Construction Of Agrobacterium-mediated Transformation System Of Scutellaria Baicalensis Georgi And The Introduction Of Insect-resistant Gene

Scutellaria baicalensis Georgi is a traditional medicine herb plant. In recent years, studies have shown that it has many important bioactive effects, such as antibacterial, anti-abnormal reaction, anti-carcinogenic, anti-oxidation, anti-HIV. eliminating free radical ect, thus receives the attention and the demand for it grows sharply, too. This paper studied the introduction of Bt gene into Scutellaria baicalensis Georgi aiming at obtain insect resistant plants, solving pest problems in the fields and increasing output of Scutellaria baicalensis Georgi.In the research, some conditions such as hormone concentration, culture conditions, concentration of Cef and Basta were studied and optimized. By optimizing experimental conditions in the process of Scutellaria baicalensis Georgi’s regeneration above, an efficient and stable transformation system of regeneration was established and a very successful result in Agrobacterium tumefaciens mediated transformation of Bt gene was got. The result as follow:The optimal medium for callus inducing is MS+3.0mg/L6-BA+0.5mg/L NAA; the optimal medium for subculture of callus is MS+l.Omg/L6-BA+0.2mg/L NAA; the optimal medium for differentiation of callus is MS+1.5mg/L6-BA+0.5mg/L NAA.The suitable concentration of Cef for Agrobacterium elimination is250mg/L. The concentration of Basta for transgenic plant screening is20-30mg/L, and which for Callus growth is3.5mg/L, and which for callus differentiation is3.0mg/LAfter Scutellaria baicalensis Georgi was transformed, bacteria-eliminated by Cef, selected by Basta, we get16To plants, among these plants10are PCR positive. The positive rate is62.5%and shoot regeneration is6.78%. It preliminary shows that regeneration plants carry the foreign genes, while Southern blot analysis is needed to confirm it.