Isolation and characterization of microsatellite markers and analysis of genetic diversity in Chinese jujube (Ziziphus jujuba Mill.)

PLoS One. 2014 Jun 16;9(6):e99842. doi: 10.1371/journal.pone.0099842. eCollection 2014.

Abstract

Chinese jujube (Ziziphus jujuba Mill, 2n = 2× = 24, Rhamnaceae) is an economically important Chinese native species. It has high nutritional value, and its medicinal properties have led to extensive use in traditional oriental medicine. The characterization of genotypes using molecular markers is important for genetic studies and plant breeding. However, few simple sequence repeat (SSR) markers are available for this species. In this study, 1,488 unique SSR clones were isolated from Z. jujuba 'Dongzao' using enriched genomic libraries coupled with a three-primer colony PCR screening strategy, yielding a high enrichment rate of 73.3%. Finally, 1,188 (80.87%) primer pairs were amplified successfully in the size expected for 'Dongzao'. A total of 350 primer pairs were further selected and evaluated for their ability to detect polymorphisms across a panel of six diverse cultivars; among these, 301 primer pairs detected polymorphisms, and the polymorphism information content (PIC) value across all loci ranged from 0.15 to 0.82, with an average of 0.52. An analysis of 76 major cultivars employed in Chinese jujube production using 31 primer pairs revealed comparatively high genetic diversity among these cultivars. Within-population differences among individuals accounted for 98.2% of the observed genetic variation. Neighbor-joining clustering divided the cultivars into three main groups, none of which correspond to major geographic regions, suggesting that the genetics and geographical origin of modern Chinese jujube cultivars might not be linked. The current work firstly reports the large-scale development of Chinese jujube SSR markers. The development of these markers and their polymorphic information represent a significant improvement in the available Chinese jujube genomic resources and will facilitate both genetic and breeding applications, further accelerating the development of new cultivars.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • China
  • DNA Primers / metabolism
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification*
  • Gene Library
  • Genetic Loci
  • Genetic Variation*
  • Microsatellite Repeats / genetics*
  • Nucleotide Motifs / genetics
  • Phylogeny
  • Polymorphism, Genetic
  • Repetitive Sequences, Nucleic Acid / genetics
  • Ziziphus / genetics*

Substances

  • DNA Primers
  • DNA, Plant

Grants and funding

Support for this work came from the following: 1.Special Fund for Forestry-Scientific Research in the Public Welfare (201004017) from the State Forestry Administration of China (www.forestry.gov.cn) to XP; 2.Grant 31372019 from the National Natural Science Foundation of China (http://www.nsfc.gov.cn/) to XP; 3. Projects in the National Science & Technology Pillar Program (2013BAD14B0302) from the Ministry of Science and Technology of China (www.most.gov.cn/) to XP; 4.the Fundamental Research Funds for the Central Universities (JD2010–5) from the Ministry of Education of China (www.moe.gov.cn) to XP; and 5. the Thousand-person Plan Award from http://www.1000plan.org/to RW. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.