Biotechnology Bulletin ›› 2019, Vol. 35 ›› Issue (11): 224-230.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0409

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Development of SNP Marker Based on the Rehmannia glutinosa Transcriptome Database and Construction of DNA Fingerprint in Rehmannia

GUO Meng-meng, ZHOU Yan-qing, DUAN Hong-ying, YANG Ke, SHAO Lu-ying   

  1. College of Life Sciences,Henan Normal University,Xinxiang 453007
  • Received:2019-05-13 Online:2019-11-26 Published:2019-11-19

Abstract: Rehmannia glutinosa possesses well-known medicinal properties and nutritional values. Accurately identifying varieties is essential for germplasm management and breeding. Using the SNP molecular markers to identify R. glutinosa germplasm and to construct fingerprints provides a new method for R. glutinosa molecular marker breeding. Rehmannia transcriptome data was applied and compared with three rehmannia transcriptome data in the SRA database to find candidate SNPs,and PCR technology amplification and sequence analysis were used to study the mutations of candidate SNP of 28 Rehmannia cultivars. Total 102 075 Unigenes were obtained from the transcriptome data of R. glutinosa,including 35 339 SNP loci with a frequency of 0.34. A 40 candidate SNPs were randomly selected,and 39 pairs of primers were designed. Seven pairs of good SNP primers were selected by PCR and sequence analysis,including 8 SNP loci with fine polymorphism. The fingerprint of R. glutinosa was constructed using these 8 SNP loci,which allowed to distinguish 17 different rehmannia germplasm,thus it can be used for the identification of interspecific and intraspecific species of R. glutinosa.

Key words: Rehmannia glutinosa, transcriptome, single nucleotide polymorphism(SNP)markers, fingerprint, variety identification