DNA damage and genetic methylation changes caused by Cd in Arabidopsis thaliana seedlings.
Amplified fragment length polymorphism (AFLP) and methylation-sensitive amplification polymorphism (MASP) techniques are sensitive to deoxyribonucleic acid (DNA) damage and genetic methylation, respectively. Using these 2 techniques, Arabidopsis thaliana cultured with 0 mg/L (control), 0.5 mg/L, 1.5 mg/L, and 5.0 mg/L Cd(2+) for 16 d was used to analyze the DNA damage and methylation changes as a result of cadmium (Cd). The DNA was amplified by 14 AFLP primer pairs and 13 MSAP primer combinations. In the AFLP experiment, 62 polymorphic sites were found in the patterns of 11 primer combinations and a total of 1116 fragments were obtained in these patterns. There were no polymorphic bands in the remaining 3 pairs. The proportions of polymorphic sites in the 0.5-mg/L Cd(2+) and 5.0-mg/L Cd(2+) treatments were significantly different. Seven polymorphic fragments were then separated and successfully sequenced, yielding 6 nucleobase substitutions and 1 nucleobase deletion. Similarly, in the MSAP experiment, the MSAP% and number of demethylated-type bands were unchanged after Cd treatment, but the number of methylated-type bands was increased significantly in the 5.0-mg/L Cd(2+) treatment group, a finding that may be associated with the AFLP results. The polymorphic bands were also sequenced and the functions of their homologous genes were determined. The DNA damage and methylation changes may be the primary cause of certain pathology changes as a result of Cd uptake in plants.
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.|University of Chinese Academy of Sciences, Beijing, China.|Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
Zhaoling
Z
Li
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.|School of Life and Environmental, Deakin University, Warrnambool, Victoria, Australia.
Zhihong
Z
Liu
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.|University of Chinese Academy of Sciences, Beijing, China.
Ruijuan
R
Chen
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.
Xiaojun
X
Li
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.
Peidong
P
Tai
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.
Zongqiang
Z
Gong
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.
Chunyun
C
Jia
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, Liaoning, China.
Wan
W
Liu
D054458:Amplified Fragment Length Polymorphism Analysis; D017360:Arabidopsis / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D001483:Base Sequence; D002104:Cadmium / Q000633:toxicity*; D004249:DNA Damage / Q000187:drug effects*; D019175:DNA Methylation / Q000187:drug effects; D017931:DNA Primers / Q000378:metabolism; D018744:DNA, Plant / Q000032:analysis / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques; D036226:Seedlings / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018744:DNA, Plant; D002104:Cadmium
Amplified fragment length polymorphism; Cadmium; DNA damage; Genetic methylation; Methylation-sensitive amplification polymorphism