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Method for obtaining drought resisting transgenic corn self-bred line through adding dropwisely in style section

A technology of transgenic corn and inbred lines, applied in the fields of plant gene improvement, botanical equipment and methods, and introduction of foreign genetic material using vectors, etc., can solve the problems of low transformation efficiency, achieve improved drought tolerance, simple operation, and easy operation high efficiency effect

Inactive Publication Date: 2011-08-31
INST OF DRY LAND FARMING SHANXI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The application of gene transfer by pollen tube passage method is becoming more and more common in the transgenic breeding of maize inbred lines, but its low transformation efficiency is still the bottleneck restricting its application

Method used

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  • Method for obtaining drought resisting transgenic corn self-bred line through adding dropwisely in style section
  • Method for obtaining drought resisting transgenic corn self-bred line through adding dropwisely in style section
  • Method for obtaining drought resisting transgenic corn self-bred line through adding dropwisely in style section

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0014] 1. Materials and methods.

[0015] 1.1 Plant materials and donors.

[0016] 1.1.1 Receptor.

[0017] The stable maize inbred lines Chang 7-2 and Zheng 58 are preserved by the Dry Farming Center of Shanxi Academy of Agricultural Sciences.

[0018] 1.1.2 The configuration of the donor introduction solution.

[0019] The transformation donor plasmid is p3UbiTPS, the target gene is TPS, and the plant selection marker is bar gene, both of which are driven by the maize Ubiquitin gene promoter. The Agrobacterium strain is EHA101, provided by Professor Lin Zhongping of Peking University. The above materials have mature preparation technology.

[0020] 1. After shaking the bacteria overnight, pour the bacteria liquid into a 2ml centrifuge tube, pour it over, 10000rpm, 1min, and settle the colony.

[0021] 2. Add 1ml STE to the sediment, shake it evenly, 10000rpm, 1min, discard the supernatant, and blot dry with paper.

[0022] 3. Add 280ul of solution I, shake and mix. Solution I: 50mM gl...

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Abstract

The invention relates to a preparation method of transgenic corns, specifically to a method for obtaining a drought resisting transgenic corn self-bred line through adding dropwisely in a style section. The method comprises the steps of employing the corn self-bred lines of Chang 7-2 and Zheng 58; selecting a sunny day for self-pollination from 14:00 to 16:00; cutting the pollination material style in the middle with a scalpel 16 to 20 hours after the pollination, i.e. 10:00 to 12:00 the next day; adding 15-20ul of introduction liquid dropwisely for three times from the cut with a dropper and simultaneously pumping air from a pollinated tassel with an injection-tube; and enveloping for self-pollination and screening for obtaining the corn self-inbred line. The introduction liquid contains TPS target gene and has Agrobacterium tumefaciens as a mediator. The method is proved by detections to be capable of transforming exogenous DNA for obtaining novel drought resisting resources of corn, thereby being a feasible and effective novel method for drought resisting breeding.

Description

technical field [0001] The invention relates to a method for preparing transgenic corn, in particular to a method for obtaining a drought-resistant transgenic corn inbred line by adding dropwise to a style section. Background technique [0002] With the scarcity of water resources, drought has become the most important abiotic factor affecting the high yield of maize. China's arid and water-scarce areas account for 52% of the country's land area, and the area affected by drought is 9 million hm 2 About 10 billion kg of direct grain income is reduced every year. Drought has become the biggest challenge for my country's agricultural production. Improving the drought tolerance of maize varieties through variety improvement is the most economical and effective method, but the drought tolerance of maize is mostly controlled by micro-effects and multiple genes, and genetic improvement is difficult and little progress has been made. Therefore, it is of great significance to use t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H1/02
Inventor 董春林张彦琴杨丽莉张明义梁改梅孙洁李洁赵巧红陈通连李强
Owner INST OF DRY LAND FARMING SHANXI ACAD OF AGRI SCI
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