Ali Akbar Heidari Zefreh1,2, Mehran E. Shariatpanahi2*,
Amir Mousaviand Sepideh Kalatejari1

1Department of Horticultural Science, College of Agricultural Science and Natural Resources, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran.
2Department of Tissue and Cell Culture, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
3Natioal Institute of Genetic Engineering and Biotechnology of Iran (NIGEB), Tehran, Iran. 

Abstract

Creation of pure lines is one of the basic requirements in plant breeding. Doubled haploid (DH) lines are completely homozygous lines and anther culture is an efficient method for DH line production. Induction of androgenic embryos in anther culture is the main restricting factor. In this study we investigated androgenic embryo induction in different F1 hybrid peppers including Cadia, Magno, Plato, King Arthur and Maratus. Excised anthers were cultured in C medium containing 2 mg L-1 2-4D and 2 mg L-1 Kinetin and incubated at 35ᵒC for eight days and followed by four days at 25ᵒC in darkness, then anthers sub cultured to R medium containing 0.1 mg L-1 Kinetin and incubated at 25ᵒC and 16h photoperiod. The effect of genotype, cold pretreatment (4ᵒ, 24 h) and heat shock on the efficiency of anther culture in pepper (Capsicum annum L.) was evaluated. Cold pretreatment applied to excised buds improved microspore embryogenesis efficiency as compared to control. Among genotypes tested, Cadia, Magno, and Maratus were shown to be more responsive than King Arthur and Plato. Plants produced in responsive genotypes were approximately three-fold higher (2.5, 2.33 and 2.33 regenerated plant per Petri dish, respectively) than King Arthur and Plato (0.83 and 0.66, respectively). Results showed that heat shock (35ᵒC) applied to cultured anthers was effective on microspore embryogenesis, as heat shock(35ᵒC) treatment for eight days had the best results in Cadia with 0.16 plant per Petri dish.

Keywords: Androgenesis; Breeding; Capsicum annum L.; cold pretreatment; Heat shock.