Exome association analysis sheds light onto leaf rust (Puccinia triticina) resistance genes currently used in wheat breeding (Triticum aestivum L.)

Resistance breeding is crucial for a sustainable control of leaf rust (Puccinia triticina) in wheat (Triticum aestivum L.) while directly targeting functional variants is the Holy Grail for efficient marker‐assisted selection and map‐based cloning. We assessed the limits and prospects of exome association analysis for severity of leaf rust in a large hybrid wheat population of 1574 single‐crosses plus their 133 parents. After imputation and quality control, exome sequencing revealed 202 875 single‐nucleotide polymorphisms (SNPs) covering 19.7% of the high‐confidence annotated gene space. We performed intensive data mining and found significant associations for 2171 SNPs corresponding to 50 different loci. Some of these associations mapped in the proximity of the already known resistance genes Lr21, Lr34‐B, Lr1 and Lr10, while other associated genomic regions, such as those on chromosomes 1A and 3D, harboured several annotated genes putatively involved in resistance. Validation with an independent population helped to narrow down the list of putative resistance genes that should be targeted by fine‐mapping. We expect that the proposed strategy of intensive data mining coupled with validation will significantly influence research in plant genetics and breeding.     

Identification of leaf rust resistance genes in wheat (Triticum aestivum L.) cultivars using molecular markers

A collection of 68 cultivars of common wheat has been screened for leaf rust resistance genes with the use of molecular markers. Markers of genes Lr1, Lr9, Lr10, Lr19, Lr20, Lr21, Lr24, and Lr26 have been used. It has been suggested that allele Xgwm295 be used as a marker for identifying the Lr34 gene. The genes originating from Triticum aestivum L., as well as the Lr26 gene contained in rye translocation 1RS, are the most frequent. Genes originating from wild wheats were rarer in the cultivars studied.     

Identification of leaf rust resistance genes in wheat (Triticum aestivum L.) cultivars using molecular markers

A collection of 68 cultivars of common wheat has been screened for leaf rust resistance genes with the use of molecular markers. Markers of genes Lr1, Lr9, Lr10, Lr19, Lr20, Lr21, Lr24, and Lr26 have been used. It has been suggested that allele Xgwm295 be used as a marker for identifying the Lr34 gene. The genes originating from Triticum aestivum L., as well as the Lr26 gene contained in rye translocation 1RS, are the most frequent. Genes originating from wild wheats were rarer in the cultivars studied.     

Endogenous gibberellins in young seedlings of wheat (Triticum aestivum) cultivars

Gibberellins A₁, A₃, A₄ and A₇ were identified by combined gas chromatography mass spectrometry (GC-MS) in leaf and stem tissues of 17-day-old seedlings of wheat ( Triticum aestivum L. ), cvs Siete Cerros (semi-dwarf, Rht1) and Møystad (tall), of F₁, hybrids from the cross Møystad × Siete Cerros and of 2 selected lines from the cross Møystad x Sonora 64 (Rht1 and Rht2). GA, and GA, were identified by full scan mass spectra separately in all 5 extracts, GA₄ and GA₇, were identified by selected ion monitoring in a bulked fraction. About 90% of the biological activity (Tan-ginbozu dwarf rice bioassay) in all 5 extracts was due to the GA₁/GA₃-fraction.     

Identification and molecular tagging of a gene from PI 289824 conferring resistance to leaf rust (Puccinia triticina) in wheat

