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Development and comparative genomic mapping of Dasypyrum villosum 6V#4S-specific PCR markers using transcriptome data

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Abstract

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Twenty-five Dasypyrum villosum 6V#4S-specific PCR markers were developed using transcriptome data and further assigned to comparative genomic maps of wheat chromosome 6A, 6B, and 6D and barley chromosome 6H contrasting their homologous genes in these genomes.

Abstract

Two Dasypyrum villosum accessions, D.v#2 and No. 1026 from England and Russia, respectively, contain Pm21 on chromosome 6V#2S and PmV on chromosome 6V#4S. Both genes confer high resistance to powdery mildew (PM) in wheat. Even though several molecular markers have been developed to detect Pm21 and PmV, only the MBH1 marker can simultaneously detect both Pm21 and PmV. In this study, we first used a high-throughput sequencing technique to obtain the transcriptome sequences of a wheat—D. villosum translocation line, Pm97033—which contains chromosome 6V#4S carrying the PmV locus, under wheat PM pathogen induction. Twenty-five 6V#4S chromosome-specific markers were developed. Three of them were able to clearly distinguish chromosomes 6V#4S and 6V#2S by product size, four amplified the product specific for chromosome 6V#4S only, and the remaining 18 markers identified chromosome 6VS in wheat backgrounds. Two different D. villosum accessions, their derived translocation lines and wheat varieties carrying different chromosome 6VS were identified using these specific markers. The 25 newly developed markers together with the known PM resistance gene Stpk-V were used to construct comparative genomic maps with the homoeologous chromosome arms of wheat and barley. The colinearity of the identified gene sequences amplified by the 25 markers among wheat chromosomes 6A, 6B, and 6D and barley chromosome 6H was not very conserved and interrupted frequently by inversion and insertion. Our markers have potential in marker assisted selection for PM resistance breeding, and for locating other potential important genes and cloning the PmV gene on chromosome 6V#4S.

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References

  • Bie TD, Zhao RH, Jiang ZJ, Gao DR, Zhang BQ, He HG (2015a) Effcient marker-assisted screening of structural changes involving Haynaldia villosa chromosome 6V using a double-distal-marker strategy. Mol Breed 35:34

    Article  Google Scholar 

  • Bie TD, Zhao RH, Zhu SY, Chen SL, Cen B, Zhang BQ, Gao DR, Jiang ZN, Chen TT, Wang L, Wu RL, He HG (2015b) Development and characterization of marker MBH1 simultaneously tagging genes Pm21 and PmV conferring resistance to powdery mildew in wheat. Mol Breed 35:189

    Article  Google Scholar 

  • Bougot Y, Lemoine J, Pavoine MT, Barloy D, Doussinault G (2002) Identification of a microsatellite marker associated with Pm3 resistance alleles to powdery mildew in wheat. Plant Breed 121:325–329

    Article  CAS  Google Scholar 

  • Bozhko M, Riegel R, Schubert R, Muller-Starck G (2003) A cyclophilin gene marker confirming geographical differentiation of Norway spruce populations and indicating viability response on excess soil-born salinity. Mol Ecol 12:3147–3155

    Article  CAS  PubMed  Google Scholar 

  • Brown GR, Kadel EE, Bassoni DL, Kiehne KL, Temesgen B, van Buijtenen JP, Sewell MM, Marshall KA, Neale DB (2001) Anchored reference loci in loblolly pine (Pinus taeda L.) for integrating pine genomics. Genetics 159:799–809

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cao AZ, Wang XE, Chen YP, Zou XW, Chen PD (2006) A sequence-specific PCR marker linked with Pm21 distinguishes chromosomes 6AS, 6BS, 6DS of Triticum aestivum and 6VS of Haynaldia villosa. Plant Breed 125:201–205

    Article  CAS  Google Scholar 

  • Cao AZ, Xing LP, Wang XY, Yang XM, Wang W, Sun YL, Qian C, Ni JL, Chen YP, Liu DJ, Wang XE, Chen PD (2011) Serine/threonine kinase gene Stpk-V, a key member of powdery mildew resistance gene Pm21, confers powdery mildew resistance in wheat. Proc Natl Acad Sci USA 108:7727–7732

