Resistance To Glyphosate Beet Gene Genetic Transformation Research

Weed in planting fields is one of the most important reasons for the sugar beetyield and sugar content decrease. Getting glyphosate herbicide resistance sugar beetvarieties by means of genetic engineering method is one of the effective solutions todeal with this problem. Sugar beet seeding petioles were used as transgene explantmaterials in this study. EPSPS gene was delivered into sugar beet explants byagrobacterium mediated method. Agrobacterium culture transform method and shootsregeneration conditions from beet explants were systematically studied in this paper.Transgenic beet regeneration system was established and optimized. transgenic sugarbeet plants were obtained.1、Establishment and optimization of sugar beet shoots regeneration systemThree different genotypes sugar beet lines (DM1-9, O-68and DY5-O) were used astransgene experiment materials in this study. The result showed that the most suitabletransgenic sugar beet lines were DM1-9and O-68according to experiments of sugarbeet different expants, different media and different seedling age. DM1-9and O-68were used as experiment materals in following experiment. Sugar beet shootsregeneration system was established list as follows. Shoots regeneration medium wasMS+1.0mg/L6-BA+0.1mg/L NAA+100mg/LKan+500mg/LCb. Subculturemedium was MS+6-BA0.5mg/L+NAA1.0mg/L+100mg/LKan+500mg/L Cb.Rooting culture medium was MS+1.0mg/L NAA. The maximum conversionefficiency was obtained by above system. The result of agrobacterium infectionexperment showed that transgenic operation optimal Agrobacterium infectionconcentration was OD600=0.3and infection time was10minute.2、The best method of agrobacterium removal from explantsThe agrobacterium-mediated trangene experiment result showed that the optimal removal method of agrobacterium removal after beet explant co-culture operation waslisted as follows:500mg/L Cb solution cleaning the explant, then sterile water rinse3times,300mg/L Cef solution washing1times, sterile water rinse explants treetimes. Water was suck dry with sterile filter paper at last.3、Getting transgene regeneration sugar beet plantsUsing Agrobacterium infection technology, sugar beet transgene experiment wasconducted in line with above optimization genetic transformation motheds. Glyphosateresistant sugarbeet planst were obtained. PCR detection was conducted with EPSPSgene among kanamycin resistance transgenic regenerated shoots plants. EPSPSfragment was amplified in some regeneration plants by PCR detection.