A method for preparing protoplasts of asparagus related wild species Asparagus fern

Abstract

A preparation method for efficiently preparing asparagus related wild species Asparagus fern tender stem protoplasts, the specific steps are: (1) select young Asparagus fern shoots with a length of 10-15cm, peel off the scales, and cut off the top The scale-coated part was cut into 2-3cm small pieces, and the skin was peeled off with a blade; (2) Gently scrubbed under tap water for 10 minutes, sterilized with 75% alcohol for 5 minutes in a sterile operating table, and rinsed twice with sterile distilled water , then sterilized with 10% sodium hypochlorite for 10 minutes, washed 4 times with sterile distilled water, cut into 0.1-0.2cm thin slices, weighed 1.0g under sterile conditions and placed in a 50mL sterile test tube; (3) Added Appropriate amount of enzymatic hydrolysis solution, oscillating enzymolysis at 100rpm on a shaker at 25°C-28°C shaded from light; (4) After the enzymolysis is complete, filter the enzymatic solution to remove the remaining tissue samples that have not been enzymatically hydrolyzed, and obtain a filter containing the protoplasts. solution, and purified protoplasts were obtained by centrifugation, washing and other steps.

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...

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 cultivars, it is necessary to conduct in-depth research on it. The protoplast fusion technology will provide great convenience for the selection of new varieties of interspecific hybridization and genetic improvement. At present, there are no relevant reports on the protoplast preparation system of asparagus close wild species Asparagus fern at home and abroad. Therefore, the method of the present invention will provide an important basis for subsequent asparagus new variety breeding and genetic improvement, and its significance is very important.