Identification of Rye Chromosome 1R Translocations and Substitutions in Hexaploid Wheats Using Storage Proteins as Genetic Markers

Grain protein compositions of 106 advanced generation backcross lines from crosses involving ‘Amigo’ (1AL.1RS), ‘Aurora’, ‘Kavkaz’, ‘Skorospelka-35’ and ‘Sunbird’ (all 1BL.1RS) and ‘Gabo’ 1DL.1RS parents and 152 cultivars with unknown pedigree were analysed by one-dimensional SDS-PAGE. Eighty seven backcross lines and 16 cultivars carried one or other of these translocations, 2 cultivars had a 1R (1B) substitution, whereas 5 backcross lines were found to be heterogeneous for the 1BL.1RS translocation. The translocation lines were easily identified by the presence of secalins (Sec-1) controlled by rye chromosome arm IRS and a simultaneous loss of the gliadin (Gli-1) and/or triticin (Tri-1) protein bands controlled by the replaced wheat chromosome arm (1AS, 1BS or 1DS). Certain gliadins, showing no allelic variation among the genotypes analysed, were identified as markers for chromosome arms 1AS (M_r= 34 kd) and IBS (M_r= 42,33 kd). The whole chromosome substitutions 1R (1B) were recognized by scoring for the presence of Sec-1 and HMW secalin bands, Sec-3 (controlled by rye chromosome arm 1RL) and the absence of Gli-B1 and HMW glutenin subunits, Glu-B1 (controlled by wheat chromosome arm 1BL). The results have shown that protein electrophoresis provides a rapid and reliable technique for screening genotypes for these translocations and substitutions in a breeding programme.     

Genetic control of crossability of triticale with rye

Limited genetic knowledge is available regarding crossability between hexaploid triticale (2n= 6x= 42, 21″, AABBRR, amphiploid Triticum turgidum L.‐Secale cereale L.) and rye (2n= 14, 7″, RR). Our objectives were to determine (1) the crossability between triticales and rye and (2) the inheritance of crossability between F₂ progeny from intertriticale crosses and rye. First, ‘8F/Corgo’, a hexaploid triticale, was crossed as a female with two landrace ryes, ‘Gimonde’ and, ‘Vila Pouca’ and two derived north European cultivars, ‘Pluto’ and ‘Breno’. These crosses produced 21.7, 20.9, 5.9, and 5.6%, seed‐set or crossability, respectively, showing that the landrace ryes produced higher seed‐set than the cultivars. Second, ‘Gimonde’ rye was crossed as a male with four triticales for 3 years. The control cross, ‘Chinese Spring’ wheat × rye, produced 80‐90% seed‐set. Of the four triticales, ‘Beagle’ produced 35.7‐56.8% seed‐set. The other three triticales produced less than 20% seed‐set, showing that the triticales differ in crossability with ‘Gimonde’ rye. Third, six FiS from intertriticale crosses (‘8F/Corgo’בBeagle’, ‘Beagle’בCachirulo’, ‘Lasko’בBeagle’, ‘8F/Corgo’בCachirulo’, ‘Lasko’בCachirulo’, ‘Lasko’ב8F/Corgo’) were crossed to ‘Gimonde’ rye. Results indicated that lower crossability trait was partially dominant in the two F₁S from crosses involving ‘Beagle’(high crossability) with‘8F/Corgo’ and ‘Cachirulo’(low crossability) and completely dominant in the ‘Beagle’בLasko’ cross, as it happens in wheat. Fourth, segregants in four F₂ populations (‘Lasko’בBeagle’, ‘8F/Corgo’בBeagle’, ‘Lasko’ב8F/Corgo’, and‘8F/Corgo’בCachirulo’) were crossed with rye. Segregation for crossability was observed, although distinct segregation classes were blurred by environmental and perhaps other factors, such as self‐incompatibility alleles in rye. Segregation patterns showed that ‘Beagle’, with high crossability to rye, carries either Kr1 or Kr2. The three triticales with low crossability with rye were most likely homozygous for Kr1 and Kr2. Therefore, it is likely that the Kr loci from A and B genomes acting in wheat also play a role in triticale × rye crosses.     

