農(nóng)桿菌介導(dǎo)CBL基因?qū)栈ㄆ贩N‘C008’的轉(zhuǎn)化
發(fā)布時(shí)間:2018-09-07 07:25
【摘要】:以菊花品種‘C008’無菌苗葉片作為外植體,在建立了菊花高效再生體系的基礎(chǔ)上,通過農(nóng)桿菌介導(dǎo)法將CBL基因?qū)刖栈ㄆ贩N‘C008’中。試驗(yàn)共獲得抗性植株41株,PCR檢測表明,陽性植株為5株,證明CBL基因已轉(zhuǎn)化到菊花‘C008’的基因組DNA中,轉(zhuǎn)化率為12.2%。遺傳轉(zhuǎn)化過程中,預(yù)培養(yǎng)2d后,將OD_(600)=0.5~0.6的農(nóng)桿菌稀釋50倍后侵染葉片8min,再共培養(yǎng)2d、延遲篩選2d,有利于獲得較高的轉(zhuǎn)化率。
[Abstract]:The leaves of aseptic seedling of chrysanthemum were used as explants. On the basis of establishing the efficient regeneration system of chrysanthemum, the CBL gene was introduced into chrysanthemum variety C008'by Agrobacterium tumefaciens. A total of 41 resistant plants were obtained. The results showed that there were 5 positive plants, which indicated that the CBL gene had been transformed into the genomic DNA of chrysanthemum C008', and the transformation rate was 12.2. In the process of genetic transformation, after 2 days of pre-culture, 50 times of Agrobacterium tumefaciens (OD_ _ (600) was diluted 50 times, then infected with leaves for 8 min, and then co-cultured for 2 days, then delayed screening for 2 days, which was beneficial to obtain higher conversion rate.
【作者單位】: 沈陽農(nóng)業(yè)大學(xué)林學(xué)院;
【基金】:遼寧省自然科學(xué)基金(2014027004) 遼寧省教育廳科學(xué)研究計(jì)劃(2015483)
【分類號】:S682.11
本文編號:2227602
[Abstract]:The leaves of aseptic seedling of chrysanthemum were used as explants. On the basis of establishing the efficient regeneration system of chrysanthemum, the CBL gene was introduced into chrysanthemum variety C008'by Agrobacterium tumefaciens. A total of 41 resistant plants were obtained. The results showed that there were 5 positive plants, which indicated that the CBL gene had been transformed into the genomic DNA of chrysanthemum C008', and the transformation rate was 12.2. In the process of genetic transformation, after 2 days of pre-culture, 50 times of Agrobacterium tumefaciens (OD_ _ (600) was diluted 50 times, then infected with leaves for 8 min, and then co-cultured for 2 days, then delayed screening for 2 days, which was beneficial to obtain higher conversion rate.
【作者單位】: 沈陽農(nóng)業(yè)大學(xué)林學(xué)院;
【基金】:遼寧省自然科學(xué)基金(2014027004) 遼寧省教育廳科學(xué)研究計(jì)劃(2015483)
【分類號】:S682.11
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