本文選題：亳菊　切入點(diǎn)：發(fā)根農(nóng)桿菌　出處：《中藥材》2017年01期

【摘要】：目的:建立亳菊EDT1抗旱基因遺傳轉(zhuǎn)化體系,并初步評價其耐旱性。方法:以亳菊莖段外植體為受體材料,抗草甘膦除草劑為篩選基因,通過發(fā)根農(nóng)桿菌介導(dǎo)法轉(zhuǎn)入EDT1抗旱基因。通過農(nóng)桿菌介導(dǎo),對影響亳菊遺傳轉(zhuǎn)化效率的四個因素進(jìn)行討論:侵染時間、農(nóng)桿菌菌液濃度、乙酰丁香酮濃度和共培養(yǎng)時間,以及草甘膦最佳篩選濃度的研究。結(jié)果:建立了農(nóng)桿菌介導(dǎo)的亳菊高效遺傳轉(zhuǎn)化體系,菌液濃度(A600)為0.6~0.8,乙酰丁香酮濃度150μmol/L,侵染時間25 min,共培養(yǎng)3 d,除菌培養(yǎng)后,可得到15 d最大轉(zhuǎn)化效率95.6%。草甘膦對亳菊組培苗致死率的研究表明其最佳篩選濃度為100 mg/L。通過聚合酶鏈?zhǔn)椒磻?yīng)(PCR)檢測,初步得到5株轉(zhuǎn)基因植株。結(jié)論:在干旱脅迫下,葉片中抗氧化酶活性和MDA含量測定表明,轉(zhuǎn)基因植株具有較高的耐旱性。
[Abstract]:Objective: to establish a genetic transformation system for drought resistant genes of Boju EDT1 and to evaluate its drought resistance. Methods: the stem explants of Boju were used as acceptor materials and glyphosate resistant herbicides were used as screening genes. Through Agrobacterium tumefaciens mediated transformation of EDT1 drought-resistant gene, four factors affecting the efficiency of genetic transformation of Boju were discussed: infection time, concentration of Agrobacterium tumefaciens, concentration of acetyl eugenone and co-culture time. Results: Agrobacterium tumefaciens mediated high efficiency genetic transformation system of Boju was established. The concentration of A600) was 0.6 渭 mol / L, the concentration of acetyl eugenone was 150 渭 mol / L, the infection time was 25 min, and the culture time was 3 days. The maximum transformation efficiency of 15 days was 95.6%. The study of glyphosate on the mortality of the plantlets in tissue culture showed that the optimum concentration of glyphosate was 100 mg / L. five transgenic plants were preliminarily obtained by polymerase chain reaction (PCR). Conclusion: under drought stress, 5 transgenic plants were obtained. The determination of antioxidant enzyme activity and MDA content in leaves showed that transgenic plants had high drought tolerance.
【作者單位】： 安徽大學(xué)資源與環(huán)境工程學(xué)院;安徽省中草藥產(chǎn)業(yè)化研發(fā)中心;
【基金】：國家自然科學(xué)基金(31471554) 安徽大學(xué)研究生創(chuàng)新強(qiáng)化項(xiàng)目(yqh100297)
【分類號】：S567.239
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本文編號：1666092