About the origin of 1RS.1BL wheat-rye chromosome translocations from Germany

In order lo investigate the origin of two of the German 1RS. 1BL wheat-rye translocations used world-wide in breeding, a number of DNA probes were considered which (a) were critical for the short arm of the rye chromosome 1 R and (b) should show a specificity for the gene pool of Petkus rye. The DNA probe CDO580 was revealed as a specific one. (1) It clearly differentiated 1RS.1AL (‘Amigo’). 1RS.lBL (‘Salmon’) and 1RS.1DL (‘Gabo’) from the two German sources. (2) Both translocation wheats deriving from the Weihenstephan (Munich) and from the Salzmünde (Halle/S.) origin showed an identical DNA fragment which was typical for the gene pool of Petkus rye. It is supposed that both German sources have one progenitor in common.     

Effects of Wheat Chromosome 1D Telosomes on Hybrid Seed Development in Wheat × Rye Crosses

Pollination of ‘Chinese Spring,’ monosome 1D plants with rye results in failure of hybrid seed development in a proportion of the F₁ seeds corresponding to the transmission rate of the nullisomic 1D egg cells. Development and viability of these hybrid seeds closely resemble that normally observed in T. aurum× rye crosses. Using ‘Chinese Spring’ chromosome ID telosomic plants in crosses with rye, it was possible to illustrate that the observed effect was associated with the long arm of this chromosome.     

Comparative study of the structure of chromosome 1R derived from Secale montanum and Secale cereale

We produced 15 dissection lines of common wheat carrying segments of chromosome 1R of wild rye (Secale montanum) (1R^m) by the gametocidal system. Using the 1R^m dissection lines and previously established 24 dissection lines of chromosome 1R from cultivated rye (Secale cereale cv. ‘Imperial’) (1Rⁱ), we conducted cytological mapping of 97 markers that were amplified in the 1Rⁱ addition line. Sixty‐eight of the 97 markers were amplified in the 1R^m addition line. To reveal what structural differentiation occurred in chromosome 1R during domestication, we compared the cytological map of chromosome 1Rⁱ with that of chromosome 1R^m, and also with the previously published cytological map of chromosome 1R from wheat cultivar ‘Burgas 2’ (1R^B). There was one discrepancy in marker order in the satellite region between chromosomes 1Rⁱ and 1R^B, while there were four discrepancies in marker order between chromosomes 1Rⁱ and 1R^m. These results suggested that during the domestication of rye, some intrachromosomal rearrangements had occurred in chromosome 1R, although this chromosome is regarded as the most stable chromosome in the rye genome.     

Intergeneric Transfer (Rye to Wheat) of a Gene(s) for Russian Wheat Aphid Resistance

An octoploid triticale was derived from the F, of a Russian wheat aphid-resistant rye, ‘Turkey 77’, and ‘Chinese Spring’ wheat. The alloploid was crossed to common wheat, and to ‘Imperial’ rye/‘Chinese Spring’ disomic addition lines. F₂, progeny from these crosses were tested for Russian wheat aphid resistance and C-banded. A resistance gene(s) was found to be associated with chromosome arm IRS of the ‘Turkey 77’ rye genome. A monotelosomic IRS (‘Turkey 77’) addition plant was then crossed with the wheat cultivar ‘Gamtoos’, which has the 1BL.1RS ‘Veery’ translocation. Unlike the IRS segment in ‘Gamtoos’, the ‘Turkey 77’-derived 1 RS telosome did not express the rust resistance genes Sr31 and Ar26, which could then be used as markers. From the F, a monotelosomic 1 RS addition plant that was also heterozygous for the 1BL. 1 RS translocation was selected and testerossed with an aphid-susceptible common wheat, ‘Inia 66’ Meiotic pairing between the rye arms resulted in the recovery of five euploid Russian-wheat-aphid-resistant plants. One recombinant also retained Sr31 and Lr26 and was selfed to produce translocation homozygotes.     

