Establishment Of Highly Efficient In Vitro Regeneration System Of Masson Ine And Larch

1、Establishment of highly efficient in vitro regeneration system of masson pine（Pinus massoniana L.）On the basic of the comparative researcher of buds forming capacity derived frommature zygotic embryos and hypocotyl explant in Masson pine (Pinus massoniana Lamb.),efficient regeneration systems were firstly developed using hypoccoty from juvenile axenicseeding, and Factors that influences on plant regeneration in vitro were studied. the resultsshowed that: The best sterilisatio method to young seedlings of masson pine were75%alcohol30s and0.1%HgCl26min；The optimal bud induction medium of hypocotylsfrom masson pine is DCR+2.0mg/L BA+0.2mg/L NAA with a high frequency(98.8%);The optimal callus differentiation medium is DCR+0.5mg/L BA+0.05mg/L NAA with ahigh frequency(91.04%),and a maximum men number of adventitious buds（18.95） perexpalnt. Shoots were elongated on medium DCR+0.1mg/L BA+0.05mg/L NAA，withthe mean length of2.22cm.The elongation shoots was produced on1/2DCR mediumsupplemented with0.4mg/L IBA with the best rooting result.The average root frequencywas94.2%.the survival rate was85.9%when transferred to soil. Theseinnovation,reproducible and high frequency regeneration system provide a basic forgenetic transformation of masson pine.2、Establishment of highly efficient in vitro regeneration system of Larch(Larixolgensis Herry.)Study on the effect of various factors of regeneration from explants hypocotyls ofLarch(Larix olgensis Herry.) and efficient regeneration systems were firstly developedusing hypoccoty from juvenile axenic seeding. The best sterilisatio method to youngseedlings of Larch(Larix olgensis Herry.) were75%alcohol30s and0.1%HgCl25min；The optimal bud induction medium of hypocotyls from Larch is DCR+2.0mg/L BA+0.2mg/LNAAwith a high frequency(85.93%), and obtain compact callus in light culture. Theoptimal callus differentiation medium isDCR+0.5mg/L BA+0.05mg/L NAA andDCR+0.7mg/L BA+0.07mg/LNAA with high frequencys71.05%and68.52%, and amaximum men number of adventitious buds（9.85and7.95） per expalnt. Shoots wereelongated on medium DCR without any plant growth regulators could get the better growthcondition of elongation shoots. The elongation shoots was produced on1/2DCR mediumsupplemented with0.4mg/L IBA with the best rooting result.