Genetic Transformation Of Soybean With Insect-Resistant Gene Cry1Iem

In our diet lives, oil has been very important ingredients, and as our country the main oil crops in soybean is an irreplaceable became an important part of food safety, in our country,pest on soybean production caused serious influence,and usually chemical control will not only increase the cost,but also causes pollution to environment and agricultural products, and through plant genetic engineeringbreeding of insect resistant soybean varieties become an effective way.At present gene engineering technology,Bt gene is the most widely used insect resistant gene,In the family of Bt gene,according to Hofte and Whiteley in 1989 according to Bt coding insecticidal crystal protein amino acid sequence homology classification.The amino acid homology of cry1 Iem gene was lower than 45%[2],belong to the first class,mainly for Lepidoptera pests,Because the gene is less studied in Soyben but the effect is good in other crops,Therefore,in this study,we putthe Plant expression vector p Cambia 3300-35s-cry1 Iem with insect resistant gene cry1 Iem into in soybean JN28 By pollen tube pathway method.In order to obtain high budworm high yield soybean transgenic soybean strains.Lay the foundation for breeding new varieties of insect resistant soybean. The main results are as follows:(1)Put The cry1 Iem gene transferred into the high yield receptor variety "JN28" soybean,transformed 800 flowers,get 200 pods,complete mature seeds 390,Select the seeds which full and complete appearance,plant 200 T1 plants.(2)Transformation of purpose genes by pollen tube pathway,introduction of plasmid concentration to be precise control in about 100ng/ Mu L. too low Concentration is not conducive to successful gene expression,the concentration is too high, will the flowers wither and die. Complex drops and protection liquid after 1h. Plasmid into and dropping liquid protection to blade or disk to do simple protective shell, reduce the risk of physical injury.(3)Detection of the selection markers bar gene,detection of transformed plants of T1 generation by herbicide basta,42 positive plants were obtained,accounted for 21% of the transformed plants;40 positive plants were detected by PCR detection test,accounted for 20% of the transformed plants.(4)T1 generation of transformed were detected by Southern blotting which 7 strains of PCR were positive plants,there are 5 plantd appeared hybridization signal.(5)Fluorescent quantitative PCR detection results show that,The expression of the purpose gene in the leaves and the stem in the 5 samples were 5.601,3.991,6.558、2.402,3.619;1.180,0.716,1.199、0.390,1.322.(6)According to the natural identification in the field and the insect damage rate of the seed identification,The insect damage rate rate of transgenic plants was 4.5% lower than that of the non transgenic plants,there were not significantly different in other plants.