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A kind of fast transgenic method of longan

A transgenic and longan technology, applied in genetic engineering, plant genetic improvement, chemical instruments and methods, etc., can solve the problems of long embryoid body or transformed plant cycle, death of transgenic plants, loss of apical dominance, etc., and can easily induce adventitious buds. , the growth and metabolism is strong, and the effect is conducive to transformation

Inactive Publication Date: 2019-03-12
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that the disadvantage of the Agrobacterium rhizogenes-mediated method is that the transgenic plants will show abnormal phenomena such as loss of apical dominance, shrunken leaves, shortened internodes and changes in flower shape; this method also relies too much on plant tissue culture to obtain embryoid bodies or transformation The cycle of the plant is long, the cost is high, and the transformation efficiency is low. In the later stage, the process of seedling hardening and transplanting is required. The transgenic plants are easy to cause death due to this process.

Method used

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  • A kind of fast transgenic method of longan
  • A kind of fast transgenic method of longan
  • A kind of fast transgenic method of longan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1. Preparation of live seedlings of longan

[0046] Take more than 8 mature longan, you can also buy domestic and foreign longan in the market, peel off the peel and pulp, select healthy and plump seeds, wash with clean water, live broadcast in the nutrient soil of the plug tray, sow 1 seed in each hole, and Common sowing seedlings in the field are basically the same.

[0047] The nutrient soil used in this example, peat soil: stone: perlite = 2: 1: 1, covered with 1 cm thick nutrient soil after sowing, then watered and moisturized to promote germination, cultivated in the dark at 25°C for 7 days, and then After the seeds germinated out of the soil, they were cultured under light in a greenhouse at 25°C for 2 weeks until they reached the stage of 4 true leaves, and they were set aside.

[0048] 2. Preparation of the Agrobacterium tumefaciens infection solution carrying the transformation vector

[0049] Add 10 µL of pCAMBIA1301, pCAMBIA1301-MFT, and pCAMBIA1301-MFT-RN...

Embodiment 2

[0064] 1. Preparation of live seedlings of longan

[0065] Take more than 8 mature longan, you can also buy domestic and foreign longan in the market, peel off the peel and pulp, select healthy and plump seeds, wash with clean water, live broadcast in the nutrient soil of the plug tray, sow 1 seed in each hole, and Common sowing seedlings in the field are basically the same.

[0066] The nutrient soil used in this example, peat soil: stone: perlite = 2: 1: 1, covered with 1 cm thick nutrient soil after sowing, then watered and moisturized to promote germination, cultivated in the dark at 25°C for 7 days, and then After the seeds germinated out of the soil, they were cultured under light in a greenhouse at 25°C for 2 weeks until they reached the stage of 4 true leaves, and they were set aside.

[0067] 2. Preparation of the Agrobacterium tumefaciens infection solution carrying the transformation vector

[0068] Add 10 µL of pCAMBIA1301, pCAMBIA1301-MFT, and pCAMBIA1301-MFT-RN...

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Abstract

The invention provides a rapid transgenic method for longan, and belongs to the field of transgenic plant preparation. The method comprises the steps of preparation of longan seedling trees, preparation of transformation vector agrobacterium tumefaciens infection bacterial liquid, preparation, infection and cocultivation of seedling tree terminal bud removal or top-off (leaf upper portion removal) materials, screening and cultivation of resistant buds, molecular biological identification of regeneration resistant buds, and transplanting of transgenic plants. The method is completely independent from plant tissue culture to infect direct-seeding seedling tree terminal mud removal and top-off materials, and the transgenic plants are obtained; sterile test-tube plantlets, embryoid and suspension cells do not need to be cultured, no subculture through connection is needed for the transgenic resistant buds, the survival rate is high, the buds are not affected by weather or environment in a greenhouse, and conversion can be performed after a whole year. According to the method, the operation steps are simple and convenient to execute, the cost is lowered, and time is shorter.

Description

technical field [0001] The invention relates to a rapid transgenic method for longan, belongs to the field of transgenic plant preparation, and is mainly used in the innovation of germplasm resources of longan. Background technique [0002] There are few studies on transgenic methods applied in longan germplasm innovation. The laboratory mainly uses Agrobacterium tumefaciens mediated transformation method and gene gun transformation method. The explants used include suspension cell lines of embryoid body and callus , the transformation method mediated by Agrobacterium tumefaciens, the gene gun transformation method, the suspension cell line using embryoid bodies and callus as explants need aseptic culture, the cycle is long and the cost is high, and only transformation has been obtained so far. The embryoid body, these two methods did not get longan transgenic plants. Zeng Lihui et al. used the method mediated by Agrobacterium rhizogenes to infect the embryoid bodies and st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/00
CPCC07K14/415C12N15/8205C12N15/8218
Inventor 陈裕坤程春振赖钟雄林玉玲刘生财张梓浩
Owner FUJIAN AGRI & FORESTRY UNIV
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