Hybrid Purity Testing of Brassica rapa Using SSR Marker Technology

in HortScience

Thirteen Chinese cabbage (Brassica rapa) hybrid cultivars and 26 parental inbred lines were used as experimental materials to screen for primers producing hybrid and parental complementary bands and for primers with high polymorphism information contents and low genotype frequencies. A total of 18 pairs of core primers were designed to identify the purity of Chinese cabbage. There was no significant difference in the purity percentage measured between different loci of the same strain. The fingerprint obtained by the amplification of each locus could be used to identify purity to obtain an authentic purity percentage. Curve mapping and significance analyses were conducted using the purity percentage of eight different seed samples and confirmed a sampling seed number of 96. The results of the purity test were verified by comparison with the grow-out test (GOT) using molecular markers. In conclusion, the simple sequence repeat (SSR) detection system could be used for the rapid identification of the purity of the tested Chinese cabbage hybrids.

Contributor Notes

This research was supported by a Project of the Committee of Science and Technology of Beijing City (No. D131100000413001), the certification and testing and service capability promotion of the 2013–14 ISTA, the International Seed Testing Laboratory of Ascension, and the Beijing Finance Project (no item number) from 2009 to 2016. For cultivar testing for DNA fingerprint detection in the Beijing region, support was received from the National Science and Technology Support Program (No. 2011BAD35B07) from 2011 to 2015, the Beijing Academy of Agriculture and Forestry Finance Special Subject (KJCX20140111) from 2014 to 2016, and Beijing Academy of Agriculture and Forestry (KJCX20150202).

Corresponding author. E-mail: lili@nercv.org.

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    Primer pair for BRMS296 used to screen for the hybrid Beijing 68. P29 and P30 are the parents, and OV is one other cultivar mixed in Beijing 68. P30 is mixed with the hybrid. The bands with size labeled are DNA molecular ladder.

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    Primer pair for BRU07947 used to screen for the hybrid Naibai No. 1. N4 and N30 are the parents, and OV contains a mixture of other cultivars present in Naibai No. 1. N4 is mixed with the hybrids. The bands with size labeled are DNA molecular ladder.

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    Primer pair for cnu-m149a used to screen for the hybrid Naibai No. 1. N4 and N30 are the parents, and lanes marked “OV” show other cultivars mixed in Naibai No. 1. N30 is mixed with the hybrids. The bands with size labeled are DNA molecular ladder.

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    Primer pair for BRU07947 used to screen for the hybrid Jingguan No. 3. N13 and N24 are the parents, and OV is one other cultivar mixed in Jingguan No. 3. M is the DNA molecular ladder.

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    (A) Curve analysis of the purity percentage for seven samples. (B) Curve analysis of the purity percentage for six samples. Regarding the data obtained from the four groups of 96 seeds, the fluctuation range of the purity percentage was obvious, and the purity percentage tended to be similar.

Article References

  • ChoiS.R.TeakleG.R.PlahaP.KimJ.H.AllenderC.J.BeynonE.PiaoZ.Y.SoengasP.HanT.H.KingG.J.BarkerG.C.HandP.LydiateD.J.BatleyJ.EdwardsD.KooD.H.BangJ.W.ParkB.S.LimY.P.2007The reference genetic linkage map for the multinational Brassica rapa genome sequencing projectTheor. Appl. Genet.115777792

    • Search Google Scholar
    • Export Citation
  • DongY.Y.YuM.N.LiX.B.XuP.F.CuiH.R.ZhangM.L.2008Diversity analysis of Brassica napus by EST-SSR and RAPD markersJ. Nucl. Agr. Sci.22611616

    • Search Google Scholar
    • Export Citation
  • HuangW.P.ZhengX.Y.1994Peroxidase isozyme analysis of F1 hybrid and parent seedling in Chinese cabbage (Brassica rapa L. ssp. pekinensis)Seed711315

    • Search Google Scholar
    • Export Citation
  • JiaX-H.2008SSR molecular maps of hybrid maize varieties. China Agr. Press Beijing China

  • KeG-L.2010Chinese cabbage breeding. China Agr. Press Beijing China

  • LiL.HeW.M.MaL.P.LiuP.Y.XuH.M.XuJ.B.ZhengX.Y.2009Construction Chinese cabbage (Brassica rapa L.) core collection and its EST-SSR fingerprint database by EST-SSR molecular markersGenomics Appl. Biol.287688

    • Search Google Scholar
    • Export Citation
  • LiL.JianY.C.MaL.P.LiX.Q.DingY.H.2011Cabbage (Brassica oleracea) hybrid purity quotient test by EST-SSR molecular makersGenomics Appl. Biol.3011611164

    • Search Google Scholar
    • Export Citation
  • LiL.ZhangW.Q.LiuL.WuP.MaL.P.2015Application of SSR markers in melon hybrid purity test and variety specificity identificationMol. Plant Breed.1325222530

    • Search Google Scholar
    • Export Citation
  • LiL.ZhengX.Y.2009The development of multiplex EST-SSR markers to identification Chinese cabbage (Brassica campestris L. chinensis (L.) Makino and Brassica campestris L. pekinensis (Lour.) Olsson) cultivarActa Hort. Sin.3616271634

    • Search Google Scholar
    • Export Citation
  • LiY.F.LiuZh.Y.WangY.Sh.YangN.XinX.F.YangSh.FengH.2012Identification of quantitative trait loci for yellow inner leaves in Chinese cabbage (Brassica rapa L. ssp. pekinensis) based on SSR and SRAP markersSci. Hort.1331017

