农杆菌GV3101中反式玉米素合成基因促进烟草再生

发布时间：2018-07-26 19:56

【摘要】：胭脂碱型农杆菌GV3101已经被广泛用于植物遗传转化研究。已有的研究结果证明,农杆菌GV3101株系含有的反式玉米素合成(trans-zeatin synthesizing,tzs)基因编码产物会影响烟草细胞器的形态及细胞的生理状态。然而,有关tzs基因对遗传转化过程外植体再生的影响研究却少有报道。本文在前期研究工作的基础上,以2种烟草、4个农杆菌株系为组培实验材料,验证了胭脂碱型农杆菌tzs基因产物的生理活性。结果表明:以外源添加生长调节物质的外植体为阳性对照,在不添加任何生长调节剂的培养基上,与GV3101菌株共培养的烟草外植体能分化再生,并发育成完整植株;外植体再生与GV3101携带的质粒种类无关;外植体与农杆菌GV3101培养液共培养24 h,烟草再生效果较好;与GV3101株系共培养24 h,将外植体烟草叶片匀浆,经亲和柱分离纯化后,检测出烟草外植体叶片中高达0.78 ng/g FW-1的反式玉米素含量。菌落PCR扩增结果证实,农杆菌GV3101株系有tzs基因序列。以上结果表明,农杆菌GV3101株系内的tzs基因的表达产物有生理学活性,能够促进烟草外植体再生,调节细胞生长。
[Abstract]:Cochineal Agrobacterium tumefaciens GV3101 has been widely used in plant genetic transformation. It has been shown that the trans- Zein synthesis (trans-zeatin synthesizingtzs) gene encoding product of Agrobacterium tumefaciens GV3101 strain may affect the morphology of organelle and the physiological state of tobacco cells. However, there are few reports on the effect of tzs gene on explant regeneration during genetic transformation. On the basis of previous research work, the physiological activity of tzs gene product of Agrobacterium tumefaciens was verified by using 2 kinds of tobacco and 4 strains of Agrobacterium tumefaciens as tissue culture materials. The results showed that the explants co-cultured with GV3101 strain could differentiate and regenerate and develop into intact plants on the medium without any growth regulator. The regeneration of explants was not related to the plasmids carried by GV3101, the regeneration effect of tobacco was better when the explants co-cultured with Agrobacterium tumefaciens GV3101 for 24 h, and cocultured with GV3101 for 24 h, the explants were homogenized and purified by affinity column. The content of trans zeatin in the leaves of tobacco explants was determined to be as high as 0. 78 ng/g FW-1. The results of colony PCR amplification confirmed that the GV3101 strain of Agrobacterium tumefaciens had tzs gene sequence. The results showed that the expression product of tzs gene in Agrobacterium GV3101 strain had physiological activity, which could promote the regeneration of tobacco explants and regulate cell growth.
【作者单位】：西北农林科技大学生命科学学院生物技术系;辽宁科技学院;
【基金】：西北农林科技大学基本科研创新项目(Z.109021715) 国家自然科学基金(No 31400233)资助~~
【分类号】：Q943.2

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