| The bad environment,such as low temperature,drought,salt,is a very important nonbiological force factor influence agricultural production and ecological environment. our country is a nation that water resources is in scarcity, dry and half dry farmland accounts for 38% of total national farmland;secondly,the low temperature is a disaster to northern agriculture in south in our country which take place regularly;In addition, our country still contain a lot of salt soil needed to be exploited.How to make use of the land of these big areas and improve the tolerance of crop is an importance research subject in bioscience. teraploid black locust is the vanguard tree of the mountain forestation to grow,keeping the livestock husbandry and planting trees in ameliorative environment,plain and mountain area development turns forestation etc. can produce the good ecosystem performance and the economic performance.If tolerance can be increased,which will having very important meaning for improving of the ecosystem environment and developping a market for keeping the livestock husbandry.so in this paper a set of representative system of teraploid black locust of the stem was founded,which provide the dependable basis of turn the genetic engineering.The gene rd29A has the anti-resisting function of dry, salt alkali, cold, therefore, rd29A gene was cloned from Arabidolpsis thalian in this research, and the plant expression vector of rd29A was constructed.the breeding of anti-drought and anti-salt crop contain gene rd29A has the urgent meaning for the increasing of unit area yield and enlargement of planting area, the breeding of anti-drought nonbiology diversidy trees and grasses can take a visible ecological and economic benefit to environmental protection and beautified.so the complete of this paper have the important meaning for the cultivars amending, primarily of the research contents has below two aspects: (1)Total RNA was isolated from the Arabidolpsis thalian leaves .By using the specific primers designed from the cDNA sequence of the Arabidolpsis thalian's rd29A gene in Genebank,a fragment was produced by reverse transcription polymerase chain reaction (RT-PCR).The amplified fragment was cloned to pMD18-T vector, and it was confirmed by sequencing.Clone vector was digested with BamHIand KpnI,and the acquired rd29A was ligated to middle vector pRT101 .The recimbinant was digested with restriction endonucleases HindIII,The expression vector pCAMBIA3301 and the designed fragment (rd29A and CaMV35S) were directively ligated with T4 DNA ligase ,and the plant expressionvector of rd29A was constructed. (2)The methods building the regenerative system of teraploid black locust were studied,Which would supply basic means and theoretical basis for its improvement and breeding andmechanism of inhibitory . In this paper, dorm ant axillary bud were used as initial explant.After sterile shoots were obtained, different explants from them were cultured and regeneratedinto plantlets. The results indicated that stem were suitable for culture invitro with the shootproduction frequency over 80 %. Optimal shoot induction medium for Stem and leaf of tenderwas MS salts+B5vitamins +3mg/L6BA+2 mg/L ZT+0.05 mg/L2.4-D, and as BA,ZT,2.4-Dconcent rations increased, the ability of shoot formation decreased. Optimal shoot induction medium for root was MS+1.0mg/L6BA+2.0mg/LNAA ,The capacity of shoot regenerationwas also related to the age of sterile explant. Optimal root induction medium was 1/2MS+0.5mg/LIBA.
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