Host-plant resistance is the most economically viable and environmentally responsible method of control for Puccinia triticina, the causal agent of leaf rust in wheat (Triticum aestivum L.). The identification and utilization of new resistance sources is critical to the continued development of improved cultivars as shifts in pathogen races cause the effectiveness of widely deployed genes to be short lived. The objectives of this research were to identify and tag new leaf rust resistance genes. Forty landraces from Afghanistan and Iran were obtained from the National Plant Germplasm System and evaluated under field conditions at two locations in Texas. PI 289824, a landrace from Iran, was highly resistant under field infection. Further evaluation revealed that PI 289824 is highly resistant to a broad spectrum of leaf rust races, including the currently prevalent races of leaf rust in the Great Plains area of the USA. Eight F₁ plants, 176 F₂ individuals and 139 F_2:3 families of a cross between PI 289824 and T112 (susceptible) were evaluated for resistance to leaf rust at the seedling stage. Genetic analysis indicated resistance in PI 289824 is controlled by a single dominant gene. The AFLP analyses resulted in the identification of a marker (P39 M48-367) linked to resistance. The diagnostic AFLP band was sequenced and that sequence information was used to develop an STS marker (TXW₂₀₀) linked to the gene at a distance of 2.3 cM. The addition of microsatellite markers allowed the gene to be mapped to the short arm of Chromosome 5B. The only resistance gene to be assigned to Chr 5BS is Lr52. The Lr52 gene was reported to be 16.5 cM distal to Xgwm443 while the gene in PI 289824 mapped 16.7 cM proximal to Xgwm443. Allelism tests are needed to determine the relationship between the gene in PI 289824 and Lr52. If the reported map positions are correct, the gene in PI 289824 is unique.     

Leaf rust resistance genes of wheat: identification in cultivars and resistance sources

Thirty-seven wheat cultivars originating from seven European countries were examined by using sequence tagged site (STS) markers for seven Lr (leaf rust = brown rust) resistance genes against the fungal pathogen of wheat Puccinia recondita f. sp. tritici (Lr9, Lr10, Lr19, Lr24, Lr26 and Lr37). Additionally, 22 accessions with various Lr genes from two germplasm collections were tested. A Scar (sequence-characterized amplified region) marker for Lr24 and a CAPS (Cleaved Amplified Polymorphic Sequence) marker for Lr47 were also used to identify those genes in the wheat accessions. Each marker amplified one specific DNA fragment. Three Lr gene markers were identified in wheat cultivars (Lr10, Lr26 and Lr37). Another four markers (Lr9, Lr19, Lr24 and Lr47) were found in breeding lines carrying leaf rust resistance genes. The results were compared with leaf rust resistance gene postulations made in previous studies, based on multipathotype testing. Markers for Lr10, Lr26 and Lr37 may be useful in marker-assisted breeding.     

Population divergence in the wheat leaf rust fungus Puccinia triticina is correlated with wheat evolution

Co-evolution of fungal pathogens with their host species during the domestication of modern crop varieties has likely affected the current genetic divergence of pathogen populations. The objective of this study was to determine if the evolutionary history of the obligate rust pathogen on wheat, Puccinia triticina, is correlated with adaptation to hosts with different ploidy levels. Sequence data from 15 loci with different levels of polymorphism were generated. Phylogenetic analyses (parsimony, Bayesian, maximum likelihood) showed the clear initial divergence of P. triticina isolates collected from Aegilops speltoides (the likely B genome donor of modern wheat) in Israel from the other isolates that were collected from tetraploid (AB genomes) durum wheat and hexaploid (ABD genomes) common wheat. Coalescence-based genealogy samplers also indicated that P. triticina on A. speltoides, diverged initially, followed by P. triticina isolates from durum wheat in Ethiopia and then by isolates from common wheat. Isolates of P. triticina found worldwide on cultivated durum wheat were the most recently coalesced and formed a clade nested within the isolates from common wheat. By a relative time scale, the divergence of P. triticinia as delimited by host specificity appears very recent. Significant reciprocal gene flow between isolates from common wheat and isolates from durum wheat that are found worldwide was detected, in addition to gene flow from isolates on common wheat to isolates on durum wheat in Ethiopia.     

Analysis of the wheat and Puccinia triticina (leaf rust) proteomes during a susceptible host-pathogen interaction

Wheat leaf rust is caused by the fungus Puccinia triticina. The genetics of resistance follows the gene-for-gene hypothesis, and thus the presence or absence of a single host resistance gene renders a plant resistant or susceptible to a leaf rust race bearing the corresponding avirulence gene. To investigate some of the changes in the proteomes of both host and pathogen during disease development, a susceptible line of wheat infected with a virulent race of leaf rust were compared to mock-inoculated wheat using 2-DE (with IEF pH 4-8) and MS. Up-regulated protein spots were excised and analyzed by MALDI-QqTOF MS/MS, followed by cross-species protein identification. Where possible MS/MS spectra were matched to homologous proteins in the NCBI database or to fungal ESTs encoding putative proteins. Searching was done using the MASCOT search engine. Remaining unmatched spectra were then sequenced de novo and queried against the NCBInr database using the BLAST and MS BLAST tools. A total of 32 consistently up-regulated proteins were examined from the gels representing the 9-day post-infection proteome in susceptible plants. Of these 7 are host proteins, 22 are fungal proteins of known or hypothetical function and 3 are unknown proteins of putative fungal origin.     