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen PD, Qi LL, Zhou B, Zhang SZ, Liu DJ (1995) Development and molecular cytogenetic analysis of wheat Haynaldia villosa 6VS/6AL translocation lines specifying resistance to powdery mildew. Theor Appl Genet 91:1125–1128

    CAS  PubMed  Google Scholar 

  • Chen X, Shi AN, Shang LM, Leath S, Murphy JP (1997) The resistance reaction of H.villosa to powdery mildew isolates and its expression in wheat background. Acta phytopathol 27(1):17–22

    Google Scholar 

  • Chen YP, Wang HZ, Cao AZ, Wang CM, Chen PD (2006) Cloning of a resistance gene analog from wheat and development of a codominant PCR marker for Pm21. J Integr Plant Biol 48:715–721

    Article  CAS  Google Scholar 

  • Chen F, Zhang FY, Morris C, He ZH, Xia XC, Cui DQ (2010) Molecular characterization of the Puroindoline a-D1b allele and development of an STS marker in wheat (Triticum aestivum L.). J Cereal Sci 52:80–82

    Article  CAS  Google Scholar 

  • Chen PD, You CF, Hu Y, Chen SW, Zhou B, Cao AZ, Wang XE (2013) Radiation-induced translocations with reduced Haynaldia villosa chromatin at the Pm21 locus for powdery mildew resistance in wheat. Mol Breed 31:477–484

    Article  CAS  Google Scholar 

  • Cordeiro GM, Casu R, McIntyre CL, Manners JM, Henry RJ (2001) Microsatellite markers from sugarcane (Saccharum spp.) ESTs cross transferable to erianthus and sorghum. Plant Sci 160:1115–1123

    Article  CAS  PubMed  Google Scholar 

  • Cui Y, Bao YG, Wang HG, Li XF (2016) Development of specific molecular markers for Thinopyrum intermedium using RNA-seq data. J Triticeae Crop 36:699–707

    CAS  Google Scholar 

  • Dai C, Zhang JP, Wu XY, Yang XM, Li XQ, Liu WH, Gao AN, Li LH (2012) Development of EST markers specific to Agropyron cristatum chromosome 6P in common wheat background. Acta Agron Sin 38:1791–1801

    Article  CAS  Google Scholar 

  • Erayman M, Turktas M, Akdogan G, Gurkok T, Inal B, Ishakoglu E, Ilhan E, Unver T (2015) Transcriptome analysis of wheat inoculated with Fusarium graminearum. Front in Plant Sci 6:867

    Article  Google Scholar 

  • Forster BP, Miller TE, Law CN (1988) Salt tolerance of two wheat Agropyron-junceum disomic addition lines. Genome 30:559–564

    Article  Google Scholar 

  • Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng QD, Chen ZH, Mauceli E, Hacohen N, Gnirke A, Rhind N, Palma FD, Birren BW, Nusbaum C, Kerstin LT, Friedman N, Regev (2011) A full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotech 29:644–652

    Article  CAS  Google Scholar 

  • He HG, Zhu SY, Sun WH, Gao DR, Bie TD (2013) Efficient development of Haynaldia villosa chromosome 6VS-specific DNA markers using a CISP-IS strategy. Plant Breed 132:290–294

    Article  CAS  Google Scholar 

  • Huang XQ, Roder MS (2004) Molecular mapping of powdery mildew resistance genes in wheat: a review. Euphytica 137:203–223

    Article  CAS  Google Scholar 

  • Jia CL, Zhang Y, Zhu L, Zhang R (2015) Application progress of transcriptome sequencing technology in biological sequencing. Mol Plant Breed 10:2388–2394

    Google Scholar 

  • King IP, Purdie KA, Rezanoor HN, Koebner RMD, Miller TE, Reader SM, Nicholson P (1993) Characterization of Thinopyrum bessarabicum chromosome segments in wheat using random amplified polymorphic DNAs (RAPDs) and genomic in situ hybridization. Theor Appl Genet 86:895–900

    CAS  PubMed  Google Scholar 

  • Lan TH, DelMonte TA, Reischmann KP, Hyman J, Kowalski SP, McFerson J, Kresovich S, Paterson AH (2000) An EST-enriched comparative map of Brassica oleracea and Arabidopsis thaliana. Genome Res 10:776–788

