Abstract
Obtaining doubled haploids of triticale is urgently needed to reduce the time and cost of new varieties' development. Therefore, the acquisition of doubled haploid lines in tissue culture should be based on efficient regeneration methods and lead to highly homozygous materials. Therefore, in this study, we investigated the effect of Cu2+ (0.1, 5, 10 µM) and Ag+ (0, 2, 10 µM) ions and the time (35, 42, 49 days) of explant incubation on induction media on tissue culture-induced variation, which may affect the genetic homogeneity of the regenerants. And also, it was checked whether manipulating the in vitro conditions can affect the level of variation in the obtained regenerants. The methylation sensitive Amplified Fragment Length Polymorphism results revealed a high level of tissue culture-induced variation (51%). At the same time, the average sequence variation was 43%, demethylation was 5%, and de novo methylation was 3%. Among the tested nine various conditions (T1–T9), different levels of variation were observed depending on the modification of the medium. The test to determine the level of changes depending on the methylation context showed that the most changes are located in the areas of symmetric contexts CG and CHG. Still, the asymmetric context (CHH) has the slightest changes. Our study revealed a relationship between medium composition and the changes observed between triticale donor plants and their regenerants. Both changes in DNA sequence and its methylation may be dependent on the culture conditions. It seems that the results obtained may give rise to studies aimed at obtaining regenerants with the desired direction of variation (lowest, highest) generated by plant tissue culture.
Key message
The gaining triticale haploids under modified in vitro culture conditions have shown a relationship between the composition of the medium and the changes observed between triticale donor plants and their regenerants.
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Abbreviations
- DH:
-
Doubled haploid
- DMV:
-
Demethylation
- DNMV:
-
De novo methylation
- metAFLP:
-
Methylation sensitive amplified fragment length polymorphism
- PIC:
-
Polymorphic index content
- SV:
-
Sequence variation
- TCIV:
-
Tissue culture-induced variation
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This research was funded by the Ministry of Agriculture and Rural Development, Poland (Grant No. HORhn-801-PB-22/15-18).
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RO and PTB conceived the project and designed the research. KAP performed the experiments. KAP and RO prepared the plant material. KAP, RO and PTB analyzed the data. KAP, RO and PB wrote the first draft of the manuscript. KAP, RO and PTB, revised and finalized the manuscript. All authors contributed to the article and approved the final version of the manuscript.
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Pachota, K.A., Orłowska, R. & Bednarek, P.T. Medium composition affects the tissue culture-induced variation in triticale regenerants. Plant Cell Tiss Organ Cult 151, 35–46 (2022). https://doi.org/10.1007/s11240-022-02327-z
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DOI: https://doi.org/10.1007/s11240-022-02327-z