Abstract
We screened 626 pairs of EST-SSR primers from the radish marker database using 37 different cultivars. Of the primer pairs, 89.3 % (559) were polymorphic. The average polymorphism information content (PIC) value per primer was 0.39, ranging from 0.07 to 0.73. Fifty EST-SSR markers were designated core markers according to PCR band reproducibility and stability, high polymorphism levels, easily discriminated alleles, and an even distribution of molecular markers over each radish chromosome. Genetic diversity and the evolutionary relationships of 93 radish accessions, representing nearly all typical Raphanus germplasms, were assessed with 50 core EST-SSR markers. A total of 254 alleles at 52 loci (4.88 alleles per locus on average), with a mean PIC value of 0.55, were detected. The wild radish had the largest range of variation, followed by the East Asian big long radish and the European small radish. Unweighted pair group method with arithmetic mean cluster analysis classified the 93 germplasms into four groups. The first group included most accessions belonging to R. sativus, while seven wild Raphanus strains were separated into the remaining groups. In the R. sativus group, the subspecies var. hortensis Becker, var. raphanistroides Makino, var. caudatus Hooker and Anderson, var. sativus L., var. niger Kerner, and var. oleiferus Metzg were sorted into separate subgroups. Population genetics structure analyses show that the accessions were attributed to three populations: (1) Asia cultivated and East Asian wild, (2) Europe cultivated, and (3) wild Raphanus species. Herein, we discuss the origin and ancestry of cultivated radishes based on these results.
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References
Aoba T (1993) Vegetables in Japan. Yasaka Shobo, Tokyo, p 311 (in Japanese)
Botstein D, White RL, Skolnick M, Davis RW (1980) Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet 32(3):314–331
Budahn H, Peterka H, Mousa MA, Ding Y, Zhang S, Li J (2009) Molecular mapping in oil radish (Raphanus sativus L.) and QTL analysis of resistance against beet cyst nematode (Heterodera schachtii). Theor Appl Genet 118:775–782
Cheng DD, Zhang FJ, Liu LW, Xu L, Chen YL, Wang XL, Limera C, Yu RG, Gong YQ (2013) TRAP markers generated with resistant gene analog sequences and their application to genetic diversity analysis of radish germplasm. Sci Hortic 161:153–159
Cui N, Qiu Y, Li XX, Shen D, Wang HP, Song JP (2012) Data mining for SSRs in EST rescources and EST-SSR markers development in radish. Acta Hortic Sin 39(7):1303–1312 (in Chinese)
Curtis IS (2011) Genetic engineering of radish: current achievements and future goals. Plant Cell Rep 30:733–744
De Candolle A (1886) Origin of cultivated plants, 2nd edn (reprinted in 1959 by Hafner, New York, pp 29–33)
Earl DA, vonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conserv Genet Resour 4(2):359–361. doi:10.1007/s12686-011-9548-7
Ellstrand NC, Marshall DL (1985) Interpopulation gene flow by pollen in wild radish, Raphanus sativus. Am Nat 126:606–616
Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620
Falush D, Stephens M, Pritchard JK (2003) Inference of population structure using multilocus genotype data: linked loci and correlated allele frequencies. Genetics 164:1567–1587
Grzebelus D, Iorizzo M, Senalik D, Ellison S, Cavagnaro P, Macko-Podgorni A, Heller-Uszynska K, Kilian A, Nothnagel T, Allender C, Simon PW, Baranski R (2014) Diversity, genetic mapping, and signatures of domestication in the carrot (Daucus carota L.) genome, as revealed by diversity arrays technology (DArT) markers. Mol Breed 33:625–637
Henslow G (1898) The history of the radish. Gard Chron 23:389
Huh MK, Huh HW (2001) Genetic diversity of Raphanus sativus var. hortensis f. raphanistroides in Korea using AFLP markers. Kor J Genet 23:45–53
Huh MK, Ohnishi O (2002) Genetic diversity and genetic relationships of East Asian natural populations of wild radish revealed by AFLP. Breed Sci 52:79–88
