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Method for increasing genetic conversion rate of transgenic soybean by using graft technology

A technology of genetic transformation efficiency and transgenic soybean, applied in the field of plant transgenic, can solve the problems of restricting the research and development of soybean genetic transformation, low survival rate, etc.

Inactive Publication Date: 2007-03-14
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the differentiation efficiencies of these two systems can basically meet the needs of soybean transgenics, after the two sets of systems go through the two stages of rooting and seedling hardening, the survival rates of transplanting are very low, about 20% and 10%, respectively. Seriously restricted the research and development of soybean genetic transformation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment one: Utilize the adventitious bud grafting of cotyledon node transformation to obtain regenerated plant

[0014] 1-Experimental material:

[0015] 1) Materials: Soybean materials Jilin 35 and Jilin 47 were purchased from Jilin Academy of Agricultural Sciences.

[0016] 2) Strains and plasmids: Agrobacterium strain EHA105 was preserved by the School of Life Sciences of Northeast Normal University; the transformation vector pSBHO-3 contained the inverted repeat sequence of the soybean phosphoenolpyruvate carboxylase gene; Gene.

[0017] 2- Acquisition of Adventitious Buds

[0018] 1) Germination medium for planting sterile soybean seeds: B5 + 3.0% sucrose + 0.58% agar, pH 5.8, 25°C, cultured in low light for 5 days.

[0019] 2) When the cotyledons turn green, separate the cotyledons with a scalpel, remove the true leaves, and keep the hypocotyl of 0.5-1cm, and then lightly cut 5 times along the direction of the veins, about 3-4mm long and 0.5mm deep.

[002...

Embodiment 2

[0039] Embodiment 2: Utilize the adventitious bud grafting of embryo tip transformation to obtain regenerated plant

[0040] 1-Experimental material:

[0041] 1) Materials: Soybean materials Jilin 35 and Jilin 47 were purchased from Jilin Academy of Agricultural Sciences.

[0042] 2) Strains and plasmids: Agrobacterium strain EHA105 was preserved by the School of Life Sciences of Northeast Normal University; the transformation vector pSBHO-3 contained the inverted repeat sequence of the soybean phosphoenolpyruvate carboxylase gene; Gene.

[0043] 2- Acquisition of Adventitious Buds

[0044] 1) Explant preparation: soak the sterilized mature soybean seeds overnight for about 20 hours, remove the cotyledons, and scratch the true leaves. Collect embryo tips and insert into medium A: 1 / 2Ms salt + 1 / 2B5 organic + 3.5mg / L BAP + 3% sucrose + 0.56% agar, pH5.8, pre-cultivate for 1 day, the culture condition is 25 ℃, light The period is 18 / 6h.

[0045] 2) Preparation of transformed ...

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Abstract

The invention relates to a method for improving the genetic transfer efficiency of gene-transferred soybean. Wherein, it uses agricillin soybean cotyledon transfer system and germ sharp transfer system to obtain transfer adventitious bud; said adventitious buds without growing root and seed are directly made into graft, via cleft grafting method to be grafted on the soybean seed stock; the graft and the stock via combine period, recover period, fusion period and survival period can improve the gene transfer efficiency of soybean. The grafting can improve the survival rate of plant more than 90%, to shorten the gene transfer period and improve the gene transfer research.

Description

technical field [0001] The invention belongs to the technical field of plant transgenesis, and in particular relates to a method for improving the genetic transformation efficiency of transgenic soybeans. Background technique [0002] Soybean genetic transformation has become a model plant for many difficult-to-transform plants. At present, the soybean genetic transformation methods that have been widely recognized and applied mainly include Agrobacterium tumefaciens-mediated cotyledon node transformation system and embryo tip transformation system. Among them, the cotyledon node transformation system is relatively mature, and several transgenic soybean varieties that have been commercialized are mainly derived from this system. The embryo tip transformation system has higher differentiation efficiency and greater application potential. Although the differentiation efficiencies of these two systems can basically meet the needs of soybean transgenics, after the two sets of ...

Claims

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Application Information

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IPC IPC(8): A01H1/00A01G1/06A01C1/00A01H4/00C12N5/04
Inventor 刘立侠李望丰王德利刘宝陈伟蔡一荣许守民
Owner NORTHEAST NORMAL UNIVERSITY
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