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Mulberry transgenic hairy root induction and reproduction method

A transgenic, hairy root technology, applied in the field of plant genetic engineering, to overcome the long cycle, broad development and application prospects, and improve the efficiency of transgenic mulberry trees

Inactive Publication Date: 2013-01-16
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the transfer of AP2 / ERF transcription factors or other target genes into Agrobacterium rhizogenes to efficiently induce and reproduce transgenic hairy roots of mulberry trees, and to establish a transgenic system for mulberry hairy roots

Method used

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  • Mulberry transgenic hairy root induction and reproduction method
  • Mulberry transgenic hairy root induction and reproduction method
  • Mulberry transgenic hairy root induction and reproduction method

Examples

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Embodiment 1

[0031] Induction of MaERF1 Transcription Factor Gene Hairy Roots of Morus alba

[0032] (1) Mulberry explant culture: The mulberry species selected was Shaer×Lun 109 mulberry seeds, or the top buds of Shaer×Lun 109 mulberry grown in the wild were cut, and all were provided by the National Germplasm Zhenjiang Mulberry Nursery. Wash the seeds or mulberry buds clipped from the field with sterile water for 3-5 times; disinfect with 75%wt alcohol for 2 minutes, and then wash with sterile water for 2 times; Place in MS medium, grow at 26°C-28°C, and cultivate aseptic seedlings; one month later, when the mulberry aseptic seedlings grow to a height of 7-8cm, cut the young leaves at the top of the aseptic seedlings at 2-3cm Or stem sections are used as explants, wherein the leaves are cut into 0.5cm×0.5cm, and the stem sections are cut into 1cm.

[0033] (2) Pre-cultivation of explants: place the above explants in pre-configured MS+2.4-D 0.5mg / L+KT1.5mg / L+CaCl 2 Culture on 4.4g / L pre...

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Abstract

Disclosed is a mulberry transgenic hairy root induction and reproduction method which includes steps of culturing mulberry aseptic seedling; pre-culturing explant; preparing a transgenic infection bacterial liquid; performing infection and co-culture; performing explant degerming culturing; culturing a mulberry transgenic hairy root; and transferring the hairy root which is subjected to solid state expanding cultivation to a B5 fluid nutrient medium with heteroauxin for expanding cultivation to finish reproduction. According to the mulberry transgenic hairy root induction and reproduction method, the step of redifferentiation of mulberry callus cells can be omitted, and problems that the period is long and transformation efficiencies are low when the callus cells are used for genetic transformation are solved; induction and reproduction of the mulberry transgenic hairy root are not limited by factors of external environments, seasons, mulberry flower sexes and the like, so that transgenic efficiencies of a mulberry are greatly improved; researches of response mechanisms and secondary metabolism substances of mulberries under adversity stress are facilitated by transformed MaERF1 transcription factor genes; and the mulberry transgenic hairy root induction and reproduction method has great significance on mulberry function gene identification and researches and molecular breeding and has wide development application prospects.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and relates to a method for infecting mulberry explants after transferring a target gene into Agrobacterium rhizogenes, inducing transgenic hairy roots of mulberry trees, and establishing a high-efficiency induction and propagation system for transgenic hairy roots of mulberry trees. Background technique [0002] Mulberry (Morus L.) is a perennial woody dicotyledonous plant. It is an important economic crop used in the sericulture industry and the only source of feed for silkworms. Mulberry also plays an important role in forestry and food industries. Its fruit, root, stem, leaf, etc. all have high medicinal value and health value; at the same time, mulberry can produce a variety of secondary metabolites with important medicinal value, such as quercetin, etc. Flavonoids and a variety of active substances with antiviral, antibacterial, antioxidative, antiaging, and tumor prevention functio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/06
Inventor 方荣俊赵卫国童伟程嘉翎刘利张林杨永华戚金亮
Owner JIANGSU UNIV OF SCI & TECH
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