TdERF1, an ethylene response factor associated with dehydration responses in durum wheat (Triticum turgidum L. subsp. durum)

Water deficit and increasing salinization reduce productivity of wheat, the leading crop for human diet. While the complete genome sequence of this crop has not been deciphered, a BAC library screening allowed the isolation of TdERF1, the first ethylene response factor gene from durum wheat. This gene is putatively involved in mediating salt stress tolerance and its characterization provides clues toward understanding the mechanisms underlying the adaptation/tolerance of durum wheat to suboptimal growth conditions. TdERF1 expression is differentially induced by high salt treatment in 2 durum wheat varieties, the salt-tolerant Grecale (GR) and the salt-sensitive Om Rabiaa (OR). To further extend these findings, we show here that the expression of this ERF is correlated with physiological parameters, such as the accumulation of osmo-regulators and membrane integrity, that discriminate between the 2 contrasted wheat genotypes. The data confirm that GR and OR are 2 contrasted wheat genotypes with regard to salt-stress and show that TdERF1 is also induced by water stress with an expression pattern clearly discriminating between the 2 genotypes. These findings suggest that TdERF1 might be involved in responses to salt and water stress providing a potential genetic marker discriminating between tolerant and sensitive wheat varieties.     

Biofortification of durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husnot) grains with nutrients

Durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husnot) was grown under conditions to promote mineral biofortification at the grain level. Along plant development, biomass accumulation and the kinetics of nutrients accumulation were assessed, identifying the nutrient fluxes of roots and shoots, and the timescale constraints of crop biofortification. Plants were grown under environmentally controlled conditions, submitted to four increasing concentrations of nutrient solutions (1-, 2-, 4- and 6-fold) of micro- (Fe, Zn, Cu and Mn) and macronutrients (Ca, K, P and Mg). The threshold of mineral toxicity was not reached as evaluated through plant biomass accumulation, but considering grain yield, the twofold nutrient concentration was the best treatment for biofortification. In the different treatments, the contents and the mineral unrests of roots uptake and shoots translocation varied, at different magnitudes and trends, before the onset of booting and from the physiological maturity onwards. Except for Cu, all mineral nutrients were mainly detected in the bran and embryo of the grains; therefore, the production of biofortified pasta for human consumption requires the use of integral semolina.     

AFLP and pedigree-based genetic diversity estimates in modern cultivars of durum wheat [ Triticum turgidum L. subsp. durum (Desf.) Husn.

A substantial amount of between and within cultivar genetic variation was detected in all the 13 registered modern Canadian durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husn.) cultivars based upon amplified restriction fragment polymorphism (AFLP). Of the approximately 950 detected AFLP markers, only 89 were polymorphic, with 41 between cultivars whereas the remaining 48 showed polymorphism within at least one cultivar. The ancestry of Canadian durum wheat cultivars was traced back to 125 cultivars, selections, and breeding lines including 17 landraces. Mean pair-wise genetic distance based on the kinship coefficient was 0.76. On the other hand, AFLP-based mean pair-wise genetic distance was 0.40. Even though there was a large difference between the means of the two diversity measures, a moderate positive correlation (r=0.457, p<0.002) was detected between the two distance matrices. Cluster analysis with the entire AFLP data divided all cultivars into three major groups reflecting their breeding origins. One group contained ’Pelissier’ alone, which was a selection from a landrace introduced into the US from Algeria. On the other hand such groupings among cultivars were not evident when KIN was used for genetic diversity measures instead. The level of genetic variation among individuals within a cultivar at the breeders’ seed level was estimated based on an inter-haplotypic distance matrix derived from the AFLP data. We found that the level of genetic variation within the most-developed cultivars is fairly substantial despite rigorous selection pressure aimed at cultivar purity in breeding programs. Comparison of AFLP and pedigree-based genetic diversity estimates in crop species such as durum wheat can provide important information for plant improvement. Received: 26 January 2001 / Accepted: 31 May 2001     

Targeted mapping of Cdu1, a major locus regulating grain cadmium concentration in durum wheat (Triticum turgidum L. var durum)

