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Method for constructing genetic transformation systems of catalpa bungei

A construction method and technology of catalpa, which is applied in the field of genetic transformation system construction, can solve problems such as limiting molecular genetic improvement of catalpa, achieve the effects of low genetic transformation efficiency, strong differentiation and regeneration ability, and improved genetic transformation efficiency

Pending Publication Date: 2017-08-18
CHINA THREE GORGES UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The construction of the genetic transformation system of Catalpa has not been reported yet, which limits the development of molecular genetic improvement of Catalpa

Method used

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  • Method for constructing genetic transformation systems of catalpa bungei
  • Method for constructing genetic transformation systems of catalpa bungei
  • Method for constructing genetic transformation systems of catalpa bungei

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Experimental program
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Effect test

Embodiment 1

[0060] MS basic medium can be prepared according to the literature (Murashige T and Skoog F., 1962). The formula of GUS staining solution was prepared according to literature (Jefferson et.al., 1987).

[0061] The method for constructing the genetic transformation system of Catalpa tree of the present invention includes the following steps:

[0062] (1) Acquisition of embryogenic callus of Catalpa tree

[0063] Take the 60-day-old immature seeds of Catalpa tree, and then sterilize them with 75% alcohol and 2% sodium hypochlorite solution, and then strip the young embryos as explants; place the explants in a series of Induced in solid medium to obtain pale yellow embryogenic callus ( figure 1 (1-A, 1-B)), as the recipient material for genetic transformation.

[0064] (2) Screening of critical lethal concentration of hygromycin

[0065] The obtained embryogenic callus of Catalpa tree was inoculated into 1 / 2MS solid-state culture with 10-80mg / L hygromycin at a culture temperature of 25±2...

Embodiment 2

[0076] MS basic medium can be prepared according to the literature (Murashige T and Skoog F., 1962). The formula of GUS staining solution was prepared according to literature (Jefferson et.al., 1987).

[0077] The method for constructing the genetic transformation system of Catalpa tree of the present invention includes the following steps:

[0078] (1) Acquisition of embryogenic callus of Catalpa tree

[0079] Take the 60-day-old immature seeds of Catalpa tree, and then sterilize them with 75% alcohol and 2% sodium hypochlorite solution, and then strip the young embryos as explants; place the explants in a series of Induced in solid medium to obtain pale yellow embryogenic callus ( figure 1 ), as the recipient material for genetic transformation.

[0080] (2) Screening of critical lethal concentration for hygromycin selection

[0081] The obtained embryogenic callus of Catalpa tree was inoculated into 1 / 2MS solid-state culture with 10-80mg / L hygromycin at a culture temperature of 25±...

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Abstract

The invention discloses a method for constructing genetic transformation systems of catalpa bungei. The method includes procedures of screening the critical lethal concentration of hygromycin by the aid of embryogenic calluses of the catalpa bungei; carrying out infection and transformation by the aid of agrobacterium-mediated processes assisted by ultrasonic treatment; detecting resistant regenerated embryogenic calluses and regenerated plants and the like. The embryogenic calluses of the catalpa bungei are used as transformation receptors. The method has the advantages that the embryogenic calluses of the catalpa bungei are used as the receptors by the aid of the method for the first time, accordingly, infection materials are wide in transformation population, and embryogenic cells are high in regeneration capacity; annual genetic transformation requirements can be met by long-term stable subculture storage of the embryogenic calluses; the embryogenic calluses of the catalpa bungei are infected and transformed under the assistance effect of ultrasonic treatment, and accordingly the method is high in transformation efficiency; T-DNA [thymine DNA (deoxyribonucleic acid)] regions of vectors are successfully integrated into genomes of the catalpa bungei by the aid of the method, and specific primers are designed for detecting transgenic catalpa bungei regenerated plants; the method is applicable to molecular genetic improvement and detection on the catalpa bungei, and transgenic catalpa bungei obtained from transformation events can be used as a donor, so that diversified novel transgenic varieties can be obtained.

Description

Technical field [0001] The invention belongs to the field of construction methods for plant genetic transformation systems, and relates to a construction method for catalpa tree heritage transformation systems. Background technique [0002] Catalpa bungei (Catalpa bungei C.A. Mey) is a deciduous tree of the Catalpa genus Bignoniaceae. It is one of the precious timber species in China. It plays an important role in industrial timber, garden viewing, ecological protection, and greening the city. It is called As the "Wood King". Catalpa has a long history of utilization. It is a precious timber species and famous garden ornamental tree species (Tang Ling et al., 2007). Its wood has the characteristics of non-cracking, corrosion resistance, easy processing, and beautiful texture. It is specially used for processing high-end commodities and special products. . Catalpa tree has a beautiful tree shape, densely hairy leaves, dense skin and dense branches. It is good for sound insulatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/00
CPCA01H4/00C12N15/8205C12N15/8206
Inventor 梁宏伟刘佳陈发菊张德春沈祥陵刘文
Owner CHINA THREE GORGES UNIV
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