Peter Stamp

Quantitative trait loci for resistance against powdery mildew in a segregating wheat × spelt population

Abstract Powdery mildew is one of the major diseases of wheat in regions with a maritime or semi-continental climate and can strongly affect grain yield. The attempt to control powdery mildew with major resistance genes (Pm genes) has not provided a durable resistance. Breeding for quantitative resistance to powdery mildew is more promising, but is difficult to select on a phenotypic basis. In this study, we mapped and characterised quantitative trait loci (QTLs) for adult-plant powdery mildew resistance in a segregating population of 226 recombinant inbred lines derived from the cross of the Swiss wheat variety Forno with the Swiss spelt variety Oberkulmer. Forno possibly contains the Pm5 gene and showed good adult-plant resistance in the field. Oberkulmer does not have any known Pm gene and showed a moderate susceptible reaction. Powdery mildew resistance was assessed in field trials at two locations in 1995 and at three locations in 1996. The high heritability (h2=0.97) for powdery mildew resistance suggests that the environmental influence did not affect the resistance phenotype to a great extent. QTL analysis was based on a genetic map containing 182 loci with 23 linkage groups (2469 cM). With the method of composite interval mapping 18 QTLs for powdery mildew resistance were detected, explaining 77% of the phenotypic variance in a simultaneous fit. Two QTLs with major effects were consistent over all five environments. One of them corresponds to the Pm5 locus derived from Forno on chromosome 7B. The other QTL on 5A, was derived from the spelt variety Oberkulmer and did not correspond to any known Pm gene. In addition, five QTLs were consistent over three environments, and six QTLs over two environments. The QTL at the Pm5 locus showed a large effect, although virulent races for Pm5 were present in the mixture of isolates. Molecular markers linked with QTLs for adult-plant resistance offer the possibility of simultaneous marker-assisted selection for major and minor genes.

Effect of growing season on the photosynthetic apparatus and leaf antioxidative defenses in two maize genotypes of different chilling tolerance

Abstract The effect of the growing season on the photosynthetic apparatus and on leaf antioxidants was investigated in a chilling-tolerant genotype (Z7) and a chilling-sensitive genotype (Penjalinan) of maize (Zea mays). Maize was sown in the field in Switzerland in 3 consecutive weeks, so plants sown early were exposed to chilling conditions, whereas those sown later developed under more favorable growth conditions. Measurements of the quantum efficiency of CO2 fixation (ΦCO2) and the photosystem II (ΦPSII) were made simultaneously on the third fully expanded leaves. The activity of scavenging enzymes and the content of pigments and antioxidants were also determined. Leaves that developed under chilling conditions showed typical chill-induced alterations, namely low photosynthetic capacity and efficiency, reduction in the pigment contents, and a decrease in catalase activity. These alterations were more pronounced in the chilling-sensitive than in the chilling-tolerant genotype. Determining the ΦPSII/ΦCO2 ratio indicated that it was very unlikely that alternative electron sinks, such as the Mehler reaction, were activated to a significant extent in either of the genotypes. This was supported by the measurements of the activity of enzymes involved in the Mehler ascorbate peroxidase reaction. However, a comparison of the genotypes showed that chilling tolerance might be correlated with an increase in the α-tocopherol and glutathione contents as well as in the activity of glutathione reductase.

Quantitative trait loci for lodging resistance in a segregating wheat×spelt population

Abstract Lodging can strongly affect both the grain yield and the quality of wheat. Lodging represents a quantitative trait and is difficult to assess on a phenotypic basis. Marker-assisted selection (MAS) could therefore become an important tool in breeding for lodging resistance. In this study, we mapped and characterised quantitative trait loci (QTLs) for lodging resistance, as well as morphological traits correlated with lodging, in a segregating population of 226 recombinant inbred lines derived from the cross of the lodging-resistant wheat variety Forno with the susceptible spelt variety Oberkulmer. Lodging, plant height, leaf width, leaf-growth habit, culm stiffness, culm swinging, culm thickness, days to ear emergence and days to flowering were assessed in field trials at two locations in 1996 and at one location in 1997. Additionally, at one location weight and length parameters were also assessed. Plant height and culm stiffness explained 77% of the phenotypic variance of lodging in a multiple regression model over all three environments. QTL analysis of lodging and morphological parameters was based on a genetic map containing 230 loci with 23 linkage groups (2469 cM). With the method of composite interval mapping nine QTLs for lodging resistance were detected, explaining 63% of the phenotypic variance in a simultaneous fit. Seven of these QTLs coincided with QTLs for morphological traits, reflecting the correlations between these traits and lodging. In our population the most efficient way to improve lodging resistance would be by a combination of indirect selection on plant height and culm stiffness together with MAS on the two QTLs for lodging resistance which did not coincide with QTLs for morphological traits.

