【摘要】：以芥藍品種小香菇為試材,分別采用基因槍法和農(nóng)桿菌介導(dǎo)轉(zhuǎn)化法將Bt抗蟲基因Cry2Aa2轉(zhuǎn)化到芥藍中,并對兩種轉(zhuǎn)化方法進行了優(yōu)化和比較。結(jié)果表明:基因槍法在轟擊距離為6cm、轟擊次數(shù)為1次時轉(zhuǎn)化效率最高,為0.43%;農(nóng)桿菌介導(dǎo)轉(zhuǎn)化法以帶子葉下胚軸為外植體、預(yù)培養(yǎng)2d、侵染10min、共培養(yǎng)2d、抑菌培養(yǎng)5d后轉(zhuǎn)入篩選培養(yǎng)基為最佳參數(shù),轉(zhuǎn)化效率為0.37%。經(jīng)PCR和PCR-Southern鑒定,基因槍法和農(nóng)桿菌介導(dǎo)轉(zhuǎn)化法均成功將目的基因?qū)虢嫠{基因組中。經(jīng)抗蟲性表型鑒定,T_0代轉(zhuǎn)基因芥藍植株對甘藍夜蛾幼蟲的抗性高于非轉(zhuǎn)基因植株。
[Abstract]:The Bt insect-resistant gene Cry2Aa2 was transformed into Brassica oleracea by gene gun method and Agrobacterium tumefaciens-mediated transformation. The two transformation methods were optimized and compared. The results showed that the transformation efficiency was the highest when the bombardment distance was 6 cm and the number of bombardment was 1, and the transformation efficiency was 0.43, and Agrobacterium tumefaciens mediated transformation was based on the hypocotyls of scallop leaves. The optimum parameters were pre-culture for 2 days, infection for 10 minutes, co-culture for 2 days, inhibition of bacteria for 5 days, and then transferred to the screening medium. The transformation efficiency was 0.37%. The results of PCR and PCR-Southern showed that the target gene was successfully introduced into the genome of kale by gunshot method and Agrobacterium tumefaciens-mediated transformation. The resistance of transgenic kale plants in generation 0 to larvae of Spodoptera spp. was higher than that of non-transgenic plants.
【作者單位】： 華南農(nóng)業(yè)大學園藝學院;
【基金】：廣東省教育廳特色創(chuàng)新項目(2014KTSCX026) 廣東省教育廳青年人才項目(2014KQNCX035) 華南農(nóng)業(yè)大學校長基金項目(K13012)
【分類號】：S635.9
，

本文編號：2279044