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Populus bolleana genetic transformation method

A genetic transformation method and technology of Xinjiang poplar, applied in the field of transgenic engineering, can solve the problems of complex cultivation procedure, long cultivation period, and influence on the research of poplar gene function, and achieve the effect of omitting the cultivation procedure and shortening the cultivation period.

Pending Publication Date: 2020-09-18
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0004] At present, the genetic transformation conditions of poplar transgenic research in Xinjiang are difficult to unify, and the transformation efficiency is not high, which affects the research of poplar gene function, and generally must go through the callus induction stage, the cultivation procedure is complicated, and the cultivation period is long, which is not conducive to large-scale replication. genetic transformation

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  • Populus bolleana genetic transformation method

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Embodiment

[0027] Embodiment: the invention provides a kind of Xinjiang poplar genetic transformation method, comprises the steps:

[0028] (1) Counting down from the first leaf on the top of the tissue culture seedlings, select leaves that are fully expanded, dark green in color, and basically consistent in size, shape, and color. Use a scalpel to cut 2-3 wounds transversely along the main veins, place the leaves of Populus xinjiang on the medium for inducing adventitious buds and pre-cultivate them for 2 days. The medium for leaf induction of adventitious buds is: MS+TDZ 0.05 mg / L+IBA 0.3 mg / L.

[0029] (2) Agrobacterium activation and preparation of bacterial solution

[0030] ①Purification of Agrobacterium: Use an inoculation loop to pick a ring from the bacterial solution preserved at -80°C, then streak on the LB medium plate with kanamycin added, and place the culture dish upside down in a constant temperature incubator , After 2-3 days of culture at 28°C, isolated single colonie...

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Abstract

The invention belongs to the technical field of transgenic engineering, and particularly relates to an agrobacterium tumefaciens-mediated populus bolleana genetic transformation method using leaves asreceptors. The method comprises the following steps of carrying out genetic transformation by using aseptic seedling leaves as explants, processing T-DNA carrying a target gene in agrobacterium in aco-culture method by utilizing an agrobacterium-mediated method, transforming into plant cells of seedling leaves, and integrating into a plant genome. The transformation method has the advantages that a complex culture procedure is omitted, the culture period is shortened, an efficient genetic transformation system of populus bolleana is established by utilizing the agrobacterium-mediated method,a new populus variety is cultured, the related gene can be functionally researched, and the method is of great significance for further establishing a gene function rapid verification system.

Description

technical field [0001] The invention belongs to the technical field of transgenic engineering, and specifically relates to a genetic transformation method of Populus xinjiang mediated by Agrobacterium using leaves as receptors. Background technique [0002] Xinjiang poplar is an important tree species for afforestation, urban greening and industrial timber in my country. It occupies an irreplaceable position in the ecological construction and commercial forest construction in my country, especially in the northwest. Due to climate drought, soil salinization and other environmental impacts in Northwest China, the growth, metabolism and productivity of poplar are affected, and drought is one of the main factors that inhibit plant growth and yield. It occupies the first place among all abiotic limiting factors, and its loss to trees and crops is second only to the loss of biotic stress such as pests and diseases. Cultivating drought- and saline-alkali-tolerant forest and gras...

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/00
CPCC12N15/8205
Inventor 白玉娥代金玲托亚桑萨尔珠拉
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY