RESEARCH PAPER
Estimation of genetic divergence within androgenic
regenerants of Capsicum annuum L. ATZ1 × C. frutescens L. F1
plants using random amplified polymorphic DNA markers
More details
Hide details
1
Department of Genetics and Plant Biotechnology, University of Technology and Life Sciences, Bydgoszcz, Poland
Submission date: 2017-01-31
Final revision date: 2017-06-01
Acceptance date: 2017-07-05
Publication date: 2018-01-02
BioTechnologia 2017;98(3):175-182
KEYWORDS
TOPICS
ABSTRACT
To prove the androgenic origin of the regenerants obtained from the anther cultures of interspecific F1 C. annuum L. ATZ1 × C. frutescens L.) hybrid, Random Amplified Polymorphic DNA (RAPD) method was applied. Thirty decamer primers were used in the experiment. The size of the amplified products ranged from 138 to 2369 bp, and their number from 3 to 25 per primer. About 274 loci, 47 of which were polymorphic, were analyzed. The results proved the hybrid status of anther donor plants detected by 20 of the applied primers. Of the 25 analyzed androgenic regenerants, 18 were diploids. The RAPD reactions confirmed their polymorphism in comparison with that of the F1 hybrid generation, which excluded their origin from somatic cells of the anthers. Moreover, the results demonstrated the existence of a genetic variation among the obtained haploids, which illustrated genetic diversity of the microspores developing in hybrid anther cultures. The results of the experiments proved the utility of the RAPD method in detecting polymorphisms between closely related pepper plants, proving at the same time the effectiveness of androgenesis, regeneration of haploid plants, and spontaneously doubled haploids (DH lines).
REFERENCES (25)
1.
Ballester J., Vicente M.C. (1998) Determinarion of F1 hybrid seed purity in pepper using PCR-based markers. Euphytica 103: 223-226.
2.
Chambonnet D. (1988) Production of haploid pepper plants. Bulletin Interne de la Station d’Amelioration des Plantes Maraicheres, d’Avignon-Montfavet, France 1-10.
3.
Dolcet-Sanjuan R., Claveria E., Huera A. (1997) Androgenesis in Capsicum annuum L.: effect of carbohydrate and carbon dioxide enrichment. J. Amer. Soc. Hort. Sci. 122: 468-475.
4.
Dumas de Vaulx R., Chambonnet D., Pochard E. (1981) Culture in vitro d’anthéres de piment (Capsicum annuum L.) amélioration des taux d’obtention de plantes chez différents génotypes par des traitements á +35EC. Agronomie 1: 859-864.
5.
Ghislain M., Zhang D., Fajardo D., Huaman Z., Hijmans. (1999) Marker-assisted sampling of the cultivated Andean potato Solanum phureja collection using RAPD markers. Genet. Resour. Crop Evol. 46: 547-555.
6.
Ilbi H. (2003) RAPD markers assisted varietal identification and genetic purity test in pepper, Capsicum annuum. Sci. Hort. 97: 211-218.
7.
Ince A.G., Karaca M., Onus A.N. (2010) Polymorphic microsatellite markers transferable across Capsicum species. Plant Mol. Biol. Rep. 28: 285-291.
8.
Irikova T., Grozeva S., Rodeva V. (2011) Anther culture in pepper (Capsicum annuum L.) in vitro. Acta Physiol. Plant. 33: 1559-1570.
9.
Jang I., Moo J.H., Yoon J.B., Yoo J.H., Yang T.J., Kim Y.J., Park H.G. (2004) Application of RAPD and SCAR markers for purity testing of F1 hybrid seed in chili pepper (Capsicum annuum.). Mol. Cells, 18: 295-299.
10.
Lanteri S., Acquardo A., Quagliotti L., Portis E. (2003) RAPD and AFLP assessment of genetic variation in a landrace of pepper (Capsicum annuum L.), grown in North-West Italy. Genet. Resour. Crop Evol. 50: 723-735.
11.
Lefebvre V., Goffinet B., Chauvet J.C., Caromel B., Signoret P., Brand R., Palloix A. (2001) Evaluation of genetic distances between pepper inbred lines for cultivar protection purpose: comparison of AFLP, RAPD and phenotypic data. Theor. Appl. Genet. 102: 741-750.
12.
Mongkolporn O., Dokmaihom Y., Kanchana-Udomkan C., Pakdeezaraporn P. (2004) Genetic purity test of F1 hybrid Capsicum using molecular analysis. J. Hort. Sci. Biotech. 79: 449-451.
13.
Munyon I.P., Hubstenberger J.F., Philips C. (1989) Origin of plantlets and callus obtained from chile pepper anther culture. In vitro Cell. Develop. Biol. 25(3): 293-296.
14.
Nei M., Li W.H. (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl Acad. Sci. USA. 5269-5273.
15.
Odeigah P.G.C., Oboh B., Ahhalokpe I.O. (1999) The characterization of Nigerian varieties of pepper, Capsicum annuum, and Capsicum fructescens by SDS-polyacrylamide gel electrophoresis of seed proteins. Genet. Resour. Crop Evol. 46: 127-131.
16.
Olszewska D., Kisiała A., Nowaczyk, P. (2011) The assessment of doubled haploid lines obtained in pepper (Capsicum annuum L.) anther culture. Folia Hort. 23(2): 93-99.
17.
Olszewska D., Kisiała A., Niklas-Nowak A., Nowaczyk P. (2014) Study of in vitro anther culture in selected genotypes of genus Capsicum. Turk. J. Biol. 38: 118-124.
18.
Paran I., Aftergoot E., Shifriss C. (1998) Variation in Capsicum annuum revealed by RAPD and AFLP markers. Euphytica 99: 167-173.
19.
Paran I., Voort J.R., Lefebvre V., Jahn M., Landry L., Schriek M., Tanyolac B., Caranta C., Chaim A.B., Livingstone K., Palloix A., Peleman J. (2004) An integrated genetic linkage map of pepper (Capsicum spp.). Mol. Breed. 13: 251-261.
20.
Prasad B., Khan R.G., Radha T., Ravi Ch., Venkataiah P., Subhash K., Reuben T. Ch. (2013) DNA profiling of commercial chili pepper (Capsicum annuum L.) varieties using random amplified polymorphic DNA (RAPD) markers. Afr. J. Biotech. 12(30): 4730-4735.
21.
Sitthiwong K., Matsui T., Sukorakarn S., Okuda N., Kosugi Y. (2005) Classification of pepper (Capsicum annuum L.) accessions by RAPD analysis. Biotechnology 4(4): 305- 309.
22.
Sliwinska E. (2003) Cell cycle and germination of fresh, dried and deteriorated sugarbeet seeds as indicators of optimal harvest time. Seed Sci. Res. 13: 131-138.
23.
Sugita T., Kinoshita T., Kawano T., Yuji K., Yamaguchi K., Nagata R., Shimizu A., Chen L., Kawasaki S., Todoroki A. (2005) Rapid construction of a linkage map using highefficiency genome scanning/AFLP and RAPD, based on an intraspecific, doubled-haploid population of Capsicum annuum. Breed. Sci. 55: 287-295.
24.
Thul S.T., Lal R.K., Shasany A.K., Darokar M.P., Gupta A.K., Gupta M.M., Verma R.K., Khanuja S.P.S. (2009) Estimation of phenotypic divergence in a collection of Capsicum species for yield-related traits. Euphytica 168: 189-196.
25.
Van de Peer Y., De Wachter Y. (1994) TREECON for Windows: a software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Comp. Applic. Biosci. 10: 569-570.