• Justyna Zbieszczyk
Stanowisko: Biolog
Jednostka: Biuro ds. Infrastruktury Badawczo-Dydaktycznej WNP
Adres: 40-032 Katowice, ul. Jagiellońska 28
Piętro: II
Numer pokoju: C-250
Telefon: (32) 2009 562
E-mail: justyna.zbieszczyk@us.edu.pl
Spis publikacji: Spis wg CINiBA
Spis publikacji: Spis wg OPUS
Scopus Author ID: 6602800908
Publikacje z bazy Scopus
2024
Nowak, K.; Wójcikowska, B.; Gajecka, M.; Elżbieciak, A.; Morończyk, J.; Wójcik, A. M.; Żemła, P.; Citerne, S.; Kiwior-Wesołowska, A.; Zbieszczyk, J.; Gaj, M. D.
The improvement of the in vitro plant regeneration in barley with the epigenetic modifier of histone acetylation, trichostatin A Journal Article
In: Journal of Applied Genetics, vol. 65, no. 1, pp. 13-30, 2024, ISSN: 12341983.
@article{2-s2.0-85176574335,
title = {The improvement of the in vitro plant regeneration in barley with the epigenetic modifier of histone acetylation, trichostatin A},
author = { K. Nowak and B. Wójcikowska and M. Gajecka and A. Elżbieciak and J. Morończyk and A.M. Wójcik and P. Żemła and S. Citerne and A. Kiwior-Wesołowska and J. Zbieszczyk and M.D. Gaj},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85176574335&doi=10.1007%2fs13353-023-00800-9&partnerID=40&md5=aba91f468acebb6e7ac6030f68dc49f7},
doi = {10.1007/s13353-023-00800-9},
issn = {12341983},
year = {2024},
date = {2024-01-01},
journal = {Journal of Applied Genetics},
volume = {65},
number = {1},
pages = {13-30},
publisher = {Springer Science and Business Media Deutschland GmbH},
abstract = {Genotype-limited plant regeneration is one of the main obstacles to the broader use of genetic transformation in barley breeding. Thus, developing new approaches that might improve responses of in vitro recalcitrant genotypes remains at the center of barley biotechnology. Here, we analyzed different barley genotypes, including “Golden Promise,” a genotype commonly used in the genetic transformation, and four malting barley cultivars of poor regenerative potential. The expression of hormone-related transcription factor (TF) genes with documented roles in plant regeneration was analyzed in genotypes with various plant-regenerating capacities. The results indicated differential expression of auxin-related TF genes between the barley genotypes in both the explants and the derived cultures. In support of the role of auxin in barley regeneration, distinct differences in the accumulation of free and oxidized auxin were observed in explants and explant-derived callus cultures of barley genotypes. Following the assumption that modifying gene expression might improve plant regeneration in barley, we treated the barley explants with trichostatin A (TSA), which affects histone acetylation. The effects of TSA were genotype-dependent as TSA treatment improved plant regeneration in two barley cultivars. TSA-induced changes in plant regeneration were associated with the increased expression of auxin biosynthesis-involved TFs. The study demonstrated that explant treatment with chromatin modifiers such as TSA might provide a new and effective epigenetic approach to improving plant regeneration in recalcitrant barley genotypes. © 2023, The Author(s).},
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}
Genotype-limited plant regeneration is one of the main obstacles to the broader use of genetic transformation in barley breeding. Thus, developing new approaches that might improve responses of in vitro recalcitrant genotypes remains at the center of barley biotechnology. Here, we analyzed different barley genotypes, including “Golden Promise,” a genotype commonly used in the genetic transformation, and four malting barley cultivars of poor regenerative potential. The expression of hormone-related transcription factor (TF) genes with documented roles in plant regeneration was analyzed in genotypes with various plant-regenerating capacities. The results indicated differential expression of auxin-related TF genes between the barley genotypes in both the explants and the derived cultures. In support of the role of auxin in barley regeneration, distinct differences in the accumulation of free and oxidized auxin were observed in explants and explant-derived callus cultures of barley genotypes. Following the assumption that modifying gene expression might improve plant regeneration in barley, we treated the barley explants with trichostatin A (TSA), which affects histone acetylation. The effects of TSA were genotype-dependent as TSA treatment improved plant regeneration in two barley cultivars. TSA-induced changes in plant regeneration were associated with the increased expression of auxin biosynthesis-involved TFs. The study demonstrated that explant treatment with chromatin modifiers such as TSA might provide a new and effective epigenetic approach to improving plant regeneration in recalcitrant barley genotypes. © 2023, The Author(s).
