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Genetic transformation method for mung beans

A genetic transformation method and mung bean technology, which are applied in the fields of biochemical equipment and methods, horticultural methods, and botany equipment and methods, etc., can solve problems such as complex relationships and no obvious rules.

Pending Publication Date: 2019-10-22
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

However, the success of genetic transformation is related to many factors such as the genotype of the recipient plant, Agrobacterium strain, explant selection, medium formula and hormone ratio, and the relationship is complex without obvious rules.

Method used

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  • Genetic transformation method for mung beans
  • Genetic transformation method for mung beans
  • Genetic transformation method for mung beans

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Embodiment Construction

[0026] Below in conjunction with specific embodiment, further illustrate the present invention.

[0027] A mung bean genetic transformation method, comprising the following steps:

[0028] (1) Seed germination: select about 100 mung bean Jilv No. 2 seeds with full grains, smooth seed coat and no cracks, place them in a sterile wide-mouth glass bottle and sterilize them by soaking them in 75% ethanol (mass fraction) for 2 minutes. Soak in 0.1% HgCl solution for 15 minutes, and finally rinse with sterile water for 3-4 times; the sterilized seeds are planted in 1 / 2MS medium containing 1mg / LTDZ, at 25°C, 16h light / 8h dark conditions cultured for 4 days ( figure 1 ).

[0029] (2) Explant preparation: get the seedling in step (1), peel off the seed coat with long-nose tweezers, and carefully tear off two cotyledons, then use a scalpel along the 1-2 mm above the cotyledon insertion position and the following 3- Transect at 4 mm, and retain the hypocotyl segment containing the coty...

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Abstract

The invention discloses a genetic transformation method for mung beans. The genetic transformation method comprises the steps: taking a hypocotyl segment containing cotyledonary nodes as explants to carry out infection and co-culture on the explants by an agrobacterium-mediated method, and inducing and regenerating transgenic plants after the infection is finished, wherein the infection liquid isan MSB liquid culture medium containing agrobacterium, 0.15-0.25 mg / L of 6-BA and 100-200 mg / L of acetosyringone, and OD is 0.5-0.6; during co-culturing, the co-culture medium is an MSB solid culturemedium containing 0.15-0.25 mg / L of 6-BA and 100-200 mg / L of acetosyringone. A technical system for genetic mung bean transformation is established, and the genetic transformation method can be used for genetic improvement of mung beans and innovative cultivation of new germplasms and provides a basis for functional gene verification and genome editing character improvement of the mung beans.

Description

technical field [0001] The invention relates to plant genetic transformation, in particular to a mung bean genetic transformation method. Background technique [0002] As an important legume crop, mung bean is rich in digestible protein, dietary fiber, vitamins and minerals. However, the current production of mung bean is generally maintained at a relatively low level of 1000-1500 kg / ha. In order to improve the production of mung bean and the ability to resist biotic and abiotic stress, artificial recombination of exogenous target genes into mung bean or direct Precisely editing the mung bean genome to complement and improve traditional hybrid breeding can not only greatly shorten the breeding cycle, but also make the breeding direction clearer. The realization of these visions relies on fast and efficient mung bean transformation technology, which is the basis for mung bean transgenic breeding, gene editing breeding, gene cloning and functional research. [0003] The gene...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H4/00A01H5/00A01H6/54
CPCA01H4/00C12N15/8205
Inventor 闫强张勤雪袁星星陈新
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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