Some durum wheat (Triticum turgidum L. var durum) cultivars have the genetic propensity to accumulate cadmium (Cd) in the grain. A major gene controlling grain Cd concentration designated as Cdu1 has been reported on 5B, but the genetic factor(s) conferring the low Cd phenotype are currently unknown. The objectives of this study were to saturate the chromosomal region harboring Cdu1 with newly developed PCR-based markers and to investigate the colinearity of this wheat chromosomal region with rice (Oryza sativa L.) and Brachypodium distachyon genomes. Genetic mapping of markers linked to Cdu1 in a population of recombinant inbred substitution lines revealed that the gene(s) associated with variation in Cd concentration resides in wheat bin 5BL9 between fraction breakpoints 0.76 and 0.79. Genetic mapping and quantitative trait locus (QTL) analysis of grain Cd concentration was performed in 155 doubled haploid lines from the cross W9262-260D3 (low Cd) by Kofa (high Cd) revealed two expressed sequence tag markers (ESMs) and one sequence tagged site (STS) marker that co-segregated with Cdu1 and explained >80% of the phenotypic variation in grain Cd concentration. A second, minor QTL for grain Cd concentration was also identified on 5B, 67 cM proximal to Cdu1. The Cdu1 interval spans 286 kbp of rice chromosome 3 and 282 kbp of Brachypodium chromosome 1. The markers and rice and Brachypodium colinearity described here represent tools that will assist in the positional cloning of Cdu1 and can be used to select for low Cd accumulation in durum wheat breeding programs targeting this trait. The isolation of Cdu1 will further our knowledge of Cd accumulation in cereals as well as metal accumulation in general.     

Molecular linkage map for an intraspecific recombinant inbred population of durum wheat (Triticum turgidum L. var. durum)

Durum wheat (Triticum turgidum L. var. durum) is an economically and nutritionally important cereal crop in the Mediterranean region. To further our understanding of durum genome organization we constructed a durum linkage map using restriction fragment length polymorphisms (RFLPs), simple sequence repeats (SSRs) known as Gatersleben wheat microsatellites (GWMs), amplified fragment length polymorphisms (AFLPs), and seed storage proteins (SSPs: gliadins and glutenins). A population of 110 F₉ recombinant inbred lines (RILs) was derived from an intraspecific cross between two durum cultivars, Jennah Khetifa and Cham 1. The two parents exhibit contrasting traits for resistance to biotic and abiotic stresses and for grain quality. In total, 306 markers have been placed on the linkage map – 138 RFLPs, 26 SSRs, 134 AFLPs, five SSPs, and three known genes (one pyruvate decarboxylase and two lipoxygenases). The map is 3598 cM long, with an average distance between markers of 11.8 cM, and 12.1% of the markers deviated significantly from the expected Mendelian ratio 1:1. The molecular markers were evenly distributed between the A and B genomes. The chromosome with the most markers is 1B (41 markers), followed by 3B and 7B, with 25 markers each. The chromosomes with the fewest markers are 2A (11 markers), 5A (12 markers), and 4B (15 markers). In general, there is a good agreement between the map obtained and the Triticeae linkage consensus maps. This intraspecific map provides a useful tool for marker-assisted selection and map-based breeding for resistance to biotic and abiotic stresses and for improvement of grain quality. Received: 14 February 2000 / Accepted: 28 April 2000     

Assessment of genetic diversity among Syrian durum (Triticum turgidum ssp. durum) and bread wheat (Triticum aestivum L.) using SSR markers

Genetic diversity among 49 wheat varieties (37 durum and 12 bread wheat) was assayed using 32 microsatellites representing 34 loci covering almost the whole wheat genome. The polymorphic information content (PIC) across the tested loci ranged from 0 to 0.88 with average values of 0.57 and 0.65 for durum and bread wheat respectively. B genome had the highest mean number of alleles (10.91) followed by A genome (8.3) whereas D genome had the lowest number (4.73). The correlation between PIC and allele number was significant in all genome groups accounting for 0.87, 074 and 0.84 for A, B and D genomes respectively, and over all genomes, the correlation was higher in tetraploid (0.8) than in hexaploid wheat varieties (0.5). The cluster analysis discriminated all varieties and clearly divided the two ploidy levels into two separate clusters that reflect the differences in genetic diversity within each cluster. This study demonstrates that microsatellites markers have unique advantages compared to other molecular and biochemical fingerprinting techniques in revealing the genetic diversity in Syrian wheat varieties that is crucial for wheat improvement.     

