Tissue culture-induced genetic and epigenetic variation in triticale (× Triticosecale spp. Wittmack ex A. Camus 1927) regenerants.
Plant regeneration via in vitro culture can induce genetic and epigenetic variation; however, the extent of such changes in triticale is not yet understood. In the present study, metAFLP, a variation of methylation-sensitive amplified fragment length polymorphism analysis, was used to investigate tissue culture-induced variation in triticale regenerants derived from four distinct genotypes using androgenesis and somatic embryogenesis. The metAFLP technique enabled identification of both sequence and DNA methylation pattern changes in a single experiment. Moreover, it was possible to quantify subtle effects such as sequence variation, demethylation, and de novo methylation, which affected 19, 5.5, 4.5% of sites, respectively. Comparison of variation in different genotypes and with different in vitro regeneration approaches demonstrated that both the culture technique and genetic background of donor plants affected tissue culture-induced variation. The results showed that the metAFLP approach could be used for quantification of tissue culture-induced variation and provided direct evidence that in vitro plant regeneration could cause genetic and epigenetic variation.
Department of Plant Physiology and Biochemistry, Plant Breeding and Acclimatization Institute-National Research Institute, 05-870, Błonie, Radzików, Poland.
Joanna
J
Machczyńska
Department of Plant Biotechnology and Cytogenetics, Plant Breeding and Acclimatization Institute-National Research Institute, 05-870, Błonie, Radzików, Poland.
Janusz
J
Zimny
Department of Plant Physiology and Biochemistry, Plant Breeding and Acclimatization Institute-National Research Institute, 05-870, Błonie, Radzików, Poland. p.bednarek@ihar.edu.pl.
Piotr Tomasz
PT
Bednarek
D019976:Cloning, Organism; D016000:Cluster Analysis; D044127:Epigenesis, Genetic; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D005838:Genotype; D021141:Nucleic Acid Amplification Techniques; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D046509:Tissue Culture Techniques; D000066889:Triticale / Q000235:genetics*
D010940:Plant Proteins
Androgenesis; De novo methylation; Demethylation; Doubled haploid; In vitro culture; Sequence variation; Somatic embryogenesis