Glutathione synthesis in maize genotypes with different sensitivities to chilling

The effect of chilling on enzymes, substrates and products of sulfate reduction, gultathione synthesis and metabolism was studied in shoots and roots of maize (Zea mays L.) genotypes with different chilling sensitivity. At full expansion of the second leaf, chilling at 12 °C inhibited dry weight increase in shoots and roots compared to controls at 25 °C and induced an increase in adenosine 5′-phosphosulfate sulfotransferase and γ-glutamylcysteine synthetase (EC 6.3.2.2) activity in the second leaf of all genotypes tested. Glutathione synthetase (EC 6.3.2.3) activity was about one order of magnitude higher than γ-glutamylcysteine synthetase activity, but remained unchanged during chilling except for one genotype. During chilling, cysteine and glutathione content of second leaves increased to significantly higher levels in the two most chilling-tolerant genotypes. Comparing the most tolerant and most sensitive genotype showed that chilling induced a greater incorporation of35S from [35S]sulfate into cysteine and glutathione in the chilling-tolerant than in the sensitive genotype. Chilling decreased the amount of35S-label incorporated into proteins in shoots of both genotypes, but had no effect on this incorporation in the roots. Glutathione reductase (EC 1.6.4.2) and nitrate reductase (EC 1.6.6.1) activity were constitutively higher in the chilling-tolerant genotypes, but showed no changes in most examined genotypes during 3 d at 12 °C. Our results indicate that in maize glutathione is involved in protection against chilling damage.

Genetic analysis of cold-tolerance of photosynthesis in maize

The genetic basis of cold-tolerance was investigated by analyzing the quantitative trait loci (QTL) of an F2:3 population derived from a cross between two lines bred for contrasting cold-tolerance using chlorophyll fluorescence as a selection tool. Chlorophyll fluorescence parameters, CO2 exchange rate, leaf greenness, shoot dry matter and shoot nitrogen content were determined in plants grown under controlled conditions at 25/22 °C or 15/13 °C (day/night). The analysis revealed the presence of 18 and 19 QTLs (LOD > 3.5) significantly involved in the variation of nine target traits in plants grown at 25/22 °C and 15/13 °C, respectively. Only four QTLs were clearly identified in both temperatures regimes for the same traits, demonstrating that the genetic control of the performance of the photosynthetic apparatus differed, depending on the temperature regime. A major QTL for the cold-tolerance of photosynthesis was identified on chromosome 6. This QTL alone explained 37.4 of the phenotypic variance in the chronic photoinhibition at low temperature and was significantly involved in the expression of six other traits, including the rate of carbon fixation and shoot dry matter accumulation, indicating that the tolerance to photoinhibition is a key factor in the tolerance of maize to low growth temperature. An additional QTL on chromosomes 2 corresponded to a QTL identified previously in another population, suggesting some common genetic basis of the cold-tolerance of photosynthesis in different maize germplasms.

Growth of axile and lateral roots of maize: I development of a phenotying platform

The objective of this study was to develop a phenotyping platform for the non-destructive, digital measurement of early root growth of axile and lateral roots and to evaluate its suitability for identifying maize (Zea mays L.) genotypes with contrasting root development. The system was designed to capture images of the root system within minutes and to batch process them automatically. For system establishment, roots of the inbred line Ac7729/TZSRW were grown until nine days after germination on the surface of a blotting paper in pouches. An A4 scanner was used for image acquisition followed by digital image analysis. Image processing was optimized to enhance the separation between the roots and the background and to remove image noise. Based on the root length in diameter-class distribution (RLDD), small-diameter lateral roots and large-diameter axile roots were separated. Root systems were scanned daily to model the growth dynamics of these root types. While the axile roots exhibited an almost linear growth, total lateral root length increased exponentially. Given the determined exponential growth, it was demonstrated that two plants, germinated one day apart but with the same growth rates differed in root length by 100%. From the growth rates we were able to identify contrasting genotypes from 236 recombinant inbred lines (RILs) of the CML444 x SC-Malawi cross. Differences in the growth of lateral roots of two selected RILs were due to differences in the final length and linear density of the primary lateral roots, as proven by the manual reanalysis of the digital images. The high throughput makes the phenotyping platform attractive for routine genetic studies and other screening purposes.

