R. L. Conner

Molecular characterization of the genome composition of partial amphiploids derived from Triticum aestivum×Thinopyrum ponticum and T. aestivum×Th. intermedium as sources of resistance to wheat streak mosaic virus and its vector, Aceria tosichella

Abstract Wheat streak mosaic virus (WSMV), vectored by the wheat curl mite (WCM), is one of the most important viral diseases of wheat (Triticum aestivum) in the world. Genetic resistance to WSMV and the WCM does not exist in wheat. Resistance to WSMV and the WCM was evaluated in five different partial amphiploids namely Agrotana, OK7211542, ORRPX, Zhong 5 and TAF 46, which were derived from hybrids of wheat with decaploid Thinopyrum ponticum or with hexaploid Th. intermedium. Agrotana was shown to be immune to WSMV and the WCM; the other four partial amphiploids were susceptible to the WCM. Genomic in situ hybridization (GISH) using genomic DNA probes from Th. elongatum (EE, 2n=14), Th. bessarabicum (JJ, 2n=14), Pseudoroegneria strigosa (SS, 2n=14) and T. aestivum (AABBDD, 2n=42) demonstrated that three of the partial amphiploids, Agrotana, OK7211542 and ORRPX, have almost identical alien genome constitutions: all have 16 alien chromosomes, with 8 chromosomes being closely related to the Js genome and 8 chromosomes belonging to the E or J genomes and no evidence of any S-genome chromosomes. GISH confirmed that the alien genomes of Agrotana and OK7211542, like ORRPX, were all derived from Th. ponticum, and not from Th. intermedium. However, in contrast to Agrotana, ORRPX and OK7211542 were susceptible to the WCM and WSMV. The partial amphiploid Zhong 5 had a reconstituted alien genome composed of 4 S-and 4 Js-genome chromosomes of Th. intermedium with 6 translocated chromosomes between the S and Js genomes. This line was highly resistant to WSMV, but was susceptible to the WCM. TAF 46, which contained a synthetic genome consisting of 3 pairs of S-genome chromosomes and 4 pairs of E- or J-genome chromosomes in addition to the 21 pairs of wheat chromosomes, was susceptible to the WCM, but moderately resistant to WSMV. Agrotana offers great potential for transferring WSMV and WCM resistance into wheat.

Molecular cytogenetic characterization of Thinopyrum intermedium-derived wheat germplasm specifying resistance to wheat streak mosaic virus

Abstract  Thinopyrum intermedium is a promising source of resistance to wheat streak mosaic virus (WSMV), a devastating disease of wheat. Three wheat germplasm lines possessing resistance to WSMV, derived from Triticum aestivum×Th. intermedium crosses, are analyzed by C-banding and genomic in situ hybridization (GISH) to determine the amount and location of alien chromatin in the transfer lines. Line CI15092 was confirmed as a disomic substitution line in which wheat chromosome 4A was replaced by Th. intermedium chromosome 4Ai?2. The other two lines, CI17766 and A29-13-3, carry an identical Robertsonian translocation chromosome in which the complete short arm of chromosome 4Ai?2 was transferred to the long arm of wheat chromosome 4A. Fluorescence in situ hybridization (FISH) using ABD genomic DNA from wheat as a probe and S genomic DNA from Pseudoroegneria stipifolia as the blocker, and vice versa, revealed that the entire short arm of the translocation was derived from the short arm of chromosome 4Ai?2 and the breakpoint was located at the centromere. Chromosomal arm ratios (L/S) of 2.12 in CI17766 and 2.15 in A29-13-3 showed that the translocated chromosome is submetacentric. This translocated chromosome is designated as T4AL ⋅ 4Ai?2S as suggested by Friebe et al. (1991).

