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Journal of Integrative Agriculture  2018, Vol. 17 Issue (08): 1720-1726    DOI: 10.1016/S2095-3119(17)61866-5
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Development of EST-PCR markers specific to the long arm of chromosome 6V of Dasypyrum villosum
SUN Hao-jie, SONG Jing-jing, XIAO Jin, XU Tao, WEI Xing, YUAN Chun-xia, CAO Ai-zhong, XING Liping, WANG Hai-yan, WANG Xiu-e
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Science and Technology/Cytogenetics Institute, Nanjing Agricultural University/Jiangsu Collaborative Innovation Center for Modern Crop Production (JCIC-MCP), Nanjing 210095, P.R.China
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摘要  Received  1 September, 2017    Accepted  13 December, 2017


Expressed sequence tags-derived polymerase chain reaction (EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provide the evidence of the syntenic relationship between homoeologous chromosomes.  In the present study, in order to develop high density and evenly distributed molecular markers specific for chromosome 6VL of Dasypyrum villosum, 297 primer pairs were designed based on the expressed sequence tags (EST) sequences, which were previously mapped in different bins of the long arms of wheat homoeologous 6AL, 6BL, and 6DL.  By using the Triticum aestivum, D. villosum, T. durum-D. villosum amphiploid, and T. aestivum-D. villosum alien chromosome lines involving chromosome 6V, it was found that 32 (10.77%) primers could amplify specific bands for chromosome 6V, and 31 could be allocated to chromosome arm 6VL.  These 6VL specific markers provided efficient tools for the characterization of structural variation involving the chromosome 6VL in common wheat background as well as for the selection of useful genes located on 6VL in breeding programs.
Keywords:  Dasypyrum villosum        molecular marker        common wheat        alien chromosome line  
Received: 01 September 2017   Accepted:
Fund: This research was supported by the National Key Research and Development Program of China (2016YFD0102001), the National Natural Science Foundation of China (31571653, 31771782, 31201204), the Technology Support Program of Jiangsu Province, China (BE2013439), the Fundamental Research Funds for the Central Universities, China (KYZ201403), the Jiangsu Agricultural Science and Technology Innovation Fund, China (CX151001), the Program of Introducing Talents of Discipline to Universities, China (B08025), the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China (PAPD), and the Six Talent Peaks Project in Jiangsu Province, China.
Corresponding Authors:  Correspondence WANG Hai-yan, Tel: +86-25-84395308, Fax: +86-25-84395344, E-mail:; WANG Xiu-e, E-mail:    

Cite this article: 

SUN Hao-jie, SONG Jing-jing, XIAO Jin, XU Tao, WEI Xing, YUAN Chun-xia, CAO Ai-zhong, XING Liping, WANG Hai-yan, WANG Xiu-e. 2018. Development of EST-PCR markers specific to the long arm of chromosome 6V of Dasypyrum villosum. Journal of Integrative Agriculture, 17(08): 1720-1726.

Ayala-Navarrete L, Bariana H S, Singh R P, Gibson J M, Mechanicos A A, Larkin P J. 2007. Trigenomic chromosomes by recombination of Thinopyrum intermedium and Th. ponticum translocations in wheat. Theoretical and Applied Genetics, 116, 63–75.
Ayala-Navarrete L, Tourton E, Mechanicos A A, Larkin P J. 2009. Comparison of Thinopyrum intermedium derivatives carrying barley yellow dwarf virus resistance in wheat. Genome, 52, 537–546.
Banks P M, Larkin P J , Bariana H S, Lagudah E S, Appels R, Waterhouse P M, Brettell R I S, Chen X, Xu H J, Xin Z Y, Qian Y T, Zhou X M, Cheng Z M, Zhou G H. 1995. The use of cell culture for subchromosomal introgressions of barley yellow dwarf virus resistance from Thinopyrum intermedium to wheat. Genome, 38, 395–405.
Bassam B J, Gresshoff P M. 2007. Silver staining DNA in polyacrylamide gels. Nature Protocols, 2, 2649–2654.
Bie T D, Cao Y P, Chen P D. 2007. Mass production of intergeneric chromosomal translocations through pollen irradiation of Triticum durum-Haynaldia villosa amphiploid. Journal of Integrative Plant Biology, 49, 1619–1626.
