Genotype-Specific Androgenic Responses of Spring Barley Lines to Cold Pretreatment for Efficient Doubled Haploid Regeneration and Accelerated Breeding Cycles
Sonia Mansouri1,*, Yassmine Abidi1,2, Leila Riahi3, Ali Ltifi2
1 Field Crop Laboratory, National Institute of Agricultural Research of Tunisia, University of Carthage, Tunis, Tunisia
2 Laboratory of Biotechnology Applied to Agriculture, National Institute of Agricultural Research of Tunisia, University of Carthage, Tunis, Tunisia
3 Laboratory of Biotechnology and Bio-Geo Resources Valorization BVBGR-LR11ES31, University of Manouba, ISBST, Ariana, Tunisia
* Corresponding Author: Sonia Mansouri. Email: 
(This article belongs to the Special Issue: Genetic Resources and Breeding Strategies for Sustainable Crop Yield Enhancement)
Phyton-International Journal of Experimental Botany https://doi.org/10.32604/phyton.2026.083697
Received 08 April 2026; Accepted 01 June 2026; Published online 29 June 2026
Abstract
Doubled haploid technology is an important tool for accelerating barley breeding by enabling the rapid development of fully homozygous lines. However, the efficiency of androgenesis in barley remains highly genotype-dependent and is strongly influenced by pretreatment conditions, particularly cold exposure. Despite advances in barley anther culture protocols, the optimal duration of cold pretreatment to enhance androgenic response and green plant regeneration remains poorly understood for many breeding materials. This study therefore aimed to examine the effects of genotype and cold pretreatment duration on androgen induction and regeneration efficiency in spring barley lines. Fifteen barley lines, including the control variety rihane, were evaluated for anther culture response. Ears containing microspores at the appropriate developmental stage were subjected to two cold pretreatments at 4°C (3 and 5 weeks) before being cultured on BAC3 induction medium. The androgenic response was assessed by the induction rate, regeneration efficiency, and the frequencies of green and albino plant production. Significant variation dependent on genotype and genotype × treatment interactions was observed. Short-term cold pretreatment promoted embryogenic induction in some genotypes, notably OR 1-10, while prolonged exposure enhanced responses in others, particularly OR 1-6 and OR 1-5. Regeneration efficiency and green plant production were similarly influenced by genotype and pretreatment duration. Prolonged cold treatment improved regeneration and chlorophyllous plant production in several genotypes but reduced performance in others. Albino plant formation also varied considerably among genotypes, highlighting its persistent impact on androgenetic efficiency. These findings provide valuable insights into genotype-dependent androgenic responses and establish a basis for optimizing cold pretreatment strategies in barley anther culture. Such genotype-specific protocols could significantly enhance doubled haploid production efficiency and facilitate the rapid development of improved barley cultivars adapted to future breeding challenges.
Keywords
Hordeum vulgare L.; anther culture; doubled haploid; genotype; cold pretreatment;
in vitro regeneration
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