Plant genetic stability of Ipomoea batatas evaluated through RAPD markers
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Abstract
sprouts, belonging to a sweet potato (Ipomoea batatas (L.)
Lam.) clone, INIVIT B 93-1, were placed in flasks containing
water under the semicontrolled conditions of a laboratory to
induce bud break, as well as vitroplants coming from the somatic
embryos of leaf limb calluses; in both cases, leaf limbs were
used to perform the molecular studies of Random Amplification
Polymorphic DNA (RAPD), to identify the material and evaluate
the genetic stability of plants obtained from somatic embryos.
The total genomic DNA was extracted from the material
mentioned before, through a homogeneous mixture of leaf limb
tissue (1,0 g) and liquid nitrogen. DNA concentration was
determined by means of spectrophotometry and materials were
evaluated for 10 arbitrary primers, following Operon
Technologies�?? protocol recommendations, which were:
OPF-15, OPF-14, OPA-13, OPF-13, OPF-04, OPF-07, OPF-01,
OPF-03, OPA-12 and OPF-05. PCR products were
electrophoretically separated at 1.5 % agarose gel in a TBE
buffer, and they were stained with etidium bromide before being
observed on an UV transiluminator. The amplification patterns
obtained were compared by evaluating bands, both for its
presence (1) and absence (0) per each of the treatments, then
observing a complete monomorphism in the donating material
bands, compared to the one coming from somatic embryo
vitroplants.
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