A novel double T-DNA system for producing stack and marker-free transgenic plants

作者：Wang, X. J.; Su, Y. Y.; Dong, Y. F.; Tang, Q. L.; Wang, Z. X.^*

来源：Biologia Plantarum, 2016, 60(4): 767-773.

DOI：10.1007/s10535-016-0653-z

• Agrobacterium tumefaciens
• isocaudamer technique
• multi-gene stack
• Nicotiana tabacum

This study aimed to develop a new vector system to remove selection genes and to introduce two or more genes of interest into plants in order to express them in a coordinated manner. A multigene expression vector was established based on pCamBIA2300 using a selectable marker gene (SMG)-free system based on the combination of the isocaudamer technique and double T-DNA. The vector DT7 containing seven target genes was constructed and introduced into tobacco using Agrobacterium-mediated transformation. Twenty-one of 27 positive transgenic plants contained both T-DNA regions. The co-transformation frequency was 77.8 %. The frequency of unlinked integration of two intact T-DNAs was 22.22 % (6/27). The frequency of removal of SMG from transgenic T1 plants was 19.10 %. These results suggest that this vector system was functional and effective for multigene expression and SMG-free transgenic plant cultivation. At least seven target genes can be co-expressed using this system. Overall, these findings provide a new and highly effective platform for multigene and marker-free transgenic plant production.

• 出版日期2016-12
• 单位中国农业科学院