    • Search Google Scholar
    • Export Citation
  • LiuG.LiuL.GongY.WangY.YuF.ShenH.GuiW.2007Seed genetic purity testing of F1 hybrid cabbage (Brassica oleracea var. capitata) with molecular marker analysisSeed Sci. Technol.35477486

    • Search Google Scholar
    • Export Citation
  • McCallumJ.ThomsonS.MeeghanP.J.FernandK.2008Genetic diversity analysis and single-nucleotide polymorphism marker development in cultivated bulb onion based on expressed sequence tag-simple sequence repeat markersJ. Amer. Soc. Hort. Sci.133810818

    • Search Google Scholar
    • Export Citation
  • PallaviH.M.GowdaR.ShadakshariY.G.BhanuprakashK.VishwanathK.2011Identification of SSR markers for hybridity and seed genetic purity testing in sunflower (Helianthus annuus L.)Seed Sci. Technol.39259264

    • Search Google Scholar
    • Export Citation
  • SaxenaR.K.SaxenaK.VarshneyR.K.2010Application of SSR markers for molecular characterization of hybrid parents and purity assessment of ICPH 2438 hybrid of pigeonpea (Cajanus cajan L. Millspaugh)Mol. Breed.26371380

    • Search Google Scholar
    • Export Citation
  • ShiL.LiL.ZhengX.Y.2007Optimization of SSR-PCR system for identification in Chinese cabbage (Brassica campestris L. ssp. pekinensis)Mol. Plant Breed.5110116

    • Search Google Scholar
    • Export Citation
  • ShiP.H.LiuZ.G.ZhangY.H.SunW.C.KongD.J.LuM.H.YangN.N.2014Analysis on antioxidant enzyme activities and peroxidase isozymes in 23 rape cultivarsActa Agr. Boreali-occidentlis Sin.23113119

    • Search Google Scholar
    • Export Citation
  • SundaramR.M.NaveenkumarB.BiradarS.K.BalachandranS.M.MishraB.IlyasAhmedM.ViraktamathB.C.RameshaM.S.SarmaN.P.2008Identification of informative SSR markers capable of distinguishing hybrid rice parental lines and their utilization in seed purity assessmentEuphytica163215224

    • Search Google Scholar
    • Export Citation
  • SuwabeK.IketaniH.NunomeT.KageT.HiraiM.2002Isolation and characterization of microsatellites in brassica rapa LTheor. Appl. Genet.10410921098

    • Search Google Scholar
    • Export Citation
  • SuwabeK.TsukazakiH.IketaniH.HatakeyamaK.KondoM.FujimuraM.NunomeT.FukuokaH.HiraiM.MatsumotoS.2006SSR-based comparative genomics between Brassica rapa and Arabidopsis thaliana: The genetic origin of clubroot resistanceGenetics173309319

    • Search Google Scholar
    • Export Citation
  • WangX.Y.YuSh.C.ZhangF.L.YuY.J.ZhaoX.Y.ZhangD.Sh.2008SSR fingerprinting and genetic distinctness of Pak-choi (Brassica rapa L. ssp. chinensis Makino)Acta Agr. Boreali Sin.2397103

    • Search Google Scholar
    • Export Citation
  • WangY.SunS.L.LiuB.WangH.DengJ.LiaoY.C.WangQ.ChengF.WangX.W.WuJ.2011A sequence-based genetic linkage map as a reference for Brassica rapa pseudo chromosome assemblyBMC Genomics12239

    • Search Google Scholar
    • Export Citation
  • WeiL.Y.ZhaoC.X.LiJ.N.LiJ.N.QianW.FuD.H.2012Screening of core simple sequence repeat primer pairs and establishment of a multiplex polymerase chain reaction system for brassica genomes A and CPlant Breed.131457459

    • Search Google Scholar
    • Export Citation
  • YuS.C.ZhangF.L.YuR.B.ZouY.M.QiJ.N.ZhaoX.Y.YuY.J.ZhangD.S.LiL.2009Genetic mapping and localization of a major QTL for seedling resistance to downy mildew in Chinese cabbage (Brassica rapa ssp. pekinensis)Mol. Breed.23573590

    • Search Google Scholar
    • Export Citation
  • YuanY.X.ZhangX.W.SunR.F.WangX.W.ZhangH.JiangW.S.YaoQ.J.GengJ.F.2010Construction of a genetic linkage of Chinese map anchoring to chromosomes cabbageActa Agr. Boreali-Sin.258086

    • Search Google Scholar
    • Export Citation
  • ZhangW.LiL.2013Construction of SSR fingerprint database of Chinese cabbage varieties (Brassica campestris L. ssp. pekinensis)Mol. Plant Breed.11843857

    • Search Google Scholar
    • Export Citation
  • ZhaoZ.GuH.ShengX.YuH.WangJ.CaoJ.2012Genetic purity testing of loose-curd cauliflower hybrids using SSR markers and grow out testSeed Sci. Technol.40209214

    • Search Google Scholar
    • Export Citation
  • ZhouX.D.WangF.Ch.ZhouG.L.ZhuX.F.LinM.ChenL.L.WangH.B.2012Testing of genetic purity of watermelon F1 varieties by SSR markersChina Cucurb. Veg.251316

    • Search Google Scholar
    • Export Citation

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