The Preliminary Analysis Of Genetic Transmission On Target Gene In Transgenic Tomato And Lettuce Of Anti-caries

Objectives:In access the seeds of T0, T1, T2, T3 generation of transgenic tomato and To generation of transgenic lettuce which including target gene pacA-ctxB and pacP-ctxB, PCR and histochemical staining for GUS activity were used to detect transgenic tomatoes progenies in different growth stages, to explore the genetic stability of the pacA-ctxB and pacP-ctxB in transgenic tomato plants,and to analysis the genetic linkage of the reporter gene gus with target gene. Identification and selection T1 generation of transgenic lettuce plants which containing target gene.Methods:The study included three partsThe first part:Genetic stability of the foreign gene pacA-ctxB,pacP-ctxB in transgenic tomato progenies of anti-caries. extract genomic DNA from T1, T2, T3, T4 generation of the transgenic tomato plants in period of young seedling, flowering, fruit maturity.11 plants of each generation are used to identify transgenic tomato plants which including the foreign gene pacA-ctxB and pacP-ctxB by PCR. The second part:Genetic linkage of the reporter gene gus with target gene in progeny of the transgenic tomato plants of anti-caries. collect young roots from the T1, T2, T3, T4 generation of the transgenic tomato plants. histochemical staining for GUS activity were used to detect the reporter gene gus.The third part:Detection of target gene in T1 generation transgenic lettuce of anti-caries. extract genomic DNA T1 generation of the transgenic lettuce plants. PCR were used to identify T1 generation of transgenic lettuce plants which containing target gene.Results:①The transgenic tomato plants genomic DNA in period of young seedling which carrying pacA-ctxB was amplified by PCR. after electrophoresis, approximately 450bp amplified banding appeared,corresponding to positive control group. The positive transgenic tomato plants are the size of the respective 9,9,8,9 strains in each generation. The positive detection rate was 81.8%,81.8%,72.7%,81.8%. Test results of the period of flowering and fruit maturity consistent with the period of young seedling.②The transgenic tomato plant's genomic DNA in period of young seedling which carrying pacP-ctxB was amplified by PCR.after electrophoresis, approximately 506bp amplified banding appeared,corresponding to positive control group. The positive transgenic tomato plants are the size of the respective 9,8, 8,7 strains in each generation. The positive detection rate was 81.8%,72.7%,72.7%,63.6%. Test results of the period of flowering and fruit maturity consistent with the period of young seedling.③In the T1,T2,T3,T4 generation of the transgenic tomato plants which carrying pacA-ctxB there were 9,9,8,9 positive strains respectively by histochemical staining for GUS activity. The coincidence rate with PCR and histochemical staining for GUS activity was as high as 100%. In the T1,T2,T3,T4 generation of the transgenic tomato plants which carrying pacP-ctxB, there are 9.8,8,7positive strains respectively by histochemical staining for GUS activity. The coincidence rate with PCR and histochemical staining for GUS activity is as high as 100%.①Ten transfer pacA-ctxB lettuce plants' genomic DNA amplify by PCR.after electrophoresis,6 strains can see approximately 450bp amplified banding,correspond ing to positive control group. Ten transfer pacP-ctxB lettuce plants' genomic DNA amplify by PCR.after electrophoresis,6 strains can seeapproximately 506bp amplified banding,corresponding to positive control group.Conclusions:①The foreign gene pacA-ctxB and pacP-ctxB were inserted into the trans-genic tomato genome.They can stablely inherit to the transgenic tomato progenies.②The reporter gene gus and foreign gene pacA-ctxB,pacP-ctxB were closely linked in the T-DNA.They were linked and inherited stably in different generations. gus can be used to study the inheritance on pacA-ctxB and pacP-ctxB.③T1 generation of transgenic lettuce plants carrying pacA-ctxB,pacP-ctxB were obtained by PCR.