2021
Gajecka, M.; Marzec, M.; Chmielewska, B.; Jelonek, J.; Zbieszczyk, J.; Szarejko, I.
Changes in plastid biogenesis leading to the formation of albino regenerants in barley microspore culture Journal Article
In: BMC Plant Biology, vol. 21, no. 1, 2021, ISSN: 14712229, (5).
@article{2-s2.0-85098859776,
title = {Changes in plastid biogenesis leading to the formation of albino regenerants in barley microspore culture},
author = { M. Gajecka and M. Marzec and B. Chmielewska and J. Jelonek and J. Zbieszczyk and I. Szarejko},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85098859776&doi=10.1186%2fs12870-020-02755-z&partnerID=40&md5=d815420ef3c84b515da6b97e036cb855},
doi = {10.1186/s12870-020-02755-z},
issn = {14712229},
year = {2021},
date = {2021-01-01},
journal = {BMC Plant Biology},
volume = {21},
number = {1},
publisher = {BioMed Central Ltd},
abstract = {Background: Microspore embryogenesis is potentially the most effective method of obtaining doubled haploids (DH) which are utilized in breeding programs to accelerate production of new cultivars. However, the regeneration of albino plants significantly limits the exploitation of androgenesis for DH production in cereals. Despite many efforts, the precise mechanisms leading to development of albino regenerants have not yet been elucidated. The objective of this study was to reveal the genotype-dependent molecular differences in chloroplast differentiation that lead to the formation of green and albino regenerants in microspore culture of barley. Results: We performed a detailed analysis of plastid differentiation at successive stages of androgenesis in two barley cultivars, ‘Jersey’ and ‘Mercada’ that differed in their ability to produce green regenerants. We demonstrated the lack of transition from the NEP-dependent to PEP-dependent transcription in plastids of cv. ‘Mercada’ that produced mostly albino regenerants in microspore culture. The failed NEP-to-PEP transition was associated with the lack of activity of Sig2 gene encoding a sigma factor necessary for transcription of plastid rRNA genes. A very low level of 16S and 23S rRNA transcripts and impaired plastid translation machinery resulted in the inhibition of photomorphogenesis in regenerating embryos and albino regenerants. Furthermore, the plastids present in differentiating ‘Mercada’ embryos contained a low number of plastome copies whose replication was not always completed. Contrary to ‘Mercada’, cv. ‘Jersey’ that produced 90% green regenerants, showed the high activity of PEP polymerase, the highly increased expression of Sig2, plastid rRNAs and tRNAGlu, which indicated the NEP inhibition. The increased expression of GLKs genes encoding transcription factors required for induction of photomorphogenesis was also observed in ‘Jersey’ regenerants. Conclusions: Proplastids present in microspore-derived embryos of albino-producing genotypes did not pass the early checkpoints of their development that are required for induction of further light-dependent differentiation of chloroplasts. The failed activation of plastid-encoded RNA polymerase during differentiation of embryos was associated with the genotype-dependent inability to regenerate green plants in barley microspore culture. The better understanding of molecular mechanisms underlying formation of albino regenerants may be helpful in overcoming the problem of albinism in cereal androgenesis. © 2021, The Author(s).},
note = {5},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Background: Microspore embryogenesis is potentially the most effective method of obtaining doubled haploids (DH) which are utilized in breeding programs to accelerate production of new cultivars. However, the regeneration of albino plants significantly limits the exploitation of androgenesis for DH production in cereals. Despite many efforts, the precise mechanisms leading to development of albino regenerants have not yet been elucidated. The objective of this study was to reveal the genotype-dependent molecular differences in chloroplast differentiation that lead to the formation of green and albino regenerants in microspore culture of barley. Results: We performed a detailed analysis of plastid differentiation at successive stages of androgenesis in two barley cultivars, ‘Jersey’ and ‘Mercada’ that differed in their ability to produce green regenerants. We demonstrated the lack of transition from the NEP-dependent to PEP-dependent transcription in plastids of cv. ‘Mercada’ that produced mostly albino regenerants in microspore culture. The failed NEP-to-PEP transition was associated with the lack of activity of Sig2 gene encoding a sigma factor necessary for transcription of plastid rRNA genes. A very low level of 16S and 23S rRNA transcripts and impaired plastid translation machinery resulted in the inhibition of photomorphogenesis in regenerating embryos and albino regenerants. Furthermore, the plastids present in differentiating ‘Mercada’ embryos contained a low number of plastome copies whose replication was not always completed. Contrary to ‘Mercada’, cv. ‘Jersey’ that produced 90% green regenerants, showed the high activity of PEP polymerase, the highly