Culture medium and method for improving genetic transformation efficiency of tobacco Honghuadajinyuan

A technology of genetic transformation efficiency and safflower jinyuan is applied in the field of medium for improving the genetic transformation efficiency of tobacco safflower jinyuan, which can solve the problems of low rooting rate and low conversion efficiency of tobacco safflower jinyuan, and achieves Good differentiation, improve the efficiency of differentiation buds, and ensure the effect of selection

Inactive Publication Date: 2020-08-18
CHINA TOBACCO YUNNAN IND
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Problems solved by technology

Although the current plant transgenic technology is relatively mature, the transformation efficiency o

Method used

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  • Culture medium and method for improving genetic transformation efficiency of tobacco Honghuadajinyuan
  • Culture medium and method for improving genetic transformation efficiency of tobacco Honghuadajinyuan
  • Culture medium and method for improving genetic transformation efficiency of tobacco Honghuadajinyuan

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Embodiment 1

[0037] Embodiment 1: as figure 1 As shown, the method for improving the genetic transformation efficiency of tobacco safflower Dajinyuan in the present embodiment comprises the following steps:

[0038] 1. Preparation of Tobacco Safflower Dajinyuan Leaf Disc:

[0039] Take plump tobacco seeds, sterilize them in 75% alcohol for 30 seconds, then sterilize them with 10% sodium hypochlorite solution for 8-10 minutes, wash them with sterile water for 4 times, and plant the sterilized seeds on MS medium. Cultivate for 45-60 days at 25°C under 16h light / 8h dark conditions to obtain sterile tobacco seedlings;

[0040] Take the middle and upper leaves of the aseptic tobacco seedlings, under aseptic conditions, use a puncher to obtain a leaf disc with a size of 5 mm × 5 mm, soak the leaf disc in MS medium, and the MS medium is MS basic medium, Sucrose 30g / L, pH=5.75;

[0041]2. Infection of Agrobacterium: Cultivate a single colony of Agrobacterium with successfully constructed knock...

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Abstract

The invention discloses a culture medium and a method for improving genetic transformation efficiency of tobacco Honghuadajinyuan. The culture medium comprises a co-culture medium and a selective culture medium; the co-culture medium and the selective culture medium each comprise an MS basic culture medium, NAA and 6-BA, the final concentration of the NAA is 0.41 mg/L, and the concentration ratioof the NAA to the 6-BA is 1:5; the method comprises the following steps: preparing a tobacco Honghua Dajinyuan leaf disc; infecting with agrobacterium tumefaciens; carrying out selective culture; andcarrying out rooting culture. According to the method, the budding rate (differentiated buds/leaf discs) of tobacco calluses reaches 127%, and the rooting rate (rooted seedlings/differentiated buds) reaches 66%; compared with an original method (the budding rate is 62%, and the rooting rate is 10%), the budding rate is increased by about 2 times, and the rooting rate is increased by about 6 times;and the requirement for large-scale genetic transformation is met.

Description

technical field [0001] The invention belongs to the technical field of genetic transformation of tobacco safflower and golden element, and in particular relates to a culture medium and a method for improving the genetic transformation efficiency of tobacco safflower and golden element. Background technique [0002] cultivated tobacco ( Nicotiana tabacum ) is an allotetraploid belonging to the family Solanaceae, and is considered an important model organism for verifying gene functions in other plants due to its easy genetic transformation. Compared with the model plant Arabidopsis thaliana, there is still a big gap in the functional research of tobacco genes. In recent years, the emergence of CRISPR / Cas9 technology has provided an effective tool for the study of tobacco gene function. In order to quickly obtain more tobacco mutants, a knockout plasmid library based on CRISPR / Cas9 can be used for mixed transfection to speed up the pace of tobacco gene function research. ...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N15/82A01H4/00A01H5/00A01H6/82
CPCA01H4/001A01H4/008C12N5/0025C12N15/8205C12N2500/30C12N2500/40
Inventor 蒋佳芮向海英曾婉俐高茜许力张建铎邓乐乐杨文武宋春满贾凌李雪梅杨光宇陈章玉夏庆友
Owner CHINA TOBACCO YUNNAN IND
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