Abstract

Plantlets derived from in vitro culture might exhibit somoclonal variation which is often heritable, and molecular variations may be generated in vitro. Since the direction of most studies is toward nuclear genome, there is a little known about the DNA of organelles. This study was conducted to test the genetic stability of wheat organelles genomes for plantlets produced by anther culture using restriction fragment length polymorphism (RFLP) analyses. One of the intergenic regions of cpDNA and one of mtDNA introns were amplified with polymerase chain reaction (PCR). The PCR products were then sequenced and digested with four restriction endonucleases (EcoR1, BamH1, NdeI and Sac1). The amplified product from cpDNA was 1000 bp in size, and digested only with NdeI into two bands with 650 and 350 bp. The amplified product from mtDNA was 1550 bp in size, and digested only with Sac1 into two bands with 1220 and 330 bp. The results obtained showed that no noticeable difference can be detected between doubled haploid plantlets and parental plants at the level of ctDNA and mtDNA organization. It can be concluded that in vitro culture by itself does not systematically generate a cytoplasmic variation in plant cells.

Key words: RFLP analysis, wheat plantlets, wheat anther culture, doubled haploids, genetic stability, mitochondria and chloroplast genome.