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Transgenic method for orange

A transgenic and citrus technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of great damage to plants, long operation time, difficult operation, etc., and achieve firm wound healing and rapid germination Quick, easy peeling effect

Inactive Publication Date: 2009-11-11
CITRUS RES INST OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the actual operation process, the operation of wrapping the wound with paraffin tape is difficult and takes a long time, so the damage to the plant is great. However, if the wound is not wrapped with paraffin tape, a high concentration of Cytokinin (10mg / l), then the wound is easy to lose water and form callus, it is difficult to differentiate and form buds again; meanwhile, this method is to use the first generation of grafted seedlings as the starting material for transformation, while grafting In the process, lignified or semi-lignified citrus adult branches are needed, so the materials are easily restricted by seasons, and it is impossible to carry out genetic transformation operations every year

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0024] 1. Preparation of test tube rootstock.

[0025] Fruit preparation: select ripe and intact citrus fruits, wash the surface with tap water, and disinfect the surface with alcohol on the ultra-clean workbench;

[0026] Preparation of sterile absorbent cotton: Use scissors to cut a piece of absorbent cotton about 10 cm in length and width, soak it in 75% alcohol for about 10 minutes, wring it out, and set it aside;

[0027] Seed inoculation: cut the fruit with a sterilized scalpel under aseptic conditions, take out the seeds, put the seeds on the absorbent cotton, hold the scalpel in one hand and the absorbent cotton in the other, peel off the seed coat, and inoculate in a sterile MS In a test tube of solid medium; cultivate in the dark at 28°C±2°C, and the rootstock grows to 7-10cm long for later use.

[0028] The MS solid medium in this example is based on the MS medium with the addition of 30g / l sucrose and 8.5g / l agar, pH 5.8. The formula of MS medium: (1) macroelemen...

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Abstract

The invention relates to a transgenic method for orange, which comprises preparation of test-tube rootstocks and explants, preparation of grafted seedlings, wound infection of the grafted seedlings, grafting of sprouts to be transformed as well as molecular biological identification of plants to be transformed so as to obtain transgenic plants, and the grafting of the transgenic plants; and the characteristics lie in that the preparation of the grafted seedlings in the method selects the grafted seedlings of the second generation and the third generation as initial materials. The invention relates to a transgenic method capable of effectively solving annually implement genetic transformation operation of the oranges.

Description

technical field [0001] The invention relates to a method for transgenic citrus. It is mainly used in the variety improvement of citrus fruit trees. Background technique [0002] There are many transgenic methods used in the improvement of citrus varieties, mainly including the direct transformation method of exogenous DNA and the transformation method mediated by Agrobacterium tumefaciens. [0003] The explants used in the direct transformation method of exogenous DNA are protoplasts, and the protoplasts of many varieties of citrus fruit trees have a long culture cycle and difficult regeneration, and the cost is also high, so few laboratories currently use this method . [0004] There are many explants used in the Agrobacterium tumefaciens-mediated method, including epicotyl sections of aseptic seedlings, cotyledon sections, adult tree stem sections, and adult axillary buds in the field. The frequency of genetic transformation using epicotyl sections and cotyledon section...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01G1/06
Inventor 陈善春何永睿彭爱红许兰珍邹修平彭祝春
Owner CITRUS RES INST OF CHINESE ACAD OF AGRI SCI
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