Analysis of ploidy and the patterns of amplified fragment length polymorphism and methylation sensitive amplified polymorphism in strawberry plants recovered from cryopreservation.
Shoot-tips of 10 strawberry genotypes were successfully cryopreserved using a modified encapsulation-dehydration method. All genotypes survived cryopreservation with high survival and regeneration rates. Eight Joho single-bud sibling lines were established as a model system for genetic analysis. Although cytological examination found chromosomal variation in both non-cryopreserved and cryopreserved samples, the ploidy constitution remained relatively stable after cryopreservation. DNA samples digested with MseI and PstI were used for amplified fragmentation length polymorphism (AFLP) assay. In 16 primer combinations, only one, namely, PCCA-MCAG, detected one site where band pattern changed after cryopreservation, which might be contributed to the change in DNA methylation status at PstI recognition site. Methylation sensitive amplified polymorphism (MSAP) assay was carried out for further investigation on the influence of cryopreservation on DNA methylation status. It was found that cryopreservation induced a significant change in DNA methylation status.
Hao
Yu-Jin
YJ
National Key laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, P.R. China. yjcxhao@hotmail.com
You
Chun-Xiang
CX
Deng
Xui-Xin
XX
D015925:Cryopreservation / Q000379:methods*; D019175:DNA Methylation; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D018520:Plant Shoots; D011003:Ploidies; D012150:Polymorphism, Restriction Fragment Length; D020127:Recovery of Function; D027824:Rosaceae / Q000235:genetics*; D015996:Survival Rate