Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
권호 표지

Identification of Different Species and Dultivars of Brassica by SDS-PAGE, Isozyme and Molecular Marker

  • Mukhlesur Rahman Md. (Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Department of Genetics and Plant Breeding, Bangladesh Agricultural University) ;
  • Hirata Yutaka (Graduate School of Agriculture, Tokyo University of Agriculture and Technology)
  • Published : 2005.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.

Keywords

References

  1. Anand IJ, Reddy WR, Rawat DS (1985) Inheritance of seed color in mustard. Indian J Genet 45: 34-37
  2. Chauhan YS, Kumar K (1987) Genetics of seed color in mustard (Brassica juncea L. Czern and Coss). Cruciferae Newsl 12: 22-23
  3. Chen BY, Heneen WK, Simonsen V (1989) Comparative and genetic studies of isozymes in resynthesized and cultivated Brassica napus L., B. campestris L. and Brassica alboglabra Bailey. Theor Appl Genet 77: 673-679 https://doi.org/10.1007/BF00261242
  4. Chevre AM, Delourme R, Eber F, Margale E, Quiros CF, Arus P (1994) Genetic analysis and nomenclature for seven isozyme systems in Brassica nigra, B. oleracea and B. campestris. Plant Breeding 114: 473-480 https://doi.org/10.1111/j.1439-0523.1995.tb00839.x
  5. Das S, Rajagopal J, Bhatia S, Srivastava PS, Lakshmikumaran M (2000) Assessment of genetic variation within Brassica campestris cultivars using amplified fragment length polymorphism and random amplification of polymorphic DAN markers. Plant Molecular Biology Division, Tata Energy Research Institute, Lodhi Road, New Delhi, India
  6. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA mini preparation: Version II. Plant Mol Biol Rpt 1: 19-21 https://doi.org/10.1007/BF02712670
  7. Eber F, Chevre AM, Baranger A, Vallee P, Tanguy X, Renard M (1994) Spontaneous hybridization between a male sterile oilseed rape and two weeds. Theor Appl Genet 88: 362-368 https://doi.org/10.1007/BF00223646
  8. Kumar R, Gupta VP (1985) Isozyme studies in Indian mustard (Brassica juncea L.). Theor Appl Genet. 69: 1-4
  9. Moriyuki S, Nagamine T, Terauchi T, Akamine F, Sugimoto A (1999) Isozyme application for variety identification and progeny hybridity in Japanese sugercane. Breeding Science 49: 89-95 https://doi.org/10.1270/jsbbs.49.89
  10. Negi MS, Devic M, Delseny M, Lakshmikumaran M (2000) Identification of AFLP fragments linked to seed coat color in Brassica juncea and conversion to a SCAR marker for rapid selection. Theor Appl Genet 101: 146-152 https://doi.org/10.1007/s001220051463
  11. Nozaki, T., Anji, M., Takahashi, T. and Ikehashi, H. (1995) Analysis of isozyme loci and their linkage in Brassica campestris L. Breeding Science 45: 57-64
  12. Person K, Fait AS, von Bothmer R (2001) Genetic diversity of allozymes in turnip (Brassica rapa L. var. rapa) from the Nordic area. Hereditas 134: 43-52 https://doi.org/10.1111/j.1601-5223.2001.00043.x
  13. Quiros CF, Ochoa O, Kianian SF, Douches D (1988) Exploring the role of x = 7 species in Brassica evolution: hybridization with B. nigra and B. oleracea. J Heredity 79: 351-358 https://doi.org/10.1093/oxfordjournals.jhered.a110527
  14. Rahman MH (2001) Introgression of alleles of the isozymic locus glucose phosphatase isomerase-2 (GPI-2) from the CC genome of Brassica carinata to the CC genome of Brassica alboglabra and their independent segregation from seed color. Plant Breeding 120: 363-364 https://doi.org/10.1046/j.1439-0523.2001.00619.x
  15. Ripley V, Thrope L, Iler S, Mizier K, Beversdorf WD (1992) Isozyme analysis as a tool for introgression of Sinapis alba germplasm into B. napus. Theor Appl Genet 84: 403-410 https://doi.org/10.1007/BF00229500
  16. Robbelen G, 1960. Beitrage zur Analyse des Brassica-Ge-noms. Chromosome 11: 205-228 https://doi.org/10.1007/BF00328652
  17. Takehisa I, Noda S, Mori S, Yamashita M, Nakanishi H, Inoue M, Kamijima O (2001) Genetic variation and geographical distribution of Azuki bean (Vigna angularis) land races based on the electrophoregram of seed storage proteins. Breeding Sciences 51: 225-230 https://doi.org/10.1270/jsbbs.51.225
  18. Tanksley SO, Jones RA (1986) Application of alcohol bedydrogenase allozymes in testing the genetic purity of F1 hybrids of tomato. Hort Sci 16: 179-181
  19. Thanh VC, Hirata Y (2002) Seed storage protein diversity of three rice species in the Mekong Delta. Biosphere Conservation 4: 59-67