Molecular characterization of Haynaldia villosa chromatin in wheat lines carrying resistance to wheat curl mite colonization

Wheat-Haynaldia villosa (L.) Schur, hybrid lines were tested as potential sources of resistance to colonization by the wheat curl mite, the vector of wheat streak mosaic virus. Two lines, Add 6V-1 and Sub 6V-1, were found to be mite-resistant. Fluorescence in situ hybridization using total genomic DNA, from H. villosa in the presence of unlabelled wheat DNA, confirmed that Add 6V-1 is a disomic wheat-H. villosa chromosome addition line. Sub 6V-1 turned out to be a homoeologous wheat-H. villosa chromosome translocation line rather than a substitution. The translocation in Sub 6V-1 occurred between a wheat chromosome and a chromosome from H. villosa through Robertsonian fusion of misdivided centromeres. Only the short arm of the group 6 chromosome of H. villosa was involved in the genetic control of mite resistance, a conclusion based on the genomic in situ hybridization signal and specific DNA fragments obtained by polymerase chain reaction.

Impact of powdery mildew on the yield of soft white spring wheat cultivars

The effect of powdery mildew (Blumeria graminis f. sp. tritici) on the grain yield and protein content of one susceptible, Springfield, and three moderately resistant cultivars, Fielder, AC Reed and AC Nanda, of soft white spring wheat (Triticum aestivum) was examined at two field locations near Lethbridge and Vauxhall, Alberta, in 1999 and 2000. At the start of heading, powdery mildew development was suppressed in half of the plots of each cultivar by a single spray application of the fungicide Tilt (propiconazole). Severe powdery mildew infection of the susceptible cultivar Springfield resulted in yield reductions ranging from 11.4 to 19.9%. The grain yield of the moderately resistant cultivar Fielder was significantly reduced at both sites in 1999 by 7.6–10.5% while AC Reed suffered a significant yield loss (7.6–9.1%) at Lethbridge in both years. The moderately resistant cultivar AC Nanda consistently had the lowest powdery mildew ratings and its yield was unaffected by the disease. A single fungicide ...

The Stripe Rust Resistance Gene Yr10 Encodes an Evolutionary-Conserved and Unique CC–NBS–LRR Sequence in Wheat

The first seedling or all-stage resistance (R) R gene against stripe rust isolated from Moro wheat (Triticum aestivum L.) using a map-based cloning approach was identified as Yr10. Clone 4B of this gene encodes a highly evolutionary-conserved and unique CC-NBS-LRR sequence. Clone 4E, a homolog of Yr10, but lacking transcription start site (TSS) and putative TATA-box and CAAT-box, is likely a non-expressed pseudogene. Clones 4B and 4E are 84% identical and divergent in the intron and the LRR domain. Gene silencing and transgenesis were used in conjunction with inoculation with differentially avirulent and virulent stripe rust strains to demonstrate Yr10 functionality. The Yr10 CC-NBS-LRR sequence is unique among known CC-NBS-LRR R genes in wheat but highly conserved homologs (E = 0.0) were identified in Aegilops tauschii and other monocots including Hordeum vulgare and Brachypodium distachyon. Related sequences were also identified in genomic databases of maize, rice, and in sorghum. This is the first report of a CC-NBS-LRR resistance gene in plants with limited homologies in its native host, but with numerous homologous R genes in related monocots that are either host or non-hosts for stripe rust. These results represent a unique example of gene evolution and dispersion across species.

Characterization of Wheat-Triticale Lines Resistant to Powdery Mildew, Stem Rust, Stripe Rust, Wheat Curl Mite, and Limitation on Spread of WSMV