Allelopathy in wheat (Triticum aestivum)

Wheat (Triticum aestivum) allelopathy has potential for the management of weeds, pests and diseases. Both wheat residue allelopathy and wheat seedling allelopathy can be exploited for managing weeds, including resistant biotypes. Wheat varieties differ in allelopathic potential against weeds, indicating that selection of allelopathic varieties might be a useful strategy in integrated weed management. Several categories of allelochemicals for wheat allelopathy have been identified, namely, phenolic acids, hydroxamic acids and short‐chain fatty acids. Wheat allelopathic activity is genetically controlled and a multigenic model has been proposed. Research is underway to identify genetic markers associated with wheat allelopathy. Once allelopathic genes have been located, a breeding programme could be initiated to transfer the genes into modern varieties for weed suppression. The negative impacts of wheat autotoxicity on agricultural production systems have also been identified when wheat straws are retained on the soil surface for conservation farming purposes. A management package to avoid such deleterious effects is discussed. Wheat allelopathy requires further study in order to maximise its allelopathic potential for the control of weeds, pests and diseases, and to minimise its detrimental effects on the growth of wheat and other crops.     

Kinetic properties of cell wall bound superoxide dismutase in leaves of wheat (Triticum aestivum L.) following stripe rust (Puccinia striiformis) infection

Stripe rust (Puccinia striiformis f.sp. tritici) is the most devastating disease of wheat (Triticum aestivum L.) accounting huge economical losses to the industry worldwide. HD 2329 was a widely grown wheat cultivar which had become highly susceptible to stripe rust and was used to understand the biochemical aspects of the host pathogen interaction through characterization of superoxide dismutase (SOD). In the present study, two types of SOD, ionically or covalently bound to the particulate fraction were found in the stripe rust infected and uninfected wheat leaves of susceptible cultivar HD 2329. Cell walls of leaves contained a high level of SOD, of which 41-44% was extractable by 2 M NaCl and 10-13% by 0.5% EDTA in infected and uninfected leaves. The NaCl-released SOD constituted the predominant fraction. It exhibited maximum activity at pH 9.0, had a Km value of 1.82-2.51 for uninfected and 1.77-2.37 mM for infected, respectively with pyrogallol as the substrate, and a Vmax of 9.55-21.4 and 12.4-24.1 delta A min(-1)g(-1)FW. A temperature optimum of 20 degrees C was observed for SOD of both uninfected and infected leaves. SOD showed differential response to metal ions, suggesting their distinctive nature. Inhibition of wall bound SOD by iodine and its partial regeneration of activity by mercaptoethanol suggested the involvement of cysteine in active site of the enzyme. These two forms showed greater differences with respect to thermodynamic properties like energy of activation (Ea) and enthalpy change (delta H), while entropy change (delta S) and free energy change (delta G) were similar. The results further showed that pathogen infection of the leaves of susceptible wheat cultivar induced a decrease in the SOD activity and kinetics which might be critical during the response of plant cells to the infection.     

Genetic and physical characterization of theLR1 leaf rust resistance locus in wheat (Triticum aestivum L.)

The objective of this study was to characterize the leaf rust resistance locusLr1 in wheat. Restriction fragment length polymorphism (RELP) analysis was performed on the resistant lineLr1/6^*Thatcher and the susceptible varieties Thatcher and Frisal, as well as on the segregating F₂ populations. Seventeen out of 37 RFLP probes mapping to group 5 chromosomes showed polymorphism betweenLr1/6^*Thatcher and Frisal, whereas 11 probes were polymorphic between the near-isogenic lines (NILs)Lr1/6^*Thatcher and Thatcher. Three of these probes were linked to the resistance gene in the segregating F₂ populations. One probe (pTAG621) showed very tight linkage toLr1 and mapped to a single-copy region on chromosome 5D. The map location of pTAG621 at the end of the long arm of chromosome 5D was confirmed by the absence of the band in the nulli-tetrasomic line N5DT5B of Chinese Spring and a set of deletion lines of Chinese Spring lacking the distal part of 5DL. Twenty-seven breeding lines containing theLr1 resistance gene in different genetic backgrounds showed the same band asLr1/6^*Thatcher when hybridized with pTAG621. The RFLP marker was converted to a sequence-tagged-site marker using polymerase chain reaction (PCR) amplification. Sequencing of the specific fragment amplified from both NILs revealed point mutations as well as small insertion/deletion events. These were used to design primers that allowed amplification of a specific product only from the resistant lineLr1/6^*Thatcher. This STS, specific for theLr1 resistance gene, will allow efficient selection for the disease resistance gene in wheat breeding programmes. In addition, the identification of a D-genome-specific probe tightly linked toLr1 should ultimately provide the basis for positional cloning of the gene.     