increased expression of Sig2, plastid rRNAs and tRNAGlu, which indicated the NEP inhibition. The increased expression of GLKs genes encoding transcription factors required for induction of photomorphogenesis was also observed in ‘Jersey’ regenerants. Conclusions: Proplastids present in microspore-derived embryos of albino-producing genotypes did not pass the early checkpoints of their development that are required for induction of further light-dependent differentiation of chloroplasts. The failed activation of plastid-encoded RNA polymerase during differentiation of embryos was associated with the genotype-dependent inability to regenerate green plants in barley microspore culture. The better understanding of molecular mechanisms underlying formation of albino regenerants may be helpful in overcoming the problem of albinism in cereal androgenesis. © 2021, The Author(s).
2020
Gajecka, M.; Marzec, M.; Chmielewska, B.; Jelonek, J.; Zbieszczyk, J.; Szarejko, I.
Plastid differentiation during microgametogenesis determines green plant regeneration in barley microspore culture Journal Article
In: Plant Science, vol. 291, 2020, ISSN: 01689452, (8).
@article{2-s2.0-85075546169,
title = {Plastid differentiation during microgametogenesis determines green plant regeneration in barley microspore culture},
author = { M. Gajecka and M. Marzec and B. Chmielewska and J. Jelonek and J. Zbieszczyk and I. Szarejko},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85075546169&doi=10.1016%2fj.plantsci.2019.110321&partnerID=40&md5=e6fa2af008d23476f81f00614719a9af},
doi = {10.1016/j.plantsci.2019.110321},
issn = {01689452},
year = {2020},
date = {2020-01-01},
journal = {Plant Science},
volume = {291},
publisher = {Elsevier Ireland Ltd},
abstract = {Developing plants from in vitro culture of microspores or immature pollen grains (androgenesis) is a highly genotype-dependent process whose effectiveness in cereals is significantly reduced by occurrence of albino regenerants. Here, we examined a hypothesis that the molecular differentiation of plastids in barley microspores prior to in vitro culture affects the genotype ability to regenerate green plants in culture. At the mid-to-late uninucleate (ML) stage, routinely used to initiate microspore culture, the expression of most genes involved in plastid transcription, translation and starch synthesis was significantly higher in microspores of barley cv. ‘Mercada’ producing 90% albino regenerants, than in cv. ‘Jersey’ that developed 90% green regenerants. The ML microspores of cv. ‘Mercada’ contained a large proportion of amyloplasts filled with starch, while in cv. ‘Jersey’ there were only proplastids. Using additional spring barley genotypes that differed in their ability to regenerate green plants we confirmed the correlation between plastid differentiation prior to culture and albino regeneration in culture. The expression of GBSSI gene (Granule-bound starch synthaseI) in early-mid (EM) microspores was a good marker of a genotype potential to produce green regenerants during androgenesis. Initiating culture from EM microspores that significantly improved regeneration of green plants may overcome the problem of albinism. © 2019 The Author(s)},
note = {8},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Developing plants from in vitro culture of microspores or immature pollen grains (androgenesis) is a highly genotype-dependent process whose effectiveness in cereals is significantly reduced by occurrence of albino regenerants. Here, we examined a hypothesis that the molecular differentiation of plastids in barley microspores prior to in vitro culture affects the genotype ability to regenerate green plants in culture. At the mid-to-late uninucleate (ML) stage, routinely used to initiate microspore culture, the expression of most genes involved in plastid transcription, translation and starch synthesis was significantly higher in microspores of barley cv. ‘Mercada’ producing 90% albino regenerants, than in cv. ‘Jersey’ that developed 90% green regenerants. The ML microspores of cv. ‘Mercada’ contained a large proportion of amyloplasts filled with starch, while in cv. ‘Jersey’ there were only proplastids. Using additional spring barley genotypes that differed in their ability to regenerate green plants we confirmed the correlation between plastid differentiation prior to culture and albino regeneration in culture. The expression of GBSSI gene (Granule-bound starch synthaseI) in early-mid (EM) microspores was a good marker of a genotype potential to produce green regenerants during androgenesis. Initiating culture from EM microspores that significantly improved regeneration of green plants may overcome the problem of albinism. © 2019 The Author(s)
2018
Szurman-Zubrzycka, M. E.; Zbieszczyk, J.; Marzec, M.; Jelonek, J.; Chmielewska, B.; Kurowska, M. M.; Krok, M.; Daszkowska-Golec, A.; Guzy-Wróbelska, J.; Gruszka, D.; Gajecka, M.; Gajewska, P.; Stolarek, M.; Tylec, P.; Sega, P.; Lip, S.; Kudełko, M.; Lorek, M.; Gorniak-Walas, M.; Malolepszy, A.; Podsiadlo, N.; Szyrajew, K. P.; Keisa, A.; Mbambo, Z.; Todorowska, E.; Gaj, M.; Nita, Z.; Orlowska-Job, W.; Maluszynski, M.; Szarejko, I.