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li HJ, Conner RL, Chen Q, Jia X, Li H, Graf RJ, Laroche A, Kuzyk AD (2002) Different reactions to the wheat curl mite and wheat streak mosaic virus in various wheat–Haynaldia villosa 6V and 6VS lines. Plant Dis 86:423–428

    Article  Google Scholar 

  • Li H, Chen X, Xin ZY, Ma YZ, Xu HJ, Chen XY, Jia X (2005) Development and identification of wheat–Haynaldia villosa T6DL.6VS chromosome translocation lines conferring resistance to powdery mildew. Plant Breed 124:203–205

    Article  CAS  Google Scholar 

  • Li XB, Cui HR, Zhang ML (2006) Molecular markers derived from EST: their development and applications in comparative genomics. Biodiversity Sci 14:541–547

    Article  CAS  Google Scholar 

  • Li AL, Zhang RZ, Pan L, Tang LC, Zhao GY, Zhu MZ, Chu JF, Sun XH, Wei B, Zhang XQ, Jia JZ, Mao L (2011) Transcriptome analysis of H2O2-treated wheat seedlings reveals a H2O2-responsive fatty acid desaturase gene participating in powdery mildew resistance. PLoS One 6:e28810

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lin ZS, Zhang YL, Wang MJ, Li JR, Wang K, Chen X, Xu QF, Zhang XS, Ye XG (2013) Isolation and molecular analysis of genes Stpk-V2 and Stpk-V3 homologous to powdery mildew resistance gene Stpk-V in a Dasypyrum villosum accession and its derivatives. J Appl Genet 54:417–426

    Article  CAS  PubMed  Google Scholar 

  • Liu RH, Meng JL (2003) MapDraw: a Microsoft Excel macro for drawing genetic linkage maps based on given genetic linkage data. Heraditas (Beijing) 25:317–321

    Google Scholar 

  • Liu Z, Sun Q, Ni Z, Yang T, McIntosh RA (1999) Development of SCAR markers linked to the Pm21 gene conferring resistance to powdery mildew in common wheat. Plant Breed 118:215–219

    Article  CAS  Google Scholar 

  • Liu C, Ye XG, Wang MJ, Li SJ, Lin ZS (2017) Genetic behavior of Triticum aestivumDasypyrum villosum translocation chromosomes T6V#4S·6DL and T6V#2S·6AL carrying powdery mildew resistance. J Integr Agr. doi:10.1016/S2095-3119(16)61568-X

    Google Scholar 

  • Luo MC, Dubcovsky J, Dvorak J (1996) Recognition of homeology by the wheat ph1 locus. Genetics 144:1195–1203

    CAS  PubMed  PubMed Central  Google Scholar 

  • Qi LL, Cao MS, Chen PD, Li WL, Liu DJ (1996) Identification, mapping, and application of polymorphic DNA associated with resistance gene Pm21 of wheat. Genome 39:191–197

    Article  CAS  PubMed  Google Scholar 

  • Qi ZJ, Du P, Qian BL, Zhuang LF, Chen HF, Chen TT, Shen J, Guo J, Feng YG, Pei ZY (2010) Characterization of a wheat-Thinopyrum bessarabicum (T2JS-2BS.2BL) translocation line. Theor Appl Genet 121:589–597

    Article  CAS  PubMed  Google Scholar 

  • Rubio M, Rodriguez-Moreno L, Ballester AR, de Moura MC, Bonghi C, Candresse T, Martinez-Gomez P (2015) Analysis of gene expression changes in peach leaves in response to Plum pox virus infection using RNA-Seq. Mol Plant Pathol 16:164–176

    Article  CAS  PubMed  Google Scholar 

  • Schmolke M, Mohler V, Hartl L, Zeller FJ, Hsam KSL (2012) A new powdery mildew resistance allele at the Pm4 wheat locus transferred from einkorn (Triticum monococcum). Mol Breed 29:449–456

    Article  CAS  Google Scholar 

  • Shen YF, Shen J, Dawadondup Zhuang LF, Wang YZ, Pu J, Feng YG, Chu CG, Wang XE, Qi ZJ (2013) Physical localization of a novel blue-grained gene derived from Thinopyrum bessarabicum. Mol Breed 31:195–204

