Culture medium and genetic transformation method of Hongyang kiwi fruits

A medium and liquid medium technology, applied in the field of kiwifruit genetic engineering, can solve the problems of low genetic transformation efficiency and long transformation time of Hongyang kiwifruit, and achieve the effect of preventing bacterial growth, efficient screening, and shortening the time of genetic transformation

Active Publication Date: 2020-05-15
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the genetic transformation efficiency of Hongyang kiwifruit published in some literatures is not high, and the transformation time is relatively long.

Method used

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  • Culture medium and genetic transformation method of Hongyang kiwi fruits
  • Culture medium and genetic transformation method of Hongyang kiwi fruits
  • Culture medium and genetic transformation method of Hongyang kiwi fruits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Preparation of leaf pre-culture medium (M1): add 30g / L sucrose, 2.2g / L MS (without vitamins), 103.1mg / L vitamins, the solvent is double distilled water, measure the pH with a pH meter after the solute is dissolved, and use hydrogen Sodium oxide solution to adjust the pH to 5.8, add 2.5g / L plant gel, sterilize at 121°C for 15min, cool to 65°C, add 50μM / L acetosyringone, 1mg / L 6-benzyl adenine, 0.1mg / L L naphthaleneacetic acid, 3mg / L zeatin.

Embodiment 2

[0065] Co-cultivation solid medium (M2) preparation: add 30g / L sucrose, 2.2g / L MS (without vitamins), 103.1mg / L vitamins, the solvent is double distilled water, measure the pH with a pH meter after the solute is dissolved, and measure the pH with hydrogen Sodium oxide solution to adjust the pH to 5.8, add 2.5g / L plant gel, sterilize at 121°C for 15min, cool to 65°C, add 50μM / L acetosyringone, 1mg / L 6-benzyl adenine, 0.1mg / L L naphthaleneacetic acid, 3mg / L zeatin.

Embodiment 3

[0067] Co-cultivation liquid medium (M3) preparation: add 20g / L sucrose, 2.2g / L MS medium (without vitamins), the solvent is double distilled water, measure the pH with a pH meter after the solute dissolves, and use sodium hydroxide solution to dissolve Adjust the pH to 6.0, sterilize at 121°C for 15 minutes, cool to 65°C, and add 100 μM / L acetosyringone.

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Abstract

The invention belongs to the technical field of kiwi fruit genetic engineering, and particularly relates to a culture medium and a genetic transformation method of Hongyang kiwi fruits. The genetic transformation method sequentially comprises the following steps: agrobacterium tumefaciens culturing, leaf pre-culture, infecting the leaves by the agrobacterium tumefaciens, callus screening and culturing, carrying out bud elongation culture, carrying out bud rooting culture and positive seedlings transplanting; steps of leaf pre-culture, leaf infection by agrobacterium tumefaciens, callus screening and culturing, bud elongation culture and bud rooting culture adopt the corresponding culture medium provided by the invention. According to the method, the genetic transformation time of the kiwifruits can be shortened, the genetic transformation efficiency can be improved, particularly, a culture medium formula for rapidly and efficiently obtaining positive seedlings is found, and the methodplays an important role in shortening the genetic transformation time of the Hongyang kiwi fruits and improving the genetic transformation efficiency.

Description

technical field [0001] The invention belongs to the technical field of kiwifruit genetic engineering, and in particular relates to a culture medium and a genetic transformation method of Hongyang kiwifruit. Background technique [0002] Kiwi fruit is deeply loved by people because of its unique flavor, rich in vitamin C, dietary fiber and various mineral nutrients. It has become one of the most successful domesticated fruit species in the past century, and is praised as the "king of fruits". my country's kiwi industry is developing rapidly. According to statistics, in 2017, the total area of ​​kiwifruit in my country has reached more than 2 million mu, and the output is 2.37 million tons, both of which have leapt to the first place in the world. Commercially grown kiwifruit are mainly Chinese kiwifruit and gourmet kiwifruit. 'Hongyang' kiwifruit is one of the main varieties of Chinese kiwifruit in my country. It is diploid, has red flesh, tender flesh, and excellent flavor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N15/82
CPCA01H4/001A01H4/008C12N15/8205
Inventor 曾云流徐哲明祝曼孙娟李昂刘普程运江
Owner HUAZHONG AGRI UNIV
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