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A co-transformation method based on double-strain/double-plasmid genetic transformation of citrus

A technology of genetic transformation and co-transformation, applied in the field of dual-strain/dual-plasmid co-transformation of citrus, to achieve the effect of improving the genetic traits of citrus

Active Publication Date: 2019-01-29
CITRUS RES INST OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention fills up the blank of multi-gene co-transformation of citrus at home and abroad, and solves the problem of how to realize double-strain / double-plasmid co-transformation of citrus

Method used

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  • A co-transformation method based on double-strain/double-plasmid genetic transformation of citrus
  • A co-transformation method based on double-strain/double-plasmid genetic transformation of citrus
  • A co-transformation method based on double-strain/double-plasmid genetic transformation of citrus

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Experimental program
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Effect test

Embodiment 1

[0048] Embodiment 1 Double strain / double plasmid method cotransforms golden orange

[0049] 1. Preparation of explants

[0050] Remove the seeds from the ripe fruit of the Jincheng. The seeds were rinsed with tap water, surface-sterilized with alcohol on an ultra-clean workbench, disinfected with 0.1% mercury liter for 10 minutes, and rinsed with sterile water 4 times. Peel off the seed coat and inoculate in a sterilized test tube filled with MS solid medium. Cultivate in the dark at 28° C. for 2 weeks, and then switch to light for 1 week under a photoperiod of 16 hours to obtain golden orange seedlings. The golden orange seedlings are cut transversely into about 1 cm stem segments under aseptic conditions.

[0051] MS Basic Medium Formula: (1) Macroelement: NH 4 NO 3 1650, KNO 3 1900, CaCl 2 2H 2 O 440, MgSO 4 ·7H 2 O 370, KH 2 PO 4 1700; (2) Trace elements: KI 0.83, H 3 BO 3 6.2. MnSO 4 4H 2 O 22.3, ZnSO 4 ·7H 2 O 8.6, Na 2 MnO 4 2H 2 O 0.25, CuSO 4...

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Abstract

The invention provides a co-transformation method of orange based on double-strain / double-plasmid genetic transformation. The co-transformation method comprises the steps of explant pre-cultivation and agrobacterium impregnation, and is characterized in that a basic medium of the pre-cultivation is an MS culture medium, and 6-BA (6-benzylaminopurine) with the concentration of 4 mg / L, IAA (indole acetic acid) with the concentration of 1 mg / L and AS (acetosyringone) of 100 [mu]M are added. According to the co-transformation method, the blank of orange polygene co-transformation at home and abroad is filled up, and the problem about how to realize double-strain / double-plasmid co-transformation of orange is solved.

Description

technical field [0001] The invention relates to a co-transformation method for citrus transgene research, which is mainly applicable to co-transformation of double strain / double plasmid citrus mediated by Agrobacterium. Background technique [0002] At present, most of the transgenic technologies are to build a single target gene on a carrier to genetically transform the recipient material. Many transgenic plants with new traits have been cultivated by using this strategy. However, many traits in organisms are coordinated by multiple genes, and the genetic transformation of a single gene can no longer meet the needs of crop variety improvement. In order to obtain transgenic crops with excellent traits, it will become a new development trend to introduce genes with multiple functions into plants using multi-gene co-transformation technology for plant variety improvement. [0003] Agrobacterium-mediated multi-gene co-transformation method includes mixed strain method and sing...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/00
Inventor 彭爱红邹修平何永睿许兰珍雷天刚陈善春
Owner CITRUS RES INST OF CHINESE ACAD OF AGRI SCI