Influence of bacterial density during preculture on Agrobacterium-mediated transformation of tomato
Influence of bacterial density during preculture on Agrobacterium-mediated transformation of tomato
复制标题
预培养过程中细菌密度对农杆菌介导番茄转化的影响
DOI：
10.1007/s11240-009-9566-2
复制
发表时间：
2009-09-01
影响因子：
3
通讯作者：
Shi, Weiming
中科院分区：
文献类型：
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作者：
Gao, Nan;Shen, Weishou;Shi, Weiming
以经济番茄品种Solanum lycopersicum L。CV.中薯4号）。瞬时基因表达和稳定转化的频率受农杆菌预培养的密度而不是用于感染的农杆菌的密度的影响。本研究中提出的改进方案依赖于使用过夜生长的农杆菌预培养物，其OD 600 nm = 1.0，用Luria-Bertani（LB）液体培养基稀释1/10，并再生长4 h。收集培养物并重悬于液体共培养基-I中，调节至OD 600 nm = 0.1。利用该改良农杆菌制剂，β-葡萄糖醛酸酶的瞬时表达率可达90%以上，转化效率达44.7%。这种改进的转化是简单的、可重复的，不需要进料层，并且最值得注意的是，转化频率是稳定的和高效的。
An improved bacterial preculture protocol for Agrobacterium-mediated genetic transformation was developed for an economic tomato cultivar (Solanum lycopersicum L. cv. Zhongshu No. 4). Frequencies of transient gene expression and stable transformation were influenced by the density of Agrobacterium preculture and not the density of Agrobacterium used for infection. The improved protocol presented in this study depends on the use of an overnight-grown Agrobacterium preculture density of OD600 nm = 1.0, diluted 1/10th with Luria-Bertani (LB) liquid medium, and grown for an additional 4 h. Cultures are collected and resuspended in a liquid cocultivation medium-I, adjusted to OD600 nm = 0.1. Using this modified Agrobacterium preparation, transient β-glucuronidase expression was higher than 90%, and transformation efficiency reached 44.7%. This improved transformation is simple, repeatable, does not require a feeder layer, and most notably, the transformation frequency is stable and highly efficient.