Genetic Resources and Crop Evolution

, Volume 54, Issue 4, pp 767–777 | Cite as

Genetic variation and species relationships in Himalayan buckwheats as revealed by SDS PAGE of endosperm proteins extracted from single seeds and RAPD based DNA fingerprints

Original paper


SDS PAGE of endosperm proteins and␣RAPD profiles from different accessions of Himalayan buckwheat were studied to determine their genetic variation and phylogenetic relationships. Comparisons based on Jaccard’s coefficient and UPGMA clustering revealed interrelationships broadly in conformity with conventional treatments. Cluster analysis of the endosperm protein profiles of the selected accessions revealed three broad clusters. A moderate level of intraspecific variability was detected in the endosperm protein profile of different accessions of Fagopyrum esculentum, Moench. Three subgroups were detected in cluster 1. Subgroup 1 included varieties designated as Local, Kamroo local, OC-2 and␣VL-7 which were collected from VPKAS, Almora. Local, Kamroo local and OC-2 showed a similarity coefficient of 1.0 inspite of their being identified as different accessions. VL-7 emerged out separately from the rest of the three accessions. Accessions having winged grains and those having striations on the seed coat formed a 2nd and 3rd subgroup, respectively. IC-13145 which been identified as “F. himalianum”, showed 100%␣similarity in endosperm protein profile with IC-13376 (F. esculentum) and 85–90% similarity which other accessions of F. esculentum. Our results indicate that “F. himalianum” belongs to the esculentum group and should not be regarded as a different species. Cluster 2 included all the accessions of F. tataricum (L.) Gaertn. with Sangla 1, 2, 3, 5, 6, 7 and IC-412863 showing 100% similarity. F. cymosum emerged as a separate group distinct from both esculentum and tataricum. Accessions of F. tataricum and F. cymosum did not show significant intraspecific variation in the SDS PAGE profile of endosperm proteins. Out of the 20 primers used, 3 generated robust, easily interpretable amplified products. While a 1490 bp and a 300 bp RAPD was detected only in F. tataricum, a 1154 bp RAPD was detected in all accessions of F. tataricum except in Shimla B-1. This variety is early maturing and has high seed yields.


Buckwheat accessions Fagopyrum spp. Genetic diversity RAPD SDS PAGE Soluble proteins 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



The authors are thankful to the Head, Dept. of Botany for the use of laboratory facilities. Financial assistance provided by DST, New Delhi in the form of a research project to NKC is gratefully acknowledged.