Root distribution and morphology of maize seedlings as affected by tillage and fertilizer placement

Suboptimal soil conditions are known to result in poor early growth of maize (Zea mays L.) in no-tillage (NT) systems in contrast with conventional tillage (CT) systems. However, most studies have generally focused on maize roots at later growth stages and/or do not give details on root morphology. In a 2-year field study at two locations (silt loam and loam soils) in the Swiss midlands, we investigated the impacts of tillage intensity, NT vs. CT, and NP-fertilizer sidebanding on the morphology, vertical and horizontal distribution, and nutrient uptake of maize roots at the V6 growth stage. The length density (RLD) and the length per diameter-class distribution (LDD) of the roots were determined from soil cores taken to a depth of 0.5 m and at distances of 0.05 and 0.15 m from both sides of the maize row. The temperature of the topsoil was lower, and the bulk density and penetration resistance were greater in the topsoil of NT compared with CT. The growth and the development of the shoot were slower in NT. RLD was greater and the mean root diameter smaller in CT than in NT, while the vertical and horizontal distribution of roots did not differ between CT and NT. RLD increased in the zone enriched by the sidebanded fertilizer, independent of the tillage system, but LDD did not change. The poorer growth of the roots and shoots of maize seedlings was presumably caused by the lower topsoil temperature in NT rather than by mechanical impedance. The placement of a starter fertilizer at planting under NT is emphasized.

Low temperature stress in maize (Zea mays L.) induces genes involved in photosynthesis and signal transduction as studied by suppression subtractive hybridization

Unfavourable environmental conditions such as cold induce the transcription of a range of genes in plants in order to acclimate to these growth conditions. To better understand the cold acclimation of maize (Zea mays L.) it is important to identify components of the cold stress response. For this purpose, cold-induced genes were analysed using the PCR-select cDNA subtraction method. We identified several novel genes isolated from maize seedling exposed for 48h to 6 degrees C. Of 18 Zea mays cold-induced genes (ZmCOI genes) characterized, the majority share similarities with proteins with known function in signal transduction and photosynthesis regulation. RT-PCR was conducted for a selected group of genes, namely ZmCOI6.1, ZmACA1, ZmDREB2A and ZmERF3, confirming the induction by low temperature. In addition, it was found that their expression was strongly induced by other abiotic stresses such as drought and high salt concentration, by stress signalling molecules such as jasmonic acid, salicylic acid and abscisic acid, and by membrane rigidification. These results suggest that this group of genes is involved in a general response to abiotic stresses.

QTLs for the elongation of axile and lateral roots of maize in response to low water potential.

Changes in root architecture and the maintenance of root growth in drying soil are key traits for the adaptation of maize (Zea mays L.) to drought environments. The goal of this study was to map quantitative trait loci (QTLs) for root growth and its response to dehydration in a population of 208 recombinant inbred lines from the International Maize and Wheat Improvement Center (CIMMYT). The parents, Ac7643 and Ac7729/TZSRW, are known to be drought-tolerant and drought-sensitive, respectively. Roots were grown in pouches under well-watered conditions or at low water potential induced by the osmolyte polyethylene glycol (PEG 8000). Axile root length (LAx) increased linearly, while lateral root length (LLat) increased exponentially over time. Thirteen QTLs were identified for six seedling traits: elongation rates of axile roots (ERAx), the rate constant of lateral root elongation (kLat), the final respective lengths (LAx and LLat), and the ratios kLat/ERAx and LLat/LAx. While QTLs for lateral root traits were constitutively expressed, most QTLs for axile root traits responded to water stress. For axile roots, common QTLs existed for ERAx and LAx. Quantitative trait loci for the elongation rates of axile roots responded more clearly to water stress compared to root length. Two major QTLs were detected: a QTL for general vigor in bin 2.02, affecting most of the traits, and a QTL for the constitutive increase in kLat and kLat/ERAx in bins 6.04–6.05. The latter co-located with a major QTL for the anthesis-silking interval (ASI) reported in published field experiments, suggesting an involvement of root morphology in drought tolerance. Rapid seedling tests are feasible for elucidating the genetic response of root growth to low water potential. Some loci may even have pleiotropic effects on yield-related traits under drought stress.

Inheritance of resistance to leaf and glume blotch caused by Septoria nodorum Berk, in winter wheat

Abstract Sixteen crosses between eight winter wheat cultivars were screened for resistance to Septoria nodorum leaf and glume blotch in the F1 and F4 generations using artificial inoculation in the field. The F1 of most crosses showed dominance for susceptibility on both ear and leaf. The effects of general combining ability were of similar magnitude as the effects for specific combining ability. On the basis of the phenotypic difference of the parents, no prediction was possible about the amount and the direction of genetic variance in the segregating populations. The variation observed in this study both within and among the segregating populations suggests a quantitative inheritance pattern influencing the expression of the two traits. The components of variance between F2 families within a population were as high as (for S. nodorum blotch on the ear) or higher (for S. nodorum blotch on the leaf) than those between populations. Therefore, strong selection within a few populations may be as effective to obtain new resistant genotypes as selection in a large number of populations. In almost all crosses, progenies were found that were more resistant than the better parent. Thus transgression breeding may be a tool to breed for higher levels of resistance to S. nodorum blotch. Highly resistant genotypes were found even in combination with two susceptible parents. The genetic source for Septoria resistance is probably broader than is generally assumed and could be used to improve S. nodorum resistance by combination breeding followed by strong selection in large populations.