Inheritance and expression of stripe rust resistance in common wheat (Triticum aestivum) transferred from Aegilops tauschii and its utilization

Stripe rust is one of the most destructive diseases for wheat crops in China. Two stripe rust physiological strains, i.e. CYR30 (intern. name: 175E191) and CYR31 (intern. name: 293E175) have been the dominant and epidemic physiological strains since 1994. One Aegilops tauschii accession (SQ-214) from CIMMYT was found immune from or highly resistant to Chinese new stripe rust races CYR30 and CYR31 at adult stage. SQ-214 was crossed with a highly susceptible Ae. tauschii accession As-80. Analysis of data from F1-F2 populations of SQ-214/As-80 revealed that the resistance was controlled by a single dominant gene. To exploit the resistance for wheat breeding, SQ-214 was crossed with Chinese Spring (CS) and backcrossed by two Chinese commercial wheat varieties MY26 and SW3243. The resistance from SQ-214 was suppressed in the F1 hybrids (CS/SQ-214) and the F2 population of CS/SQ-214//MY26. However, the resistance of SQ-214 was expressed in several F2 individuals of CS/SQ-214//SW3243. Eleven advanced lines with high level of resistance to the Chinese stripe rust CYR30 and CYR31 have been developed. This result suggested that SW3243 does not suppress the expression of the Chinese stripe rust and should be used as wheat germplasm for exploiting resistance of Ae. tauschii in wheat breeding. The gliadin electrophoretic pattern of the eleven advanced lines with high stripe rust resistances was compared with their parents SQ-214, CS and SW3243 by acid polyacrylamide gel electrophoresis. The omega-gliadin bands of Gli-Dt1 in Ae. tauschii SQ-214 were transferred to some advanced lines and freely expressed in common wheat genetic background. One of advanced lines possesses a null Gli-D1 allele, where the omega-gliadin bands encoding by the Gli-D1 allele were absent. The potential utilization of this advanced line for wheat quality and stripe rust resistance breeding is also discussed in this paper.     

In vitro selection for Russian wheat aphid (Diuraphis noxia) resistance in wheat (Triticum aestivum)

The Russian wheat aphid (Diuraphis noxia) is a pest on wheat (Triticum aestivum) in many regions of the world. The aphid injects a phytotoxin when it feeds. Identification of somaclonal variants with phytotoxin resistance may shorten development time for resistance. Wheat calli from the susceptible cultivar Stephens were exposed to an extract from the aphid. Five plants were regenerated from 100 treated calli. Resistance to the aphid was observed in both the R₂ and R₃ generations. One of the six R₃ populations had improved resistance for leaf curling and leaf folding, while another had improved response for chlorosis damage. These results indicate that the use of aphid extract on wheat callus offers an alternate method for development of resistance to the Russian wheat aphid.     

