Inter-simple Sequence Repeat Molecular Markers In Ilex And The Tissue Culture System Of Ilex Cornuta Lindl Ex Paxt

China is a big country in the holly plant resources, but the ornamental aspects of Ilex in development and utilization is still in a preliminary stage, so carryingr out parental molecular marker-assisted selection, promoting horticultural varietal breeding in order to expedite the development and utilization of Ilex resources become more and more important.Inter species relationship of 36 species in Ilex was studied by ISSR markers.12 primers were screened out from 100 random primers. 235 DNA bands were amplified ranging from 400-1 500 bp, the polymorphic rate of 232 was accounted for 98.7%. The average number of DNA bands amplified by each primer was 19.6. Genetic similarity coefficient was calculated by NTSYS and the dendrogram were constructed by UPMGA.Most sameness between the classify result by marker and former morphological taxon is discovered through clustering analysis in 36 Ilex species : 36 Ilex plants were classified into 2 groups that was deciduous plant species and evergreen plant species at similarity cofeecient 0.64. I.macrocapar,I.asprella,I.aculeolata and I.micrococca were clustered into one group .The rest of 32 species were clustered into another group. The latter growp was divided into 2 sets at similarity cofeecicent 0.66.One contained I.cornuta,,Ilex×altaclerensis'Lawsoniana'and Ilex aquifolium etc 6 species. Ilex opaca and I.rotuda etc 26 species were divided into the other set. The preliminary proof proved that it was feasible to analysis the system evolution and genetic relationship of Ilex plants species by using ISSR..During the experiment of the tissue culture system of Ilex cornuta by seed embody, evrery facters influence on it have been studied.The substratum of each stage for tissue culture and rapid propagation of Ilex cornuta has been determined preliminarily through check experiment, which will be helpful for spreading Ilex cornuta.It is shown that the suitable medium for embody genmination is 1/2MS+GA3 1.0 mg.L^-1, sound seedling culture medium is MS+BA 1.5 mg.L^-1+NAA 0.5 mg.L^-1; The best adventitious buds induced best medium is MS+BA 2.0 mg.L^-1+NAA 0.1 mg.L^-1;The better multiplication and subculture medium is MS+BA 3.0 mg.L^-1+KT 0.5 mg.L^-1+IBA 0.5 mg.L^-1;the better growth of roots is in 1/2MS+IBA 0.2 mg.L^-1+NAA 0.2 mg.L^-1.