The Identification of a Gibberellic-Acid-Insensitive Gene in Secale cereale*

Among 16 dwarfing genes identified in wheat (Triticum aestiuvm L. em Thell.), four are known to be associated with insensitivity to the externally-supplied growth hormone gibberellin (GA). Rht1 and Rht2 (Reduced height 1 and 2, respectively) have been the most extensively used, because of their positive effect on yield. To increase the germplasm pool for dwarfism, a spring rye (Secale cereale) population (UC-90, CI-174) was selected because it contains high variability and any useful genes would benefit triticale and wheat as well. Seedlings of the CI-174 rye population were treated with 50 ppm of GA to identify any insensitive types. GA-insensitive and -sensitive seedlings were identified and, after three generations of selfing, GA-insensitive and -sensitive lines were fixed. Rye insensitive was crossed to a sensitive wheat and to rye and, reciprocally, insensitive wheat was crossed to sensitive rye. The results indicated that a GA-insensitivity dwarfism system similar to that originally found in wheat also operates in rye and appears to be under simple inheritance. Rye GA-insensitivity was expressed in triticale. Therefore, it is possible to transfer this new source of insensitivity and dwarfism into triticale and wheat.     

Gene Localization of Aspartate Aminotransferase and Endopeptidase Isozymes in Wheat and Rye Using Developmental and Organ-Specific Patterns

Wheat, rye and wheat-rye addition lines have been investigated regarding their developmental and organ-specific isozyme patterns of aspartate amino-transferase (AAT) and endopeptidase (EP). Evidence is given, that development-specific isozymes of AAT are encoded by chromosomes 3R and 4R of ‘Imperial’ rye which can be used as biochemical markers up to leaf age of 14 days. Organ-specific increase of intensity of bands for AAT in stems could be assigned to genes or alleles of chromosome 3A of ‘Chinese Spring’ wheat. For EP new markers were localized on chromosomes 4R and 6R of ‘Imperial’ rye showing variability. Utilization of these markers is possible at all developmental stages of the leaves. Mechanisms of gene regulation are discussed.     

Anther-Culture ability in Secale cereale L.

Success in anther culture of rye (Secale cereale L.) has mainly been restricted to genotypes whose genome is partly determined by S. vavilovii Grossh. The aim of this study was to identify true S. cereale genotypes with anther-culture ability. The plant material used included the semi-wild standard genotype SC35, six single crosses of the ‘Petkus’ and 10 of the ‘Carsten’ gene pool, plus a double cross between SC35 and one of the ‘Carsten’ single crosses. Three anther-culture experiments were performed. Several single crosses within the ‘Carsten’ gene pool produced green plants. The ‘Petkus’ single crosses tested did not show anther-culture ability. As expected, the induction and regeneration rates of SC35 were higher than those of all other genotypes tested. The performance of the double cross was intermediate between its parents. The results demonstrate for the first time that anther culture in rye is a valuable tool in plant breeding. The data also show the possibility of transferring anther-culture ability from high- to low-responding genotypes.     

Identification of Rye Chromosomes Involved in Tolerance to Barley Yellow Dwarf Virus Disease in Wheat × Triticale Hybrids

Common wheat × hexaploid triticale hybrids were produced and evaluated for tolerance to barley yellow dwarf virus disease (BYD). The BYD tolerance expression varied with wheat × triticale combination. The selection for BYD tolerance increased the recovery of tolerant genotypes in the next generations. Homozygous tolerant and susceptible lines were obtained in advanced generations. The rye chromosomes 1R, 2R, and 4R with 7R were transmitted as disomic or monosomic, disomic, and double disomic substitution to the late generations of ‘Musala’ (common wheat) בMuskox 658’ (triticale), ‘Encruzilhada’ (common wheat) בNord Kivu’ (triticale) and ‘Encruzilhada’× 12th. International Triticale Screening Nursery 267 (12ITSN267) (triticale), respectively. A clear association was established between the 1R chromosome of the ‘Muskox 658’ triticale line and the tolerance to BYDV. Results suggest that the 2R chromosome may be involved in BYD tolerance of ‘Nord Kivu’ triticale line.     