High yield potential and the wide adaptability of wheat-rye T1BL·1RS translocation lines are attractive to breeders. The wheat-rye lines Lankao 1, 3, 4, and 5 were resistant to a wide spectrum of wheat powdery mildew (Blumeria graminis f. sp. tritici) isolates from both China and Canada. They also were resistant to a mixture of wheat stem rust (Puccinia graminis f. sp. tritici) pathotypes (98WSR) and wheat stripe rust (P. striiformis f. sp. tritici) races from western Canada and China. Colonization of wheat curl mite (WCM) (Aceria tosichella) resulted in slower development of rolling and trapping leaves in the Lankao lines than in the WCM-susceptible check cultivars. The delayed development of Wheat streak mosaic (WSM) symptoms on Lankao lines was observed when transmitted by viruliferous WCM, even though they were susceptible to Wheat streak mosaic virus (WSMV). This effect of Lankao lines on limiting the spread of WSM was comparable with other known sources of WCM resistance. Sequential C-banding and genomic in situ hybridization analyses revealed the presence of a pair of T1BL·1RS translocated chromosomes in the Lankao lines. Segregation analysis of the F2 progeny plants derived from crosses between Lankao 4 and the susceptible wheat cvs. Mingxian 169 and Lovrin 13 indicated that a single dominant gene was responsible for the isolate-specific resistance against wheat powdery mildew in Lankao 4. Polymerase chain reaction analysis using an STS marker amplified rye chromatin in powdery mildew-resistant and -susceptible F2 plants of the Mingxian 169 × Lankao 4 cross demonstrated that the resistance of Lankao 4 was not controlled by a gene or genes located on the rye chromosome arm of T1BL·1RS. The resistance of the Lankao lines to diseases and limitation of the spread of WSMV, in combination with good quality and high yield potential, makes them useful for wheat improvement and production.

Virulence of Puccinia striiformis f. sp. tritici, cause of stripe rust on wheat, in western Canada from 1984 to 2002

Fifty-seven isolates of Puccinia striiformis f. sp. tritici, causing stripe rust on wheat, were collected in western Canada from 1984 to 2002. The virulence of these isolates was tested on the 17 World and European differentials as well as 7 supplemental differentials. Thirty-nine races were identified. Before 2000, races virulent on wheat ‘Lee’ (Yr7, Yr22, Yr23) and ‘Owens’ mainly originated from Creston, British Columbia, while most of the races from the prairies were avirulent on these two cultivars. New races detected in Canada since 2000 were virulent on both ‘Compair’ (Yr8, Yr19) and ‘Clement’ (Yr9, YrCle). Race 33 (238E150) was repeatedly detected from 2000 to 2002, and other new races had a virulence spectrum that was similar to that of race 33. Some of the prevalent races of stripe rust in the prairie provinces were also detected in the Creston area of British Columbia, suggesting a possible link in the source of inoculum between these two areas. None of the races were virulent on Triticum spelta var. album (Yr5). ‘Chinese 166’ (Yr1), ‘Moro’ (Yr10, YrMor), ‘Hybrid 46’ (Yr3b, Yr4b), Triticum dicoccoides selection G-25 (Yr15), and ‘Spalding Prolific’ (YrSP) were resistant to more than 90% of the races detected in western Canada, and all of them were resistant to the new races detected since 2000. Climatic conditions and cultivar resistance play an important role in the survival and propagation of these races on the Canadian prairies.

Different reactions to the Wheat curl mite and Wheat streak mosaic virus in various wheat-Haynaldia villosa 6V and 6VS lines

Wheat curl mite (WCM), Aceria tosichella, is the vector of Wheat streak mosaic virus (WSMV), a destructive viral pathogen in wheat (Triticum aestivum). Genetic resistance to WCM colonization can reduce the incidence of wheat streak mosaic. Chromosome 6V in Hay-naldia villosa is a new source of WCM resistance. We compared variation in resistance among different sources of H. villosa chromosome 6V and 6VS lines to WCM and WSMV and their effectiveness in controlling the incidence of WSMV following exposure to viruliferous WCM. WCM resistance varied among the 6V and 6VS lines depending on the H. villosa parent. The 6V substitution lines Yi80928, GN21, and GN22 derived from an accession of H. villosa from China, and the 6VS translocation lines 92R137, 92R178, and Sub6V from an H. villosa accession collected from the United Kingdom were uniformly resistant to WCM colonization. In contrast, the 6V substitution line RW15 and a 6VS translocation line Pm33 developed from an H. villosa collection from the former Union of Soviet Socialist Republics were susceptible to WCM. All 6V and 6VS lines were susceptible to WSMV when manually inoculated. However, symptom expression was delayed in the WCM-resistant 6V and 6VS lines after exposure to viruliferous WCM. The 6V and 6VS lines differed in their ability to control WSMV infection. WCM-susceptible lines RW15 and Pm33 had no effect on controlling the infection by WSMV. Lines GN21 and GN22 were the most effective of the three H. villosa sources in limiting the spread of WSMV. Their high yield potential and protein content, in combination with resistance to stripe rust (Puccinia striiformis f. sp. tritici) and powdery mildew (Erysiphe graminis f. sp. tritici), make GN21 and GN22 promising sources of WCM resistance.