  1. Arora RK, Baniya BK, Joshi BD (1995) Buckwheat genetic resources in the Himalayas: their diversity, conservation and use. In: Matano T, Ujihara A (eds) Current advances in buckwheat research. Shinshu University Press, Asahi Matsumoto City, Japan, pp 39–46Google Scholar
  2. Baniya BK (1990) Buckwheat in Nepal. Fagopyrum 10:23–32Google Scholar
  3. Baniya BK, Riley KW, Dongal DMS, Sherchand K (1992) Characterization of Nepalese hill crop landraces (barley, buckwheat, finger millet, grain amaranth, foxtail, proso and barnyard millets). NHCRP-NARC, Nepal, p 51Google Scholar
  4. Bharali S (2002) Isolation, cloning and molecular analysis of the legumin gene of common buckwheat (Fagopyrum esculentum). PhD Thesis, North Eastern Hill University, Shillong (India)Google Scholar
  5. Campbell CG (1997) Buckwheat Fagopyrum esculentum Moench. Promoting the conservation and use of underutilized and neglected crops. 19. IPGRI, RomeGoogle Scholar
  6. Doll H, Brown ADH (1979) Hordein variation in wild (Hordeum spontaneum) and cultivated (H. vulgare) barley. Can J Genet Cytol 21:239–404Google Scholar
  7. Duarte JM, Santos JB- dos, Melo LC (1999) Genetic diversity among common bean cultivar from different races based on RAPD markers. Genet Mol Biol 22:419–426Google Scholar
  8. Hammer K (2001) Polygonaceae. In: Hanelt P, Institute of Plant Genetics and Crop Plant Research (eds.), Mansfeld’s Encyclopedia of Agricultural and Horticultural Crops. Springer, Berlin, pp 285–308Google Scholar
  9. Javornik B, Kump B (1993) Random amplified polymorphic DNA (RAPD) markers in buckwheat. Fagopyrum 13:35–39Google Scholar
  10. Jiang J, Xing J (1992) Dalianshan region in Sichuan province one of the habitats of tatary buckwheat. In: Proceedings of 5th international symposium on Buckwheat, 20–26 August 1992, Taiyuan, China. Agriculture Publishing House, Taiwan, pp 17–18Google Scholar
  11. Joshi BD, Paroda RS (1991) Buckwheat in India. Science Monograph 2, NBPGR, Shimla, p 117Google Scholar
  12. KishimaYK, Mizukami OK, Mikami T, Adachi T (1995) Chloroplast DNA analysis in buckwheat species: phylogenetic relationships, origin of reproductive systems and extended inverted repeats. Plant Sci 108:173–179CrossRefGoogle Scholar
  13. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  14. Marshall HG, Pomeranz Y (1982) Buckwheat description, breeding, production and utilization. In: Y. Pomeranz (ed) Advances in cereal science and technology. Am Assoc Cereal Chem, St. Paul, MN, pp 157–212Google Scholar
  15. Mignouna HD, Ng NQ, Ikea J, Thotapilly G (1998) Genetic diversity in cowpea as revealed by ramdom amplified polymorphic DNA. J Genet Plant Breeding 2:151–159Google Scholar
  16. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucl Acid Res 8:4321–4325CrossRefGoogle Scholar
  17. Nishiyama K, Lachman S, Miura M (1991) Electrophoretic property of buckwheat seed protein. In: Proc ICOBB, Miyazaki, pp 215–225Google Scholar
  18. Ohnishi O (1998a) Search of the wild ancestors of buckwheat. III The wild ancestor of cultivated common buckwheat and of tatary buckwheat. Econ Bot 52:123–133Google Scholar
  19. Ohnishi O. (1998b) Search of the wild ancestors of buckwheat. I Description of new Fagopyrum species and their distribution in China. Fagopyrum 15:18–28Google Scholar
  20. Ohnishi O (2004) Wild Buckwheat species in the San Jiang (Three rivers) area of Southwestern China. In: Proceedings of 9th international symposium on Buckwheat, 18–22 August 2004, Prague, Czech Republic. Research Institute of Crop Production, Prague, pp␣226–232Google Scholar
  21. Ohnishi O, Asona N (1999) Genetic diversity of Fagopyrum homotropicum, a wild species related to common buckwheat. Genet Resour Crop Evol 46:389–398CrossRefGoogle Scholar
  22. Ohnishi T, Matsuoka Y (1996) Search for the wild ancestor of Buckwheat. II Taxonomy of the Fagopyrum (Polygonaceae) species based on morphology, isozyme and chloroplast DNA variability. Genes Genet Syst 71:383–390CrossRefGoogle Scholar
  23. Ohsako T, Ohnishi O (1998) New Fagopyrum species revealed by morphological and molecular analysis. Genes Genet Syst 73:85–94CrossRefGoogle Scholar
  24. Ohsako T, Ohnishi O (2000) Intra and inter-specific phylogeny of the wild Fagopyrum (Polygonaceae) species based on nucleotide sequences of non coding regions of chloroplast DNA. Am J Bot 87:573–582PubMedCrossRefGoogle Scholar
  25. Potokina E, Vaughan DA, Eggi EF, Tomooka N (2000) Population diversity of the Vicia sativa (Fabaceae) in the flora of former USSR deduced from RAPD and seed protein analysis. Genet Resour Crop Evol 47:171–183CrossRefGoogle Scholar
  26. Rabbani MA, Iwabuchi A, Murakami Y, Suzuki T, Takayanagi K (1998) Genetic diversity in mustard (Brassica juncea L.) germplasm from Pakistan as determined by RAPDs. Euphytica 103:235–242CrossRefGoogle Scholar
  27. Rohlf FJ (1993) NTSYS-PC: Numerical taxonomy and multivariate analysis system. Version 2.1. Exeter Publishing, Setauket NYGoogle Scholar
  28. Rout MK, Chrungoo NK (1996) Partial characterization of the lysine rich 13S globulin from buckwheat: its antigenic homology with seed proteins of some other crops. Biochem Mol Biol Int 40:587–595PubMedGoogle Scholar
  29. Sharma TR, Jana S (2002) Species relationships in Fagopyrum revealed by PCR-based DNA fingerprinting. Theor Appl Genet 105:306–312PubMedCrossRefGoogle Scholar
  30. Svetek S (1994) Electrophoretic analysis of buckwheat (Fagopyrum esculentum Moench). In: Proc IPBA, Rogla, Slovenia. pp 161–171Google Scholar
  31. Thompson JA, Nelson RL, Vodkin LO (1998) Identification of diverse soybean germplasm using RAPD markers. Crop Sci 38:1348–1355CrossRefGoogle Scholar
  32. Tsuji K, Ohnishi O (1998) Phylogenetic relationships among cultivated landraces and natural populations of tartary buckwheat (Fagopyrum tataricum) revealed by RAPD analysis. In: Proc 7th Int Symp Buckwheat, pp 41–49Google Scholar
  33. Yasui Y, Ohnishi O (1998a) Interspecific relationship among Fagopyrum species revealed by nucleotide sequences of the rbcL and accD genes and their intergeneric region. Am J Bot 85:1134–1142CrossRefGoogle Scholar
  34. Yasui Y, Ohnishi O (1998b) Interspecific relationship among Fagopyrum species revealed by nucleotide sequences of the ITS region of the nuclear rRNA gene. Genes Genet Syst 3:201–210CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2007

Authors and Affiliations

  1. 1.Department of BotanyNorth Eastern Hill UniversityShillongIndia

Personalised recommendations