To prove the androgenic origin of the regenerants obtained from the anther cultures of interspecific F 1 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 F 1 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).

In research on androhaploids in the progeny of interspecific hybrids within the Capsicum genus, three genetically stable lines of F 7 generation, selected from the C. annuum L. 9 C. chinense Jacq. hybrid, were used. In the first line, only callus tissue was formed as a reaction to the conditions of culture. Cytometric analysis of this tissue revealed the presence of cells with DNA content in nuclei at the level of 1C to 16C. The tissue was mixoploid and non-embryogenic. Anthers of the other line did not respond. In the third one, nine embryos were obtained, and they developed into plants. By means of cytometric analysis, the 1C DNA level was found in eight of them and these were androgenic plants. The origin of one of the diploid plant was not established due to the homozygous character of the donor plants. The experiment results confirm the already known diversity of genotype reaction to the conditions of culture. It moreover point to the possibility of selection of the forms with an androgenic potency from interspecific hybrids.

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Summary In the research on induced androgenesis, eight groups of plants, the progeny of interspecific hybrid of Capsicum frutescens L. x C. annuum L., were used. Half of them were standard hard-flesh forms, while the others have represented SS or Ss genotype conditioning the soft flesh of ripe fruit. Anthers from the plants of six groups produced mixoploid callus and the DNA content ranged from 1C to 16C. Three groups formed embryos which converted into plants. The total number of regenerants was 19, included both androgenic haploids (13) and diploids (6). The results did not allow to present the simple relationship between the physiological feature of soft-flesh and the effectiveness of androgenesis. Pungent, soft-flesh genotypes appeared as a poor responsive in anther culture. The R2 and R3 generation of two soft-flesh diploids, evaluated in two-year experiments, showed full phenotypic uniformity, proofing the androgenic origin of diploids. These DH lines were different with regard to capsaicinoid profiles. The content of capsaicinoids ranged from 0.056 to 2.170 and from 0.019 to 1.610 g*kg-1 for capsaicin and dihydrocapsaicin, respectively. The highest concentration of the compounds was observed in the placenta. Although the androgenic effectiveness was not fully satisfactory, the doubled haploid technology can be used for the rapid genetic stabilization of soft-flesh Capsicum spp. recombinants and the special attention should be paid to the spontaneous androdiploids.