Huh MK, Ohnishi O (2003) Genetic diversity and relationships among natural and cultivated populations of radish in Korea revealed by RAPD. Kor J Genet 25:119–125
Jiang L, Wang L, Liu L, Zhu X, Zhai L, Gong Y (2012) Development and characterization of cDNA library based novel EST-SSR marker in radish (Raphanussativus L.). Sci Hortic 140:164–172
Kaneko Y, Kimizuka-Takagi C, Bang SW, Matsuzawa Y (2007) Radish. In: Kole C (ed) Genome mapping and molecular breeding in plants, vol 5. Vegetables. Springer, Berlin, pp 141–160
Kitamura S (1958) Varieties of radish and their transition. In: Nishiyama I (ed) Japanese radish. The Japan Science Society Press, Tokyo, pp 1–19
Kitashiba H, Li F, Hirakawa H, Kawanabe T, Zou Z, Hasegawa Y, Tonosaki K, Shirasawa S, Fukushima A, Yokoi S, Takahata Y, Kakizaki T, Ishida M, Okamoto S, Sakamoto K, Shirasawa K, Tabata S, Nishio T (2014) Draft sequences of the radish (Raphanus sativus L.). Genome DNA Res 21(5):481–490
Kobabe G (1959) Naturliche Einkreuzung von Hederich (Raphanus raphanistrum L.) in Radies (Raphanus sativus var. radicula DC.) und das Verhalten von Knollenform und Farbe in den nachfolgenden F-und R-Generationen. Zeitschrift fur Pflanzenzuchtung 42:1–10
Kong Q, Li X, Xiang C, Wang H, Song J, Zhi H (2011) Genetic diversity of radish (Raphanus sativus L.) germplasm resources revealed by AFLP and RAPD markers. Plant Mol Biol Rep 29:217–223
Kumazawa S (1961) Horticulture, detailed discussion of crops. Yokendo, Tokyo, p 637 (in Japanese)
Lewis-Jones LJ, Thorpe JP, Wallis GP (1982) Genetic divergence in four species of the genus Raphanus: implications for the ancestry of the domestic radish R. sativus. Biol J Linn Soc 18:35–48
Li S (1989) The origin and resources of vegetable crops in China. International symposium on horticultural germplasm, cultivated and wild, Beijing, China, September 1988. Chinese Society for Horticultural Science, International Academic Publishers, Beijing, pp 197–202
Li XX, Shen D et al (2008) Descriptors and data standards for radish (Raphanus sativus L.). China Agriculture Press, Beijing (in Chinese)
Li F, Hasegawa Y, Saito M, Shirasawa S, Fukushima A, Ito T, Fujii H, Kishitani S, Kitashiba H, Nishio T (2011) Extensive chromosome homoeology among Brassiceae species were revealed by comparative genetic mapping with high density EST-based SNP markers in radish (Raphanus sativus L.). DNA Res 18:401–411
Liu RH, Meng JL (2003) MapDraw: a Microsoft Excel macro for drawing genetic linkage maps based on given genetic linkage data. Heraditas (Beijing) 25(3):317–321
Liu LW, Zhao LP, Gong YQ, Wang MX, Chen LM, Yang JL, Wang Y, Yu FM, Wang LZ (2008) DNA fingerprinting and genetic diversity analysis of late-bolting radish cultivars with RAPD, ISSR and SRAP markers. Sci Hortic 116:240–247
Lü N, Yamane K, Ohnishi O (2008) Genetic diversity of cultivated and wild radish and phylogenetic relationships among Raphanus and Brassica species revealed by the analysis of trnK/matK sequence. Breed Sci 58:15–22
Madhou P, Wells A, Pang ECK, Stevenson TW (2005) Genetic variation in populations of Western Australian wild radish. Aust J Agric Res 56:1079–1087
Makino T (1961) Makino’s new illustrated flora of Japan. Hokuryukan, Tokyo (in Japanese)
Matveeva TV, Simonova AV, Lutova LA (2002) Molecular markers of inbred radish (Raphanus sativus var. radicola Pers) lines. Cell Mol Biol Lett 7:845–848
Muminović J, Merz A, Melchinger AE (2005) Genetic structure and diversity among radish varieties as inferred from AFLP and ISSR analyses. J Am Soc Hortic Sci 130(1):79–87
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325
Muuoz MC, Bermejo JEH (1978) La corola en la tribu Brassiceae. Anal del Insti Bot Cava 35:297–334
Nakatsuji R, Hashida T, Matsumoto N, Tsuro M, Kubo N, Hirai M (2011) Development of genomic and EST-SSR markers in radish (Raphanus sativus L.). Breed Sci 61:413–419
Ohsako T, Hirai M, Yamabuki M (2010) Spatial structure of microsatellite variability within and among populations of wild radish Raphanus sativus L. var. hortensis Backer f. raphanistroides Makino (Brassicaceae) in Japan. Breed Sci 60:195–202
