METHOD OF PRODUCING AND ESTABLISHING EMBRYOGENIC TISSUE FROM MULTIPLE PINE GENOTYPES (GENUS PINUS)

摘要

The invention relates to a conifer cloning method which offers the basic essentials in order to provide genetic diversity, said diversity being regarded as the opposite to cloning. The inventive method involves the production and establishment of embryogenic tissue from a wide range of genotypes of selected trees which correspond to species of the Diploxylon and Haploxylon subgenera. Said novel method comprises: the initiation of embryogenic tissue from a wide variety of genotypes, for a period of up to 5 months, of the Diploxylon subgenus; the reinitiation of more than 80 % of the polyembryogenic masses which are produced in vivo and the establishment of almost 90 % of said polyembryogenic masses; the induction of between 50 and 93 % of the embryogenic tissue from the precotyledonary embryos, said percentage being dependent on the tree selected and the quantity of seed used; and, finally, the establishment of 100 % of the genotypes that have proliferated from the Haploxylon subgenus. All of the above is obtained by exploiting the genetic survival capacity of the immature embryos from both subgenera, which enables same to proliferate in any culture medium. The inventive method is based on the principle of not exhausting said capacity in one medium, but rather maintaining the suitable, periodic stimulus when interchanging the embryogenic masses between the culture media used. The advantage of said method lies in the different components from each culture medium used which can be applied alternatively or simultaneously. The media are modified with different ratios and sources of ammonium nitrate, carbon sources and, optionally, a chemical adsorbent or a maturation promoter which are generally used for other purposes.