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A method for Agrobacterium-mediated infection and transformation of Mongolian willow

An Agrobacterium-mediated, Agrobacterium-mediated technology, applied in the field of plant genetic engineering, can solve problems such as no reports on the establishment of a Mongolian willow regeneration system.

Inactive Publication Date: 2019-04-02
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still no report on the establishment of the genetic transformation receptor regeneration system of willow mongolica

Method used

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  • A method for Agrobacterium-mediated infection and transformation of Mongolian willow
  • A method for Agrobacterium-mediated infection and transformation of Mongolian willow
  • A method for Agrobacterium-mediated infection and transformation of Mongolian willow

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specific Embodiment approach 1

[0034] Specific embodiment one: the method of the Agrobacterium-mediated infection transformation Mongolian willow of the present embodiment is carried out according to the following steps:

[0035] One, the induction of the leaf callus of artificially cultivated Mongolian willow aseptic seedlings: Mongolian willow seeds are disinfected and sterilized by 69% to 71% alcohol and 2% to 4% sodium hypochlorite by volume fraction, and then in 1 After germination on the / 2MS medium, grow to the aseptic seedlings of the 4-leaf stage or the 5-leaf stage, and cut the leaves of the aseptic seedlings whose length×width is (0.5cm±0.1cm)×(0.5cm±0.1cm) in MSYD1 Induce callus on the culture medium; at 27.5°C-28.5°C, culture under light conditions for 14 hours, then culture under dark conditions for 10 hours, so that light and dark conditions are alternately cultured for 20-25 days, and then replace the medium once, 60-65 days callus induced;

[0036] 2. Agrobacterium infection transformation...

specific Embodiment approach 2

[0039] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that step 1 MSYD1 medium is composed of MS dry powder medium 4.73g / L, sucrose 30g / L, 2,4-D 3mg / L, 6-benzyl Aminopurine 0.5mg / L, naphthaleneacetic acid 0.2mg / L, agar powder 8g / L and distilled water, pH=5.8±0.2. Other steps and parameters are the same as those in the first embodiment.

specific Embodiment approach 3

[0040] Embodiment 3: This embodiment differs from Embodiment 1 in that the YEP liquid medium in step 2 consists of 10 g of tryptone, 10 g of yeast extract, 5 g of sodium chloride and 1000 mL of water, with a pH of 7.0±0.2. Other steps and parameters are the same as those in the first embodiment.

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Abstract

The invention discloses a method using agrobacterium tumefaciens mediated infection to transform salix mongolica, relates to the field of plant genetic engineering, and aims to solve the problem that no building methods of the genetic transformation receptor regeneration system of the salix mongolica exist in the prior art. The method is characterized in that the agrobacterium tumefaciens mediated infection is used to transform callus with leaf-induced explant, the callus is subjected to co-culture and screening transformation, the callus is induced to differentiate resistance multiple shoots, a plant is regenerated on a rooting culture medium, and GUS staining detection of the transformed regenerated plant shows that the plant has beta-galactosidase activity under 35S promoter and proves that the method can transform the salix mongolica by using the agrobacterium tumefaciens mediated infection. The method is applicable to the field of plant genetic engineering.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a method for bacteria-mediated infection and transformation of Mongolian willow. Background technique [0002] Mongolia willow (Salix mongolica), Salicaceae (Salicaceae), Salix (Salix), genus and species name: Salix mongolica f.sericea Chang.et Skv. Native shrub species on Songnensoda saline-alkaline meadow grasslands, slender branchlets, linear or linear-lanceolate leaves, 5-15 cm long, 5-9 mm wide, young leaves with downy hairs, back glabrous, green surface , the back is pale, with glandular serrations on the margin; the petiole is 8-12mm long. Flowers open first, nearly simultaneously, without peduncle, often with 3 leaflets at the base; male inflorescences 2-3 cm long, 2-3 mm thick; 2 stamens fully connate, female inflorescences 2.5-3.0 cm long, ca. 4 mm; ovary hairy, sessile, style absent or very short, stigma capitate; bracts with male flowers. Distribution: Heil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/00
CPCC12N15/8205C12N15/8273
Inventor 管清杰汪振娟马海燕柳参奎
Owner NORTHEAST FORESTRY UNIVERSITY
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