is ideal for producing hybrid wheat seeds when fertility restoration lines with strong fertility restoration ability are available. To develop PCR-based molecular markers for use in marker-assisted selection of fertility restorer lines, two F₂ populations derived from crosses R18/ND36 and R9034/ND36 were used to map fertility restoration genes in the two elite fertility restorer lines (R-lines) R18 and R9034. Over 678 SSR markers were analyzed, and markers closely linked to fertility restoration genes were identified. Using SSR markers, a major fertility restoration gene, Rf3, was located on the 1B chromosome in both populations. This gene was partially dominant in conferring fertility restoration in the two restorer lines. SSR markers Xbarc207, Xgwm131, and Xbarc61 are close to this gene. These markers may be useful in marker-assisted selection of new restorer lines with T. timopheevii cytoplasm. Two minor QTL conferring fertility restoration were also identified on chromosomes 5A (in R18) and 7D (in R9034) in two R-lines.     

Molecular mapping and identification of QTL's associated to oat crown rust partial resistance

Summary Molecular mapping is a promising strategy for studying and understanding traits with complex genetic control, such as partial resistance to oat crown rust. The objectives of this research were to develop molecular maps from the progenies of the cross UFRGS7 (susceptible) × UFRGS910906 (partially resistant) and to identify QTLs (quantitative trait loci) associated to partial resistance to oat crown rust in two generations of that population.DNA of 86 genotypes of the F₂ and 90 genotypes of the F₆ UFRGS7 × UFRGS910906 population were used to generate AFLP markers. Molecular maps were constructed using Mapmaker Exp. 3.0 and QTLs for partial resistance to oat crown rust were identified with Mapmaker/QTL software. Five hundred and fifty seven markers in the F₂ and 243 markers in the F₆ generations were identified. The F₂ map integrated 250 markers in 37 linkage groups. The F₆ map integrated 86 markers in 17 linkage groups.Five QTLs were identified for partial resistance to oat crown rust in the F₂ generation and three QTLs in the F₆. The QTL identified on F₆ through the PaaaMctt340 AFLP marker showed consistency across two environments and two generations (F₄ and F₆), and appear to have potential for marker-assisted selection in oat.     

Breeding tomato for pollen tolerance to low temperatures by gametophytic selection

Haploid selection for traits related to pollen cold tolerance in tomato was performed in segregating populations derived from a Lycopersicon esculentum × L. pennellii hybrid. BC₁ populations were obtained by combining normal and low temperature treatments on two stages of pollen development: pollen formation, and germination and pollen tube growth. F₁ hybrids were cultivated under low and normal temperatures and their pollen was used to pollinate L. esculentum plants at low and normal temperatures. The four BC₁ populations obtained were tested for the quality and quantity of pollen produced at low temperatures. The population obtained by cold treatment at both stages had a significantly improved pollen germination ability at low temperatures. The two other coldselected BC₁ populations showed no differences compared with the unselected BC population. A second cycle of pollen selection, corresponding to BC₂, was applied in order to test its persistence in the subsequent generations and the possibility to further improve the character. This second cycle showed no improvement although some plants retained the high pollen germination ability at low temperatures that was observed in the first cycle. Hence, gametophytic selection of some characters related with tomato pollen performance may be feasible, at least for the first cycle of selection.     

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.