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Preparation method of sorghum protoplast, and conversion method of prepared protoplast

A protoplast and sorghum technology, applied in the field of plant cytology, can solve the problems of time-consuming, low-efficiency and labor-intensive protoplasts, and achieve the effect of a simple method

Inactive Publication Date: 2018-09-18
GUIZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a method for preparing sorghum protoplasts to solve the problems of time-consuming, laborious and inefficient preparation of protoplasts by using suspension cells

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  • Preparation method of sorghum protoplast, and conversion method of prepared protoplast

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Embodiment Construction

[0020] Below in conjunction with accompanying drawing and specific embodiment the invention is further introduced:

[0021] 1. Cultivation of sorghum seeds

[0022] Soak the sorghum seeds in water for 24 hours to make the seeds fully absorb water, change the water every 12 hours; pour off the water after 24 hours, cover the seeds with a damp cloth, and continue for 24 hours until the seeds grow white buds. Then it is sown in a seedling tray, and placed under 25 degrees of light after sowing until the seedlings grow together (about 1-2 days), and cultivated in the dark at 25 degrees for 8 days.

[0023] 2. Pretreatment of sorghum etiolated seedlings

[0024] Cut well-grown sorghum etiolated seedlings (about 50 plants), pretreat them in 0.6M mannitol for 10 minutes, take the middle leaf sheath, and cut them into 0.5mm thin strips with a blade (the thinner the better, try to chop them with one knife, Do not cut repeatedly to avoid damage to the cells).

[0025] 3. Enzymatic tr...

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Abstract

The invention discloses a preparation method of a sorghum protoplast, and a conversion method of the prepared protoplast. The preparation method comprises the following steps: (1) taking sorghum seedsinto water, accelerating germination, and sowing and cultivating obtained buds to form etiolated seedlings; (2) shearing the well-grown sorghum etiolated seedling, pretreating the sheared etiolated seedlings with mannitol, taking middle leaf sheaths, and cutting the sheaths by a blade to form thin strips; (3) performing enzymatic digestion on the cut leaf sheaths by using a protoplast enzymatic hydrolysate to cleave sorghum protoplasts; and (4) performing solid-liquid separation on the cleaved sorghum protoplasts to obtain the sorghum protoplast. A preparation and conversion system for the sorghum protoplast is established for the first time, the methods are simple and effective, and the sorghum protoplast can be used for studying sorghum protein subcellular localization, sorghum proteininteraction and protein purification and luciferase-labeled gene expression.

Description

technical field [0001] The invention relates to a preparation method of sorghum protoplasts and a transformation method of the prepared protoplasts, belonging to the field of plant cytology. Background technique [0002] Sorghum is an annual herbaceous crop of Poaceae. The grains of sorghum can be used for food and wine making; the ears can be used to make brooms; the young leaves can be used as silage when dried in the shade; In recent years, the research on sorghum has mainly focused on the cultivation of disease-resistant, cold-resistant and stress-resistant sorghum varieties by means of genetic engineering. However, due to the immaturity of sorghum transformation and regeneration system technology, the use of transgenic technology to study sorghum gene function is restricted, and the use of protoplasts can quickly verify plant gene function. Therefore, the research on sorghum protoplast preparation and transformation is of great significance. [0003] Since Klercker fi...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N5/10
CPCC12N5/04C12N2510/00
Inventor 谢鑫任明见王勇蒋君梅
Owner GUIZHOU UNIV
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