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CaCl2 induced soybean pollen tube channel gene transfer method

A pollen tube channel and transgene technology, which is applied in the field of transgene in the field of biotechnology, can solve the problem of low transgene efficiency of pollen tube channel, achieve the effect of reducing DNA damage and improving work efficiency

Inactive Publication Date: 2010-07-21
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a CaCl 2 Induced Soybean Pollen Tube Transgenic Method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] The experimental variety is autumn soybean Shanghai local green, 501 flowers;

[0015] (1) Prepare DNA solution. The DNA was diluted with TE buffer to prepare a solution with a DNA concentration of 300 μg / m, pH 6.5, and the transformed DNA fragment was 3 kb (including the GUS gene).

[0016] (2) Preparation containing inducer CaCl 2 DNA solution: add inducer CaCl to the DNA solution obtained above 2 , so that the solution CaCl 2 The final concentration of 0.001mol / L.

[0017] (3) Select qualified flower buds and the time of transformation: at 10:00 every day, select the flowers in the upper part of the soybean plant, the corolla is higher than the highest calyx 0.5mm, brightly colored flower buds, that is, 4 hours after soybean self-pollination conduct.

[0018] (4) Inducing soybean pollen tube channels to transform soybeans: removing the flowers and young pods of the next node of the selected flower bud and other unqualified flowers and pods on the same node. The...

Embodiment 2

[0022] The experimental variety is autumn soybean Shanghai local green, 489 flowers;

[0023] (1) Prepare DNA solution. The DNA was diluted with TE buffer to prepare a solution with a DNA concentration of 500 μg / m, pH 8.0, and the transformed DNA fragment was 10 kb (including the GUS gene).

[0024] (2) Preparation containing inducer CaCl 2 DNA solution: add inducer CaCl to the DNA solution obtained above 2 , so that the solution CaCl 2 The final concentration of 0.01mol / L.

[0025] (3) Select qualified flower buds and the time of transformation: at 19:00 every day, select the flowers in the upper part of the soybean plant, the corolla is higher than the highest calyx 1.0mm, brightly colored flower buds, that is, 8 hours after soybean self-pollination conduct.

[0026] (4) Inducing soybean pollen tube channels to transform soybeans: removing the flowers and young pods of the next node of the selected flower bud and other unqualified flowers and pods on the same node. The...

Embodiment 3

[0030] The experimental variety is autumn soybean Shanghai local green, 539 flowers;

[0031] (1) Prepare DNA solution. The DNA was diluted with TE buffer to prepare a solution with a DNA concentration of 400 μg / m, pH 7.3, and the transformed DNA fragment was 5 kb (including the GUS gene).

[0032] (2) Preparation containing inducer CaCl 2 DNA solution: add inducer CaCl to the DNA solution obtained above 2 , so that the solution CaCl 2 The final concentration of 0.005mol / L.

[0033] (3) Select qualified flower buds and the time of transformation: at 15:00 every day, select the flowers in the upper part of the soybean plant, the corolla is higher than the highest calyx 0.8mm, brightly colored flower buds; carry out 6 hours after the soybean flower self-pollination transform.

[0034] (4) Inducing soybean pollen tube channels to transform soybeans: removing the flowers and young pods of the next node of the selected flower bud and other unqualified flowers and pods on the s...

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Abstract

The invention discloses a transgenic method for soybean tube pollen tube channel induced by CaCl2, and it is used for field of biotechnology and modern agricultural technique. The method comprises following steps: preparing DNA solution, adding CaCl2 solution to produce DNA solution containing inducer CaCl2, selecting flower bulbs and time for tranversion, inducing soybean tube pollen tube channel to transverse into soybean with said CaCl2- containing DNA solution, and managing. The CaCl2- containing DNA solution includes target gene, and the DNA damage is reduced; CaCl2 of proper concentration can induce pollen tube to extend and grow, increase rate of outer gene entering into ovary, the gradient of calcium ion aslo accelerates the speed of outer gene into ovary. The transversion rate isincreased by 1- 2 times in this invention comparing with current pollen tube channel.

Description

technical field [0001] The invention relates to a transgenic method in the field of biotechnology, in particular to a CaCl 2 Induced Transgenic Method for Pollen Tube Channels in Soybean. Background technique [0002] The use of genetic engineering to improve crops has become an important technical approach of modern agriculture. The pollen tube passage method is one of several main genetic transformation methods. The limiting factor in the scope of application of the pollen tube passage method is the transformation efficiency. The conversion efficiency of the pollen tube passage method is relatively low, generally only 0.3% to 0.6%. [0003] Found through the document search of prior art, Yang Qingkai etc. in " Journal of Oil Crops of China ", 2002 24 (3): 17~20, author " the research that utilizes pollen tube channel technology to import insect resistance gene into soybean " this article utilizes The pollen tube channel technology introduces the target gene into soybea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/87
Inventor 李卉武天龙马晓红
Owner SHANGHAI JIAOTONG UNIV
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