Blanco A, Resta P, Simeone R, Parmar S, Shewry P R, Sabelli P, La?andra D. 1991. Chromosomal location of seed storage protein genes in the genome of Dasypyrum villosum (L.) Candargy. Theoretical and Applied Genetics, 82, 358–362.
Blanco A, Simeone R, Resta P. 1987. The addition of Dasypyrum villosum (L.) Candargy chromosomes to durum wheat (Triticum durum Desf.). Theoretical and Applied Genetics, 74, 328–333.
Bossolini E, Wicker T, Knobel P A, Keller B. 2007. Comparison of orthologous loci from small grass genomes Brachypodium and rice: Implications for wheat genomics and grass genome annotation. The Plant Journal, 49, 704–717.
Chen P D, Liu D J. 1982. Cytogenetic studies of hybrid progenies between Triticum aestivum and Haynaldia villosa. Journal of Nanjing Agricultural University, 4, 1–15. (in Chinese)
Chen P D, Liu W X, Yuan J H, Wang X E, Zhou B, Wang S L, Zhang S Z, Feng Y G, Yang B J, Liu D J, Qi L L, Zhang P, Friebe B, Gill B S. 2005. Development and characterization of wheat-Leymus racemosus translocation lines with resistance to Fusarium Head Blight. Theoretical and Applied Genetics, 111, 941–948.
Chen P D, Qi L L, Zhou B, Zhang S Z, Liu D J. 1995. Development and molecular cytogenetic analysis of wheat-Haynaldia villosa 6VS/6AL translocation lines specifying resistance to powdery mildew. Theoretical and Applied Genetics, 91, 1125–1128.
Crasta O R, Francki M G, Bucholtz D B, Sharma H C, Zhang J, Wang R C, Ohm H W, Anderson J M. 2000. Identification and characterization of wheat-wheatgrass translocation lines and localization of barley yellow dwarf virus resistance. Genome, 43, 698–706.
Devos K M, Gale M D. 1992. The use of random amplified polymorphic DNA markers in wheat. Theoretical and Applied Genetics, 84, 567–572.
Feuillet C, Keller B. 2002. Comparative genomics in the grass family: Molecular characterization of grass genome structure and evolution. Annals of Botany, 89, 3–10.
Feuillet C, Travella S, Stein N, Albar L, Nublat A, Keller B. 2003. Map-based isolation of the leaf rust disease resistance gene Lr10 from the hexaploid wheat (Triticuma estivum L.) genome. Proceedings of the National Academy of Sciences of the United States of America, 100, 15253–15258.
Gale M D, Devos K M. 1998. Comparative genetics in the grasses. Proceedings of the National Academy of Sciences of the United States of America, 95, 1971–1974.
Gradzielewska A. 2006. The genus Dasypyrum - Part 2. Dasypyrum villosum - A wild species used in wheat improvement. Euphytica, 152, 441–454.
Hagras A A A, Kishii M, Sato K, Tanaka H, Tsujimoto H. 2005. Extended application of barley EST markers for the analysis of alien chromosomes added to wheat genetic background. Breeding Science, 55, 335–341.
Li H F, Gill B S, Wang X E, Chen P D. 2011. A Tal-PhI wheat genetic stock facilitates efficient alien introgression. Genetic Resources and Crop Evolution, 58, 667–678.
Liu W X, Rouse M, Friebe B, Jin Y, Gill B, Pumphrey M O. 2011. Discovery and molecular mapping of a new gene conferring resistance to stem rust, Sr53, derived from Aegilops geniculata and characterization of spontaneous translocation stocks with reduced alien chromatin. Chromosome Research, 19, 669–682.
Lu H J, Fellers J P, Friesen T L, Meinhardt S W, Faris J D. 2006. Genomic analysis and marker development for the Tsn1 locus in wheat using bin-mapped ESTs and banking BAC contigs. Theoretical and Applied Genetics, 112, 1132–1142.
Niu Z X, Klindworth D L, Friesen T L, Chao S M, Jin Y, Cai X W, Xu S S. 2011. Targeted introgression of a wheat stem rust resistance gene by DNA marker-assisted chromosome engineering. Genetics, 187, 1011–1021.
