Intra-specific genetic relationship analyses of Elaeagnus angustifolia based on RP-HPLC biochemical markers


Elaeagnus angustifolia Linn. has various ecological, medicinal and economical uses. An approach was established using RP-HPLC (reversed-phase high-performance liquid chromatography) to classify and analyse the intra-specific genetic relationships of seventeen populations of E. angustifolia, collected from the Xinjiang areas of China. Chromatograms of alcohol-soluble proteins produced by seventeen populations of E. angustifolia, were compared. Each chromatogram of alcohol-soluble proteins came from a single seed of one wild plant only. The results showed that when using a Waters Delta Pak. C18, 5 μm particle size reversed phase column (15 mm×3.9 mm), a linear gradient of 25%:_60% solvent B with flow rate of 1 ml/min and run time of 67 min, the chromatography yielded optimum separation of E. angustifolia alcohol-soluble proteins. Representative peaks in each population were chosen according to peak area and occurrence in every seed. The converted data on the elution peaks of each population were different and could be used to represent those populations. GSC (genetic similarity coefficients) of 41% to 62% showed a medium degree of genetic diversity among the populations in these eco-areas. Cluster analysis showed that the seventeen populations of E. angustifolia could be divided into six clusters at the GSC=0.535 level and indicated the general and unique biochemical markers of these clusters. We suggest that E. angustifolia distribution in these eco-areas could be classified into six variable species. RP-HPLC was shown to be a rapid, repeatable and reliable method for E. angustifolia classification and identification and for analysis of genetic diversity.

This is a preview of subscription content, access via your institution.


  1. Batey, I.L., 1984. Wheat varietal identification by rapid ion-exchange chromatography of gliadins. Journal of Cereal Science, 2:241–248.

    CAS  Google Scholar 

  2. Bietz, J.A., 1983. Separation of cereal proteins by reversed-phase high-performance liquid chromatography. Journal of Chromatography, 155:219–238.

    Google Scholar 

  3. Bietz, J.A., Cobb, L.A., 1985. Improved procedures for rapid wheat varietal identification by reversed-phase high-performance liquid chromatography of gliadin. Cereal Chemistry, 62:332–339.

    CAS  Google Scholar 

  4. Brink, D.E., Price, S.C., Nguyen, H., Martinez, C., 1989. Genetic purity assessment of commercial single cross maize hybrids: isoelectric focusing of zein. Seed Science and Technology, 17:91–98.

    Google Scholar 

  5. Dinelli, G., Bonetti, A., 1992. Capillary electrophoresis in species and cultivar determination. Seed Science and Technology, 20:241–249.

    Google Scholar 

  6. FLORA of China, 1983. Reipublicae Popularis Sinicae. Tomus, Science Press, Beijing (in Chinese).

    Google Scholar 

  7. Hosseinzadeh, H., Ramezani, M., Namjo, N., 2003. Muscle relaxant activity of Elaeagnus angustifolia L. fruit seeds in mice. Journal of Ethnopharmacology, 84(2–3):275–278. [doi:10.1016/S0378-8741(02)00331-8]

    PubMed  Google Scholar 

  8. Huebner, F.R., Bietz, J.A., Webb, B.D., Juliano, B.O., 1990. Rice cultivar identification by high-performance liquid chromatography of endosperm proteins. Cereal Chemistry, 67:129–135.

    CAS  Google Scholar 

  9. Klich, M.G., 2000. Leaf variations in Elaeagnus angustifolia related to environment heterogeneity. Environmental and Experimental Botany, 44(3):171–183. [doi:10.1016/S0098-8472(00)00056-3]

    Article  PubMed  Google Scholar 

  10. Kubiczek, R.P., Huebner, F.R., Bietz, J.A., 1993. Reversed-phase high-performance liquid chromatography of secalins: application to rye cultivar identification. Journal of Cereal Science, 17(3):191–201. [doi:10.1006/jcrs.1993.1019]

    Article  CAS  Google Scholar 

  11. Lookhart, G.L., Pomeranz, Y., 1985. Characterization of oat species by polyacrylamide gel electrophoresis and high performance liquid chromatography of their prolamin proteins. Cereal Chemistry, 62:162–166.

    CAS  Google Scholar 

  12. Marchylo, B.A., Kruger, J.E., 1984. Identification of Canadian barley cultivars by reversed-phase high-performance liquid chromatography. Cereal Chemistry, 61:295–301.

    CAS  Google Scholar 

  13. Mirhydar, H., 1998. Encyclopedia of Plants: Indications of Plants in the Prevention and Treatment of Diseases, Vol. 2. Islamic Farhang, Tehran, p.163–164.

    Google Scholar 

  14. Nei, M., 1972. Genetic distance between populations. Am. Nat., 106(949):283–292. [doi:10.1086/282771]

    Article  Google Scholar 

  15. Perry, L.M., 1980. Medicinal Plants of the East and Southeast Asia. MIT Press, London, p.131.

    Google Scholar 

  16. Ram, C., Chandgi, F.R., Bietz, J.A., 1995. Identification of Indian wheat varieties by reversed-phase high-performance liquid chromatography. Seed Science and Technology, 23:259–262.

    Google Scholar 

  17. Smith, J.S.C., Smith, O.S., 1992. Fingerprinting crop varieties. Advances in Agronomy, 47:85–140.

    CAS  Google Scholar 

  18. Wang, Q., Ruan, X., Jin, Z.H., Yan, Q.C., Tu, S.J., 2005. Identification of Rhodiola species by using RP-HPLC. Journal of Zhejiang University SCIENCE, 6B(6):477–482. [doi:10.1631/jzus.2005.B0477]

    CAS  Google Scholar 

  19. Zargari, A., 1990. Medicinal Plants, Vol. 4. Tehran University Press, Tehran, p.275–277.

    Google Scholar 

  20. Zhang, B.Z., Zhao, K.F., 1996. Study on salt tolerance in Robinia and Elaeagnus angustifolia. Shandong Science, 9(2):53–55.

    Google Scholar 

Download references

Author information



Corresponding author

Correspondence to Qiang Wang.

Additional information

Project (Nos. 30470330 and 30100126) supported by the National Natural Science Foundation of China

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wang, Q., Ruan, X., Huang, Jh. et al. Intra-specific genetic relationship analyses of Elaeagnus angustifolia based on RP-HPLC biochemical markers. J. Zhejiang Univ. - Sci. B 7, 272–278 (2006).

Download citation

Key words

  • E. angustifolia
  • Intra-specific genetic relationship
  • Genetic diversity
  • Biochemical marker

CLC number

  • Q94