用枳殼 Poncirus trifoliata Raf.實(shí)生苗的上胚軸為材料,探索以根癌農(nóng)桿菌介導(dǎo)的GUS基因在枳殼中的轉(zhuǎn)移,結(jié)果表明:抑菌劑選擇頭孢霉素較好;卡那霉素作為選擇試劑,濃度為50mg/L;外植體與農(nóng)桿菌共培養(yǎng)時間以2～3d為宜。GUS基因瞬時表達(dá)檢測,80%的外植體呈陽性反應(yīng);GUS基因穩(wěn)定表達(dá)檢測,抗性植株中,GUS反應(yīng)呈陽性所占比例為70.0%。

The transformation of GUS gene by Agrobacterium-mediation was studied.The explants used for transformation were the epicotyls from Poncirus trifoliata Raf.The results indicated that Cefotaxime used as antibiotics was better than carbenicillin.The concentration of kanamycin used as the reagent of choice was 50mg/L.Shoot regeneration frequency was high when cocultivation time was 2～3 days.Histochemical GUS assay showed that 80.0% of the explants had the transient expression of GUS gene and 70.0% of the resistant plants were GUS-positive.