Qi L, Friebe B, Zhang P, Gill B S, 2007. Homoeologous recombination, chromosome engineering and crop improvement. Chromosome Research, 15, 3–19.
Qi L L, Chen P D, Liu D J, Gill B S. 1999. Homoeologous relationships of Haynaldia villosa  chromosomes with those of Triticum aestivum as revealed by RFLP analysis. Genes & Genetic Systems, 74, 77–82.
Qi L L, Echalier B, Chao S, Lazo G R, Butler G E, Anderson O D, Akhunov E D, Dvo?ák J, Linkiewicz A M, Ratnasiri A. 2004. A chromosome bin map of 16 000 expressed sequence tag loci and distribution of genes among the three genomes of polyploid wheat. Genetics, 168, 701–712.
Qi L L, Pumphrey M O, Friebe B, Chen P D, Gill B S. 2008. Molecular cytogenetic characterization of alien introgressions with gene Fhb3 for resistance to Fusarium head blight disease of wheat. Theoretical and Applied Genetics, 117, 1155–1166.
Qi L L, Pumphrey M O, Friebe B, Zhang P, Qian C, Bowden R L, Rouse M N, Jin Y, Gill B S. 2011. A novel Robertsonian translocation event leads to transfer of a stem rust resistance gene (Sr52) effective against race Ug99 from Dasypyrum villosum into bread wheat. Theoretical and Applied Genetics, 123, 159–167.
Riley R, Chapman V, Johnson R. 1968. Introduction of yellow rust resistance of Aegilops comosa into wheat by genetically induced homoeologous recombination. Nature, 217, 383–384.
Rowland L J, Mehra S, Dhanaraj A L, Ogden E L, Slovin J P, Ehlenfeldt M K. 2003. Development of EST-PCR markers for DNA fingerprinting and genetic relationship studies in blueberry (Vaccinium, section Cyanococcus). Journal of the American Society for Horticultural Science, 128, 682–690.
Sargent D J, Rys A, Nier S, Simpson D W, Tobutt K R. 2007. The development and mapping of functional markers in Fragaria and their transferability and potential for mapping in other genera. Theoretical and Applied Genetics, 114, 373–384.
Schubert R, Mueller-Starck G, Riegel R. 2001. Development of EST-PCR markers and monitoring their intrapopulational genetic variation in Picea abies (L.) Karst. Theoretical and Applied Genetics, 103, 1223–1231.
Shen X R, Ohm H. 2007. Molecular mapping of Thinopyrum-derived Fusarium head blight resistance in common wheat. Molecular Breeding, 20, 131–140.
Wang M J, Zhang Y, Lin Z S, Ye X G, Yuan Y P, Ma W, Xin Z Y. 2010. Development of EST-PCR markers for Thinopyrum intermedium chromosome 2Ai#2 and their application in characterization of novel wheat-grass recombinants. Theoretical and Applied Genetics, 121, 1369–1380.
Zhang J J, Yuan H X, Zhang R Q, Xing X P, Dai J L, Niu J S, Li H L, Chen P D. 2012. Analysis of resistance to Heterodera filipjevi in Triticum aestivum-Dasypyrum villosum germplasm. Aata Agronomica Sinica, 38, 1969–1976. (in Chinese)
Zhang Q P, Li Q, Wang X E, Wang H Y, Lang S P, Wang Y N, Wang S L, Chen P D, Liu D J. 2005. Development and characterization of a Triticum aestivum-Haynaldia villosa translocation line T4VS·4DL conferring resistance to wheat spindle streak mosaic virus. Euphytica, 145, 317–320.
Zhang W, Gao A L, Zhou B, Chen P D. 2006. Screening and applying wheat microsatellite markers to trace individual Haynaldia villosa chromosomes. Acta Genetica Sinica, 33, 236–243.
Zhao R H, Wang H Y, Jia Q, Xiao J, Yuan C X, Zhang Y J, Hu Q S, Wang X E. 2014. Development of EST-PCR markers for the chromosome 4V of Haynaldia villosa and their application in identification of 4V chromosome structural aberrants. Journal of Integrative Agriculture, 13, 282–289.
Zhong G Y, Dvo?ák J. 1995. Evidence for common genetic mechanisms controlling the tolerance of sudden salt stress in the tribe Triticeae. Plant Breeding, 114, 297–302.
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