Method for directly generating adult seedlings by genetic transformation of septate internodes of mature seed seedlings of maize inbred line

A technology of genetic transformation and inbred lines, applied in horticultural methods, botanical equipment and methods, horticulture, etc., to achieve the effect of convenient material collection and less pollution

Inactive Publication Date: 2016-02-03
JILIN UNIV
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  • Method for directly generating adult seedlings by genetic transformation of septate internodes of mature seed seedlings of maize inbred line
  • Method for directly generating adult seedlings by genetic transformation of septate internodes of mature seed seedlings of maize inbred line
  • Method for directly generating adult seedlings by genetic transformation of septate internodes of mature seed seedlings of maize inbred line

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Embodiment Construction

[0043] A kind of corn inbred line mature seed seedling stem node genetic transformation method of the present invention directly becomes seedling comprises the following steps:

[0044] (1) Disinfecting and inoculating mature seeds of corn inbred lines, including the following steps:

[0045] 1.1 Dilute the mature seeds of corn inbred lines with 70% ethanol and 0.1% HgCL 2 Disinfect in turn for 12 minutes, rinse with sterile water 3-5 times;

[0046] 1.2 Inoculate the corn seeds treated in step 1.1 into the seed germination medium, and germinate in complete darkness at 24-26°C;

[0047] 1.3 When the height of the seedlings reaches 1cm-2cm, grow under the light;

[0048] (2) carrying out the genetic transformation of the stem nodes of corn seed germination seedlings, comprising the following steps:

[0049] 2.1 When the height of corn seedlings reaches 3-5cm, cut out the stem nodes under aseptic conditions, keep 0.5cm above the stem nodes and 0.5cm below the stem nodes, and ...

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Abstract

The invention discloses a method for directly generating adult seedlings by genetic transformation of septate internodes of mature seed seedlings of a maize inbred line, and belongs to the field of bioengineering. The method comprises the following steps: 1, disinfecting and inoculating mature seeds of the maize inbred line; 2, performing genetic transformation on seedlings obtained by germination of the maize seeds; 3, transferring maize septate internodes subjected to genetic transformation into a co-culture medium; 4, transferring the maize septate internodes from the co-culture medium into an inducing culture medium; 5, performing seedling hardening and indoor transplanting on regenerated seedlings of the maize septate internodes; 6, transplanting the regenerated seedlings of the maize septate internodes into a field. Compared with a way of somatic embryogenesis, the method disclosed by the invention has the advantages that the seedlings are directly generated in an organogenesis way, so that a large quantity of regenerated plants can be obtained quickly within a short period, without being limited by the types of genes and the seasons; materials are convenient to take and are unlikely to pollute; the planting percent is high; a strong foundation is laid for industrial research on maize genetic transformation.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a method for directly growing seedlings from the stem nodes of aseptic seedlings of corn inbred lines, and a new set of corn genetic transformation method established by combining it with the method of genetic transformation of corn mediated by Agrobacterium Transformation method, this method can quickly and efficiently obtain complete transgenic plants. Background technique [0002] Maize is an important food and feed crop, and its genetic improvement has been receiving much attention. The establishment of transgenic acceptor system is the key to transgenic operation of maize. A good receptor system should meet five conditions: high regeneration ability and stability; good genetic stability; easy access to receptor materials; Agrobacterium can transform the receptor materials; transformed materials can be screened. The callus induced by immature embryos is currently the m...

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Application Information

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IPC IPC(8): A01H4/00
Inventor 原亚萍张曼冯逸龙吴颖韩俊友单晓辉苏胜忠李世鹏刘宏魁李颖
Owner JILIN UNIV
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