Separation method of xylem parenchyma cell protoplast of rice root

A technology of parenchyma cells and protoplasts, applied in plant cells and other directions, can solve problems such as few reports

Inactive Publication Date: 2009-02-04
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Rice is an important food crop in my country. Its protoplasts have been widely studied in cell fusion, transgenic hybrid breeding, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0016] 1. Cultivation of the medium column protoplast isolation material:

[0017] Take one Petri dish of each size, spread two layers of filter paper in the small Petri dish, moisten it, spread a layer of rice seeds evenly and at appropriate intervals, cover the large Petri dish, and place it for 24 hours without light.

[0018] After the seeds germinated, they were sown in a cultivation pot filled with quartz sand, and the cultivation conditions were 255°C, 60% humidity, and 10 hours of sunlight. After a week, take out the seedlings and wash off the quartz sand.

[0019] 2. Separation of middle column:

[0020] Clean the root system of rice seedlings, take the thicker root system, remove the 1cm length of the tip and the part with root hairs on the upper part, and use the remaining 2-3cm as the source of the central column material.

[0021] Put the cut root in 0.2% CaCl2 solution, tap it gently with small tweezers, so that the root system is flat, use the tip of the tweez...

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PUM

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Abstract

The present invention provides a method for separating out rice root xylem parenchyma cell protoplasts. The method chooses the part of the extended and the mature sections of a rice Oryza sativa L. seeding root, which is 2cm to 3cm long and does not has lateral roots grown, uses hydrolytic enzyme liquids with different ingredients and finds the optimal separation conditions to successfully separate out the intact stele xylem parenchyma cell protoplasts. At present, the related report is not published yet at home and abroad.

Description

technical field [0001] The invention relates to a method for isolating protoplasts of xylem parenchyma cells of plant roots, in particular to a method for isolating protoplasts of xylem parenchyma cells of rice roots. Background technique [0002] In 1960, Cocking isolated active protoplasts for the first time by enzymatic method. In 1971, Takebe et al. isolated protoplasts from tobacco leaves and obtained regenerated plants through cultivation. The research and application of protoplasts entered a new stage. Protoplasts are viable "naked cells" that have had their cell walls removed and surrounded by a plasma membrane, making them ideal recipients for gene transformation. Plant protoplast culture is a prerequisite for cell fusion and can be used to overcome certain obstacles in distant hybridization and more broadly combine various favorable genetic traits of plants. And it can be used for organelle transplantation and cell mutant screening, which is an important part and...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 赵福庚李隼张豊
Owner NANJING UNIV
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