Study On Introduction Of Mustar Trypsin Inhibitor Gene Mti2 Into Arabidopsis Thaliana

The pests is a main factor that restrain the yield and quality of crops, so it is great importance for agriculture production that breeding new cullivars which can resistance to pest, but the common methods of pest-resistance breeding are restrained greatly because the breeding week is too long or resource is limited and the pest-resistance mechanism are not clear. But the quickly development of plant genetic engineering provides a new and powerful method for breeding pest-resistance cultivars. Mustar trypsin inhibitor has been isolated from white mustar seeds, it can significantly affected the activity of overall digestive protease in the gut of Lepidopteran pest larvals.Therefore, mti2 is thought to be a good candidate for enhancing pest resitancc in plant.In this reaserch,the mti2 encoding cDNA sequence that recombinated in PKYLX-71 vector has been transfer into arabidopsis by in planta infiltration to study the character of expression and pest resistance in transgenic plants. The main results in this experiment were as follows:1. The mti2 gene has been transferred into arabidopsis thaliana(Columbia ecotype)by in planta infiltration, the final transformation frequency is about 0.2%~2.8%. in the course of this transformation, the surfactants Tween 20 has been used as effective substitutes for Silwet L-77.the final Tween 20 concentration is 0.02%.2. The kanamycin-resist plants had been test by PCR and PCR-Southern bloting, the positive frequency of PCR test is about 98.9% and the positive frequency of PCR-Southern bloting is 100%.it can be confirmed that the target gene has recombinated into the genomic of arabidopsis plants.3. One-step RT-PCR test had been done in 80 plants that have passed the PCR test, a special bind had been acquired in every plant, it can be confirmed that the target gene which recombinated into the genomic of plant had expressed in plant.