Plant Diversity ›› 2013, Vol. 35 ›› Issue (1): 48-54.DOI: 10.7677/ynzwyj201312023

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Start Codon Region-related Polymorphism (SCRP): A Novel DNA Marker Technique for Assessing Genetic Diversity in Alfalfa Germplasm Collections

 HE  Qing-Yuan-1、2, LI  Zheng-Peng-1, WU  Ping-1, YANG  Hong-Yan-2, WANG  Song-Hua-1   

  1. 1 Life Science College of Anhui Science and Technology University, Fengyang 233100, China; 2 National Center for Soybean Improvement,
    National Key Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2012-02-17 Online:2013-01-25 Published:2012-09-13


Mostly PCRbased molecular markers amplified noncoding regions, or the whole genome randomly, the locus is far away from the gene of targeted trait. We developed a novel marker technique called start codon regionrelated polymorphism (SCRP) aimed for the amplification of gene start condon regions. It is based on the use of two primers of 18 nucleotides. The forward primer is designed from the targeted “Kozak” sequence; and the other primer, the reverse primer is an arbitrary sequence with an ATrich core to anneal with an intron. PCR amplification is applied for the first 5 cycles with an annealing temperature of 35℃, followed by 35 cycles with an annealing tempe-rature of 50℃. In the present study, we utilized SCRP to study genetic diversity of 34 alfalfa cultivars (varieties). Each PCR reaction has generated as many as 3 to 16 fragments of 50 to 2000bp in size. The successful genetic diversity of 34 alfalfa cultivars by SCRP was achieved. This new technique should be useful in genotyping germplasm collections and in tagging genes govern desirable agronomic traits of crop plants.

Key words: Genetic diversity, Medicago sativa, Start Codon Region-related Polymorphism

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