A rice vacuum infiltration genetic transformation device

Abstract

The invention relates to a rice vacuum permeation genetic transformation device. The middle part of a supporting table top of the device is provided with a through hole; a bottom plate is arranged on the supporting table; the middle parts of the upper and lower surfaces of the bottom plate are respectively provided with an upper cylinder and a lower cylinder; a magnet ring II and a magnet ring I are respectively sheathed on the upper cylinder and lower cylinder; the adjacent magnetic poles of the magnet ring I and magnet ring II are homopolar; the lower cylinder and magnet ring I penetrate through the through hole and are positioned under the table top; a vacuum permeation genetic transformation chamber is arranged on the bottom plate; the upper cylinder is positioned in the chamber; a bracket is arranged on the magnet ring II and provided with a permeation solution container; the upper inner wall of the chamber is provided with an annular loading platform, an air vent pipe with a valve, and a vacuum air exhaust connecting pipe; and the bottom end port and top end port of the chamber are respectively provided with a seal ring. The device can perform transformation on the plant to be transformed in the vacuum state immediately after the plant starts contacting the permeation solution, implements in-situ genetic transformation on the rice in the vacuum state in deed, and enhances the infecting potential of agrobacterium for the plant. The device is applicable to both smaller and bigger crop individuals.

Description

technical field [0001] The invention relates to a rice vacuum infiltration genetic transformation device, which belongs to the technical field of genetic engineering and molecular biology experiments. Background technique [0002] Rice is the main food crop for more than half of the world's population. However, rice production is limited by more and more unfavorable factors (such as pests and diseases, etc.), so the improvement of rice varieties is very urgent. Transgenic technology can break through the barriers between species and realize the transfer of genes between species, thus providing a new way for crop genetic improvement. The success of genetic improvement of rice by transgenic technology largely depends on the development and application of effective genetic transformation methods. Now, known rice genetic transformation methods are generally divided into two categories: one is direct gene transformation (including biolistic method, electric shock method, PEG met...

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Problems solved by technology

[0003] However, the traditional Agrobacterium-mediated method obviously has many deficiencies: the realization of transformation must go through callus induction on explants such as mature embryos, subculture of embryogenic callus, co-cultivation of callus and Agrobacterium, and resistant callus. In the tissue culture process of at least four months, such as wound screening and resistant callus differentiation and seedling emergence, the transformation operation is cumbersome and has a long cycle, the cost of human resources and property is high, and the operator's technical requirements are high; secondly, in the tissue culture process of nearly four months During the process, problems such as somatic cell variation, morphological abnormality, chromosome number change and loss of fertility are prone to occur, resulting in complex genetic background; moreover, the obtained transformed plants often have gene silencing, and the transformation efficiency is low

The existing vacuum infiltration transformation device has the following deficiencies: (1) In the vacuum infiltration transformation carried out by the existing vacuum dryer, the plants have been soaked in the Agrobacterium infiltration solution (permeation solution) in the container before vacuuming and decompression , although the plants and the infiltrate in the container are still under normal pressure, the genetic transformation of the plants has already begun when the plants are soaked in the Agrobacterium infiltrate in the container, and the transformation process can be said to include part of the osmotic transformation under normal pressure and part of the vacuum The osmotic transformation of the osmotic transformation has not really realized the vacuum infiltration genetic transformation, and the invasiveness of the Agrobacterium to the plant is reduced, and the transformation efficiency is also reduced.

This may also be the main reason for the low conversion efficiency of the current vacuum permeation conversion method.

(2) The existing vacuum dryers are limited by factors such as small size, lack of corresponding supporting parts suitable for living plants, and inconvenient operation, so that individual large crops such as rice, flowers, cotton, and wheat cannot be planted in the original place. The osmotic genetic transformation is carried out under the condition of the living body (that is, the in situ condition), and the operation method of digging out the recipient plant, putting it directly upside down in the container for transformation, and transplanting after the transformation is not a real method on the one hand. On the other hand, it will affect the growth state, robustness and seed setting rate of the transformed plants, resulting in low vacuum infiltration transformation efficiency.

(3) Even in the vacuum infiltration genetic transformation operation of small living plants like Arabidopsis, many side branches are crushed, resulting in a low transformation effect

In summary, the existing vacuum infiltration transformation device and its transformation method are not suitable for the vacuum infiltration genetic transformation of rice and other crops. Therefore, a simple, fast and efficient vacuum infiltration method suitable for rice and other crops was designed and developed Genetic transformation device, which will greatly shorten the transformation cycle, save scientific research costs, improve transformation efficiency, and provide strong technical support for genetic improvement of crop varieties and gene function research

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