Genetic variation of salt-stressed durum wheat (Triticum turgidum subsp. durum Desf.) genotypes under field conditions and gynogenetic capacity

Agriculture has new challenges against the climate change: the preservation of genetic resources and the rapid creation of new varieties better adapted to abiotic stress, specially salinity. In this context, the agronomic performance of 25 durum wheat (Triticum turgidum subsp. durum Desf.) genotypes (nineteen landraces and six improved varieties), cultivated in two semi-arid regions in the center area of Tunisia, were assessed. These sites (Echbika, 2.2?g?l^?1; Barrouta, 4.2?g?l^?1) differ by their degree of salinity of the water irrigation. The results showed that most of the agronomic traits (e.g. spike per meter square, thousand kernels weight and grain yield) were reduced by salinity. Durum wheat landraces, Mahmoudi and Hmira, and improved varieties, Maali and Om Rabia showed the widest adaptability to different quality of irrigation water. Genotypes including Jneh Kotifa and Arbi were estimated as stable genotypes under adverse conditions. Thereafter, salt-tolerant (Hmira and Jneh Khotifa) and the most cultivated high-yielding (Karim, Razzak and Khiar) genotypes were tested for their gynogenetic ability to obtain haploids and doubled haploid lines. Genotypes with good induction capacity had not necessarily a good capacity of regeneration of haploid plantlets. In our conditions, Hmira and Khiar exhibited the best gynogenetic ability (3.1% and 2.9% of haploid plantlets, respectively).     

Copper phytotoxicity affects root elongation and iron nutrition in durum wheat (Triticum turgidum durum L.)

This work investigated how copper (Cu) phytotoxicity affected iron (Fe) nutrition and root elongation in hydroponically grown durum wheat (Triticum turgidum durum L., cv Acalou) in order to establish the critical level of Cu concentration in roots above which significant Cu phytotoxicity occurs. This was assessed at two levels of Fe supply (2 and 100 μM). Severe symptoms of Cu phytotoxicity were observed at Cu²⁺ concentration above 1 μM, i.e. interveinal chlorosis symptoms and global root growth alteration. Total root Cu concentration of about 100, 150 and 250–300 mg kg^?1 corresponded to 10%, 25% and 50% reduction in root elongation, respectively. Copper and Fe concentrations as well as amounts of Cu and Fe accumulated in shoots varied inversely which suggested an antagonism between Cu and Fe leading to Fe deficiency. In addition, the root-induced release of complexing compounds increased significantly with increasing Cu concentration in nutrient solution and was positively correlated with Cu uptake without significant difference between the two Fe treatments (high and low Fe supply). This work suggests that total root Cu concentration might be a simple, sensitive indicator of Cu rhizotoxicity. It also indicated that Cu phytotoxicity which may have resulted in Fe deficiency and significant increase in root-induced release of complexing compounds (presumably phytosiderophores) was independent of the level of Fe supply provided that the threshold values of phytotoxicity were based on the free Cu-ion concentration.     

Accumulation of cadmium in near-isogenic lines of durum wheat (Triticum turgidum L. var durum): the role of transpiration

Concentrations of cadmium in the grain of durum wheat (Triticum turgidum L. var durum) are often above the internationally acceptable limit of 0.2 mg kg⁻¹. Cultivars that vary in concentrations of cadmium in the grain have been identified but the physiology behind differential accumulation has not been determined. Three pairs of near-isogenic lines (isolines) of durum wheat that vary in aboveground cadmium accumulation (8982-TL ‘high’ and ‘low’, W9260-BC ‘high’ and ‘low’, and W9261-BG ‘high’ and ‘low’) were used to test the hypothesis that the greater amounts of cadmium in shoots of the ‘high’ isolines are correlated with greater volumes of water transpired. In general, cadmium content was positively correlated with transpiration only in the ‘low’ isolines. Although shoots of the ‘high’ isolines of W9260-BC and W9261-BG contained higher concentrations of cadmium than did their corresponding ‘low’ isolines, they did not transpire larger volumes of water. In addition, isolines of 8982-TL transpired less water than did the other pairs of isolines yet both ‘high’ and ‘low’ isolines of 8982-TL contained higher amounts of cadmium than did the other pairs. The difference between ‘high’ and ‘low’ isolines appears to be related to the relative contribution of transpiration to cadmium translocation to the shoot. Increased transpiration was associated with increased cadmium content in the ‘low’ isolines but in the ‘high’ isolines increased cadmium in the shoot occurred independently of the volume of water transpired.     

