Efficient transformation method for transforming agrobacterium into setaria viridis seeds under assistance of carborundum

Abstract

The invention belongs to the field of biotechnology and relates to an efficient transformation method for transforming agrobacterium into setaria viridis seeds under assistance of carborundum. The efficient transformation method for transforming the agrobacterium into the setaria viridis seeds under assistance of the carborundum comprises the processes of explant treatment and activation, carborundum wound treatment and agrobacterium impregnation, co-culture, bacteriostasis and screening culture, resistant callus seedling culture, rooting culture and so on. The efficient transformation method for transforming the agrobacterium into the setaria viridis seeds under assistance of the carborundum, provided by the invention, has the advantages that the steps of the method are simple; the method is easy to operate; as the mature setaria viridis seeds are used as explants and wound are formed on the seeds under the assistance of the carborundum to carry out agrobacterium transformation after the seeds germinate and are activated, the transformation efficiency of the setaria viridis is greatly enhanced, the transformation period is reduced, the transformation procedure is simplified and the transformation strain range is enlarged; and a flexible and efficient transgenic technology system with short period is established for the setaria viridis.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a high-efficiency transformation method of Agrobacterium setariae, in particular to a high-efficiency transformation method for Agrobacterium carborundum to transform Setaria seeds. Background technique [0002] C4 photosynthesis drives the productivity of several major food crops and bioenergy grasses, including maize (Zeamays), sugarcane (Saccharum officinarum), sorghum (Sorghum bicolor), Miscanthus giganteus, and switchgrass (Panicum virgatum), among others. Setaria viridis is a true diploid with many characteristics suitable for genetic analysis, including a relatively small genome (510Mb), short stature, short life cycle and large seed production, etc. An excellent model for C4 photosynthesis, but also for studying abiotic stress tolerance and bioenergy feedstocks. Morphologically similar to most grasses, including major bioenergy crops switchgrass (Panicum virgatum) and ...

AI Technical Summary

Problems solved by technology

[0003] At present, the transgenic technology of Setaria mainly adopts the method of inducing embryogenic callus from the meristems of the young bud nodes after mature seeds germinate to carry out Agrobacterium transformation. The biggest problem with this method is that the induction rate of embryogenic callus is low. Different from the condition of seeds, the ratio of the number of induced embryogenic callus to the number of seeds is generally less than 10%. Secondly, the induction time of embryogenic callus is longer, and a small amount of embryogenic callus in the first stage is obtained from the inoculation to the medium. It takes about one month for the wounded tissue, and then it needs to be subcultured twice (once every half month) before there is enough embryogenic callus for Agrobacterium transformation, and the whole transformation cycle needs about 4 months; meanwhile, this Embryogenic callus is not resistant to subculture and cannot be subcultured for long-term preservation for transformation. The transformation efficiency of embryogenic callus subcultured more than 3 times is significantly reduced, and it is not suitable for continued transformation; in addition, embryogenic callus The selection requires some experience

These all reduce the flexibility and simple practicality of this conversion technology

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