| Soybean (Glycine max (L.) Merr) is an important source of oil and protein. Isoflavonesis a part of secondary meTableolites flavonoids compounds.It plays an important role inhuman health. Therefore, esTablelish an efficient genetic transformation system and the useof transgenic technology to improve soy isoflavones is very important. Chalcone synthase(CHS) is one of the key enzymes controlling the synthesis of flavonoids. MYB transcriptionfactor is one of the most versatile kind of transcription factors. It involved in the regulationof plant secondary meTableolites phenylpropanoid pathway, which is a branch of flavonoidpathway.We cloned two genes CHS8and MYB12b2. Then, we constructed plant expressionvectors: pCB35SR1R2-GFP-CHS8and pCB35SR1R2GFP-FAD3. Agrobacterium strain ofEHA105was transformed with the two plant expression vectors respectively for plantinfections.Because of the lower regeneration frequency of soybean Regeneration System,genotype-dependent strength, poor reproducibility, the system of soybean tissue culture stillexist some problems. In order to esTablelish an efficient system of genetic transformation,the paper based on the study of soybean embryonic tips regeneration system of Jilin35. Thekey factors that may affect the conversion rate is the choice of pre-culture medium,pre-incubation time, the concentration of hormone, ultrasonic time of infection, screeningconcentration. Through this series of key elements of optimization, we expect an efficientembryos tip genetic transformation system of soybean Jilin35. The results showed that thechoice of germination of soybean Jilin35is the liquid medium, and add2mg/L2,4-D,germination of16-20h. After ultrasound treatment6s, vacuum infection15min. The bestBasta concentration in the bud induction period is0.6mg/L. In this paper, use the optimizedof embryos tip genetic transformation system, the average of conversion rate is31.3%,Significantly improved than before. It proves that these optimization measures of Jilin35embrponic tip genetic transformation system are effective.We gained76T0transgenic plants,37lines of T1generation,14lines of T2generation,two lines of T3generation which confirmed by PCR molecular analysis and bar gene test strip; Glufosinate smear proved that positive transgenic plants resistant to herbicidessignificantly. Isoflavones measurement results of T2indicate the isoflavone content ofpositive transgenic plants than the wild type and negative transgenic plants has increased. |
