Establishing The Regeneration System Of Trifolium Repens Ladino And Through The Agrobacterium-mediated Approach Transfer The TPS Into The Trifolium Repens Ladino

White clover (Trifolium repens L.) is a kind of forage legume which has been developed for many years both at home and abroad. Because it has stolon, clever appearance and could adjust every kind of climate it has been used in afforestation of city, soil and water conservation and ecological construction. But because white clover cannot endure in the environment of drought and high salt, its production capacity is limited seriously. Recently the climate changing has caused rainfall reduction and rainfall uneven distribution which make people focus on how to improve the capability of white clover to endure in the drought and high temperature environment. This research use Latin white clover (T. repens Ladino) as the receptor which has poor capability to endure in the drought environment through the agrobacterium-mediated method, transfer the TPS into the plant in order to improve its capability of drought stress endurance. The main result is as below:(1) Stage of callus induction:During the stage of callus induction we had set5levels of hormone. As the concentration added the rate of callus induction increased but at the highest two levels the calluses were too wet and too soft to continue the experiment. So at this stage it is necessary to test the quality of the calluses. Finally with3.0mg/L2,4-D the calluses had the highest quality.(2) Stage of callus subculture:At the subculture stage the concentration of plant growth regulator was3.0mg/L2,4-D.30ds latter the calluses grown bigger then separated into small pieces.(3) Stage of callus differentiation:At callus differentiation stage we used the6-BA and NAA as the plant growth regulators. Both of them were separated into3concentration level. By orthogonal experiment we have9processing, during them the NO.8processing (6-BA2.0mg/L+NAA0.3mg/L) got the highest rate of differentiation.(4) Stage of callus rooting:At this stage every adventitious bud could be used in next stage. At the adventitious bud rooting stage we used the IBA and NAA as the plant growth regulators. Both of them were separated into3concentration level. By orthogonal experiment we have9processing. During them the NO.9processing (IBA0.3mg/L+NAA0.3mg/L) got the highest rate of adventitious bud rooting. At this stage every shoot could be transplanted into field and every one of them were same as the normal Trifolium repens.Ladino.(5) Stage of transformation:At this stage, when The infecting time>20min the calluses were infected seriously and the tissues were destroyed seriously. Finally when the infecting time was at20min the calluses were differentiated successfully. Then we use recognition sequences to test the DNA of the plant by PCR. The result was positive.(6) Stage of drought stress testing:At last we test the capacity of the drought resistance with normal Trifolium repens Ladino as the contrast. The result showed that the drought resistance capacity of the plant with TPS was little more strong than the normal one. Finally, we have summarized the normal process of gene transferring into Trifolium repens Ladino.