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Molecular Cytogenetic Characterization and Stem Rust Resistance of Five Wheatlthinopyrum Ponticum Partial Amphiploids

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Molecular Cytogenetic Characterization and Stem Rust Resistance of Five Wheatlthinopyrum Ponticum Partial Amphiploids Available online at www.sciencedirect.com ScienceDirect JGG Journal of Genetics and Genomics 41 (2014) 591e599 ORIGINAL RESEARCH Molecular Cytogenetic Characterization and Stem Rust Resistance of Five WheatLThinopyrum ponticum Partial Amphiploids Qi Zheng a, Zhenling Lv a, Zhixia Niu b, Bin Li a, Hongwei Li a, Steven S. Xu b, Fangpu Han a, Zhensheng Li a,* a State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China b United States Department of Agriculture, Agricultural Research Service, Cereal Crops Research Unit, Fargo, ND 58102-2765, USA Received 24 January 2014; revised 27 May 2014; accepted 9 June 2014 Available online 14 June 2014 ABSTRACT Partial amphiploids created by crossing common wheat (Triticum aestivum L.) and Thinopyrum ponticum (Podp.) Barkworth & D. R. Dewey are important intermediates in wheat breeding because of their resistance to major wheat diseases. In this study, we examined the chromosome compositions of five Xiaoyan-series wheatÀTh. ponticum partial amphiploids (Xiaoyan 68, Xiaoyan 693, Xiaoyan 784, Xiaoyan 7430, and Xiaoyan 7631) using GISH, multicolor-GISH, and multicolor-FISH. We found several chromosome changes in these lines. For example, wheat chromosomes 1B and 2B were added in Xiaoyan 68 and Xiaoyan 7430, respectively, while wheat chromosome 6B was eliminated from Xiaoyan 693 and Xiaoyan 7631. Chromosome rearrangements were also detected in these amphiploids, including an interspecific translocation involving chromosome 4D and some intergenomic translocations, such as AeB and AeD translocations, among wheat genomes. Analysis of the Th. ponticum chromosomes in the amphiploids showed that some lines shared the same alien chromosomes. We also evaluated these partial amphiploids for resistance to nine races of stem rust, including TTKSK (commonly known as Ug99). Three lines, Xiaoyan 68, Xiaoyan 784, and Xiaoyan 7430, exhibited excellent resistance to all nine races, and could therefore be valuable sources of stem rust resistance in wheat breeding. KEYWORDS: Thinopyrum ponticum; Triticum aestivum; Partial amphiploid; Multicolor-genomic in situ hybridization (mc-GISH); Multicolor-fluorescent in situ hybridization (mc-FISH); Stem rust INTRODUCTION that is important for wheat improvement because of its resistance to a number of wheat diseases, as well as its stress Wild relatives of common wheat (Triticum aestivum L.) have tolerance and high crossability with various Triticum species been used extensively to transfer beneficial alien genes into (Shannon, 1978; Sharma et al., 1989; Cox, 1991; McIntosh, wheat (Friebe et al., 1996; Bommineni and Jauhar, 1997; 1991). To transfer resistance genes from Th. ponticum to Sepsi et al., 2008). Tall wheatgrass, Thinopyrum ponticum wheat, creating partial amphiploids from crosses between (Podp.) Barkworth and D. R. Dewey [ ¼ Agropyron elon- common wheat and tall wheatgrass is an important interme- gatum ssp. ruthenicum Beldie; Elytrigia pontica (Podp.) diate step. In the past several decades, a number of stable Holub; Lophopyrum ponticum (Podp.) A´ Lo¨ve] wheateTh. ponticum partial amphiploids have been devel- (2n ¼ 10x ¼ 70) is a perennial species in the tribe Triticeae oped that carry useful agronomic traits, such as resistance to wheat streak mosaic virus, barley yellow dwarf virus, com- mon root rot, Fusarium head blight, tan spot, and Stagono- þ þ * Corresponding author. Tel: 86 10 6480 6607, fax: 86 10 6480 6605. spora nodorum blotch (Chen et al., 1998a, 1998b; Thomas E-mail address: [email protected] (Z. Li). http://dx.doi.org/10.1016/j.jgg.2014.06.003 1673-8527/Copyright Ó 2014, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Limited and Science Press. All rights reserved. 592 Q. Zheng et al. / Journal of Genetics and Genomics 41 (2014) 591e599 et al., 1998; Fedak et al., 2000; Li et al., 2004; Oliver et al., hybridization signals on chromosomes, the A-, B-, and D- 2006). genome from common wheat and the genomes from Th. pon- Understanding of the compositions and structures of the ticum could be distinguished at the same time. The A-, B-, and introgressed Th. ponticum chromosomes in partial amphi- D-genome chromosomes were labeled with yellowegreen, ploids is useful for further transferring desirable genes into gray, and red florescences, respectively, while the Th. ponticum common wheat. Therefore, the chromosome compositions of chromosomes were labeled with green florescences. many wheateTh. ponticum partial amphiploids have been A number of intergenomic rearrangements were detected in characterized. Chen et al. (1998a, 1998b) reported two octo- the wheat genome by mc-GISH analysis. As illustrated in ploid Agrotriticum partial amphiploids, Agrotana and Fig. 1A, Xiaoyan 68 had 12 green-fluorescing chromosomes OK7211542, which had eight Js-genome chromosomes and that originated from tall wheatgrass and two interspecific eight E- or J-genome chromosomes. Oliver