Studies On Aloe Vera L.Transformation System And IGF-Iintroduction Into Aloe Vera L.

Cloning of IGF-I gene and constructing of it's plant expression vectors. Moncotyledon promoter Ubiquitin was inserted into the vector PUCAMBIA1301 , PUKH-35S-GUS-PA for the construction of PUC1301-U and PUKH-U. A pair of primers PI and P2 were synthesized , in which , partial codons of plant were chosen for correcting the codons behind ATG, and two restrict sites of Bam H I and Bst E II were designed. The fragment IGF-I and the promoter Ubiquitin were inserted into PCAMBIA1301 vector for the construction of PC1301-U-I.The construction of Aloe vera L.transformation system . Using MS solid basal medium supplemented with 2.0mg/L 6-BA,2.0 mg/L 2,4-D and 30g/L sucrose, the explants were cultured at 25±2 centigrade and dark condition, After about 2 weeks, calli and abduction buds were got at the same. But it needs 3-4 weeks to get the calli.It was a rapid and high efficiency approach that the abuction buds produced from the transgenes explants directly.Genetic transformation system of Aloe vera L. mediated by Agrobacterium trmefaciens. Use the sturdy individual aloe vera L. stem basal part as the explant; 3 days preculture of the explants before infected by the Agrobacterium tumefaciens had a positive effect on transformation. The bacterium density was OD600O. 3 and the infection time was 15~30mins. The concentration of hygromycin for screen of transgenic plant and carbenicillin of cefotaxime for controlthe increase of Agrobacterium tumefaciens was 50-70mg/L and 250-300mg/L respectively. It was necessary to add acetosyringone in coculture medium.Test and analysis of transgenic plants. Introduced the IGF-I gene into the aloe vera L. with the optimizing transformation system of electrical transformation .