    Article  CAS  Google Scholar 

  • Song W, Xie C, Du J, Xie H, Liu Q, Ni Z, Yang T, Sun Q, Liu Z (2009) A ‘‘one-marker-for-two-genes’’ approach for efficient molecular discrimination of Pm12 and Pm21 conferring resistance to powdery mildew in wheat. Mol Breed 23:357–363

    Article  CAS  Google Scholar 

  • Song XM, Liu GF, Huang ZN, Duan WK, Tan HW, Li Y, Hou XL (2016) Temperature expression patterns of genes and their coexpression with LncRNAs revealed by RNA-Seq in non-heading Chinese cabbage. BMC Genom 17:297

    Article  Google Scholar 

  • Varshney RK, Sigmund R, Borner A, Korzun V, Stein N, Sorrells ME, Langridged P, Graner A (2005) Interspecific transferability and comparative mapping of barley EST-SSR markers in wheat, rye and rice. Plant Sci 168:195–202

    Article  CAS  Google Scholar 

  • Wang CM, Bie TD, Chen QZ, Cao AZ, Chen PD (2007) Development and application of molecular markers specific to chromosome 6VS of Haynaldia villosa (in Chinese). Acta Agron Sin 33:1595–1600

    CAS  Google Scholar 

  • Xue F, Ji WQ, Wang CY, Zhang H, Yang BJ (2012) High-density mapping and marker development for the powdery mildew resistance gene PmAS846 derived from wild emmer wheat (Triticum turgidum var. dicoccoides). Theor Appl Genet 124:1549–1560

    Article  CAS  PubMed  Google Scholar 

  • Zeng XY, Zhang ZY, Du LP, Xin ZY, Chen X (2005) Development of wheat germplasms with multi-resistance to powdery mildew, stripe rust and yellow dwarf virus by molecular marker-assisted selection. Sci Agric Sin 38:2380–2386

    CAS  Google Scholar 

  • Zhang YL, Wang MJ, Zhang Y, Chu CP, Lin ZS, Xu QF, Ye XG, Chen X, Zhang XS (2012) Development and application of functional markers specific to powdery mildew resistance on chromosome arm 6VS from different origins of Haynaldia villosa. Acta Agron Sin 38:1827–1832

    Article  CAS  Google Scholar 

  • Zhang H, Yang YZ, Wang CY, Liu M, Li H, Fu Y, Wang YJ, Nie YB, Liu XL, Ji WQ (2014) Large-scale transcriptome comparison reveals distinct gene activations in wheat responding to stripe rust and powdery mildew. BMC Genom 15:898

    Article  Google Scholar 

  • Zhang H, Hu WG, Hao JL, Lv SK, Wang CY, Tong W, Wang YJ, Wang YZ, Liu XL, Ji WQ (2016a) Genome-wide identification and functional prediction of novel and fungi-responsive lincRNAs in Triticum aestivum. BMC Genom 17:238

    Article  Google Scholar 

  • Zhang JC, Zheng HY, Li YW, Li HJ, Liu X, Qin HJ, Dong LL, Wang DW (2016b) Coexpression network analysis of the genes regulated by two types of resistance responses to powdery mildew in wheat. Sci Rep 6:23805

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We are grateful to Prof. Wenxiang Zhang and Ms. Yiping Yan at ICS-CAAS for providing powdery mildew pathogens and managing wheat plants in the growth chamber and the field. We would like to express our gratitude to Mr. Chunkun Fan at Agricultural Institute of Tibet Academy of Agricultural and Animal Husbandry Sciences, China, for collecting wheat samples post PM infection. We are indebted to Prof. Hongjie Li at ICS-CAAS for critical reading of this manuscript.

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Correspondence to Xingguo Ye.

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This research was financially supported by the National Key Research and Development Program of China (2016YFD0102001, 2016YFD0102002) and the Agricultural Science and Technology Innovation Program (ASTIP) of Chinese Academy of Agricultural Sciences.

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The authors declare that they have no conflict of interest.

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Communicated by Dr. Mark E. Sorrells.

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Li, S., Lin, Z., Liu, C. et al. Development and comparative genomic mapping of Dasypyrum villosum 6V#4S-specific PCR markers using transcriptome data. Theor Appl Genet 130, 2057–2068 (2017). https://doi.org/10.1007/s00122-017-2942-0

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  • DOI: https://doi.org/10.1007/s00122-017-2942-0

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