Plant Molecular Biology, Biological Sciences, Monsanto Company, 700 Chesterfield Village Pkwy, 63198, St. Louis, MO, USA.
Plant Mol Biol. 1988 May;10(3):225-34. doi: 10.1007/BF00027399.
We have adapted the "agroinfection" procedure of Grimsley and co-workers [4,5] to develop a simple, efficient, reproducible infectivity assay for the insect-transmitted, split-genome geminivirus, tomato golden mosaic virus (TGMV). Agrobacterium T-DNA vectors provide efficient delivery of both components of TGMV when used in mixed inoculation of wild-type host plants. A greater increase in infection efficiency can be obtained by Agrobacterium delivery of the TGMV A component to "permissive" transgenic plants. These "permissive" plants contain multiple tandem copies of the B component integrated into the host genome. An inoculum containing as few as 2000 Agrobacterium cells can produce 100% infection under these conditions. Further, our results show that there is a marked effect of the configuration of the TGMV A components within the T-DNA vector on time of symptom development. We have also found that transgenic plants carrying tandem copies of the A component do not complement the B component. Possible mechanisms to explain these results and the potential use of this system to further study the functions of the geminivirus components in infection are discussed.
我们采用了 Grimsley 及其同事的“农杆菌感染”程序[4,5]，开发了一种简单、高效、可重复的传染性测定方法，用于研究昆虫传播的分裂基因组双生病毒——番茄金黄镶嵌病毒（TGMV）。当在野生型宿主植物的混合接种中使用时，农杆菌 T-DNA 载体可有效递呈 TGMV 的两个成分。通过农杆菌向“允许”的转基因植物递呈 TGMV A 成分，可以获得更高的感染效率。这些“允许”的植物含有整合到宿主基因组中的 B 成分的多个串联拷贝。在这些条件下，含有少至 2000 个农杆菌细胞的接种物即可产生 100%的感染。此外，我们的结果表明，T-DNA 载体中 TGMV A 成分的构型对症状出现时间有明显影响。我们还发现，携带 A 成分串联拷贝的转基因植物不能互补 B 成分。这些结果的可能解释机制以及该系统在进一步研究双生病毒成分在感染中的功能方面的潜在用途正在讨论中。
