Effect of Genotype, Cold Pre-Treatment, Incubation Conditions and Media on Embryo and Plantlet Formation in Pepper Anther Culture

Authors : Perihan Durna, Naif Geboloğlu, Emine Polat, Ertan Sait Kurtar

Pages : 1-12

Doi:10.29278/azd.1662151

View : 78 | Download : 51

Publication Date : 2025-06-30

Article Type : Research Paper

Abstract :Objectives: The aim of this study was to investigate the effects of various culture conditions on embryoid and plantlet formation in pepper anther cultures. To achieve this, the effects of media compositions, vitamin B₁₂, activated charcoal, bud treatments, and incubation temperatures were evaluated. Materials and Methods: The study was conducted at the screenhouse and tissue culture laboratory of Gaziosmanpaşa University. Three pepper (Capsicum annuum L.) genotypes—Belissa F₁, Bafra F₁, and İstek F₁—and their F₂ populations were used to examine the effects of different culture conditions on embryoid formation. Donor plants were grown in a controlled environment using Hoagland’s nutrient solution and an integrated pest management approach. Anthers were cultured in DDVX medium, modified with different concentrations of vitamin B₁₂ (control, 0.03 mg L⁻¹, 0.05 mg L⁻¹), and in Double Layer (DL) medium with 0.10% and 0.20% activated charcoal. To stimulate embryoid formation, buds were subjected to cold shock at 4°C for 24 or 48 hours, and incubation temperatures ranging from 9°C to 35°C were tested. Results: In DDVX medium, 702 embryoids and 124 haploid plantlets were obtained. The highest embryoid formation (51) was observed in the Belissa F₂ genotype under conditions of 24-hour pre-treatment at 4°C, no vitamin B₁₂ supplementation, and incubation at 35°C. In the Bafra and İstek genotypes, vitamin B₁₂ (0.03–0.05 mg·L⁻¹) and 35°C incubation significantly enhanced embryoid formation. A 24-hour pre-treatment at 4°C yielded the best results, while a 48-hour pre-treatment did not provide any additional benefit. In DL medium, 274 embryoids and 49 haploid plantlets were produced. The highest number of embryoids (34) was observed in the Belissa F₂ genotype under control conditions with 0.20% activated charcoal and incubation at 9°C. In the Bafra and İstek genotypes, 0.10–0.20% activated charcoal and 9°C incubation provided the most effective results. Unlike DDVX, bud pre-treatments in DL medium did not show a clear advantage. DDVX medium was 256% more successful in embryoid formation compared to DL medium. F₂ populations produced higher embryoid numbers compared to F₁ populations. In DDVX, 35°C incubation was more effective, while in DL, 9°C incubation yielded better results. Conclusion: The study demonstrated that genotype, nutrient medium, and incubation conditions significantly influence embryoid formation in pepper. DDVX medium produced significantly more embryoids and plantlets compared to DL medium.
Keywords : DDVX ortamı, double layer ortamı, aktif kömür, vitamin B₁₂, haploid

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