Qi JJ, Liu X, Shen D, Miao H, Xie BY, Li XX et al (2013) A genomic variation map provides insights into the genetic basis of cucumber domestication and diversity. Nat Genet 12(45):1510–1518
Rohlf FJ (2008) NTSYSpc: numerical taxonomy system, ver. 2.20. Exeter, Setauket
Shen D, Sun H, Huang M, Zheng Y, Li X, Fei Z (2013a) RadishBase: a database for genomics and genetics of radish. Plant Cell Physiol 54:e3
Shen D, Sun H, Huang M, Zheng Y, Qiu Y, Li X, Fei Z (2013b) Comprehensive analysis of expressed sequence tags from cultivated and wild radish (Raphanus spp.). BMC Genomics 14:721
Shirasawa K, Oyama M, Hirakawa H, Sato S, Tabata S, Fujioka T, Kimizuka-Takagi C, Sasamoto S, Watanabe A, Kato M et al (2011) An EST-SSR linkage map of Raphanus sativus and comparative genomics of the Brassicaceae. DNA Res 18:221–232
Tsuro M, Suwabe K, Kubo N, Matsumoto S, Hirai M (2008) Mapping of QTLs controlling root shape and red pigmentation in radish, Raphanus sativus L. Breed Sci 58:55–61
Wang N, Kitamoto N, Ohsawa R, Fujimure T (2008) Genetic diversity of radish (Raphanus sativus) germplasms and relationships among worldwide accessions analyzed with AFLP markers. Breed Sci 58:107–112
Wang LX, Chang LF, Li HB, Ji W, Liu LH, Zhao CP (2010) Molecular markers for estimating distinctness, uniformity, and stability of wheat lines in regional trials. Acta Agron Sin 36(7):1114–1125 (in Chinese)
Wang QB, Zhang YY, Fang ZY, Liu YM, Yang LM, Zhuang M (2012) Chloroplast and mitochondrial SSR help to distinguish allo-cytoplasmic male sterile types in cabbage (Brassica oleracea L. var. capitata). Mol Breed 30:709–716
Xu L, Wang L, Gong Y, Dai W, Wang Y, Zhu X, Wen T, Liu L (2012) Genetic linkage map construction and QTL mapping of cadmium accumulation in radish (Raphanus sativus L.). Theor Appl Genet 125:659–670
Yamagishi H (2004) Assessment of cytoplasmic polymorphisms by PCR-RFLP of the mitochondrial orfB region in wild and cultivated radishes (Raphanus). Plant Breed 123:141–144
Yamagishi H, Terachi T (2003) Multiple origins of cultivated radishes as evidenced by a comparison of the structural variations in mitochondrial DNA of Raphanus. Genome 46:89–94
Yamagishi H, Tateishi M, Terachi T, Murayama S (1998) Genetic relationships among Japanese wild radishes (Raphanus sativus f. raphanistroides Makino), cultivated radishes and R. raphanistrum revealed by RAPD analysis. J Jpn Soc Hortic Sci 67:526–531
Yamane K, Lü N, Ohnishi O (2005) Chloroplast DNA variations of cultivated radish and its wild relatives. Plant Sci 168:627–634
Yamane K, Lü N, Ohnishi O (2009) Multiple origins and high genetic diversity of cultivated radish inferred from polymorphism in chloroplast simple sequence repeats. Breed Sci 59:55–65
Zohary D, Hopf M (2000) Domestication of plants in the old world, 3rd edn. University Press, Oxford
Acknowledgments
This work was funded by grants from National Key Technology R&D Program of China (2012BAD02B01, 2012BAD50G00) and the Technological Innovation Capacity Program of Beijing Academy of Agricultural and Forestry Sciences (KJCX20140111).
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Supplement Figure S1 Amplified bands of primer RSS2474 in 37 radish accessions and their patterns and codes for each allele. There were seven alleles at locus a, meanwhile, could not be detected polymorphism at locus b in this study (JPEG 187 kb)
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Supplement Figure S5 Compare UPGMA cluster dendrogram in this study with Neighbor-joining tree of radish in Shen et al. (JPEG 113 kb)
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Wang, Q., Zhang, L. & Zheng, P. Genetic diversity and evolutionary relationship analyses within and among Raphanus species using EST-SSR markers. Mol Breeding 35, 62 (2015). https://doi.org/10.1007/s11032-015-0261-1
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DOI: https://doi.org/10.1007/s11032-015-0261-1