Genomic DNA isolation and amplification from callus culture in succulent plants,Carpobrotus species (Aizoaceae)

Abstract

Genomic DNA of high quality and quantity is needed to analyze genetic diversity with AFLP.Carpobrotus plant species, like most succulents, contain high amounts of polysaccharides and polyphenols, making PCR amplification difficult. Our protocol eliminates contaminants before DNA isolation by using leaf callus as plant material. This simple and inexpensive technique gives an average DNA yield of 1800 ng/g of callus and high reproducible profiles in AFLP. Our results indicate that no genetic variability is associated with callus culture conditions. This technique is suitable for studying genomic polymorphism in succulents and other plants when classic DNA extraction procedures fail.