Molecular cytogenetic evidence for a high level of chromosome pairing among different genomes in Triticum aestivum - Thinopyrum intermedium hybrids

Abstract Intergeneric hybrids (ABDJJsS genomes) were made between Triticum aestivum cv. Chinese Spring (CS) and Thinopyrum intermedium. Genomic in situ hybridization (GISH) using genomic DNA probes from Pseudoroegneria libanotica (Hackel) D.R. Dewey (genome S, 2n = 14) was used to study chromosome pairing among J, Js, S and wheat ABD genomes in the hybrids. It was shown that in the hexaploid (ABDJJsS) hybrids, high pairing occurred among wheat chromosomes and among Thinopyrum chromosomes. A closer relationship was observed among the three genomes of Th. intermedium than among the three genomes of T. aestivum. It was further discerned that S genome chromosomes paired with J- and Js-genome chromosomes at a high frequency. The frequency of heterologous pairing between S and J or S and Js chromosomes was higher than those between J and Js chromosomes, indicating that the S-genome was more closely related with these two genomes. Our results provided direct molecular cytogenetic evidence for the hypothesis that S-genome chromosomes are genetically similar to the J-genome chromosomes and, therefore, genetic exchange between these genomes is possible. The discovery of a close relationship among S, J and Js genomes provides valuable markers for molecular cytogenetic analyses using S-genomic DNA probes in monitoring the transfer of useful traits from Thinopyrum species into wheat.

Promising genetic resources for resistance to wheat streak mosaic virus and the wheat curl mite in wheat-Thinopyrum partial amphiploids and their derivatives

Wheat streak mosaic virus (WSMV), vectored by the wheat curl mite (WCM),Aceria tosichella Keifer, is one of the most destructive viral diseases of wheat found in many wheat producing areas of the world. Host resistance is the most effective method for controlling this disease and its vector. Symptomatological analysis and enzyme-linked immunosorbent assay (ELISA) were used to characterize WSMV-resistance in wheat-alien partial amphiploid lines and their derivatives. The results showed that most of partial amphiploids derived fromThinopyrum ponticum andTh. intermedium were free of systemic symptoms with very low ELISA readings that were similar to that of the non-inoculated Chinese Spring control. While the partial amphiploid lines 693 and PWM706 were identified as new genetic resources of resistance to WSMV. The present study demonstrated that both symptomatological and ELISA methods efficiently assessed WSMV-resistance in the wheat-alien hybrids and systemic symptom incidence and ELISA absorbance readings were highly correlated (r2 = 0.8658−0.9323) over time following inoculation. The ELISA results also indicated that the virus did not buildup in the plant tissues of these virus-resistant partial amphiploids. Similar results were observed in chromosome translocation and substitution lines that have the geneWsm1 conferring WSMV resistance. However, the lines containing the geneWsm1 and all the partial amphiploid lines, except Agrotana, were susceptible to the WCM. One line derived from a cross of wheat and Agrotana, was effective in controlling the spread of WSMV and was highly resistant to the WCM. Another line and an accession ofTriticum dicoccoides (Koern.) Schweinf. were highly susceptible to WSMV and WCM. Early disease development was delayed in a new hard red winter cultivar McClintock. The partial WSMV-resistance of McClintock was demonstrated by initially low ELISA readings, and a lower percentage of infected plants than other WSMV-susceptible wheat. The use of the identified promising sources of resistance to WSMV and the WCM in wheat breeding is discussed.