Santarém E R, Trick H N, Essig J S, Finer J J
Department of Horticulture and Crop Science, Plant Molecular Biology and Biotechnology Program, OARDC, The Ohio State University, Wooster, OH, 44691, USA Fax: 330-263-3887 e-mail:
Plant Cell Rep. 1998 Jul;17(10):752-759. doi: 10.1007/s002990050478.
Sonication-assisted Agrobacterium-mediated transformation (SAAT) tremendously improves the efficiency of Agrobacterium infection by introducing large numbers of microwounds into the target plant tissue. Using immature cotyledons of soybean as explants, we evaluated the effects of the following parameters on transient β-glucuronidase (GUS) activity: cultivars, binary vectors, optical density of Agrobacterium during infection, duration of sonication treatment, co-culture conditions, length of explant preculture and addition of acetosyringone during co-culture. The extent of tissue disruption caused by sonication was also determined. The highest GUS expression was obtained when immature cotyledons were sonicated for 2 s in the presence of Agrobacterium (0.11 OD) followed by co-cultivation with the abaxial side of the explant in contact with the culture medium for 3 days at 27°C. The addition of acetosyringone to the co-culture medium enhanced transient expression. No differences were observed when different cultivars or binary vectors were used. Cotyledons sonicated for 2 s had moderate tissue disruption, while the longer treatments resulted in more extensive damage.
超声辅助农杆菌介导转化（SAAT）通过在目标植物组织中引入大量微伤口极大地提高了农杆菌感染效率。以大豆未成熟子叶为外植体，我们评估了以下参数对瞬时β-葡萄糖醛酸酶（GUS）活性的影响：品种、二元载体、感染期间农杆菌的光密度、超声处理持续时间、共培养条件、外植体预培养时长以及共培养期间乙酰丁香酮的添加。还测定了超声处理引起的组织破坏程度。当未成熟子叶在农杆菌（0.11 OD）存在下超声处理2秒，然后将外植体背面与培养基接触在27°C下共培养3天时，获得了最高的GUS表达。向共培养基中添加乙酰丁香酮可增强瞬时表达。使用不同品种或二元载体时未观察到差异。超声处理2秒的子叶有适度的组织破坏，而较长时间的处理会导致更广泛的损伤。
