Method for establishing agrobacterium rhizogenes-mediated vegetable pea genetic transformation system

A genetic transformation system, the technology of Agrobacterium rhizogenes, is applied in the field of plant genetic engineering to achieve the effects of saving experimental costs, efficient and faster experimental operations, and reducing pollution rates

Pending Publication Date: 2021-12-24
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims at many shortcomings of the existing two-step pea hairy root genetic transformation system, and the existing system does not optimize the rooting rate and positive rate of the hairy root of vegetable pea as well as the best cutting position, seedling age and culture time Combination and other problems, through one-step method to induce genetic transformation of pea hairy roots and optimize combination cutting position, seedling age and culture time, provide a one-step method to establish a high-efficiency genetic transformation system of vegetable pea mediated by Agrobacterium rhizogenes. Provide technical support for functional genome research of vegetable pea

Method used

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  • Method for establishing agrobacterium rhizogenes-mediated vegetable pea genetic transformation system
  • Method for establishing agrobacterium rhizogenes-mediated vegetable pea genetic transformation system
  • Method for establishing agrobacterium rhizogenes-mediated vegetable pea genetic transformation system

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Experimental program
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Embodiment

[0029] 1) preparation of the explant of the above-ground part of vegetable pea;

[0030] Select mature vegetable pea seeds that are free from diseases and insect pests, disinfect them with 75% sodium hypochlorite for 15 minutes, rinse them with sterile water for 5-6 times, and sow them in the substrate (the preparation of the substrate: mix vermiculite and peat in a ratio of 3 to 1, and heat them at 121° C. , autoclaved for 20 min, poured with sterile water), placed in a light incubator (22°C, photoperiod 16h light / 8h dark) for cultivation. After growing for about one week (5 days, 7 days, 9 days), the plants were cut off at the upper and lower ends of the cotyledon adjacent to the plants respectively, to obtain the explants of the aboveground part of the vegetable pea.

[0031] 2) Activation and cultivation of AR1193 Agrobacterium rhizogenes strain;

[0032] ① Take 200 μL of Agrobacterium rhizogenes strain AR1193 glycerol bacterium (carrying the binary expression vector pCAM...

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Abstract

The invention discloses a method for establishing an agrobacterium rhizogenes-mediated vegetable pea genetic transformation system. The method comprises the following steps: (1) preparing an explant; (2) cutting roots and infecting; (3) forming hairy roots. According to the method, agrobacterium rhizogenes mediation is adopted as mediation, the hairy roots of the vegetable peas are formed through one-step induction, the optimal cutting mode, seedling age and culture time are defined, the cost is low, the space is saved, the pollution rate is reduced, and the convenience of experimental operation is improved.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and belongs to a method for establishing a high-efficiency genetic transformation system of vegetable peas mediated by Agrobacterium rhizogenes, especially a method for establishing a high-efficiency genetic transformation system of vegetable peas mediated by Agrobacterium rhizogenes. method. Background technique [0002] Pea (Pisum.sativum L.) is the second largest edible bean crop in the world. According to its use, it can be divided into food pea (dry pea), vegetable pea (sweet pea, green pea) and edible tender pod. Vegetable peas (snow peas). The total output of green peas in my country ranks first in the world and occupies an important position in the world's pea production. Pea has the ability to fix nitrogen and is an environmentally friendly crop. It has the functions of reducing weight and increasing efficiency, improving soil, etc., and plays an important role in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/06A01H6/54A01G31/00A01G22/40
CPCC12N15/8205C12N15/8261A01G31/00A01G22/40
Inventor 冯志娟刘娜卜远鹏张古文王斌龚亚明
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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