Minimally invasive non-tissue-culture transformation method for plant seed embryos

Abstract

The invention discloses a minimally invasive non-tissue-culture transformation method for plant seed embryos, and relates to a gene transformation method. The method comprises the steps that a plurality of plump seeds without mechanical damage are selected; the seeds are soaked in mercuric chloride with the concentration of 0.1% for 2 minutes, and are rinsed for more than 3-5 times by sterile water or tap water which is boiled and cooled; the seeds are placed in a pre-sterilized container, sterile water is added until the seeds are submerged, and the seeds are cultured for 4-6 hours on a table; water-swollen seeds are placed in a clean container, an operator puts on sanitary gloves in advance, and stabs slightly in the embryo parts of the seeds by a needle; the seeds with the scratched embryos are subjected to ultrasonication, and placed in a plasmid bacterial solution with a target gene for co-culture on the table; and seedling screening is performed by antibiotics in a sprouting stage and a seedling emergence stage. The method can avoid long tissue cultivation process required by an agrobacterium-mediated method and the like, so that a large amount of time is saved; and further, expensive instruments or equipment is not needed, and the operation can be finished in a common laboratory.

Description

technical field [0001] The invention relates to a gene transformation method, in particular to a plant seed embryo minimally invasive non-tissue culture transformation method. Background technique [0002] At present, there are many transformation methods included in plant genetic engineering. Such as gene gun method, Agrobacterium-mediated method, electroporation method, laser perforation method and so on. These methods either require special expensive instruments and equipment, or the conversion period is too long, and are generally not easy to operate in ordinary laboratories. The progress of society and the rapid development of other scientific fields require appropriate technologies to drive the development of plant genetic engineering. Through the unremitting efforts of scientists from all over the world, the technical progress of the transformation method itself has been dizzying. This includes the establishment of new transformation methods and the improvement of ...

AI Technical Summary

Problems solved by technology

[0003] Sun Yi, Wang Jingxue and Liu Shaoxiang applied for the Agrobacterium-mediated plant germination seed gene transformation method invented in China. The wound area is too large and the germination rate is too low

And the germination time of the seeds is too long, which will have a greater impact on the seeds in the subsequent embryo trauma treatment.