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Method for realizing gene editing in corn by pollen tube channel

A technology of pollen tube channeling and gene editing, which is applied to other methods of inserting foreign genetic materials, genetic engineering, plant gene improvement, etc., to achieve reasonable structural design, eliminate the need for tissue culture, and facilitate large-scale gene transformation

Pending Publication Date: 2021-12-10
JILIN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Therefore, the purpose of the present invention is to provide a method for realizing gene editing in maize by means of pollen tube channels, aiming to solve the problem of certain limitations in the genetic transformation of maize by the gene introduction method of the traditional gene gun method

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  • Method for realizing gene editing in corn by pollen tube channel

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Embodiment 1

[0022] The present invention provides the following technical solutions: a method for realizing gene editing in corn by means of pollen tube channels, comprising the following steps:

[0023] S1, selection of corn ears, select 24-28h after self-pollination, the same corn ears at the silking stage;

[0024] S2. Preparation of gene editing plasmids;

[0025] S3. Pollen-mediated transformation: introduce the gene editing vector through the pollen tube channel, treat the pollen grains with the plasmid solution to be transformed, make the pollen grains carry the plasmid containing the Cas9 gene and sgRNA, and then pollinate;

[0026] S4, resistance screening of progeny materials;

[0027] S5. Study on the genetic characteristics of the offspring and the results of PCR testing, and obtain the offspring whose target gene has been edited.

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Abstract

The invention belongs to the technical field of corn transgenosis and particularly relates to a method for realizing gene editing in corn by a pollen tube channel. The method comprises the following steps: step 1, selecting corn ears, namely selecting the corn ears 24-28 h after self-pollination and having a consistent silking period; step 2, preparing a gene editing plasmid; step 3, pollen mediated transformation: introducing a gene editing vector by a pollen tube channel and treating pollen grains with a plasmid solution to be transformed to enable the pollen grains to carry plasmids containing Cas9 genes and sgRNA, and conducting pollinating; step 4, conducting resistance screening of progeny materials; and step 5, conducting genetic characteristic research of offsprings and obtaining PCR test results. Exogenous DNA is introduced into a receptor through a channel formed when a pollen tube enters an embryo sac and gene editing is achieved. The method has advantages of being free of genotype limitation and easy to achieve large-scale transformation, achieves gene editing of corn, does not need expensive instruments and complicated tissue culture, and saves a tissue culture process.

Description

technical field [0001] The invention relates to the field of corn transgenic technology, in particular to a method for realizing gene editing in corn by means of pollen tube channels. Background technique [0002] Corn is one of the crops with the highest total output in the world today, and it is an important source of food and feed, as well as an industrial raw material. Due to the continuous increase of the world's population, the continuous decrease of cultivated land and environmental changes, the demand for corn will continue to increase. How to increase the yield of corn per unit area is still an important and widely concerned topic. Maize yield is the result of the coordinated development of the number of ears per unit area, number of grains per ear and grain weight, and these traits are also controlled by many factors such as: genetic factors, hormone levels and environmental factors. [0003] In terms of genetic transformation of maize, a breakthrough was not made...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87C12N15/29A01H5/00A01H6/46
CPCC12N15/8202C12N15/8216C07K14/415
Inventor 郭嘉李楠朱旭贺红霞李毅丹
Owner JILIN ACAD OF AGRI SCI
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