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Device for screening and culturing of transgenic arabidopsis seedlings in situ and application thereof

A Arabidopsis thaliana, transgenic technology, applied in the fields of application, plant genetic improvement, cultivation, etc., can solve the problems of long adaptation time of seedlings, cumbersome generation of seeds, etc., and achieve the effect of avoiding adverse effects

Inactive Publication Date: 2010-12-08
INST OF SOIL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method screens transgenic Arabidopsis T 2 Generation of seeds is very cumbersome, mainly has the following disadvantages: 1)T 2 Progeny seeds must be screened on kanamycin growth plates under sterile conditions
At the same time, this step will also make the seedlings need a long time to adapt

Method used

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  • Device for screening and culturing of transgenic arabidopsis seedlings in situ and application thereof
  • Device for screening and culturing of transgenic arabidopsis seedlings in situ and application thereof
  • Device for screening and culturing of transgenic arabidopsis seedlings in situ and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The seedling tube and the incubator device for in-situ screening cultivation, the seedling tube is in the shape of a rounded table, the seedling tube is covered on the incubator device containing the nutrient solution, and there are a number of circular culture holes on it. The seedling tube can be inserted and fixed in the culture hole; there is an aeration pump and a pH adjustment interface at the bottom of the incubator, which can be connected to an aeration pump and a pH regulator to continuously oxygenate the nutrient solution in the incubator and regulate the nutrient solution in real time. pH value, so that the growth environment of Arabidopsis thaliana root system can have sufficient oxygen and the pH value required for growth is stable; the four sides of the incubator are made of transparent materials, each side is marked with a scale and blocked by a shading plate, when it is necessary to monitor Arabidopsis thaliana When the root system is growing, open the sh...

Embodiment 2

[0019] A device for in-situ screening of transgenic Arabidopsis seedlings, including an incubator cover 1 and a box body 2, the incubator cover 1 is provided with an array of cultivation holes 3, and the cultivation holes are provided with seedling tubes 4 ; The side wall of the box 2 is provided with a shading plate 8, and the shading plate is movably connected with the box. It also includes an oxygenation pump and a pH adjustment device, and the oxygenation pump and the pH adjustment device are respectively connected with the box body. Coordinate scales are provided on two adjacent sides of the surface of the cover plate of the incubator according to the positions of the holes. The side wall of the box body 2 is spliced ​​up and down by a light-transmitting material 5 and an opaque material 6, and the upper and lower edges of the side walls of the light-transmitting material around the box body are provided with a chute 7, and the light-shielding plate 8 slides in the chute ...

Embodiment 3

[0022] Real-time RT-PCR (real-time fluorescent quantitative PCR)

[0023] We used our own Real-time RT-PCR system for experimental analysis (Xu and Shi, Annals of Botany, 2006, 98: 965-974). The primers used are shown in Table 1. At-ACT2 is a highly conserved housekeeping gene in Arabidopsis, and we set its expression level as 100 REU (relative expression unit). In order to verify whether the total RNA we extracted was degraded, I designed two pairs of primers for the At-ACT2 gene. In addition, we chose At-L23a (another highly conserved housekeeping gene in Arabidopsis thaliana) as a control.

[0024] Table 1 Primers used in Real-time RT-PCR

[0025]

[0026] Plant Growth Analysis

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Abstract

The invention discloses a device for screening and culturing of transgenic arabidopsis seedlings in situ, which comprises an incubator cover plate and a box body. A culture hole array is arranged on the incubator cover plate; a seedling tube is arranged inside a culture hole; and a shading board is arranged on a side wall of the box body and is movably connected with the box body. Compared with the traditional methods, the novel method has the following advantages that: 1) the novel method can screen T2-generation seeds of transgenic arabidopsis strains under the natural condition, does not need the sterile condition like the traditional methods, and is time-saving and labor-saving; 2) the novel method is an in-situ screening method, does not need to transfer the resistant seedlings from a sterile growing environment to a vermiculite or other growing environments like the traditional methods, and does not cause loss of the seedlings; and 3) the growth potential of the resistant seedlings screened by the novel method is better than that of the traditional methods, and transgenic pure lines (T3-generation seeds) are obtained early.

Description

1. Technical field [0001] The invention belongs to the technical field of cultivation of transgenic Arabidopsis thaliana, in particular to a device for in-situ screening cultivation of transgenic Arabidopsis seedlings and its application. 2. Background technology [0002] Arabidopsis is a model plant for plant science research, and overexpression of exogenous genes in Arabidopsis has become one of the basic means for people to study gene functions. The flower dipping method is one of the convenient methods to obtain transgenic Arabidopsis thaliana, which does not require tissue culture. It is very convenient to obtain transgenic Arabidopsis T 1 Generation of seeds (Weigel D, Glazebrook J. 2002. Arabidopsis: A laboratory manual. Cold Spring Harbor Laboratory Press, New York.). However, this method screens transgenic Arabidopsis T 2 Generation of seeds is very cumbersome, mainly has the following disadvantages: 1)T 2 Progeny seeds must be screened on kanamycin growth plates...

Claims

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Application Information

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IPC IPC(8): A01G31/02A01H1/04
CPCY02P60/21
Inventor 许卫锋施卫明
Owner INST OF SOIL SCI CHINESE ACAD OF SCI
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