Characterization of broad‐spectrum resistance to Soybean mosaic virus in soybean [Glycine max (L.) Merr.] cultivar ‘RN‐9’

Soybean mosaic virus (SMV) can cause serious yield losses in soybean. Soybean cultivar ‘RN‐9’ is resistant to 15 of 21 SMV strains. To well‐characterize this invaluable broad‐spectrum SMV‐resistance, populations (F₁, F₂ and F_2:3) derived from resistant (R) × susceptible (S) and R × R crosses were tested for SMV‐SC18 resistance. Genetic analysis revealed that SC18 resistance in ‘RN‐9’ plus two elite SMV‐resistant genotypes (‘Qihuang No.1’ and ‘Kefeng No.1’) are controlled by independently single dominant genes. Linkage analysis showed that the resistance of ‘RN‐9’ to SMV strains SC10, SC14, SC15 and SC18 is controlled by more than one gene(s). Moreover, Rsc10‐r and Rsc18‐r were both positioned between the two simple sequence repeats markers Satt286 and Satt277, while Rsc14‐r was fine‐mapped in 136.8‐kb genomic region containing sixteen genes, flanked by BARCSOYSSR_06_0786 and BARCSOYSSR_06_0790 at genetic distances of 3.79 and 4.14 cM, respectively. Allelic sequence comparison showed that Cytochrome P450‐encoding genes (Glyma.06g176000 and Glyma.06g176100) likely confer the resistance to SC14 in ‘RN‐9’. Our results would facilitate the breeding of broad‐spectrum and durable SMV resistance in soybeans.     

Frequencies of Vertical Resistances and Virulences in the Rye-Powdery Mildew Pathosystem*

Nine populations of rye (Secale cereale L.; the cultivars ‘Kustro’, ‘Danko’ and ‘Carokurz’. a breeding population PA 14/75 and five Iranian primitive ryes) were tested with three or two pathotypes of powder)’ mildew (Erysiphe graminis DC. f. sp. secalis Marchal) to determine the frequencies of vertical resistances. Similarly, three populations of powder)’ mildew isolated from the above eultivars were tested with two rye pathodemes to estimate the frequencies of vertical virulences. Tests were carried out on leaf segments cultivated in vitro. To explain the pattern of the host-parasite interaction, a model with at least four resistance and virulence genes was required. In the rye populations the genotypes of most plants could be determined unambiguously whereas in the powdery mildew populations no unique classification of one-postule isolates was possible due to the limited number of rye differentials. Both the host and the pathogen populations were polymorphic for resistance and virulence, respectively. In all lye populations except PA 14/75 the resistance frequencies were low. In the mildew populations the virulence frequencies were high and complex races occurred rather frequently. The virulence frequencies were related to the resistance frequencies of the respective host population. Results were compared with mathematical host-parasite models accounting for gene-for-gene interaction and balancing natural selection. Observations agree well with theory.     

Rapid introduction of disease resistance from rye into common wheat by anther culture of a 6x triticale × nulli-tetrasomic wheat

Powdery mildew (caused by Erysiphe graminis) and yellow rust (caused by Puccinia striiformis) are the two most serious wheat diseases found in China. Rye chromosomes, carrying genes for resistance to these diseases, were introduced into common wheat in two generations using chromosome engineering and anther culture. The F₁ hybrids from a cross involving a hexaploid triticale (×Triticosecale Wittmack) בChinese Spring’ nulli‐tetrasomic N6DT6A wheat aneuploid line were anther cultured and doubled‐haploid plants were regenerated. Using genomic in situ hybridization, C‐banding and biochemical marker analyses, one of the anther‐cultured lines (ZH‐1)studied in detail, proved to be a doubled‐haploid with one rye chromosome pair added (1R) and a homozygous 6R/6D substitution (2n= 44). The line was tested for expression of disease resistance and found to be highly resistant to powdery mildew and moderately resistant to yellow rust.     

Quantitative trait loci for scald resistance in barley localized by a non-interval mapping procedure

Three quantitative trait loci (QTL) for scald resistance in barley were identified and mapped in relation to molecular markers using a population of chromosome doubled‐haploid lines produced from the F₁ generation of a cross between the spring barley varieties ‘Alexis’ and ‘Regatta’. Two field experiments were conducted in Denmark and two in Norway to assess disease resistance. The percentage leaf area covered with scald (Rhynchosporium secalis) ranged from 0 to 40% in the 189 doubled‐haploid (DH) lines analysed. One quantitative trait locus was localized in the centromeric region of chromosome 3H, Qryn3, using the MAPQTL program. MAPQTL was unable to provide proper localization of the other two resistance genes and so a non‐interval QTL mapping method was used. One was found to be located distally to markers on chromosome 4H (Qryn4) and the other, Qryn6, was located distally to markers on chromosome 6H. The effects of differences between the Qryn3, Qryn4 and Qryn6 alleles in two barley genotypes for the QTL were estimated to be 8.8%, 7.3% and 7.0%, respectively, of leaf covered by scald. No interactions between the QTLs were found.     

