Study On The Transformation Of Triploids Of Chinese White Poplar With Two Genes Of Insect-resistant

Triploid of Chinese white poplar is one of growing rapidly trees that are popularized in our country in recent years. However the quality of resisting insect has not been improved. With the enlargements of area of planting triploids of Chinese white poplar, many kinds of danger from the pest which eating leaves are more and more serious. It is important to cultivate poplar that can resist insect. Transgenosis technology has become main means to improve the quality of insect-resistant of present plant by creating new varieties.This research mainly about the transformation to the triploid of Chinese white poplar with the gene constructing two insect-resistant genes (BtCryI gene and arrowhead protease inhibitor gene API). The transgenic plant have been test tentatively by molecular biology measures and raises worms experiment . The results showed that two insect-resistant genes have been transfered into triploid of Chinese white poplar.1 Different hormone was supplemented in MS medium and their effects on shoot inducement multiplication were compared. The results showed that: MS + 6-BA 0.6mg L-1 + NAAO.lmg L-1 was the suitable combination for the inducement of buds. The selected media for rooting was: 1/2MS+IBA 0.4 mg L-1.2 Optimize studying have been done on the influencing of each factor when genes have been transformed. The result shows: the material of Transforming had better be selected the blade of the state for splitting. Produce the rate of healing to increase obviously after adding 100umol L-1 acetosyringone. In the optimum regeneration system, the optimum bacterium concentration in value of OD600 was 0.4, it had a best promoting effect on the leaf . the optimum time of infection and co-culture for leaf were 15 -20mins and 2 days respectively; Proved by the experiment the regeneration rate of Chinese white poplar of triploid reach 20% by this optimum regeneration system .3 Proved by the experiment that Bt's gene and protease inhibitor gene have already combined in genome DNA of triploid of Chinese white poplar according to PCR analysis and western's hybridization, ELISA. Prove gene that change over to have corresponding albumen express, toxoprotein determine most 0.0477% high person who expresses by content tentatively.4 Feeding Clostera anachoreta (Fabricius) with the young tender bladeseses of transgenosis triploid of Chinese white poplar, It shows that : Toxicity of transgenosispoplar with of a pair of genes is higher than contrast to the larva. Its death rate is more than 80% after fooding some Clostera anachoreta (Fabricius) with 5 number of this transgenosis plant. It has been proved that these 2 genes can express effectively in the cell, and what is expressed have better toxicity relatively to Clostera anachoreta (Fabricius) . It also proved that these 2 genes have different poison function in coordination to the larva by killing mechanism at the same time. It has improved the resisting worm's function of the triploid Chinese white poplar after transfering to a pair of genes.