HorTILLUS—a rich and renewable source of induced mutations for forward/reverse genetics and pre-breeding programs in barley (Hordeumvulgare L.) Journal Article
In: Frontiers in Plant Science, vol. 9, 2018, ISSN: 1664462X, (30).
@article{2-s2.0-85043326007,
title = {HorTILLUS—a rich and renewable source of induced mutations for forward/reverse genetics and pre-breeding programs in barley (Hordeumvulgare L.)},
author = { M.E. Szurman-Zubrzycka and J. Zbieszczyk and M. Marzec and J. Jelonek and B. Chmielewska and M.M. Kurowska and M. Krok and A. Daszkowska-Golec and J. Guzy-Wróbelska and D. Gruszka and M. Gajecka and P. Gajewska and M. Stolarek and P. Tylec and P. Sega and S. Lip and M. Kudełko and M. Lorek and M. Gorniak-Walas and A. Malolepszy and N. Podsiadlo and K.P. Szyrajew and A. Keisa and Z. Mbambo and E. Todorowska and M. Gaj and Z. Nita and W. Orlowska-Job and M. Maluszynski and I. Szarejko},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85043326007&doi=10.3389%2ffpls.2018.00216&partnerID=40&md5=2be0ee225baa473124ed786053f7b1c1},
doi = {10.3389/fpls.2018.00216},
issn = {1664462X},
year = {2018},
date = {2018-01-01},
journal = {Frontiers in Plant Science},
volume = {9},
publisher = {Frontiers Media S.A.},
abstract = {TILLING (Targeting Induced Local Lesions IN Genomes) is a strategy used for functional analysis of genes that combines the classical mutagenesis and a rapid, high-throughput identification of mutations within a gene of interest. TILLING has been initially developed as a discovery platform for functional genomics, but soon it has become a valuable tool in development of desired alleles for crop breeding, alternative to transgenic approach. Here we present the HorTILLUS (Hordeum—TILLING—University of Silesia) population created for spring barley cultivar “Sebastian” after double-treatment of seeds with two chemical mutagens: sodium azide (NaN3) and N-methyl-N-nitrosourea (MNU). The population comprises more than 9,600 M2 plants from which DNA was isolated, seeds harvested, vacuum-packed, and deposited in seed bank. M3 progeny of 3,481 M2 individuals was grown in the field and phenotyped. The screening for mutations was performed for 32 genes related to different aspects of plant growth and development. For each gene fragment, 3,072–6,912 M2 plants were used for mutation identification using LI-COR sequencer. In total, 382 mutations were found in 182.2Mb screened. The average mutation density in the HorTILLUS, estimated as 1 mutation per 477kb, is among the highest mutation densities reported for barley. The majority of mutations were G/C to A/T transitions, however about 8% transversions were also detected. Sixty-one percent of mutations found in coding regions were missense, 37.5% silent and 1.1% nonsense. In each gene, the missense mutations with a potential effect on protein function were identified. The HorTILLUS platform is the largest of the TILLING populations reported for barley and best characterized. The population proved to be a useful tool, both in functional genomic studies and in forward selection of barley mutants with required phenotypic changes. We are constantly renewing the HorTILLUS population, which makes it a permanent source of new mutations. We offer the usage of this valuable resource to the interested barley researchers on cooperative basis. © 2018 Szurman-Zubrzycka, Zbieszczyk, Marzec, Jelonek, Chmielewska, Kurowska, Krok, Daszkowska-Golec, Guzy-Wrobelska, Gruszka, Gajecka, Gajewska, Stolarek, Tylec, Sega, Lip, Kudełko, Lorek, Gorniak-Walas, Malolepszy, Podsiadlo, Szyrajew, Keisa, Mbambo, Todorowska, Gaj, Nita, Orlowska-Job, Maluszynski and Szarejko.},