A polymorphic microsatellite in the 3'end of 'waxy' genes of wheat, Triticum aestivum

Potential polymorphism of an (AT)_N microsatellite at the 3’end of waxy genes in bread wheat was examined. Primers were designed from a published cDNA sequence of a wheat waxy gene. Polymerase chain reaction (PCR) amplification of genomic DNA from 135 mainly Australian cultivars revealed eight alleles on chromosome 7A. This polymorphic microsatellite is a potential codominant marker for the Wx-A1 locus in breeding programmes. A distinguishable fragment was also amplified from chromosome 7D. This fragment was absent where a plant was null for the waxy gene on chromosome 7D, being a dominant marker for the Wx-D1 locus. The primers were also useful for amplifying genomic DNA from barley, rye and triticale and can be used to detect potential polymorphism in these species.     

Genetic stability and maintenance of Raphanus sativus lines with an added Brassica rapa chromosome

The genetic stability and maintenance of Raphanus sativus‐Brassica rapa monosomic chromosome addition lines (a‐h‐types MALs, 2n = 19, BC₂), developed by backcrossing the synthesized amphidiploid Raphanobrassica (Raphanus sativus × Brassica rapa, 2n = 38, RRAA) with R. sativus cv. ‘Shogoin’ (2n = 18, RR), was investigated. Transmission of the added alien chromosome through selected smaller seeds (SSS) and the inheritance of morphological traits and random amplified polymorphic DNA (RAPD)‐specific markers together with meiotic chromosome configuration and seed fertility were also investigated for three successive generations (BC₃ to BC₅). The distinctive traits and the RAPD‐specific markers of the eight types of MAL were substantially inherited and stably maintained throughout three generations, although a few variant plants (2n =18) resembling MALs (2n = 19) and hyperploidal plants (2n = 26 and 2n = 37) were generated in the earlier generations of BC₃ and BC₄ in comparison with BC₅. The average transmission rates for three generations ranged from 26% for both the b‐type and the d‐type to 44% for the e‐type through SSS. On the other hand, the transmission rates through randomly selected seeds (RSS) were lower, ranging from 6.5% for the f‐type to 23.5% for the b‐type. In meiosis, more than 90% of PMCs showed the 9II +1I pairing configuration at metaphase I throughout three generations. For seed fertility, when backcrossed with the radish cv. ‘Shogoin’, the values were approximately 180% to 500% with the mode around 300% with the seed harvested from a pod increasing with the advancing generations. Genetic recombination between the radish chromosomes and the added chromosome is probably rare, suggesting that the added chromosome is mostly maintained unaltered in the background of the radish genome.     

Isolation of a new repetitive DNA sequence from Secale africanum enables targeting of Secale chromatin in wheat background

A genome specific DNA sequence that detects Secale africanum chromatin incorporated into wheat was developed in this study. Random amplified polymorphic DNA (RAPD) analysis was used to search for genome specific DNA sequences of S. africanum in lines, R111, “mianyang11” (MY11) and wheat-rye 1RS/1BL translocations R25 and R57. A high copy rye-specific DNA segment pSaD15₉₄₀ of the S. africanum genome was obtained. The sequence of pSaD15 did not show any significant homology to other reported sequences in databases and it is therefore a new repetitive sequence of Secale. PCR primers were designed for pSaD15₉₄₀, which amplify a clear 887 bp fragment in S. africanum but not in any wheat. The primers also amplified an 887 bp fragment in other accessions of rye, Chinese Spring-Imperial rye chromosome additions and a diverse range of material carrying different rye chromosomes or chromosomal segments. In situ hybridization showed that probe pSaD15₉₄₀ was specifically hybridized throughout all rye chromosomes arms except for the terminal regions. The advantage of the rye-specific probe developed herein compared to those of previous reports is that it has been shown to be widely applicable to other Secale species. The probe will be useful as a molecular marker for the introgression of S. africanum and other rye chromosome segments into the wheat genome.     

