METHOD TO INDUCE GENETIC VARIABILITY IN VANILLA SPP.

摘要

The present disclosure is related to the process followed for in vitro induction and regeneration of friable appearance calluses; that also generates Vanilla spp. plants with high percentages of somaclonal variation. These will be used in biotechnological improvement works of this species, selecting the seedlings that present characteristics of interest. This process comprises the following stages: Stage 0. Obtaining the plant material and pre-treatment; Stage 1. Induction of the friable calluses; Stage 2. Buds regeneration; Stage 3. Elongation and rooting; Stage 4. Acclimatization; and Stage 5. Genetic characterization of the regenerated material. In stage 0, immature capsules with 7 months of maturity are collected in the field. The capsules should be healthy and vigorous. In stage 1, immature seeds are used as explant in semisolid MS medium supplemented with at least 0.5 mg L-1 of thidiazuron (TDZ) cultivating them in darkness. In stage 2, semisolid MS medium supplemented with 2.1 mg L-1 of 6-Benzylaminopurine (BAP) is used. Cultivating them in photoperiods of 16 hours of light/8 hours of darkness. In stage 5, inter-microsatellite sequences ISSR (Inter-Simple Sequence Repeat) are used. With this process plants of Vanilla spp. with a 72.2% polymorphism to each other, are obtained; in order to expand the genetic base that in this species is reduced.