Differences in developmental plasticity and growth rate among drought-resistant and susceptible cultivars of durum wheat (Triticum turgidum L. var. durum)

Understanding how growth and development of durum wheat cultivars respond to drought could provide a basis to develop crop improvement programmes in drought-affected tropical and subtropical countries. A greenhouse experiment was conducted to study the responses of five durum wheat cultivars to moisture stress at different developmental phases. Phenology, total dry matter (TDM), relative growth rate (RGR), leaf area ratio (LAR), net assimilation rate (NAR), leaf weight ratio (LWR), specific leaf area (SLA) and shoot:root ratio were compared. Pre-anthesis moisture stress delayed phenological development, whereas post-anthesis moisture stress accelerated it. TDM accumulation rate was different between drought-resistant and susceptible cultivars. RGR and its components changed with age and moisture availability. Drought-resistant cultivars had a high RGR in favourable periods of the growing season and a low RGR during moisture stress. In contrast, the drought-susceptible cultivar (Po) showed an opposite trend. LAR explained the differences in RGR (r=0.788) best, whereas the relationship between NAR and RGR was not significant. Even though both LWR and SLA were important factors determining the potential growth rate, LWR was of major importance to describe cultivar differences in LAR, and consequently in RGR. The drought-resistant cultivars Omrabi-5 and Boohai showed vigorous root development and/or a low shoot:root ratio. It is concluded that biomass allocation is the major factor explaining variation in RGR among the investigated durum wheat cultivars.     

Genotypic difference in root penetration ability by durum wheat (Triticum turgidum L. var. durum) evaluated by a pot with paraffin-Vaseline discs

Improvement of root penetration (RP) ability in durum wheat (Triticum turgidum L. var. durum) is an important breeding target to reduce yield losses due to soil compaction and drought. This study investigated the genotypic difference of RP ability in durum wheat of seven Ethiopian landraces (ET genotypes), which were presumed to adapt to soil compaction, and 17 genotypes bred in North America (NA genotypes). The pregerminated seed was planted in a pot with four paraffin-Vaseline (PV) discs of hardness 0.12, 0.24, 0.50 and 0.73 MPa. After eight weeks from planting, each plant was sampled and the number of roots penetrating through the PV discs was counted. No genotype penetrated through the PV disc of 0.73 MPa hardness. The number of roots penetrating through the PV disc of 0.50 MPa hardness showed a significant difference among the genotypes: highest in an ET genotype and lowest in a NA genotype. The mean value was also significantly higher in ET genotypes than in NA genotypes, and it significantly correlated with root weight density below the disc and plant height. The mean root diameter and root weight density above the disc showed no significant difference between NA and ET genotypes. Our results indicate that a large genotypic difference exists for RP ability in durum wheat, and that ET genotypes have a higher RP ability than NA genotypes.     

Identification of a microsatellite on chromosome 7B showing a strong linkage with yellow pigment in durum wheat (Triticum turgidum L. var. durum)

The objective of this study is to identify QTLs linked to yellow pigment content in durum wheat. A durum-dicoccoides genetic linkage map was constructed using 124 microsatellites, 149 amplified fragment length polymorphism (AFLPs), and six seed storage proteins (SSP) in a population of 114 recombinant inbred lines (F8). The population has been obtained from a cross between a durum cultivar Omrabi5 and Triticum dicoccoides600545 and backcrossed to Omrabi5. The map consists of 14-durum chromosomes plus an unknown group; and shows a good synteny to the previously published wheat maps. Yellow pigment was measured in the population in three different locations during 3 seasons. Analysis of QTLs was based on simple and simplified composite interval mapping (SIM and sCIM). Three QTLs for yellow pigment were detected on the chromosomal group 7 (7AL and 7BL telomeres) explaining 62% of the total variation. On 7BL, a major microsatellite (Xgwm344) explained by itself 53%, whereas on 7AL, the other two QTLs have contributed 13 and 6%. All determined QTLs showed a strong genetic effect and a weak QTL x E effect. The QTLs effect was consistent across all environments and showed a large effect. Consequently, promising QTLs will be used in the marker assisted breeding program to enhance the selection efficiency for yellow pigment.     

