Genetic integrity assessment of cryopreserved tomato (Lycopersicon esculentum Mill.) genotypes

Authors : ANA COSTE, DANA SUTEU, IOAN BACILA, CONSTANTIN DELIU, SERGIU VALIMAREANU, ADELA HALMAGYI

Pages : 638-648

Doi:10.3906/biy-1411-6

View : 16 | Download : 10

Publication Date : 2015-12-01

Article Type : Research Paper

Abstract :Shoot tips of five tomato insert ignore into journalissuearticles values(Lycopersicon esculentum Mill.); genotypes were successfully cryopreserved by droplet vitrification. Recovered plants were studied for genetic stability by two different assays: amplified fragment length polymorphism insert ignore into journalissuearticles values(AFLP); using 4 primer combinations and sequencing of lycopene $\beta$-cyclase gene insert ignore into journalissuearticles values(LCY-B); from leaves. The highest shoot regeneration after cryopreservation was 70% insert ignore into journalissuearticles values(Pontica); following 24 h osmoprotection in 0.5 M sucrose and 30 min dehydration in plant vitrification solution 2 insert ignore into journalissuearticles values(PVS2);. The data inferred from AFLP showed no genetic dissimilarities between in vitro regenerants derived from cryopreserved tissues compared with the noncryopreserved controls. Although a single nucleotide polymorphism, a G→T transversion, was identified in position 1139 in Capriciu and Darsirius, this was not due to the cryopreservation process itself, since it was detected in both cryopreserved and control samples. Thus, sequencing of LCY-B gene from leaves revealed no DNA damages after cryopreservation and subsequent in vitro regeneration. Our results indicate that cryostorage by droplet vitrification is an efficient and reliable technique to preserve the selected tomato genotypes and to regenerate true-to-type plants
Keywords : AFLP, droplet vitrification, genetic stability, LCY B, tomato