Genetic Relationships Among Four Pepper Genotypes Resistant to Phytophthora capsici

According to our previous investigations, resistance to Phytophthora capsid in Capsicum annuum genotypes, ‘Line 29’, ‘PI201232’, ‘PI201234’ and Serrano Criollo de Morelos 334 (‘SCM334’), seems to be controlled by three genes. In order to determine the genie relationships between these four sources of resistance, three experiments were conducted which included the four genotypes, their F₁s, F₂s, F3s and BC₁ generations together with the susceptible pepper genotype ‘Morron INI A 224’. Inoculations were made, when plants had 4—6 leaves, by irrigating the culture substrate with a zoospore suspension of P. capsici isolate ‘Bl’. Though the four genotypes showed percentages of resistance close to a 100%, none of them actually reached this level in the three experiments. ‘SCM334’ was the most resistant genotype, transmitting a high level of resistance to its F₁, F₂ and BQ generations. ‘Line 29’ was more resistant than ‘PI201232’ and ‘PI201234’. However, the F₁ F₂ and BQ generations of these three lines showed similar degrees of resistance. The four genotypes seem to have one of the three genes postulated for their resistance in common. All genes displayed a similar level of resistance, except the specific genes of ‘SCM334’, the effect of which was slightly higher. Several working procedures are suggested for breeding programmes.     

Effect of alien 5R(5A) chromosome substitution on ear-emergence time and winter hardiness in wheat-rye substitution lines

Ear emergence time and response to vernalization were investigated in 12 alien substitution lines in which a pair of chromosomes 5A of recipient spring wheat cultivars was replaced by a pair of chromosomes 5R of Siberian spring rye ‘Onokhoiskaya’. The recipients were 12 spring cultivars of common wheat, each carrying different Vrn genes. Spring rye ‘Onokhoiskaya’ had the Sp1 (now called Vrn-R1) gene for spring growth habit located on chromosome 5R, but its expression was weaker. The Vrn-R1 gene had no effect on growth habit, ear emergence time and response to vernalization in wheat-rye substitution lines. Ears emerged significantly later in the 5R(5A) alien substitution lines than in the recipient wheat cultivars with the Vrn-A1/Vrn-B1/vrn-D1 or Vrn-A1/vrn-B1/Vrn-D1 genotypes. No difference in ear emergence time was found between most of the 5R(5A) alien substitution lines and the cultivars carrying the recessive vrn-A1 gene. The presence of the Vrn2a and Vrn2b alleles at the Vrn2 (now called Vrn-B1) locus located on wheat chromosome 5B was confirmed.The replacement of chromosome 5A by chromosome 5R in wheat cultivars ‘Rang’ and ‘Mironovskaya Krupnozernaya’, which carries the single dominant gene Vrn-A1, converted them to winter growth habit. In field studies near Novosibirsk the winter hardiness of 5R(5A) wheat–rye substitution lines of ‘Rang’ and ‘Mironovskaya Krupnozernaya’ was increased by 20–47% and 27–34%, respectively, over the recurrent parents.     

Cumulative and interaction effects of prolamin allelic variation and of 1BL/1RS translocation on flour quality in bread wheat

The allelic variation of prolamin loci was studied in three F₂ progenies from three crosses between the 1BL/1RS cultivar Triana and Yécora Rojo, Pavón and Florence Aurora, cultivars without the translocation. According to the 1:2:1 theoretical proportions observed in the allelic variants of the Glu-B3/Gli-B1 loci of the parent without the translocation, the inheritance as a block of the rye chromosome arm was confirmed. A group of F₃-F₄ recombinant lines, developed from these crosses was evaluated using the SDS-sedimentation test and the mixograph and alveograph tests. The presence of the 1BL/1RS translocation was not associated with significantly lower grain protein content values or with the optimum mixing time in the mixograph of the genotypes. The effect of the 1BL/1RS translocation on most of the quality parameters was highly dependent on the genetic pool. Significant increases in gluten strength and better mixing properties associated with the presence of some alleles of the Glu-A1, Glu-A3/ Gli-A1 and Gli-D2 loci were detected. The additivity and the interaction of prolamin gene effects with the rye translocation in the 1BL/1RS lines and its possible use in plant breeding are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.