Anther culture and Hordeum bulbosum-derived barley doubled haploids: mutations and methylation.
Anther culture and Hordeum bulbosum-derived doubled haploid (DH) lines of barley (Hordeum vulgare L.) were analyzed for RFLP and RAPD polymorphisms. Polymorphisms were not detected in the anther culture- or H. bulbosum-derived DH lines among 273 RFLP and 89 polymerase chain reaction (PCR)-amplified DNA fragments assayed. It was calculated that base substitution or small deletion/insertion mutations had not been induced among 401,640 bp screened. Large deletion/insertion mutations were not observed among 33 Mb screened. Polymorphisms were observed when DNA was digested with the methylation-sensitive restriction enzymes HpaII and MspI: these RFLPs originated primarily from the anther culture-derived doubled haploids. The data indicate that heritable DNA methylation changes had occurred during DH production, particularly with the anther culture method.
Devaux
P
P
Department of Crop and Soil Sciences, and Genetics and Cell Biology, Washington State University, Pullman 99164-6420.
Kilian
A
A
Kleinhofs
A
A
D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers / Q000737:chemistry; D006238:Haploidy; D001467:Hordeum / Q000235:genetics*; D008745:Methylation; D008969:Molecular Sequence Data; D009154:Mutation; D011003:Ploidies; D012150:Polymorphism, Restriction Fragment Length; D012639:Seeds
D017931:DNA Primers