Method for obtaining hairy roots of broccoli

A technology for broccoli and hairy roots, which is applied in the field of hairy root acquisition, can solve problems such as a culture system that mediates broccoli hairy roots, and achieves good market development prospects, short infection time, and high induction rate. Effect

Inactive Publication Date: 2015-11-18
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although it has been achieved in individual plants to obtain hairy roots through plant explants mediated by A. So far, there have been no reports at home and abroad on the use of the Agrobacterium rhizogenes Ri plasmid to mediate the broccoli hairy root culture system

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 A method for obtaining broccoli hairy roots, comprising the steps of:

[0017] ⑴On the ultra-clean workbench, use high-temperature sterilized scissors to cut out the same batch of rapidly propagated aseptic broccoli seedling explants, put them on sterile filter paper, and then prick 2 on the surface with a sterilized needle Wounds were inoculated in MS medium with a pH value of 5.8, and finally pre-cultured at 25° C. in the dark for 2 days to obtain pre-cultured sterile seedling explants.

[0018] Among them: the cut broccoli aseptic seedling explant is 0.5cm 2 leaves and 0.5 cm long stem segments.

[0019] ⑵Use the pre-cultivated sterile seedling explants with OD 600 Agrobacterium rhizogenes with a concentration of 0.2 were infected at 25°C for 2 minutes, and then blotted dry the residual bacterial solution on the surface with sterile filter paper, and inoculated it on the MS medium with a concentration of 50 μmol / L acetosyringone and a pH value of 5.8 , ...

Embodiment 2

[0022] Example 2 A method for obtaining broccoli hairy roots, comprising the steps of:

[0023] ⑴On the ultra-clean workbench, cut the same batch of fast-propagating aseptic broccoli seedling explants with high-temperature sterilized scissors, put them on sterile filter paper, and then pierce 5 of them on the surface with a sterilized needle Wounds were inoculated in MS medium with a pH value of 5.8, and finally pre-cultivated in the dark at 25°C for 5 days to obtain pre-cultured sterile seedling explants.

[0024] Among them: the cut broccoli aseptic seedling explant is 0.5cm 2 leaves and 0.5 cm long stem segments.

[0025] ⑵Use the pre-cultivated sterile seedling explants with OD 600 Agrobacterium rhizogenes with a concentration of 0.6 were infected at 25°C for 6 minutes, and then blotted dry the residual bacterial solution on the surface with sterile filter paper, and inoculated on the MS medium with a concentration of 150 μmol / L acetosyringone and a pH value of 5.8 , ...

Embodiment 3

[0028] Example 3 A method for obtaining broccoli hairy roots, comprising the steps of:

[0029] ⑴On the ultra-clean workbench, use high-temperature sterilized scissors to cut out the same batch of rapidly propagated aseptic broccoli seedling explants, put them on sterile filter paper, and then pierce 8 of them on the surface with a sterilized needle Wounds were inoculated in MS medium with a pH value of 5.8, and finally pre-cultivated in the dark at 25°C for 7 days to obtain pre-cultured sterile seedling explants.

[0030] Among them: the cut broccoli aseptic seedling explant is 0.5cm 2 leaves and 0.5 cm long stem segments.

[0031] ⑵Use the pre-cultivated sterile seedling explants with OD 600 Agrobacterium rhizogenes with a concentration of 0.8 were infected at 25°C for 10 minutes, and then blotted dry the residual bacterial solution on the surface with sterile filter paper, and inoculated on the MS medium with a concentration of 200 μmol / L acetosyringone and a pH value o...

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Abstract

The invention relates to a method for obtaining hairy roots of broccoli. The method comprises the step (1) of clipping explants of aseptic seedlings of rapid propagation of the same batch, putting the explants on sterile filter paper, using a sterile needle head to prick 2-8 wounds on the surfaces of the explants, inoculating the explants in an MS culture medium and performing preincubate in the dark to obtain the oreincubate sterile seedling explants; step (2) of using agrobacterium rhizogenes to infect the sterile seedling explants to be cultured, using the sterile filter paper to soak up residual bacterial liquid on the surfaces, inoculating the explants to an MS culture medium adding acetosyringone to culture the explants together in the dark to obtain receptor explants cultured together; and step (3) of transferring the receptor explants cultured together to an MS culture medium adding Carbencillin Disodium to be cultured in the dark, transferring the receptor explants once every 5 days and obtaining the aseptic hairy roots after the explants are transferred from 2 to 5 times. The method is simple and efficient and suitable for industrialized production.

Description

technical field [0001] The invention relates to a method for obtaining hairy roots, in particular to a method for obtaining broccoli hairy roots. Background technique [0002] Broccoli is rich in anti-cancer substance sulforaphane, and was rated as one of the top ten most nutritious vegetables by the US "Time Magazine". Broccoli can enhance the body's immunity, promote growth, enhance blood vessel toughness and other effects, and can prevent the risk of breast cancer, gastric cancer and other cancers. [0003] The secondary metabolite sulforaphane in broccoli has important application value in medicine because of its anti-cancer and anti-cancer effects. At present, it is usually obtained from broccoli seeds or broccoli seedlings. Due to the high cost of the seed raw materials, the breeding method of broccoli is complicated and the cycle is long, and the content of sulforaphane in broccoli seeds and seedlings is low, which cannot meet the daily needs. People's demand for su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N5/10
Inventor 马绍英李胜胡翠珍赵生琴
Owner GANSU AGRI UNIV
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