note = {30},
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TILLING (Targeting Induced Local Lesions IN Genomes) is a strategy used for functional analysis of genes that combines the classical mutagenesis and a rapid, high-throughput identification of mutations within a gene of interest. TILLING has been initially developed as a discovery platform for functional genomics, but soon it has become a valuable tool in development of desired alleles for crop breeding, alternative to transgenic approach. Here we present the HorTILLUS (Hordeum—TILLING—University of Silesia) population created for spring barley cultivar “Sebastian” after double-treatment of seeds with two chemical mutagens: sodium azide (NaN3) and N-methyl-N-nitrosourea (MNU). The population comprises more than 9,600 M2 plants from which DNA was isolated, seeds harvested, vacuum-packed, and deposited in seed bank. M3 progeny of 3,481 M2 individuals was grown in the field and phenotyped. The screening for mutations was performed for 32 genes related to different aspects of plant growth and development. For each gene fragment, 3,072–6,912 M2 plants were used for mutation identification using LI-COR sequencer. In total, 382 mutations were found in 182.2Mb screened. The average mutation density in the HorTILLUS, estimated as 1 mutation per 477kb, is among the highest mutation densities reported for barley. The majority of mutations were G/C to A/T transitions, however about 8% transversions were also detected. Sixty-one percent of mutations found in coding regions were missense, 37.5% silent and 1.1% nonsense. In each gene, the missense mutations with a potential effect on protein function were identified. The HorTILLUS platform is the largest of the TILLING populations reported for barley and best characterized. The population proved to be a useful tool, both in functional genomic studies and in forward selection of barley mutants with required phenotypic changes. We are constantly renewing the HorTILLUS population, which makes it a permanent source of new mutations. We offer the usage of this valuable resource to the interested barley researchers on cooperative basis. © 2018 Szurman-Zubrzycka, Zbieszczyk, Marzec, Jelonek, Chmielewska, Kurowska, Krok, Daszkowska-Golec, Guzy-Wrobelska, Gruszka, Gajecka, Gajewska, Stolarek, Tylec, Sega, Lip, Kudełko, Lorek, Gorniak-Walas, Malolepszy, Podsiadlo, Szyrajew, Keisa, Mbambo, Todorowska, Gaj, Nita, Orlowska-Job, Maluszynski and Szarejko.
2016
Szarejko, I.; Szurman-Zubrzycka, M. E.; Nawrot, M.; Marzec, M.; Gruszka, D.; Kurowska, M. M.; Chmielewska, B.; Zbieszczyk, J.; Jelonek, J.; Maluszynski, M.
Creation of a TILLING population in barley after chemical mutagenesis with sodium azide and MNU Book Chapter
In: pp. 91-111, Springer International Publishing, 2016, ISBN: 9783319450216; 9783319450193, (13).
