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. 1994 74 631 634 Molnár, I., Dulai, S., Molnár-Láng, M. (2008): Can the drought tolerance traits of Ae. Biuncialis

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One way of incorporating useful traits from Aegilops biuncialis (2n=4x=28, U b U b M b M b ) into wheat ( Triticum aestivum L. 2n=6x=42, AABBDD) is to develop first addition then translocation lines. The 2M b , 3M b , 7M b , 3U b , 5U b and 5U b /6U b wheat- Ae. biuncialis addition lines were produced in Martonvásár. To facilitate the exact identification of the addition lines, it was necessary to analyse the fluorescence in situ hybridisation patterns of the parental wheat genotype, Ae. biuncialis and its diploid progenitors ( Ae. umbellulata 2n=2x=14, UU and Ae. comosa 2n=2x=14, MM). The great genetic variability of the Aegilops species causes polymorphism in the fluorescence in situ hybridisation (FISH) patterns of the individual chromosomes. Due to the high level of FISH polymorphism, it is advisable to confirm the identification of the Ae. biuncialis chromosomes with the help of molecular (microsatellite, SSR) markers, so 119 wheat SSR markers were tested on Aegilops biuncialis , on Ae. geniculata (2n=4x=28, U g U g M g M g ), on five wheat- Ae. biuncialis addition lines (2M b , 3M b , 7M b , 3U b , 5U b ) and on an addition series of wheat- Ae. geniculata in order to select SSR markers specific to the U and M genomes of Ae. biuncialis and Ae. geniculata .

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The aims of the study were the optimisation of genomic in situ hybridization (GISH) and the use of a GAA satellite sequences as a fluorescence in situ hybridization (FISH) probe to discriminate Aegilops biuncialis and wheat chromosomes. The application of genomic DNA from the diploid progenitors, Ae. umbellulata and Ae. comosa , as a probe with a blocking ratio of 1:200 resulted in the clear visualization of Ae. biuncialis chromosomes. Based on the strong hybridization signals of the PCR-amplified GAA satellite sequences it was possible to identify chromosomes 1U, 2U, 4U and 5U of Ae. umbellulata . The improved GISH and GAA idiogram presented here allow the Ae. biuncialis chromatin to be traced in more detail during chromosome-mediated gene transfer to wheat.

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New wheat × barley, wheat × Aegilops biuncialis and wheat × rye hybrids were produced with the aim of alien gene transfer from these species into wheat. Amphiploids were produced with the help of colchicine treatment from the last two combinations. The new wheat × barley hybrids were multiplied in tissue culture because of the high degree of sterility and then pollinated with wheat to obtain backcross progenies. Wheat-barley chromosome pairing was detected using genomic in situ hybridization (GISH) in two combinations (Mv9 kr1 × Igri, Asakazekomugi × Manas). In vitro conditions caused an increase in chromosome arm association frequency in both combinations and in fertility in some regenerants. Five wheat-barley translocations were produced in a wheat background and characterized through the combination of cytogenetic and molecular genetic approaches (GISH, FISH and SSR markers). The following translocations were identified: 2DS.2DL-1HS, 3HS.3BL, 6BS.6BL-4HL, 4D-5HS and 7DL.7DS-5HS. Physical mapping of the SSR markers on chromosomes 1H and 5H was carried out using the intragenomic and interspecific translocation breakpoints and the centromere as physical landmarks.  Disomic wheat-Aegilops biuncialis additions were produced after backcrossing the wheat-Ae. biuncialis amphiploids. Fluorescence in situ hybridization (FISH) was carried out using two repetitive DNA clones (pSc119.2 and pAs1) on Ae. biuncialis and its two diploid progenitor species to detect chromosome polymorphism. The 7M and 3M disomic chromosome additions were selected and five more lines still need to be characterized.  The octoploid triticale (Mv9 kr1 × Lovászpatonai) produced in Martonvásár was crossed with a 1RS.1BL wheat cultivar Matador. GISH analysis detected pairing between the 1RS arm of the translocation chromosome and that of Lovászpatonai rye in 32 % of the pollen mother cells, making it possible to select recombinants from this combination. The new recombinants between the 1RS of Petkus and the 1RS of Lovászpatonai rye cultivars are being analysed with the help of microsatellite markers.

