Preparation method of protoplast of asparagus officinalis related species asparagus filicinus

A technology of Asparagus ferns and protoplasts is applied in the field of bioengineering, which can solve the problems of interspecific incompatibility, inability to pollinate and hybridize asparagus, etc., and achieve the effects of convenient material collection, saving tissue culture and propagation time, and easy operation.

Inactive Publication Date: 2013-12-25
VEGETABLE & FLOWER INST JIANGXI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, our research found that under normal conditions, it cannot perform interspecific pollination hybridization with asparagus, and there is a serious interspecific incompatibility. Therefore, it is urgent to find new methods such as protoplast fusion to try to solve the incompatibility between them. In order to transfer good genes and traits from wild resources to asparagus cultivars for breeding of new asparagus varieties
In view of the important role of Fern Tianmendong in expanding the narrow genetic background of asparagus culti...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0018] Example 1: Select 10-15cm long Xing'an Asparagus tender shoots, peel off the scales, cut off the part covered by the top scales, cut into 2-3cm small pieces, and peel off the skin with a blade. Gently scrub under tap water for 10 minutes, in a sterile operating table, sterilize with 75% alcohol for 5 minutes, rinse twice with sterile distilled water; disinfect with 10% sodium hypochlorite for 10 minutes, rinse with sterile distilled water 4 times. Blot the distilled water on the surface of the young shoots with sterile cotton cloth, cut the above young shoots into 0.1-0.2cm thin slices, weigh 1.0g under aseptic conditions and place them in a 50mL sterile test tube; add 10mL3g / L cellulase in the test tube , 1g / L pectinase and 5g / L helicase mixed enzyme solution (pH5.8), oscillating enzymolysis at 100rpm on a shading shaker at 25°C for 2.5h, using 0.1% MES + 0.2% CaCl 2 2H 2 O+0.8mol / L glucose as osmotic pressure stabilizer, the concentration can be obtained as 2.2×10 6...

example 2

[0019] Example 2: Select 10-15cm long Xing'an Asparagus tender shoots, peel off the scales, cut off the part covered by the top scales, cut into 2-3cm small pieces, and peel off the epidermis with a blade. Gently scrub under tap water for 10 minutes, in a sterile operating table, sterilize with 75% alcohol for 5 minutes, rinse twice with sterile distilled water; disinfect with 10% sodium hypochlorite for 10 minutes, rinse with sterile distilled water 4 times. Blot the distilled water on the surface of the tender bamboo shoots with sterile cotton cloth, cut the above tender shoots into 0.1-0.2 cm thin slices, weigh 1.0 g under sterile conditions and place them in a 50 mL sterile test tube; add 10 mL of 5g / L cellulase in the test tube , 3g / L pectinase and 5g / L helicase mixed enzyme solution (pH5.8), oscillating enzymolysis at 100rpm on a shading shaker at 28°C for 4.0h, using 0.1% MES + 0.2% CaCl 2 2H 2 O+0.8mol / L glucose as osmotic pressure stabilizer, the concentration can be...

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Abstract

The invention discloses a method for efficiently preparing an immature stem protoplast of asparagus officinalis related species asparagus filicinus. The method particularly comprises the steps that (1) 10-15cm long immature asparagus filicinus is selected and scaled; a top end part coated with scales is cut out; the immature asparagus filicinus is cut into 2-3cm small sections; skin is chipped by a blade; (2) the immature asparagus filicinus is lightly scrubbed for 10min in tap water, disinfected for 5min by 75% alcohol in a sterile operation platform, flushed for two times by sterile distilled water, disinfected for 10min by 10% sodium hypochlorite, flushed for four times by the sterile distilled water, and cut into 0.1-0.2cm sheets; 1.0g is weighed under the sterile condition, and placed in a 50mL sterile test tube; (3) an appropriate amount of enzymatic hydrolysate is added to the test tube; oscillation enzymolysis is performed at 100rpm on a light-proof shaking table at 25-28 DEG C; and (4) after the enzymolysis, the enzymatic hydrolysate is filtered; a residual tissue sample not enzymolyzed is removed; a filtrate containing the protoplast is obtained; and the purified protoplast is obtained by centrifugation, washing and the like.

Description

technical field [0001] The invention belongs to the field of bioengineering, and relates to a high-efficiency preparation method of plant protoplasts, in particular to a preparation method of the protoplasts of Asparagus asparagus, a wild species closely related to asparagus. Background technique [0002] Asparagus (Asparagus officinalis L.), also known as Asparagus officinalis L., is a perennial herb of Asparagus officinalis. It is a kind of health-care vegetable with high nutritional value. One", "King of Vegetables" and other reputations. Asparagus originated in the valleys and grasslands of Eurasia and North Africa, and has been cultivated for more than 2,500 years. Because it is rich in various active factors such as saponins, rutin, selenium, plant polysaccharides, free amino acids and vitamins, it has good effects on weight loss, anti-oxidation, anti-tumor, lowering blood pressure, lowering blood lipids and lowering blood sugar. At present, the asparagus planting an...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 张岳平陈光宇罗绍春周劲松汤泳萍黄燕萍谢启鑫赵萍
Owner VEGETABLE & FLOWER INST JIANGXI ACADEMY OF AGRI SCI
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