@inbook{2-s2.0-85021060188,
title = {Creation of a TILLING population in barley after chemical mutagenesis with sodium azide and MNU},
author = { I. Szarejko and M.E. Szurman-Zubrzycka and M. Nawrot and M. Marzec and D. Gruszka and M.M. Kurowska and B. Chmielewska and J. Zbieszczyk and J. Jelonek and M. Maluszynski},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85021060188&doi=10.1007%2f978-3-319-45021-6_6&partnerID=40&md5=769f75988eadba3f52354d86c8d9403e},
doi = {10.1007/978-3-319-45021-6_6},
isbn = {9783319450216; 9783319450193},
year = {2016},
date = {2016-01-01},
journal = {Biotechnologies for Plant Mutation Breeding: Protocols},
pages = {91-111},
publisher = {Springer International Publishing},
abstract = {Since the development of the Targeting Induced Local Lesions in Genome (TILLING) strategy, it has been applied in both plants and animals in many studies. The creation of an appropriate population is the first and most crucial step of TILLING. The goal is to obtain a highly mutagenized population that allows many mutations in any gene of interest to be found. Therefore, an effective method of mutation induction should be developed. A high mutation density is associated with saving time, costs, and the labor required for the development of a TILLING platform. The proper handling of the mutated generations, the establishment of a seed bank, and the development of a DNA library are essential for creating a TILLING population. The database in which all of the data from the molecular and phenotypic analyses are collected is a very useful tool for maintaining such population. Once developed, a TILLING population can serve as a renewable resource of mutations for research that uses both forward and reverse genetic approaches. In this chapter, we describe the methods for the development and maintenance of a TILLING population in barley. © International Atomic Energy Agency 2017.},
note = {13},
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Since the development of the Targeting Induced Local Lesions in Genome (TILLING) strategy, it has been applied in both plants and animals in many studies. The creation of an appropriate population is the first and most crucial step of TILLING. The goal is to obtain a highly mutagenized population that allows many mutations in any gene of interest to be found. Therefore, an effective method of mutation induction should be developed. A high mutation density is associated with saving time, costs, and the labor required for the development of a TILLING platform. The proper handling of the mutated generations, the establishment of a seed bank, and the development of a DNA library are essential for creating a TILLING population. The database in which all of the data from the molecular and phenotypic analyses are collected is a very useful tool for maintaining such population. Once developed, a TILLING population can serve as a renewable resource of mutations for research that uses both forward and reverse genetic approaches. In this chapter, we describe the methods for the development and maintenance of a TILLING population in barley. © International Atomic Energy Agency 2017.
1997
Szarejko, I.; Maluszynski, M.; Polok, K.; Zbieszczyk, J.
Gene segregation in a barley DH population Journal Article
In: Journal of Applied Genetics, vol. 38, no. 4, pp. 415-424, 1997, ISSN: 12341983, (3).
@article{2-s2.0-0347093962,
title = {Gene segregation in a barley DH population},
author = { I. Szarejko and M. Maluszynski and K. Polok and J. Zbieszczyk},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-0347093962&partnerID=40&md5=92263fa33ea3a16d30d573f568653ef6},
issn = {12341983},
year = {1997},
date = {1997-01-01},
journal = {Journal of Applied Genetics},
volume = {38},
number = {4},
pages = {415-424},
abstract = {A large population of anther culture-derived barley regenerants and their progeny was tested for allelc segregation at 1 isozyme and 8 morphological marker loci. The segregation of genetic markers was examined separately for haploid, diploid and polyploid regenerants. All the 9 analysed genes except al (albino lemma) on chromosome 3 segregated according to the expected 1:1 ratio in the microsporederived barley population. There was no difference in allele distribution between haploid and diploid regenerants. Among the limited number of 34 analysed tetraploids a significant excess of the dominant allele at locus o (orange lemma) of chromosome 6 was also observed. The recombination frequency between linked genes (n - lk2 on chromosome 1 and r - s on chromosome 7) estimated in the DH population did not differ significantly from recombination rates calculated in F2 progeny or presented in barley chromosome maps. The phenomenon of gametic selection is discussed in relation to the genotype dependency of anther culture response and procedures used for DH production in barley.},
note = {3},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
A large population of anther culture-derived barley regenerants and their progeny was tested for allelc segregation at 1 isozyme and 8 morphological marker loci. The segregation of genetic markers was examined separately for haploid, diploid and polyploid regenerants. All the 9 analysed genes except al (albino lemma) on chromosome 3 segregated according to the expected 1:1 ratio in the microsporederived barley population. There was no difference in allele distribution between haploid and diploid regenerants. Among the limited number of 34 analysed tetraploids a significant excess of the dominant allele at locus o (orange lemma) of chromosome 6 was also observed. The recombination frequency between linked genes (n - lk2 on chromosome 1 and r - s on chromosome 7) estimated in the DH population did not differ significantly from recombination rates calculated in F2 progeny or presented in barley chromosome maps. The phenomenon of gametic selection is discussed in relation to the genotype dependency of anther culture response and procedures used for DH production in barley.