Method for realizing gene transient transfection in capitulum

A flower head and transient transfection technology, applied in the field of genetic engineering, can solve the problems of chimera, low transformation efficiency, and long transformation cycle, and achieve the effect of short time, high transformation efficiency, and short transformation cycle

Pending Publication Date: 2022-01-14
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

At present, the Agrobacterium-mediated leaf disc transformation system is not yet perfect, and there are disadvantages such as long transformation cycle, low transformation efficiency, and easy occurrence of chimeras.
Transient transformation is an exogenous gene expression method in which the expression product and function can be detected within a short period of time after the introduction of the exogenous gene. There are fewer applications in inflorescences

Method used

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  • Method for realizing gene transient transfection in capitulum

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Experimental program
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Effect test

Embodiment 1

[0055] 1) Preparation of plant material:

[0056] On the adult plant of Chrysanthemum chrysanthemum, the cuttings with young bud diameters of 2.5-4mm are selected, and the length of the cuttings is 6-8cm. In plug trays, cultivate them for 20-30 days at 20° C. under humid, short-day (light / dark=8h / 16h) conditions, keep the soil moist, and wait for them to take root and bloom.

[0057] 2) Preparation of infection solution:

[0058] On the first day, pick a single colony of Agrobacterium pCAMBIA1305.2 carrier to 50mL YEB liquid medium + 50μL 50mg / mL-1 Kan + 50μL 50mg / mL-1 Rif, shake the bacteria overnight at 28°C and 200rpm;

[0059] On the second day, draw 10mL of the first-day bacterial solution to 500ml YEB liquid medium + 5ml EMS (1M) + 200μl 1% AS + 500μl 50μg / mL Kan + 500μl 50mg / ml Rif, shake the bacteria overnight at 28°C and 200rpm;

[0060] On the third day, prepare the infection solution 500mL ddH 2 O+5ml EMS(1M)+2mL 1%AS+5ml MgCl 2 . Centrifuge the overnight shake...

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Abstract

The invention provides a method for realizing gene transient transfection in capitulum, and relates to the technical field of genetic engineering. According to the method for realizing gene transient transfection in capitulum, the plant capitulum with bud cutting rooting and flowering is instantaneously transformed by adopting a vacuum infiltration effect, and transfection of a detectable amount of target genes can be realized after instantaneous transformation for 5-7 days. The method has the advantages of being simple to operate, short in transformation period, high in transformation efficiency and the like, exogenous gene transient expression in the high-efficiency capitulum can be realized, the problem that the capitulum is difficult to transform successfully is solved, a methodology foundation is laid for plant flowering gene function research, a method is provided for molecular breeding and variety improvement, and the method has high practicability and scientific research application value.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for realizing transient transfection of genes in flower heads. Background technique [0002] Flower heads are a type of infinity inflorescence. Its characteristic is that the flower axis is extremely shortened and expanded into a flat shape; the bracts at the base of the flower axis are densely integrated into involucre. This type of inflorescence mostly exists in the genus Chrysanthemum. The edge flowers of Chrysanthemum are female ray flowers, and the central disc flowers are bisexual tubular flowers. The entire inflorescence is composed of dozens or even hundreds of ray flowers and tubular flowers. combined. Its special structure makes it more difficult to transfer foreign genes into the whole flower head. At present, the Agrobacterium-mediated leaf disc transformation system has not been perfected, and there are disadvantages such as long transformatio...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/02A01H6/14
CPCC12N15/8205
Inventor 宋爱萍张佳丽余琪孙道金管志勇房伟民陈素梅陈发棣
Owner NANJING AGRICULTURAL UNIVERSITY
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