Leaf primordia initiation,leaf emergence and tillering in wheat (Triticum aestivum L.) grown under low-phosphorus conditions

In two simultaneous experiments we examined the effects of phosphorus (P) supply on leaf area development in wheat (Triticum aestivum L.) grown in sand with nutrient solutions. In Experiment 1 we studied leaf emergence, leaf elongation, tiller emergence, shoot growth, and P uptake under four levels of P supply (mM) 0.025 (P1), 0.05 (P2), 0.1 (P3), and 0.5 (P4), and. In Experiment 2 there were two levels of P supply, P1 and P4, and we examined the effects of P on leaf primordia differentiation and leaf emergence. The phyllochron was calculated as the inverse of the rate of leaf emergence calculated from the regression of number of leaf tips (PHY-Ltip), Haun index (PHY-Haun), and as the cumulated thermal time between the emergence of two consecutive leaves (PHYtt). The plastochron was calculated from the inverse of the rate of leaf primordia initiation in the apex. P deficiency delayed the emergence of leaves on the main stem and on the tiller 1. Phosphorus deficiency increased the time from emergence to double ridge and anthesis. The final number of leaves was not affected by P. The effects of P on the value of the phyllochron were attributed to both a reduced rate of leaf primordia initiation, and to a reduced leaf elongation rate. P deficiency delayed or even suppressed the emergence of certain tillers. In this work a phosphorus deficiency that reduced shoot growth by 25% at 44 days after emergence significantly modified the structure of the plants by increasing the value of the phyllochron and delaying tillering. These results suggest that any attempt to simulate leaf area development and growth of wheat plants for P-limited conditions should include the effects of the deficiency on leaf emergence.     

Marker-assisted selection for leaf rust resistance genes Lr19 and Lr24 in wheat (Triticum aestivum L.)

Leaf rust caused by Puccinia recondita f.sp. tritici is a wheat disease of worldwide importance. Wheat genotypes known to carry specific rust resistance genes and segregating lines that originated from various cross combinations and derived from distinct F2 lineage, so as to represent a diverse genetic background, were included in the present study for validation of molecular markers for Lr19 and Lr24. STS markers detected the presence of the leaf rust resistance gene Lr19 in a Thatcher NIL (Tc*Lrl9) and Inia66//CMH81A575 and of the gene Lr24 in the genotypes Arkan, Blue Boy II, Agent and CI 17907. Validation of molecular markers for Lr19 and Lr24 in parental lines, followed by successful detection of these genes in F3 lines from various cross combinations, was carried out. The molecular test corresponded well with the host-pathogen interaction test response of these lines.     

小麦化感作用研究进展

小麦是世界第一大粮食作物,在农业生产中占有重要地位.然而,由于人们为保证小麦产量往往施用大量的除草剂和杀菌剂,对环境造成了极大的危害.小麦化感作用是利用小麦活体或残体向环境中释放次生代谢物质对自身或其他生物产生作用,它克服了除草剂和杀菌剂等引起的环境污染问题,具有抑制杂草控制病害的潜力.本文对已有的小麦化感作用的研究进展情况进行了综合评述.其中小麦对杂草、虫害及病害产生防御功能的主要化感物质为异羟肟酸和酚酸类物质.小麦化感物质活性的发挥除了取决于化感物质的种类外,还由小麦自身的遗传因素、环境因素和生物因素的共同作用所决定.小麦化感物质在根际土壤中的滞留、迁移和转化过程、小麦化感作用与土壤生物的关系以及相关的作用机理是小麦化感作用研究的薄弱环节,其研究方法还需进一步探索改进.小麦化感作用在植物保护、环境保护以及作物育种等方面具有广泛的应用前景,促进了小麦抗逆性的增强以及产量和品质的提高.     

小麦化感作用研究进展

小麦是世界第一大粮食作物,在农业生产中占有重要地位．然而,由于人们为保证小麦产量往往施用大量的除草剂和杀菌剂,对环境造成了极大的危害．小麦化感作用是利用小麦活体或残体向环境中释放次生代谢物质对自身或其他生物产生作用,它克服了除草剂和杀菌剂等引起的环境污染问题,具有抑制杂草控制病害的潜力．本文对已有的小麦化感作用的研究进展情况进行了综合评述．其中小麦对杂草、虫害及病害产生防御功能的主要化感物质为异羟肟酸和酚酸类物质．小麦化感物质活性的发挥除了取决于化感物质的种类外,还由小麦自身的遗传因素、环境因素和生物因素的共同作用所决定．小麦化感物质在根际土壤中的滞留、迁移和转化过程、小麦化感作用与土壤生物的关系以及相关的作用机理是小麦化感作用研究的薄弱环节。其研究方法还需进一步探索改进．小麦化感作用在植物保护、环境保护以及作物育种等方面具有广泛的应用前景,促进了小麦抗逆性的增强以及产量和品质的提高．