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Method for obtaining brassica napus material suitable for high-density planting and mechanized harvesting

A high-density, rapeseed technology, applied in plant genetic engineering and biological fields, can solve the problems of high planting cost and high harvest loss rate, and achieve the effect of simple operation, low cost and fast speed

Inactive Publication Date: 2017-06-13
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The rapeseed varieties adapted to high-density planting and mechanized harvesting obtained by this method can effectively solve the problems of high harvest loss rate and high planting cost in current rapeseed production

Method used

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  • Method for obtaining brassica napus material suitable for high-density planting and mechanized harvesting
  • Method for obtaining brassica napus material suitable for high-density planting and mechanized harvesting
  • Method for obtaining brassica napus material suitable for high-density planting and mechanized harvesting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Transformation of Activation Tag Vector pCB260 into Wild-type Rapeseed Oil 12

[0051] 1. The activation tag vector pCB260 was transformed into Agrobacterium competent cell GV3101 (purchased from Shanghai Maiqi Biotechnology Co., Ltd.)

[0052] (1) Take about 5 μL of the activation tag vector pCB260 and add it to 50 μL of Agrobacterium competent cells GV3101

[0053] In the bacterial solution, mix slowly.

[0054] (2) Put it in an ice box for an ice bath for half an hour, quickly take it out and transfer it to liquid nitrogen to freeze for 1 minute (or place it in

[0055] -70°C refrigerator, 3-5 minutes), quickly placed in a 37°C water bath for 5 minutes.

[0056] (3) Add 800 μL LB liquid medium (without antibiotics), shake at 250 rpm at a constant temperature of 28°C

[0057] Beds were incubated for 4-5 hours. Centrifuge at 3500 rpm for 30 seconds, suck out the upper layer and discard it.

[0058] (4) Add 100 μL LB liquid medium (without antibiotics) a...

Embodiment 2

[0074] Embodiment two, rape transformed plant screening

[0075] 1. Using green fluorescent protein GFP to screen seeds harvested by flower dipping (method 1)

[0076] (1) Treatment of rapeseed seeds: select about 1000 seeds of the transformed plants obtained in Example 1, place them in a large Petri dish covered with filter paper, arrange the seeds evenly, add a small amount of water until the seeds are soaked, and then darkly treat 2- 3 days.

[0077] (2) Observation of fluorescence: After the seeds sprout, they can be placed in an optical image analysis system (purchased from Shanghai Tianneng Technology Co., Ltd.) for observation, and the exposure time is adjusted to 50s during observation. like figure 2 As shown, compared with the wild-type Ningyou 12, the mutant plants containing the pCB260 vector obviously emitted green fluorescence, that is, the white dots in the figure.

[0078] (3) Transplanting: Gently pick out the fluorescent seeds and place them in small squar...

Embodiment 3

[0095] Embodiment three, less branched rapeseed mutant sfz Phenotyping

[0096] Through the method described above, among a large number of transformed strains, a mutant of oligobranched rapeseed was found sfz, And the plant conforms to the law of Mendelian inheritance in the growth of the F2 generation, and its phenotype analysis shows that compared with the wild-type Ningyou 12 plant, the mutant at the seedling stage sfz shiny leaves ( Figure 5 ), the mature mutant sfz There are fewer branches, the effective branch height is increased, and the siliques are concentrated at the top of the plant.

[0097] Image 6 oligobranched rapeseed mutant sfz Comparison of flowering plants and wild-type plants; the inflorescences and siliques of the less-branched rapeseed mutant are concentrated at the top of the plant, and the effective branch height is significantly higher than that of the wild-type Ningyou 12 plant, which is very suitable for dense planting and mechanized har...

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Abstract

The invention relates to a method for obtaining a brassica napus material suitable for high-density planting and mechanized harvesting, belonging to the technical field of plant genetic engineering and biology. The method disclosed by the invention comprises the following steps: transforming an activation tagging vector pCB260 into brassica napus by adopting an agrobacterium flower soaking method; screening transformed plant seeds by utilizing a green fluorescent protein (GFP); screening brassica napus transformation plants by using herbicide phosphinothricin; and finally, identifying the transformed plants by using a PCR (Polymerase Chain Reaction) method. Phenotypic analysis of the transformed plants discovers that subramose brassica napus mutants are obtained and recorded as sfz. The plant accords with Mendel's law in F2-generation growth, the leaf surface at the seedling stage is bright, a few branches can grow at the maturation stage, the effective branch height is increased, and the siliques are centralized on the top of the plant. The brassica napus mutant obtained by transforming the brassica napus by utilizing the activation tagging vector pCB260 provides a novel material for large-scale mechanized production study and has important application prospect.

Description

technical field [0001] The invention relates to a method for obtaining high-density planted and mechanized harvested rapeseed materials, belonging to the fields of plant genetic engineering and biotechnology. Background technique [0002] rape( Brassica napus ) belongs to the cruciferous family and is the fifth largest crop in my country after rice, wheat, corn and soybeans. my country is a big vegetable oil consumer, and domestic rapeseed oil production can only meet 2 / 3 of people's consumption needs, and a large amount of oil consumption depends on imports. In addition, rape, as a raw material crop for biodiesel, is valued by more and more countries and is expected to become an important substitute to solve the fossil energy crisis. Therefore, increasing the output of rapeseed has important strategic significance for our country. [0003] In recent years, my country's society has developed rapidly, the rural labor force has decreased, labor prices have risen, the cost of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12Q1/68A01H5/00
CPCC12N15/8261C12N15/8205C12Q1/6895C12Q2600/13
Inventor 谭小力付正莉黄琪朱克明王政丁丽娜曹军张志燕陈克平
Owner JIANGSU UNIV
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