et al. (2006) translocation chromosomes with green terminal fragments on described four partial wheateTh. ponticum amphiploids with red short arms. Among the remaining 42 wheat chromosomes, different amounts of alien chromatin. Based on their genomic 14 fluoresced yellowegreen, 16 gray, and 12 red. Among the in situ hybridization (GISH) results, Fedak et al. (2000) 12 red-fluorescing chromosomes, one pair carried a terminal revealed that each of six wheateTh. ponticum partial amphi- yellowegreen signal, suggesting that an intergenomic rear- ploids contained a synthetic alien genome composed of rangement involving the A and D genomes had taken place in different combinations of St-, J-, or Js-genome chromosomes. the wheat genome. Therefore, in addition to 12 alien chro- Li et al. (1985) reported the production of partial amphiploids, mosomes and 2 wheateTh. ponticum translocation chromo- and designated them the Xiaoyan series, from hybrids between somes, Xiaoyan 68 also had 14 A-genome, 16 B-genome, 10 common wheat and Th. ponticum. Zhang et al. (1996) inves- D-genome, and 2 AeD translocation chromosomes. tigated the chromosome complements of this series with an St- Similarly, Xiaoyan 784 possessed 14 Th. ponticum chro- genome probe and A-, B-, D- and E-genome blocks. Xiaoyan mosomes and 42 wheat chromosomes composed of 10 A- 693 and Xiaoyan 7631 had identical St-genomes, while genome, 14 B-genome, and 12 D-genome chromosomes plus Xiaoyan 784, Xiaoyan 68, and Xiaoyan 7430 had a synthetic two pairs of AeD translocation chromosomes and one pair of alien genome, including six pairs of St-genome chromosomes AeB translocation chromosomes (Fig. 1C). Xiaoyan 7430 and one pair of Ee- (or Eb-) genome chromosomes. However, contained 12 alien chromosomes plus 44 wheat chromosomes the specific wheat genome compositions in these partial am- comprising 8 A-genome, 16 B-genome, and 10 D-genome phiploids, including the missing, substituted, and translocated chromosomes, 4 pairs of AeD translocation chromosomes, chromosomes, remain unclear. To more extensively explore and one pair of AeB translocation chromosomes (Fig. 1D). these partial amphiploids for wheat improvement, detailed Among the four pairs of AeD translocation chromosomes in information on their genomic constitutions is needed. Xiaoyan 7430, three were mainly D genome with a small A- In the present study, we used a combination of multicolor- genome segment on the distal region of the long arm, and one genomic in situ hybridization (mc-GISH) and multicolor- comprised a large part of the A genome with a small D- fluorescent in situ hybridization (mc-FISH) to examine the genome fraction on its long arm. Both Xiaoyan 693 and cytogenetic compositions of five partial amphiploids in the Xiaoyan 7631 carried 16 Th. ponticum chromosomes plus 40 Xiaoyan series (Xiaoyan 68, Xiaoyan 693, Xiaoyan 784, wheat chromosomes consisting of 12 A-genome, 12 B- Xiaoyan 7430, and Xiaoyan 7631). In addition, because stem genome, 14 D-genome, and two AeB translocation chromo- rust Ug99-lineage races are currently a major threat to global somes (Fig. 1B and E). wheat production (Singh et al., 2011) and some wheateTh. ponticum partial amphiploids have excellent resistance to Sequential GISH and mc-FISH analyses revealed the Ug99 (Xu et al., 2009; Turner et al., 2013), we evaluated chromosome compositions of the partial amphiploids seedlings of these five lines for resistance to stem rust Ug99 and eight other major races of stem rust from Africa and North Although each of the five lines consistently had 56 chromo- America and confirmed that three partial amphiploids are good somes, the number of alien chromosomes ranged from 12 to 16, breeding material for stem rust resistance. suggesting that chromosome deletion and substitution occurred from the wheat genome. Chromosome elimination and addition RESULTS can be detected by sequential GISH and mc-FISH methodology (Fig. 2AeJ). The results of mc-GISH analysis revealed that a mc-GISH analysis revealed intergenomic pair of B-genome chromosomes was absent from Xiaoyan 693 rearrangements in the partial amphiploids and Xiaoyan 7631 (Fig. 2D and J). Comparing their mc-FISH results using probes pAs1 and pSc119.2 with that of common In the mc-GISH analysis of the five partial amphiploids, the wheat (Mukai et al., 1993), the missing chromosome was mitotic chromosomes were probed by D-genomic DNA determined to be 6B, because of faint pAs1 bands on the satellite (Aegilops tauschii) with Texas-red-5-dCTP (red), and total and short arm and a terminal and an interstitial pSc119.2 band on genomic DNA of Th. ponticum and Triticum urartu was probed the long arm. with Fluorescein-12-dUTP (green) and blocked by S-genome In both Xiaoyan 68 and Xiaoyan 7430 a pair of B-genome (Aegilops speltoides) DNA. Based on the distribution of chromosomes was added (Fig. 2B and H). The added Q. Zheng et al. / Journal of Genetics and Genomics 41 (2014) 591e599 593 Fig. 1. Multicolor (mc)-GISH patterns of the five partial amphiploids. The different florescence signals show A-genome chromosomes (yellow-green), B-genome chromosomes (gray), D-genome chromosomes (red) and the chro- mosomes of Th.
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