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In this study, a new substitution line, 12-5-1, with 42 chromosomes that was derived from BC3F2 descendants of the hybridization between Triticum aestivum cv. CN19 and Aegilops biuncialis was created and reported. The 12-5-1 was immune to both powdery mildew and stripe rust and has stable fertility. Multi-color fluorescence in situ hybridization indicated that 12-5-1 was a substitution line 1Mb(1B). The seed storage protein electrophoresis showed that 12-5-1 presented high molecular weight glutenin subunits (2 + 12) of CN19 and a new subunit designated as M which apparently originated from parent Ae. biuncialis, and absent 7 + 8 subunits. Additionally, the flour quality parameters showed that the protein content, Zeleny sedimentation value, wet gluten content, and grain hardness and mixing time of 12-5-1 were signifiantly higher than those of its parent CN19. Moreover, 5 pairs of the chromosome 1Mb-specifi polymerase chain reaction-based landmark unique gene markers, TNAC1021, TNAC1026, TNAC1041, TNAC1-02 and TNAC1-04, were also obtained. The new substitution line 1Mb(1B) 12-5-1 could be a valuable source for wheat improvement, especially for wheat end product quality and resistance to disease.

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The seed germination of bread ( Triticum aestivum cv. Bezostaya 1 and Ceyhan 99) and durum ( Triticum durum cv. Diyarbakır 81 and Fırat 93) wheat species and their wild relative species ( Aegilops biuncialis and Ae. triuncialis ) was compared at two light levels, nine NaCl concentrations and three alternating temperature regimes. No seeds germinated at 675 mmol/L NaCl. The highest seed germination (100%) of cultivated wheat cultivars was noted in the control at 15/25°C and 20/30°C, and that of wild wheat species in both the control and the 150 mmol/L NaCl treatment under all temperature regimes. The seed germination of bread and durum wheat cultivars was completely inhibited at or above 450 and 375 mmol/L NaCl, respectively. No Ae. biuncialis seeds germinated at 600 mmol/L NaCl, while seeds of Ae. triuncialis germinated at this concentration (38.9%) only at 20/30°C in darkness. The inhibitory effect of light on germination in all genotypes was determined in some salinity levels at 15/25°C. The optimal germination treatment for all genotypes was 15/25°C temperature regime and darkness. The inhibitory effect of high salinity on germination was greater at 25/35°C than at 15/25°C or 20/30°C. In salinity and temperature interactions, the seeds of wild wheat species were found to be more tolerant than those of wheat cultivars.

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Growth, cell viability and heat shock proteins (HSPs) in Bezostaya-1, Çukurova-86 and Diyarbakır-81 cultivated wheat cultivars and three Aegilops species were investigated. Etiolated seedlings were exposed to 23 °C, 32 °C, 35 °C, 37 °C and 38 °C for 24 h, and 35 °C (24 h) → 50 °C(1 h) and 37 °C (24 h) → (50 °C (1 h). At the end of recovery growth periods, the shoot lengths of the genotypes generally decreased significantly at 35, 37 and 38 °C. The acquired thermal tolerance (ATT) in intact seedlings was over 50% at 35 °C → 50 °C and 37 °C → 50 °C, but in cell viability test it ranged from 2.75% (Ae. triuncialis) to 32.87 (Bezostaya-1) at 35 °C, and from 2.82% (Ae. triuncialis) to 37.82 (Bezostaya-1) at 37 °C. Ae. triuncialis was most sensitive genotype in both ATT determination. In electrophoretic profiles of proteins, while some HSPs were newly synthesized, some normal cellular proteins disappeared at 37 °C and 37 °C → 50 °C compared to 23 °C. The number of low molecular weight (LMW) HSPs were more than intermediate- (IMW) and high- (HMW) HSPs. The genotypes had both common (12 HSPs between at least two genotypes) and genotype-specific (33 HSPs) LMW HSPs. The common HSP of 19.8 kDa (pI 6.5) was synthesized in Bezostaya-1, Çukurova-86, Diyarbakır-81, Ae. biuncialis and Ae. umbellulata. Bezostaya-1 is the only genotype that synthesized 12 IMW and 2 HMW HSPs at 37 °C → 50 °C. Ae. triuncialis had only two common LMW HSPs [22.1 (pI 7.1) and 24.2 kDa (pI 6.5)].

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and their allotetraploid hybrids Ae. biuncialis and Ae. Geniculate . PLoS ONE, 6 (11), e27708. doi:10.1371/journal.pone.0027708 Doležel J. Chromosome isolation by flow sorting in

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