Production of doubled haploids in durum wheat (Triticum turgidum L.) through isolated microspore culture

The objective of this work was to produce doubled haploid plants from durum wheat through the induction of androgenesis. A microspore culture technique was developed and used to produce fertile doubled haploid plants of agronomic interest. Five cultivars, one selected line, plus a collection of 20 F₁ crosses between different genotypes of high breeding value were used. Studies on several factors such as pre-treatments and media components were carried out in order to develop a protocol to regenerate green haploid plantlets. Anthers were pre-treated in 0.7 M mannitol. Microspores, from anther maceration, were plated on a C₁₇ induction culture medium with ovary co-culture. The optimum regeneration medium J25–8 was used. From 35 microspore isolations, 407 green plantlets were obtained. With this technique mature embryos were obtained. Green plants were regenerated from all genotypes used and approximately 67% of them were spontaneously doubled haploids. Some haploids and a very few polyploids plants were obtained. From the 407 plants, 275 were completely fertile and gave enough seeds to be assayed in the field. This protocol could be used complementary to or instead of the intergeneric crossing with maize as an economically feasible method to obtain doubled haploids from most durum wheat genotypes.     

A multiparental cross population for mapping QTL for agronomic traits in durum wheat (Triticum turgidum ssp. durum)

Multiparental cross designs for mapping quantitative trait loci (QTL) provide an efficient alternative to biparental populations because of their broader genetic basis and potentially higher mapping resolution. We describe the development and deployment of a recombinant inbred line (RIL) population in durum wheat (Triticum turgidum ssp. durum) obtained by crossing four elite cultivars. A linkage map spanning 2664 cM and including 7594 single nucleotide polymorphisms (SNPs) was produced by genotyping 338 RILs. QTL analysis was carried out by both interval mapping on founder haplotype probabilities and SNP bi‐allelic tests for heading date and maturity date, plant height and grain yield from four field experiments. Sixteen QTL were identified across environments and detection methods, including two yield QTL on chromosomes 2BL and 7AS, with the former mapped independently from the photoperiod response gene Ppd‐B1, while the latter overlapped with the vernalization locus VRN‐A3. Additionally, 21 QTL with environment‐specific effects were found. Our results indicated a prevalence of environment‐specific QTL with relatively small effect on the control of grain yield. For all traits, functionally different QTL alleles in terms of direction and size of genetic effect were distributed among parents. We showed that QTL results based on founder haplotypes closely matched functional alleles at known heading date loci. Despite the four founders, only 2.1 different functional haplotypes were estimated per QTL, on average. This durum wheat population provides a mapping resource for detailed genetic dissection of agronomic traits in an elite background typical of breeding programmes.     

Transformation of pasta wheat (Triticum turgidum L. var. durum) with high-molecular-weight glutenin subunit genes and modification of dough functionality

Particle bombardment has been used to transform three cultivars (L35, Ofanto, Svevo) and one breeding line (Latino × Lira) of durum wheat (Triticum turgidum L. var. durum). These varieties were co-transformed with plasmids containing selectable and scorable marker genes (bar and uidA) and plasmids containing one of two high-molecular-weight (HMW) glutenin subunit genes (encoding subunits 1Ax1 or 1Dx5). Ten independent transgenic lines were recovered from 1683 bombarded scutella (transformation efficiency thus 0.6%). Five lines expressed either subunit 1Dx5 or 1Ax1 at levels similar to those of endogenous subunits encoded on chromosome 1B. To identify the effects of the transgenes on the functional properties of grain, three lines showing segregation for transgene expression were used to isolate sibling T₂ plants which were null or positive for the transgene product. Analysis of these plants using a small-scale mixograph showed that expression of the additional subunits resulted in increased dough strength and stability, demonstrating that transformation can be used to modify the quality of durum wheat for bread and pasta making.